Patent Publication Number: US-2021180096-A1

Title: Engineered biosynthetic pathways for production of histamine by fermentation

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims the benefit of U.S. provisional application No. 62/660,875, filed Apr. 20, 2018, which is hereby incorporated by reference in its entirety. 
    
    
     STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT 
     This invention was made with Government support under Agreement No. HR0011-15-9-0014, awarded by DARPA. The Government has certain rights in the invention. 
    
    
     INCORPORATION BY REFERENCE OF THE SEQUENCE LISTING 
     This application includes a sequence listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. This ASCII copy, created on Apr. 17, 2019, is named ZMGNP011WO_Seq_List_ST25.txt and is 312,107 bytes in size. 
     FIELD OF THE DISCLOSURE 
     The present disclosure relates generally to the area of engineering microbes for production of histamine by fermentation. 
     BACKGROUND 
     Biogenic amines are organic bases endowed with biological activity, which are frequently found in fermented foods and beverages. Histamine is known to exist in nature in fermented foods such as yogurt (13-36 mg/kg) [1], miso (24 mg/kg) [2], and red wine (24 mg/L) [3]. Some bacteria that live in the human gut also make histamine, and it functions to regulate the immune system by an anti-inflammatory effect [4]. Production of histamine in fermented foods relies on a source of proteins that contain histidine and microbes that histidine decarboxylase. Histamine is the decarboxylation product of histidine that is catalyzed specifically by the enzyme histidine decarboxylase (EC 4.1.1.22). Production of histamine in an industrial fermentation from simple, non-protein, carbon and nitrogen sources requires assembly of a pathway with improved biosynthesis of the amino acid precursor histidine and a highly active histidine decarboxylase. 
     SUMMARY 
     The disclosure provides engineered microbial cells, cultures of the microbial cells, and methods for the production of histamine, including the following: 
     Embodiment 1: An engineered microbial cell that expresses a non-native histidine decarboxylase, wherein the engineered microbial cell produces histamine. 
     Embodiment 2: The engineered microbial cell of embodiment 1, wherein the engineered microbial cell includes increased activity of one or more upstream histamine pathway enzyme(s), said increased activity being increased relative to a control cell. 
     Embodiment 3: The engineered microbial cell of embodiment 2, wherein the one or more upstream histamine pathway enzyme(s) are selected from the group consisting of an ATP phosphoribosyltransferase, a phosphoribosyl-ATP pyrophosphatase, a phosphoribosyl-AMP cyclohydrolase, a 5′ProFAR isomerase, an imidazole-glycerol phosphate synthase, an imidazole-glycerol phosphate dehydratase, a histidinol-phosphate aminotransferase, a histidinol-phosphate phosphatase, histidinol dehydrogenase, and a ribose phosphate pyrophosphokinase. 
     Embodiment 4: The engineered microbial cell of any one of embodiments 1-3, wherein the engineered microbial cell includes reduced activity of one or more enzyme(s) that consume one or more histamine pathway precursors, said reduced activity being reduced relative to a control cell. 
     Embodiment 5: The engineered microbial cell of embodiment 4, wherein the one or more enzyme(s) that consume one or more histamine pathway precursors are selected from the group consisting of an enolase, a pyruvate dehydrogenase, a pentose phosphate pathway sugar isomerase, a transaldolase, a transketolase, a ribulose-5-phosphate epimerase, and a ribulose-5-phosphate isomerase. 
     Embodiment 6: The engineered microbial cell of embodiment 4 or embodiment 5, wherein the reduced activity is achieved by replacing a native promoter of a gene for said one or more enzymes with a less active promoter. 
     Embodiment 7: The engineered microbial cell of any one of embodiments 1-6, wherein the engineered microbial cell additionally expresses a feedback-deregulated glucose-6-phosphate dehydrogenase or a feedback-deregulated ATP phosphoribosyltransferase. 
     Embodiment 8: An engineered microbial cell, wherein the engineered microbial cell includes means for expressing a non-native histidine decarboxylase, wherein the engineered microbial cell produces histamine. 
     Embodiment 9: The engineered microbial cell of embodiment 8, wherein the engineered microbial cell includes means for increasing the activity of one or more upstream histamine pathway enzyme(s), said increased activity being increased relative to a control cell. 
     Embodiment 10: The engineered microbial cell of embodiment 9, wherein the one or more upstream histamine pathway enzyme(s) are selected from the group consisting of an ATP phosphoribosyltransferase, a phosphoribosyl-ATP pyrophosphatase, a phosphoribosyl-AMP cyclohydrolase, a 5′ProFAR isomerase, an imidazole-glycerol phosphate synthase, an imidazole-glycerol phosphate dehydratase, a histidinol-phosphate aminotransferase, a histidinol-phosphate phosphatase, a histidinol dehydrogenase, and a ribose phosphate pyrophosphokinase. 
     Embodiment 11: The engineered microbial cell of any one of embodiments 8-10, wherein the engineered microbial cell includes means for reducing the activity of one or more enzyme(s) that consume one or more histamine pathway precursors, said reduced activity being reduced relative to a control cell. 
     Embodiment 12: The engineered microbial cell of embodiment 11, wherein the one or more enzyme(s) that consume one or more histamine pathway precursors are selected from the group consisting of an enolase, a pyruvate dehydrogenase, pentose phosphate pathway sugar isomerase, a transketolase, a translaldolase, a ribulose-5-phosphate epimerase, and a ribulose-5-phosphate isomerase. 
     Embodiment 13: The engineered microbial cell of embodiment 11 or embodiment 12, wherein the reduced activity is achieved by means for replacing a native promoter of a gene for said one or more enzymes with a less active promoter. 
     Embodiment 14: The engineered microbial cell of any one of embodiments 8-13, wherein the engineered microbial cell additionally includes means for expressing glucose-6-phosphate dehydrogenase or a feedback-deregulated ATP phosphoribosyltransferase. 
     Embodiment 15: The engineered microbial cell of any one of embodiments 1-14, wherein the engineered microbial cell includes a fungal cell. 
     Embodiment 16: The engineered microbial cell of embodiment 15, wherein the engineered microbial cell includes a yeast cell. 
     Embodiment 17: The engineered microbial cell of embodiment 16, wherein the yeast cell is a cell of the genus  Saccharomyces  or  Yarrowia.    
     Embodiment 18: The engineered microbial cell of embodiment 17, wherein the yeast cell is a cell of the genus  Saccharomyces  and of the species  cerevisiae.    
     Embodiment 19: The engineered microbial cell of embodiment 17, wherein the yeast cell is a cell of the genus  Yarrowia  and of the species  lipolytica.    
     Embodiment 20: The engineered microbial cell of any one of embodiments 1-19, wherein the non-native histidine decarboxylase includes a histidine decarboxylase having at least 70% amino acid sequence identity with a histidine decarboxylase from  Chromobacterium  sp. LK1 or from  Acinetobacter baumannii  strain AB0057. 
     Embodiment 21: The engineered microbial cell of any one of embodiments 1 and 16-20, wherein the engineered microbial cell includes increased activity of one or more upstream histamine pathway enzyme(s), said increased activity being increased relative to a control cell, wherein the one or more upstream histamine pathway enzyme(s) comprise an ATP phosphoribosyltransferase. 
     Embodiment 22: The engineered microbial cell of embodiment 21 wherein the increased activity of the ATP phosphoribosyltransferase is achieved by heterologously expressing it. 
     Embodiment 23: The engineered microbial cell of embodiment 22, wherein the heterologous ATP phosphoribosyltransferase has at least 70% amino acid sequence identity with an ATP phosphoribosyltransferase from  S. cerevisiae.    
     Embodiment 24: The engineered microbial cell of any one of embodiments 16-23, wherein the engineered microbial cell includes a feedback-deregulated variant of a  Corynebacterium glutamicum  ATP phosphoribosyltransferase. 
     Embodiment 25: The engineered microbial cell of any one of embodiments 1-14, wherein the engineered microbial cell is a bacterial cell. 
     Embodiment 26: The engineered microbial cell of embodiment 25, wherein the bacterial cell is a cell of the genus  Corynebacteria  or  Bacillus.    
     Embodiment 27: The engineered microbial cell of embodiment 26, wherein the bacterial cell is a cell of the genus  Corynebacteria  and of the species  glutamicum.    
     Embodiment 28: The engineered microbial cell of embodiment 26, wherein the bacterial cell is a cell of the genus  Bacillus  and of the species  subtilis.    
     Embodiment 29: The engineered microbial cell of any one of embodiments 25-28, wherein the non-native histidine decarboxylase includes a histidine decarboxylase having at least 70% amino acid sequence identity with a histidine decarboxylase from  Acinetobacter baumannii  or from  Lactobacillus  sp. (strain 30a). 
     Embodiment 30: The engineered microbial cell of any one of embodiments 1 and 25-29, wherein the engineered microbial cell includes increased activity of one or more upstream histamine pathway enzyme(s), said increased activity being increased relative to a control cell, wherein the one or more upstream histamine pathway enzyme(s) comprise an ATP phosphoribosyltransferase and an imidazole-glycerol phosphate dehydratase. 
     Embodiment 31: The engineered microbial cell of embodiment 30, wherein the increased activity of the ATP phosphoribosyltransferase or the imidazole-glycerol phosphate dehydratase is achieved by heterologously expressing it. 
     Embodiment 32: The engineered microbial cell of embodiment 31, wherein the heterologous ATP phosphoribosyltransferase has at least 70% amino acid sequence identity with an ATP phosphoribosyltransferase from  Saccharomyces cerevisiae  S288c or from  Salmonella typhimurium  LT2, or the heterologous imidazole-glycerol phosphate dehydratase has at least 70% amino acid sequence identity with an imidazole-glycerol phosphate dehydratase from  Corynebacterium glutamicum.    
     Embodiment 33: The engineered microbial cell of any one of embodiments 25-32, wherein the engineered microbial cell includes a feedback-deregulated variant of a  Salmonella typhimurium  ATP phosphoribosyltransferase. 
     Embodiment 34: The engineered microbial cell of any one of embodiments 1-33, wherein, when cultured, the engineered microbial cell produces histamine at a level of at least 20 mg/L of culture medium. 
     Embodiment 35: The engineered microbial cell of embodiment 34, wherein, when cultured, the engineered microbial cell produces histamine at a level of at least 300 mg/L of culture medium. 
     Embodiment 36: A culture of engineered microbial cells according to any one of embodiments 1-35. 
     Embodiment 37: The culture of embodiment 36, wherein the engineered microbial cells are present in a concentration such that the culture has an optical density at 600 nm of 10-500. 
     Embodiment 38: The culture of any one of embodiments 36-37, wherein the culture includes histamine. 
     Embodiment 39: The culture of any one of embodiments 36-38, wherein the culture includes histamine at a level at least 20 mg/L of culture medium. 
     Embodiment 40: A method of culturing engineered microbial cells according to any one of embodiments 1-35, the method including culturing the cells under conditions suitable for producing histamine. 
     Embodiment 41: The method of embodiment 40, wherein the method includes fed-batch culture, with an initial glucose level in the range of 1-100 g/L, followed controlled sugar feeding. 
     Embodiment 42: The method of any one of embodiments 40-41, wherein the fermentation substrate includes glucose and a nitrogen source selected from the group consisting of urea, an ammonium salt, ammonia, and any combination thereof. 
     Embodiment 43: The method of any one of embodiments 40-42, wherein the culture is pH-controlled during culturing. 
     Embodiment 44: The method of any one of embodiments 40-43, wherein the culture is aerated during culturing. 
     Embodiment 45: The method of any one of embodiments 40-44, wherein the engineered microbial cells produce histamine at a level at least 20 mg/L of culture medium. 
     Embodiment 46: The method of any one of embodiments 40-45, wherein the method additionally includes recovering histamine from the culture. 
     Embodiment 47: A method for preparing histamine using microbial cells engineered to produce histamine, the method including: (a) expressing a non-native histidine decarboxylase in microbial cells; (b) cultivating the microbial cells in a suitable culture medium under conditions that permit the microbial cells to produce histamine, wherein the histamine is released into the culture medium; and isolating histamine from the culture medium. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1 : Biosynthetic pathway for histamine. 
         FIG. 2 : Histamine titers measured in the extracellular broth following fermentation by the first-round engineered host  Corynebacteria glutamicum . (See also Example 1, Table 1.) 
         FIG. 3 : Histamine titers measured in the extracellular broth following fermentation by the first-round engineered host  Saccharomyces cerevisiae . (See also Example 1, Table 1.) 
         FIG. 4 : Histamine titers measured in the extracellular broth following fermentation by the second-round engineered host  Corynebacteria glutamicum . (See also Example 1, Table 2.) 
         FIG. 5 : Histamine titers measured in the extracellular broth following fermentation by the second-round engineered host  Saccharomyces cerevisiae . (See also Example 1, Table 2.) 
         FIG. 6 : Histamine titers measured in the extracellular broth following fermentation by the first-round engineered host  Yarrowia lipolytica . (See also Example 2, Table 4.) 
         FIG. 7 : Histamine titers measured in the extracellular broth following fermentation by the first-round engineered host  Bacillus subtilis.    
         FIG. 8 : Histamine acid titers measured in the extracellular broth following fermentation of  Saccharomyces cerevisiae  expressing the host evaluation designs. 
         FIG. 9 : Histamine acid titers measured in the extracellular broth following fermentation of  Corynebacteria glutamicum  expressing the host evaluation designs. 
         FIG. 10 : Histamine titers measured in the extracellular broth following fermentation by the third-round engineered host  Saccharomyces cerevisiae . (Improvement round.) 
         FIG. 11 : Integration of Promoter-Gene-Terminator into  Saccharomyces cerevisiae  and  Yarrowia lipolytica.    
         FIG. 12 : Promoter replacement in  Saccharomyces cerevisiae  and  Yarrowia lipolytica.    
         FIG. 13 : Targeted gene deletion in  Saccharomyces cerevisiae  and  Yarrowia lipolytica.    
         FIG. 14 : Integration of Promoter-Gene-Terminator into  Corynebacteria glutamicum  and  Bacillus subtilis.    
     
    
    
     DETAILED DESCRIPTION 
     This disclosure describes a method for the production of the small molecule histamine via fermentation by a microbial host from simple carbon and nitrogen sources, such as glucose and urea, respectively. This objective can be achieved by introducing a non-native metabolic pathway into a suitable microbial host for industrial fermentation of large-scale chemical products. Illustrative hosts include  Saccharomyces cerevisiae, Yarrowia lypolytica, Corynebacteria glutamicum , and  Bacillus subtilis . The engineered metabolic pathway links the central metabolism of the host to a non-native pathway to enable the production of histamine. The simplest embodiment of this approach is the expression of an enzyme, a non-native histidine decarboxylase enzyme, in a microbial host strain that can produce histidine. Further engineering of the metabolic pathway by modification of the microbial host central metabolism through overexpression and mutation of a key upstream pathway enzyme, ATP phosphoribosyltransferase, enabled titers of 505 mg/L histamine to be achieved. 
     The following disclosure describes how to engineer a microbe with the necessary characteristics to produce industrially feasible titers of histamine from simple carbon and nitrogen sources. Active histidine decarboxylases have been identified, and it has been found that feedback-deregulated ATP phosphoribosyltransferase and/or constitutive expression of native ATP phosphoribosyltransferase improve the titers of histidine by fermentation. 
     Definitions 
     Terms used in the claims and specification are defined as set forth below unless otherwise specified. 
     The term “fermentation” is used herein to refer to a process whereby a microbial cell converts one or more substrate(s) into a desired product (such as histamine) by means of one or more biological conversion steps, without the need for any chemical conversion step. 
     The term “engineered” is used herein, with reference to a cell, to indicate that the cell contains at least one targeted genetic alteration introduced by man that distinguishes the engineered cell from the naturally occurring cell. 
     The term “native” is used herein to refer to a cellular component, such as a polynucleotide or polypeptide, that is naturally present in a particular cell. A native polynucleotide or polypeptide is endogenous to the cell. 
     When used with reference to a polynucleotide or polypeptide, the term “non-native” refers to a polynucleotide or polypeptide that is not naturally present in a particular cell. 
     When used with reference to the context in which a gene is expressed, the term “non-native” refers to a gene expressed in any context other than the genomic and cellular context in which it is naturally expressed. A gene expressed in a non-native manner may have the same nucleotide sequence as the corresponding gene in a host cell, but may be expressed from a vector or from an integration point in the genome that differs from the locus of the native gene. 
     The term “heterologous” is used herein to describe a polynucleotide or polypeptide introduced into a host cell. This term encompasses a polynucleotide or polypeptide, respectively, derived from a different organism, species, or strain than that of the host cell. In this case, the heterologous polynucleotide or polypeptide has a sequence that is different from any sequence(s) found in the same host cell. However, the term also encompasses a polynucleotide or polypeptide that has a sequence that is the same as a sequence found in the host cell, wherein the polynucleotide or polypeptide is present in a different context than the native sequence (e.g., a heterologous polynucleotide can be linked to a different promotor and inserted into a different genomic location than that of the native sequence). “Heterologous expression” thus encompasses expression of a sequence that is non-native to the host cell, as well as expression of a sequence that is native to the host cell in a non-native context. 
     As used with reference to polynucleotides or polypeptides, the term “wild-type” refers to any polynucleotide having a nucleotide sequence, or polypeptide having an amino acid, sequence present in a polynucleotide or polypeptide from a naturally occurring organism, regardless of the source of the molecule; i.e., the term “wild-type” refers to sequence characteristics, regardless of whether the molecule is purified from a natural source; expressed recombinantly, followed by purification; or synthesized. The term “wild-type” is also used to denote naturally occurring cells. 
     A “control cell” is a cell that is otherwise identical to an engineered cell being tested, including being of the same genus and species as the engineered cell, but lacks the specific genetic modification(s) being tested in the engineered cell. 
     Enzymes are identified herein by the reactions they catalyze and, unless otherwise indicated, refer to any polypeptide capable of catalyzing the identified reaction. Unless otherwise indicated, enzymes may be derived from any organism and may have a native or mutated amino acid sequence. As is well known, enzymes may have multiple functions and/or multiple names, sometimes depending on the source organism from which they derive. The enzyme names used herein encompass orthologs, including enzymes that may have one or more additional functions or a different name. 
     The term “feedback-deregulated” is used herein with reference to an enzyme that is normally negatively regulated by a downstream product of the enzymatic pathway (i.e., feedback-inhibition) in a particular cell. In this context, a “feedback-deregulated” enzyme is a form of the enzyme that is less sensitive to feedback-inhibition than the native enzyme native to the cell. A feedback-deregulated enzyme may be produced by introducing one or more mutations into a native enzyme. Alternatively, a feedback-deregulated enzyme may simply be a heterologous, native enzyme that, when introduced into a particular microbial cell, is not as sensitive to feedback-inhibition as the native, native enzyme. In some embodiments, the feedback-deregulated enzyme shows no feedback-inhibition in the microbial cell. 
     The term “histamine” refers to 2-(1I-Imidazol-4-yl)ethanamine (CAS #51-45-6). 
     The term “sequence identity,” in the context of two or more amino acid or nucleotide sequences, refers to two or more sequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection. 
     For sequence comparison to determine percent nucleotide or amino acid sequence identity, typically one sequence acts as a “reference sequence,” to which a “test” sequence is compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence relative to the reference sequence, based on the designated program parameters. Alignment of sequences for comparison can be conducted using BLAST set to default parameters. 
     The term “titer,” as used herein, refers to the mass of a product (e.g., histamine) produced by a culture of microbial cells divided by the culture volume. 
     As used herein with respect to recovering histamine from a cell culture, “recovering” refers to separating the histamine from at least one other component of the cell culture medium. 
     Engineering Microbes for Histamine Production 
     Histamine Biosynthesis Pathway 
     Histamine is typically derived from the amino acid histidine. The histamine biosynthesis pathway is shown in  FIG. 1 . The first enzyme of the amino acid biosynthesis pathway, ATP phosphoribosyltransferase, is subject to feedback inhibition by histidine. Histamine production is enabled by the addition of a single non-native enzymatic step in  Saccharomyces cerevisiae, Yarrowia lypolytica, Corynebacteria glutamicum , and  Bacillus subtilis  hosts, which is catalyzed by histidine decarboxylase (EC 4.1.1.22). 
     Engineering for Microbial Histamine Production 
     Any histidine decarboxylase that is active in the microbial cell being engineered may be introduced into the cell, typically by introducing and expressing the gene(s) encoding the enzyme(s)s using standard genetic engineering techniques. Suitable histidine decarboxylase may be derived from any source, including plant, archaeal, fungal, gram-positive bacterial, and gram-negative bacterial sources. Exemplary sources include, but are not limited to:  Aeromonas salmonicida  subsp.  pectinolytica  34mel,  Acinetobacter baumannii  (strain AB0057),  Chromobacterium haemolyticum, Chromobacterium  sp. LK1 , Citrobacter pasteurii, Drosophila melanogaster, Lactobacillus aviarius  DSM 20655,  Lactobacillus fructivorans, Lactobacillus reuteri, Lactobacillus  sp. (strain 30a),  Methanosarcina barkeri  (strain Fusaro/DSM804),  Methanosarcina barkeri  str. Wiesmoor,  Morganella psychrotolerans, Mus musculus, Oenococcus oeni  ( Leuconostoc oenos ),  Pseudomonas putida  ( Arthrobacter siderocapsulatus ),  Pseudomonas rhizosphaerae, Pseudomonas  sp. bs2935,  Solanum lycopersicum, Oryza sativa, Penicillium marneffei, Streptomyces hygroscopicus, Pseudomonas putida, Arabidopsis thaliana  (Mouse-ear cress),  Glycine soja  (Wild soybean),  Solanum lycopersicum  (Tomato) ( Lycopersicon esculentum ),  Clostridium perfringens, Lactobacillus buchneri, Drosophila melanogaster  (Fruit fly),  Morganella morganii  ( Proteus morganii ),  E. coli, Bos taurus  (Bovine),  Raoutella planticol  ( Klebsiella planticola ),  Acinetobacter baumannii, Acinetobacter haemolyticus, Photobacterium damselae, Tetragenococcus muriaticus, Moritella  sp JT01,  Streptococcus thermophilus, Enterobacter aerogenes, Citrobacter youngae, Raoultella omithinolytica , and  Raoultella planticola.    
     One or more copies of histidine decarboxylase gene can be introduced into a selected microbial host cell. If more than one copy of a gene is introduced, the copies can have the same or different nucleotide sequences. In some embodiments, one or both of the heterologous gene(s) is/are expressed from a strong, constitutive promoter. In some embodiments, the heterologous histidine decarboxylase gene(s) is/are expressed from an inducible promoter. The heterologous gene(s) can optionally be codon-optimized to enhance expression in the selected microbial host cell. Illustrative codon-optimization tables for hosts used in the Examples are as follows:  Bacillus subtilis  Kazusa codon table: www.kazusa.or.jp/codon/cgi-bin/showcodin.cgi?species=1423&amp;aa=1&amp;style=N;  Yarrowia lipolytica  Kazusa codon table: www.kazusa.or.jp/codon/cgi-bin/showcodoon/cgi?species=4952&amp;aa=1&amp;style=N;  Corynebacteria glutamicum  Kazusa codon table: www.kazusa.or.jp/codon/cgi-bin/showcodoon/cgi?species=340322&amp;aa=1&amp;style=N;  Saccharomyces cerevisiae  Kazusa codon table: www.kazusa.or.jp/codon/cgi-bin/showcodoon/cgi?species=4932&amp;aa=1&amp;style=N. Also used, was a modified, combined codon usage scheme for  S. cereviae  and  C. glutamicum , which is reproduced below. 
     
       
         
           
               
            
               
                   
               
               
                 Modified Codon Usage Table for Sc and Cg 
               
            
           
           
               
               
               
            
               
                 Amino 
                   
                   
               
               
                 Acid 
                 Codon 
                 Fraction 
               
               
                   
               
            
           
           
               
               
               
            
               
                 A 
                 GCG 
                 0.22 
               
               
                 A 
                 GCA 
                 0.29 
               
               
                 A 
                 GCT 
                 0.24 
               
               
                 A 
                 GCC 
                 0.25 
               
               
                 C 
                 TGT 
                 0.36 
               
               
                 C 
                 TGC 
                 0.64 
               
               
                 D 
                 GAT 
                 0.56 
               
               
                 D 
                 GAC 
                 0.44 
               
               
                 E 
                 GAG 
                 0.44 
               
               
                 E 
                 GM 
                 0.56 
               
               
                 F 
                 TTT 
                 0.37 
               
               
                 F 
                 TTC 
                 0.63 
               
               
                 G 
                 GGG 
                 0.08 
               
               
                 G 
                 GGA 
                 0.19 
               
               
                 G 
                 GGT 
                 0.3 
               
               
                 G 
                 GGC 
                 0.43 
               
               
                 H 
                 CAT 
                 0.32 
               
               
                 H 
                 CAC 
                 0.68 
               
               
                 I 
                 ATA 
                 0.03 
               
               
                 I 
                 ATT 
                 0.38 
               
               
                 I 
                 ATC 
                 0.59 
               
               
                 K 
                 MG 
                 0.6 
               
               
                 K 
                 AAA 
                 0.4 
               
               
                 L 
                 TTG 
                 0.29 
               
               
                 L 
                 TTA 
                 0.05 
               
               
                 L 
                 CTG 
                 0.29 
               
               
                 L 
                 CTA 
                 0.06 
               
               
                 L 
                 CTT 
                 0.17 
               
               
                 L 
                 CTC 
                 0.14 
               
               
                 M 
                 ATG 
                 1 
               
               
                 N 
                 MT 
                 0.33 
               
               
                 N 
                 MC 
                 0.67 
               
               
                 P 
                 CCG 
                 0.22 
               
               
                 P 
                 CCA 
                 0.35 
               
               
                 P 
                 CCT 
                 0.23 
               
               
                 P 
                 CCC 
                 0.2 
               
               
                 Q 
                 CAG 
                 0.61 
               
               
                 Q 
                 CM 
                 0.39 
               
               
                 R 
                 AGG 
                 0.11 
               
               
                 R 
                 AGA 
                 0.12 
               
               
                 R 
                 CGG 
                 0.09 
               
               
                 R 
                 CGA 
                 0.17 
               
               
                 R 
                 CGT 
                 0.34 
               
               
                 R 
                 CGC 
                 0.18 
               
               
                 S 
                 AGT 
                 0.08 
               
               
                 S 
                 AGC 
                 0.16 
               
               
                 S 
                 TCG 
                 0.12 
               
               
                 S 
                 TCA 
                 0.13 
               
               
                 S 
                 TCT 
                 0.17 
               
               
                 S 
                 TCC 
                 0.34 
               
               
                 T 
                 ACG 
                 0.14 
               
               
                 T 
                 ACA 
                 0.12 
               
               
                 T 
                 ACT 
                 0.2 
               
               
                 T 
                 ACC 
                 0.53 
               
               
                 V 
                 GTG 
                 0.36 
               
               
                 V 
                 GTA 
                 0.1 
               
               
                 V 
                 GTT 
                 0.26 
               
               
                 V 
                 GTC 
                 0.28 
               
               
                 W 
                 TGG 
                 1 
               
               
                 Y 
                 TAT 
                 0.34 
               
               
                 Y 
                 TAC 
                 0.66 
               
               
                   
               
            
           
         
       
     
     Increasing the Activity of Upstream Enzymes 
     One approach to increasing histamine production in a microbial cell that is capable of such production is to increase the activity of one or more upstream enzymes in the histamine biosynthesis pathway. Upstream pathway enzymes include all enzymes involved in the conversions from a feedstock all the way to into the last native metabolite (histidine, in the illustrative microbial cells described in the Examples below). Such enzymes include an ATP phosphoribosyltransferase, a phosphoribosyl-ATP pyrophosphatase, a phosphoribosyl-AMP cyclohydrolase, a 5′ProFAR isomerase, an imidazole-glycerol phosphate synthase, an imidazole-glycerol phosphate dehydratase, a histidinol-phosphate aminotransferase, a histidinol-phosphate phosphatase, histidinol dehydrogenase, and a ribose phosphate pyrophosphokinase. Suitable upstream pathway genes encoding these enzymes may be derived from any source, including, for example, those discussed above as sources for a histidine decarboxylase gene. 
     In some embodiments, the activity of one or more upstream pathway enzymes is increased by modulating the expression or activity of the native enzyme(s). For example, native regulators of the expression or activity of such enzymes can be exploited to increase the activity of suitable enzymes. 
     Alternatively, or in addition, one or more promoters can be substituted for native promoters using, for example, a technique such as that illustrated in  FIG. 12 . In certain embodiments, the replacement promoter is stronger than the native promoter and/or is a constitutive promoter. 
     In some embodiments, the activity of one or more upstream pathway enzymes is supplemented by introducing one or more of the corresponding genes into the histidine decarboxylase-expressing microbial host cell. An introduced upstream pathway gene may be from an organism other than that of the host cell or may simply be an additional copy of a native gene. In some embodiments, one or more such genes are introduced into a microbial host cell capable of histamine production and expressed from a strong constitutive promoter and/or can optionally be codon-optimized to enhance expression in the selected microbial host cell. 
     Example 1 describes the successful engineering of  C. glutamicum  to express a heterologous histamine decarboxylase from  Acinetobacter baumannii  (SEQ ID NO:1) and to constitutively express a heterologous  C. glutamicum  imidazoleglycerol-phosphate dehydratase (SEQ ID NO:2). This strain resulted from two rounds of genetic engineering and produced histamine at a titer of 24 mg/L of culture medium. This titer was increased to 68 mg/L in a  C. glutamicum  strain engineered to express a histamine decarboxylase from  Acinetobacter baumannii  (strain AB0057) (SEQ ID NO:1) and an ATP phosphoribosyltransferase from  S. cerevisiae  S288c (SEQ ID NO:3). 
     Example 2 describes the successful engineering of  Y. lypolytica  to express a histidine decarboxylase from  Acinetobacter baumannii  (strain AB0057) (SEQ ID NO:1) and an ATP phosphoribosyltransferase from  S. cerevisiae  S288c (SEQ ID NO:3) to give a histamine titer of 505 mg/L. Example 2 also describes the engineering  B. subtilis  to express a histamine decarboxylase from  Lactobacillus  sp. (strain 30a) (SEQ ID NO:4) and an ATP phosphoribosyltransferase from  Salmonella typhimurium  LT2 (SEQ ID NO:5) to give a histamine titer of 18 mg/L. Also in Example 2,  S. cerevisiae  was engineered to express a histamine decarboxylase from  Chromobacterium  sp. LK1 (SEQ ID NO:6) and an ATP phosphoribosyltransferase  S. cerevisiae  S288c (SEQ ID NO:3) to give a histamine titer of 111 mg/L. 
     In various embodiments, the engineering of a histamine-producing microbial cell to increase the activity of one or more upstream pathway enzymes increases the histamine titer by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90 percent or by at least 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 5.5-fold, 6-fold, 6.5-fold, 7-fold, 7.5-fold, 8-fold, 8.5-fold, 9-fold, 9.5-fold, 10-fold, 11-fold, 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, 20-fold, 21-fold, 22-fold, 23-fold, 24-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65-fold, 70-fold, 75-fold, 80-fold, 85-fold, 90-fold, 95-fold, or 100-fold. In various embodiments, the increase in histamine titer is in the range of 10 percent to 100-fold, 2-fold to 50-fold, 5-fold to 40-fold, 10-fold to 30-fold, or any range bounded by any of the values listed above. (Ranges herein include their endpoints.) These increases are determined relative to the histamine titer observed in a histamine-producing microbial cell that lacks any increase in activity of upstream pathway enzymes. This reference cell may have one or more other genetic alterations aimed at increasing histamine production, e.g., the cell may express a feedback-deregulated enzyme. 
     In various embodiments, the histamine titers achieved by increasing the activity of one or more upstream pathway genes are at least 1, 10, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg/L or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, or 10 gm/L. In various embodiments, the titer is in the range of 10 mg/L to 10 gm/L, 20 mg/L to 5 gm/L, 50 mg/L to 4 gm/L, 100 mg/L to 3 gm/L, 500 mg/L to 2 gm/L or any range bounded by any of the values listed above. 
     Introduction of Feedback-Deregulated Enzymes 
     Since histidine biosynthesis is subject to feedback inhibition, another approach to increasing histamine production in a microbial cell engineered to produce histamine is to introduce feedback-deregulated forms of one or more enzymes that are normally subject to feedback regulation. Examples of such enzymes include glucose-6-phosphate dehydrogenase and ATP phosphoribosyltransferase. A feedback-deregulated form can be a heterologous, native enzyme that is less sensitive to feedback inhibition than the native enzyme in the particular microbial host cell. Alternatively, a feedback-deregulated form can be a variant of a native or heterologous enzyme that has one or more mutations or truncations rendering it less sensitive to feedback inhibition than the corresponding native enzyme. Examples of the latter include a variant ATP phosphoribosyltransferase (from  C. glutamicum ) containing the amino acid substitutions N215K, L231F, and T235A (SEQ ID NO:7) and a variant ATP phosphoribosyltransferase (from  Salmonella typhimurium ) containing the deletion of amino acids Q207 and E208 (SEQ ID NO:5). 
     In various embodiments, the engineering of a histamine-producing microbial cell to express a feedback-deregulated enzymes increases the histamine titer by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90 percent or by at least 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 5.5-fold, 6-fold, 6.5-fold, 7-fold, 7.5-fold, 8-fold, 8.5-fold, 9-fold, 9.5-fold, 10-fold, 11-fold, 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, 20-fold, 21-fold, 22-fold, 23-fold, 24-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65-fold, 70-fold, 75-fold, 80-fold, 85-fold, 90-fold, 95-fold, or 100-fold. In various embodiments, the increase in histamine titer is in the range of 10 percent to 100-fold, 2-fold to 50-fold, 5-fold to 40-fold, 10-fold to 30-fold, or any range bounded by any of the values listed above. These increases are determined relative to the histamine titer observed in a histamine-producing microbial cell that does not express a feedback-deregulated enzyme. This reference cell may (but need not) have other genetic alterations aimed at increasing histamine production, i.e., the cell may have increased activity of an upstream pathway enzyme resulting from some means other than feedback-insensitivity. 
     In various embodiments, the histamine titers achieved by using a feedback-deregulated enzyme to increase flux though the histamine biosynthetic pathway are at least 100, 200, 300, 400, 500, 600, 700, 800, or 900 μg/L, or at least 1, 10, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg/L or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 20, 50 g/L. In various embodiments, the titer is in the range of 50 μg/L to 50 g/L, 75 μg/L to 20 g/L, 100 μg/L to 10 g/L, 200 μg/L to 5 g/L, 500 μg/L to 4 g/L, 1 mg/L to 3 g/L, 500 mg/L to 2 g/L or any range bounded by any of the values listed above. 
     The approaches of supplementing the activity of one or more native enzymes and/or introducing one or more feedback-deregulated enzymes can be combined in histamine decarboxylase-expressing microbial cells to achieve even higher histamine production levels. For example, a histamine titer of 385 mg/L was achieved in  S. cerevisiae  in two rounds of engineering from the introduction of three genes: a histidine decarboxylase gene (from  Chromobacterium  sp. LK1) (SEQ ID NO:6), an ATP phosphoribosyltransferase (from  C. glutamicum ) containing the amino acid substitutions N215K, L231F, and T235A (SEQ ID NO:7), and a constitutively expressed ATP phosphoribosyltransferase from  S. cerevisiae  S288c (SEQ ID NO:3). (Example 1.) 
     Reduction of Precursor Consumption 
     Another approach to increasing histamine production in a microbial cell that is capable of such production is to decrease the activity of one or more enzymes that consume one or more histamine pathway precursors. In some embodiments, the activity of one or more such enzymes is reduced by modulating the expression or activity of the native enzyme(s). Illustrative enzymes of this type include an enolase, a pyruvate dehydrogenase, a pentose phosphate pathway sugar isomerase, a transaldolase, a transketolase, a ribulose-5-phosphate epimerase, and a aribulose-5-phosphate isomerase. The activity of such enzymes can be decreased, for example, by substituting the native promoter of the corresponding gene(s) with a less active or inactive promoter or by deleting the corresponding gene(s). See  FIGS. 12 and 13  for examples of schemes for promoter replacement and targeted gene deletion, respectively, in  S. cervisiae  and  Y. lipolytica.    
     In various embodiments, the engineering of a histamine-producing microbial cell to reduce precursor consumption by one or more side pathways increases the histamine titer by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90 percent or by at least 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 5.5-fold, 6-fold, 6.5-fold, 7-fold, 7.5-fold, 8-fold, 8.5-fold, 9-fold, 9.5-fold, 10-fold, 11-fold, 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, 20-fold, 21-fold, 22-fold, 23-fold, 24-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65-fold, 70-fold, 75-fold, 80-fold, 85-fold, 90-fold, 95-fold, or 100-fold. In various embodiments, the increase in histamine titer is in the range of 10 percent to 100-fold, 2-fold to 50-fold, 5-fold to 40-fold, 10-fold to 30-fold, or any range bounded by any of the values listed above. These increases are determined relative to the histamine titer observed in a histamine-producing microbial cell that does not include genetic alterations to reduce precursor consumption. This reference cell may (but need not) have other genetic alterations aimed at increasing histamine production, i.e., the cell may have increased activity of an upstream pathway enzyme. 
     In various embodiments, the histamine titers achieved by reducing precursor consumption by one or more side pathways are at least 100, 200, 300, 400, 500, 600, 700, 800, or 900 μg/L, or at least 1, 10, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg/L or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 20, 50 g/L. In various embodiments, the titer is in the range of 50 μg/L to 50 g/L, 75 μg/L to 20 g/L, 100 μg/L to 10 g/L, 200 μg/L to 5 g/L, 500 μg/L to 4 g/L, 1 mg/L to 3 g/L, 500 mg/L to 2 g/L or any range bounded by any of the values listed above. 
     The approaches of increasing the activity of one or more native enzymes and/or introducing one or more feedback-deregulated enzymes and/or reducing precursor consumption by one or more side pathways can be combined to achieve even higher histamine production levels. 
     Microbial Host Cells 
     Any microbe that can be used to express introduced genes can be engineered for fermentative production of histamine as described above. In certain embodiments, the microbe is one that is naturally incapable of fermentative production of histamine. In some embodiments, the microbe is one that is readily cultured, such as, for example, a microbe known to be useful as a host cell in fermentative production of compounds of interest. Bacteria cells, including gram positive or gram negative bacteria can be engineered as described above. Examples include, in addition to  C. glutamicum  cells,  Bacillus subtilus, B. lichenformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. clausii, B. halodurans, B. megaterium, B. coagulans, B. circulans, B. lautus, B. thuringiensis, S. albus, S. lividans, S. coelicolor, S. griseus, Pseudomonas  sp.,  P. alcaligenes, P. citrea, Lactobacilis  spp. (such as  L. lactis, L. plantarum ),  L. grayi, E. coi, E. faecium, E. gallinarum, E. casseliflavus , and/or  E. faecalis  cells. 
     There are numerous types of anaerobic cells that can be used as microbial host cells in the methods described herein. In some embodiments, the microbial cells are obligate anaerobic cells. Obligate anaerobes typically do not grow well, if at all, in conditions where oxygen is present. It is to be understood that a small amount of oxygen may be present, that is, there is some level of tolerance level that obligate anaerobes have for a low level of oxygen. Obligate anaerobes engineered as described above can be grown under substantially oxygen-free conditions, wherein the amount of oxygen present is not harmful to the growth, maintenance, and/or fermentation of the anaerobes. 
     Alternatively, the microbial host cells used in the methods described herein can be facultative anaerobic cells. Facultative anaerobes can generate cellular ATP by aerobic respiration (e.g., utilization of the TCA cycle) if oxygen is present. However, facultative anaerobes can also grow in the absence of oxygen. Facultative anaerobes engineered as described above can be grown under substantially oxygen-free conditions, wherein the amount of oxygen present is not harmful to the growth, maintenance, and/or fermentation of the anaerobes, or can be alternatively grown in the presence of greater amounts of oxygen. 
     In some embodiments, the microbial host cells used in the methods described herein are filamentous fungal cells. (See, e.g., Berka &amp; Barnett, Biotechnology Advances, (1989), 7(2):127-154). Examples include  Trichoderma longibrachiatum, T. viride, T. koningii, T. harzianum, Penicillium  sp.,  Humicola insolens, H. lanuginose, H. grisea, Chrysosporium  sp.,  C. lucknowense, Gliocladium  sp.,  Aspergillus  sp. (such as  A. oryzae, A. niger, A. sojae, A. japonicus, A. nidulans , or  A. awamori ),  Fusarium  sp. (such as  F. roseum, F. graminum F. cerealis, F. oxysporuim , or  F. venenatum ),  Neurospora  sp. (such as  N. crassa  or  Hypocrea  sp.),  Mucor  sp. (such as  M. miehei ),  Rhizopus  sp., and  Emericella  sp. cells. In particular embodiments, the fungal cell engineered as described above is  A. nidulans, A. awamori, A. oryzae, A. aculeatus, A. niger, A. japonicus, T. reesei, T. viride, F. oxysporum , or  F. solani . Illustrative plasmids or plasmid components for use with such hosts include those described in U.S. Patent Pub. No. 2011/0045563. 
     Yeasts can also be used as the microbial host cell in the methods described herein. Examples include:  Saccharomyces  sp.,  Schizosaccharomyces  sp.,  Pichia  sp.,  Hansenula polymorpha, Pichia stipites, Kluyveromyces marxianus, Kluyveromyces  spp.,  Yarrowia lipolytica  and  Candida  sp. In some embodiments, the  Saccharomyces  sp. is  S. cerevisiae  (See, e.g., Romanos et al., Yeast, (1992), 8(6):423-488). Illustrative plasmids or plasmid components for use with such hosts include those described in U.S. Pat. No. 7,659,097 and U.S. Patent Pub. No. 2011/0045563. 
     In some embodiments, the host cell can be an algal cell derived, e.g., from a green algae, red algae, a glaucophyte, a chlorarachniophyte, a euglenid, a chromista, or a dinoflagellate. (See, e.g., Saunders &amp; Warmbrodt, “Gene Expression in Algae and Fungi, Including Yeast,” (1993), National Agricultural Library, Beltsville, Md.). Illustrative plasmids or plasmid components for use in algal cells include those described in U.S. Patent Pub. No. 2011/0045563. 
     In other embodiments, the host cell is a cyanobacterium, such as cyanobacterium classified into any of the following groups based on morphology: Chlorococcales, Pleurocapsales, Oscillatoriales, Nostocales, Synechosystic or Stigonematales (See, e.g., Lindberg et al., Metab. Eng., (2010) 12(1):70-79). Illustrative plasmids or plasmid components for use in cyanobacterial cells include those described in U.S. Patent Pub. Nos. 2010/0297749 and 2009/0282545 and in Intl. Pat. Pub. No. WO 2011/034863. 
     Genetic Engineering Methods 
     Microbial cells can be engineered for fermentative histamine production using conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, and biochemistry, which are within the skill of the art. Such techniques are explained fully in the literature, see e.g., “Molecular Cloning: A Laboratory Manual,” fourth edition (Sambrook et al., 2012); “Oligonucleotide Synthesis” (M. J. Gait, ed., 1984); “Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications” (R. I. Freshney, ed., 6th Edition, 2010); “Methods in Enzymology” (Academic Press, Inc.); “Current Protocols in Molecular Biology” (F. M. Ausubel et al., eds., 1987, and periodic updates); “PCR: The Polymerase Chain Reaction,” (Mullis et al., eds., 1994); Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley &amp; Sons (New York, N.Y. 1994). 
     Vectors are polynucleotide vehicles used to introduce genetic material into a cell. Vectors useful in the methods described herein can be linear or circular. Vectors can integrate into a target genome of a host cell or replicate independently in a host cell. For many applications, integrating vectors that produced stable transformants are preferred. Vectors can include, for example, an origin of replication, a multiple cloning site (MCS), and/or a selectable marker. An expression vector typically includes an expression cassette containing regulatory elements that facilitate expression of a polynucleotide sequence (often a coding sequence) in a particular host cell. Vectors include, but are not limited to, integrating vectors, prokaryotic plasmids, episomes, viral vectors, cosmids, and artificial chromosomes. 
     Illustrative regulatory elements that may be used in expression cassettes include promoters, enhancers, internal ribosomal entry sites (IRES), and other expression control elements (e.g., transcription termination signals, such as polyadenylation signals and poly-U sequences). Such regulatory elements are described, for example, in Goeddel, Gene Expression Technology: Methods In Enzymology 185, Academic Press, San Diego, Calif. (1990). 
     In some embodiments, vectors may be used to introduce systems that can carry out genome editing, such as CRISPR systems. See U.S. Patent Pub. No. 2014/0068797, published 6 Mar. 2014; see also Jinek M., et al., “A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity,” Science 337:816-21, 2012). In Type II CRISPR-Cas9 systems, Cas9 is a site-directed endonuclease, namely an enzyme that is, or can be, directed to cleave a polynucleotide at a particular target sequence using two distinct endonuclease domains (HNH and RuvC/RNase H-like domains). Cas9 can be engineered to cleave DNA at any desired site because Cas9 is directed to its cleavage site by RNA. Cas9 is therefore also described as an “RNA-guided nuclease.” More specifically, Cas9 becomes associated with one or more RNA molecules, which guide Cas9 to a specific polynucleotide target based on hybridization of at least a portion of the RNA molecule(s) to a specific sequence in the target polynucleotide. Ran, F. A., et al., (“In vivo genome editing using  Staphylococcus aureus  Cas9,” Nature 520(7546):186-91, 2015 Apr. 9], including all extended data) present the crRNA/tracrRNA sequences and secondary structures of eight Type II CRISPR-Cas9 systems. Cas9-like synthetic proteins are also known in the art (see U.S. Published Patent Application No. 2014-0315985, published 23 Oct. 2014). 
     Example 1 describes illustrative integration approaches for introducing polynucleotides and other genetic alterations into the genomes of  C. glutamicum  and  S. cerevisiae  cells. 
     Vectors or other polynucleotides can be introduced into microbial cells by any of a variety of standard methods, such as transformation, conjugation, electroporation, nuclear microinjection, transduction, transfection (e.g., lipofection mediated or DEAE-Dextrin mediated transfection or transfection using a recombinant phage virus), incubation with calcium phosphate DNA precipitate, high velocity bombardment with DNA-coated microprojectiles, and protoplast fusion. Transformants can be selected by any method known in the art. Suitable methods for selecting transformants are described in U.S. Patent Pub. Nos. 2009/0203102, 2010/0048964, and 2010/0003716, and International Publication Nos. WO 2009/076676, WO 2010/003007, and WO 2009/132220. 
     Engineered Microbial Cells 
     The above-described methods can be used to produce engineered microbial cells that produce, and in certain embodiments, overproduce, histamine. Engineered microbial cells can have at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more genetic alterations, such as 30-100 alterations, as compared to a native microbial cell, such as any of the microbial host cells described herein. Engineered microbial cells described in the Example below have one, two, or three genetic alterations, but those of skill in the art can, following the guidance set forth herein, design microbial cells with additional alterations. In some embodiments, the engineered microbial cells have not more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, or 4 genetic alterations, as compared to a native microbial cell. In various embodiments, microbial cells engineered for histamine production can have a number of genetic alterations falling within the any of the following illustrative ranges: 1-10, 1-9, 1-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-7, 3-6, 3-5, 3-4, etc. 
     In some embodiments, an engineered microbial cell expresses at least one heterologous histamine decarboxylase, such as in the case of a microbial host cell that does not naturally produce histamine. In various embodiments, the microbial cell can include and express, for example: (1) a single heterologous histamine decarboxylase gene, (2) two or more heterologous histamine decarboxylase genes, which can be the same or different (in other words, multiple copies of the same heterologous histamine decarboxylase genes can be introduced or multiple, different heterologous histamine decarboxylase genes can be introduced), (3) a single heterologous histamine decarboxylase gene that is not native to the cell and one or more additional copies of an native histamine decarboxylase gene, or (4) two or more non-native histamine decarboxylase genes, which can be the same or different, and one or more additional copies of an native histamine decarboxylase gene. 
     This engineered host cell can include at least one additional genetic alteration that increases flux through the pathway leading to the production of histidine (the immediate precursor of histamine). These “upstream” enzymes in the pathway include: an ATP phosphoribosyltransferase, a phosphoribosyl-ATP pyrophosphatase, a phosphoribosyl-AMP cyclohydrolase, a 5′ProFAR isomerase, an imidazole-glycerol phosphate synthase, an imidazole-glycerol phosphate dehydratase, a histidinol-phosphate aminotransferase, a histidinol-phosphate phosphatase, histidinol dehydrogenase, and a ribose phosphate pyrophosphokinase, including any isoforms, paralogs, or orthologs having these enzymatic activities (which as those of skill in the art readily appreciate may be known by different names). The at least one additional alteration can increase the activity of the upstream pathway enzyme(s) by any available means, e.g., by: (1) modulating the expression or activity of the native enzyme(s), (2) expressing one or more additional copies of the genes for the native enzymes, and/or (3) expressing one or more copies of the genes for one or more non-native enzymes. 
     In some embodiments, increased flux through the pathway can be achieved by expressing one or more genes encoding a feedback-deregulated enzyme, as discussed above. For example, the engineered host cell can include and express one or more feedback-deregulated ATP phosphoribosyltransferase genes. 
     The engineered microbial cells can contain introduced genes that have a native nucleotide sequence or that differ from native. For example, the native nucleotide sequence can be codon-optimized for expression in a particular host cell. The amino acid sequences encoded by any of these introduced genes can be native or can differ from native. In various embodiments, the amino acid sequences have at least 60 percent, 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity with a native amino acid sequence. 
     In some embodiments, increased availability of precursors to histamine can be achieved by reducing the expression or activity of enzymes that consume one or more histamine pathway precursors, such as an enolase, a pyruvate dehydrogenase, a pentose phosphate pathway sugar isomerase, a transaldolase, a transketolase, a ribulose-5-phosphate epimerase, and a aribulose-5-phosphate isomerase. For example, the engineered host cell can include one or more promoter swaps to down-regulate expression of any of these enzymes and/or can have their genes deleted to eliminate their expression entirely. 
     The approach described herein has been carried out in bacterial cells, namely  C. glutamicum  and  B. subtilis  (prokaryotes) and in fungal cells, namely the yeasts  S. cerevisiae  and  Y. lypolytica  (eukaryotes). (See Examples 1 and 2.) 
     Illustrative Engineered Yeast Cells 
     In certain embodiments, the engineered yeast (e.g.,  S. cerevisiae ) cell expresses a heterologous histamine decarboxylase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity to a histamine decarboxylase from  Chromobacterium  sp. LK1 (e.g., SEQ ID NO:6). In particular embodiments, the  Chromobacterium  sp. LK1 histamine decarboxylase can include SEQ ID NO:6. The engineered yeast (e.g.,  S. cerevisiae ) cell can also express a heterologous ATP phosphoribosyltransferase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity with an ATP phosphoribosyltransferase from  S. cerevisiae  (SEQ ID NO:3). In particular embodiments, the  S. cerevisiae  ATP phosphoribosyltransferase includes SEQ ID NO:3. 
     In certain embodiments, the engineered yeast (e.g.,  Y. lipolytica ) cell expresses a heterologous histamine decarboxylase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity to a histamine decarboxylase from  Acinetobacter baumannii  strain AB0057 (e.g., SEQ ID NO:1). In particular embodiments, the  Acinetobacter baumannii  strain AB0057 histamine decarboxylase can include SEQ ID NO:1. The engineered yeast (e.g.,  Y. lipolytica ) cell can also express a heterologous ATP phosphoribosyltransferase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity with an ATP phosphoribosyltransferase from  S. cerevisiae  S288c (SEQ ID NO:3). In particular embodiments, the  S. cerevisiae  S288c ATP phosphoribosyltransferase includes SEQ ID NO:3. 
     These may be the only genetic alterations of the engineered yeast cell, or the yeast cell can include one or more additional genetic alterations, as discussed more generally above. 
     For example, in particular embodiments, the engineered yeast  S. cerevisiae  cell described above additionally expresses a feedback deregulated variant of a  C. glutamicum  ATP phosphoribosyltransferase, which typically has at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, or 95 percent amino acid sequence identity to a variant of a  C. glutamicum  ATP phosphoribosyltransferase containing the amino acid substitutions N215K, L231F, and T235A (SEQ ID NO:7) In particular embodiments, the  C. glutamicum  ATP phosphoribosyltransferase variant can include SEQ ID NO:7. 
     Illustrative Engineered Bacterial Cells 
     In certain embodiments, the engineered bacterial (e.g.,  C. glutamicum ) cell expresses a heterologous histamine decarboxylase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity to a histamine decarboxylase from  Acinetobacter baumannii  (e.g., SEQ ID NO:1). In particular embodiments, the  Acinetobacter baumannii  histamine decarboxylase can include SEQ ID NO:1. The engineered bacterial (e.g.,  C. glutamicum ) cell can also express a heterologous ATP phosphoribosyltransferase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity with an ATP phosphoribosyltransferase from  Saccharomyces cerevisiae  S288c (SEQ ID NO:3). In particular embodiments, the  S. cerevisiae  S288c ATP phosphoribosyltransferase includes SEQ ID NO:3. In some embodiments, the engineered bacterial (e.g.,  C. glutamicum ) cell expresses, instead of the ATP phosphoribosyltransferase, an imidazole-glycerol phosphate dehydratase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity to an imidazole-glycerol phosphate dehydratase from  C. glutamicum  (SEQ ID NO:2). In particular embodiments, the  C. glutamicum  imidazole-glycerol phosphate dehydratase includes SEQ ID NO:2. 
     In certain embodiments, the engineered bacterial (e.g.,  B. subtilis ) cell expresses a heterologous histamine decarboxylase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity to a histamine decarboxylase from  Lactobacillus  sp. (strain 30a) (e.g., SEQ ID NO:4). In particular embodiments, the  Lactobacillus  sp. (strain 30a) histamine decarboxylase can include SEQ ID NO:4. The engineered bacterial (e.g.,  B. subtilis ) cell can also express a heterologous ATP phosphoribosyltransferase having at least 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent or 100 percent amino acid sequence identity with an ATP phosphoribosyltransferase from  Salmonella typhimurium  LT2 (SEQ ID NO:5). In particular embodiments, the  Salmonella typhimurium  LT2 ATP phosphoribosyltransferase includes SEQ ID NO:5. 
     Culturing of Engineered Microbial Cells 
     Any of the microbial cells described herein can be cultured, e.g., for maintenance, growth, and/or histamine production. 
     In some embodiments, the cultures are grown to an optical density at 600 nm of 10-500, such as an optical density of 50-150. 
     In various embodiments, the cultures include produced histamine at titers of at least 10, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, or 900 μg/L, or at least 1, 10, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg/L or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 20, 50 g/L. In various embodiments, the titer is in the range of 10 μg/L to 10 g/L, 25 μg/L to 20 g/L, 100 μg/L to 10 g/L, 200 μg/L to 5 g/L, 500 μg/L to 4 g/L, 1 mg/L to 3 g/L, 500 mg/L to 2 g/L or any range bounded by any of the values listed above. 
     Culture Media 
     Microbial cells can be cultured in any suitable medium including, but not limited to, a minimal medium, i.e., one containing the minimum nutrients possible for cell growth. Minimal medium typically contains: (1) a carbon source for microbial growth; (2) salts, which may depend on the particular microbial cell and growing conditions; and (3) water. Suitable media can also include any combination of the following: a nitrogen source for growth and product formation, a sulfur source for growth, a phosphate source for growth, metal salts for growth, vitamins for growth, and other cofactors for growth. 
     Any suitable carbon source can be used to cultivate the host cells. The term “carbon source” refers to one or more carbon-containing compounds capable of being metabolized by a microbial cell. In various embodiments, the carbon source is a carbohydrate (such as a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide), or an invert sugar (e.g., enzymatically treated sucrose syrup). Illustrative monosaccharides include glucose (dextrose), fructose (levulose), and galactose; illustrative oligosaccharides include dextran or glucan, and illustrative polysaccharides include starch and cellulose. Suitable sugars include C6 sugars (e.g., fructose, mannose, galactose, or glucose) and C5 sugars (e.g., xylose or arabinose). Other, less expensive carbon sources include sugar cane juice, beet juice, sorghum juice, and the like, any of which may, but need not be, fully or partially deionized. 
     The salts in a culture medium generally provide essential elements, such as magnesium, nitrogen, phosphorus, and sulfur to allow the cells to synthesize proteins and nucleic acids. 
     Minimal medium can be supplemented with one or more selective agents, such as antibiotics. 
     To produce histamine, the culture medium can include, and/or is supplemented during culture with, glucose and/or a nitrogen source such as urea, an ammonium salt, ammonia, or any combination thereof. 
     Culture Conditions 
     Materials and methods suitable for the maintenance and growth of microbial cells are well known in the art. See, for example, U.S. Pub. Nos. 2009/0203102, 2010/0003716, and 2010/0048964, and International Pub. Nos. WO 2004/033646, WO 2009/076676, WO 2009/132220, and WO 2010/003007, Manual of Methods for General Bacteriology Gerhardt et al., eds), American Society for Microbiology, Washington, D.C. (1994) or Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc., Sunderland, Mass. 
     In general, cells are grown and maintained at an appropriate temperature, gas mixture, and pH (such as about 20° C. to about 37° C., about 6% to about 84% CO 2 , and a pH between about 5 to about 9). In some aspects, cells are grown at 35° C. In certain embodiments, such as where thermophilic bacteria are used as the host cells, higher temperatures (e.g., 50° C.-75° C.) may be used. In some aspects, the pH ranges for fermentation are between about pH 5.0 to about pH 9.0 (such as about pH 6.0 to about pH 8.0 or about 6.5 to about 7.0). Cells can be grown under aerobic, anoxic, or anaerobic conditions based on the requirements of the particular cell. 
     Standard culture conditions and modes of fermentation, such as batch, fed-batch, or continuous fermentation that can be used are described in U.S. Publ. Nos. 2009/0203102, 2010/0003716, and 2010/0048964, and International Pub. Nos. WO 2009/076676, WO 2009/132220, and WO 2010/003007. Batch and Fed-Batch fermentations are common and well known in the art, and examples can be found in Brock, Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc. 
     In some embodiments, the cells are cultured under limited sugar (e.g., glucose) conditions. In various embodiments, the amount of sugar that is added is less than or about 105% (such as about 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10%) of the amount of sugar that can be consumed by the cells. In particular embodiments, the amount of sugar that is added to the culture medium is approximately the same as the amount of sugar that is consumed by the cells during a specific period of time. In some embodiments, the rate of cell growth is controlled by limiting the amount of added sugar such that the cells grow at the rate that can be supported by the amount of sugar in the cell medium. In some embodiments, sugar does not accumulate during the time the cells are cultured. In various embodiments, the cells are cultured under limited sugar conditions for times greater than or about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, or 70 hours or even up to about 5-10 days. In various embodiments, the cells are cultured under limited sugar conditions for greater than or about 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 95, or 100% of the total length of time the cells are cultured. While not intending to be bound by any particular theory, it is believed that limited sugar conditions can allow more favorable regulation of the cells. 
     In some aspects, the cells are grown in batch culture. The cells can also be grown in fed-batch culture or in continuous culture. Additionally, the cells can be cultured in minimal medium, including, but not limited to, any of the minimal media described above. The minimal medium can be further supplemented with 1.0% (w/v) glucose (or any other six-carbon sugar) or less. Specifically, the minimal medium can be supplemented with 1% (w/v), 0.9% (w/v), 0.8% (w/v), 0.7% (w/v), 0.6% (w/v), 0.5% (w/v), 0.4% (w/v), 0.3% (w/v), 0.2% (w/v), or 0.1% (w/v) glucose. In some cultures, significantly higher levels of sugar (e.g., glucose) are used, e.g., at least 10% (w/v), 20% (w/v), 30% (w/v), 40% (w/v), 50% (w/v), 60% (w/v), 70% (w/v), or up to the solubility limit for the sugar in the medium. In some embodiments, the sugar levels falls within a range of any two of the above values, e.g.: 0.1-10% (w/v), 1.0-20% (w/v), 10-70% (w/v), 20-60% (w/v), or 30-50% (w/v). Furthermore, different sugar levels can be used for different phases of culturing. For fed-batch culture (e.g., of  S. cerevisiae  or  C. glutamicum ), the sugar level can be about 100-200 g/L (10-20% (w/v)) in the batch phase and then up to about 500-700 g/L (50-70% in the feed). 
     Additionally, the minimal medium can be supplemented 0.1% (w/v) or less yeast extract. Specifically, the minimal medium can be supplemented with 0.1% (w/v), 0.09% (w/v), 0.08% (w/v), 0.07% (w/v), 0.06% (w/v), 0.05% (w/v), 0.04% (w/v), 0.03% (w/v), 0.02% (w/v), or 0.01% (w/v) yeast extract. Alternatively, the minimal medium can be supplemented with 1% (w/v), 0.9% (w/v), 0.8% (w/v), 0.7% (w/v), 0.6% (w/v), 0.5% (w/v), 0.4% (w/v), 0.3% (w/v), 0.2% (w/v), or 0.1% (w/v) glucose and with 0.1% (w/v), 0.09% (w/v), 0.08% (w/v), 0.07% (w/v), 0.06% (w/v), 0.05% (w/v), 0.04% (w/v), 0.03% (w/v), or 0.02% (w/v) yeast extract. In some cultures, significantly higher levels of yeast extract can be used, e.g., at least 1.5% (w/v), 2.0% (w/v), 2.5% (w/v), or 3% (w/v). In some cultures (e.g., of  S. cerevisiae  or  C. glutamicum ), the yeast extract level falls within a range of any two of the above values, e.g.: 0.5-3.0% (w/v), 1.0-2.5% (w/v), or 1.5-2.0% (w/v). 
     Illustrative materials and methods suitable for the maintenance and growth of the engineered microbial cells described herein can be found below in Example 1. 
     Histamine Production and Recovery 
     Any of the methods described herein may further include a step of recovering histamine. In some embodiments, the produced histamine contained in a so-called harvest stream is recovered/harvested from the production vessel. The harvest stream may include, for instance, cell-free or cell-containing aqueous solution coming from the production vessel, which contains histamine as a result of the conversion of production substrate by the resting cells in the production vessel. Cells still present in the harvest stream may be separated from the histamine by any operations known in the art, such as for instance filtration, centrifugation, decantation, membrane crossflow ultrafiltration or microfiltration, tangential flow ultrafiltration or microfiltration or dead end filtration. After this cell separation operation, the harvest stream is essentially free of cells. 
     Further steps of separation and/or purification of the produced histamine from other components contained in the harvest stream, i.e., so-called downstream processing steps may optionally be carried out. These steps may include any means known to a skilled person, such as, for instance, concentration, extraction, crystallization, precipitation, adsorption, ion exchange, and/or chromatography. Any of these procedures can be used alone or in combination to purify histamine. Further purification steps can include one or more of, e.g., concentration, crystallization, precipitation, washing and drying, treatment with activated carbon, ion exchange, nanofiltration, and/or re-crystallization. The design of a suitable purification protocol may depend on the cells, the culture medium, the size of the culture, the production vessel, etc. and is within the level of skill in the art. 
     The following examples are given for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the present disclosure in any fashion. Changes therein and other uses which are encompassed within the spirit of the disclosure, as defined by the scope of the claims, will be identifiable to those skilled in the art. 
     Example 1—Construction and Selection of Strains of  Corynebacteria glutamicum  and  Saccharomyces cerevisiae  Engineered to Produce Histamine 
     Plasmid/DNA Design 
     All strains tested for this work were transformed with plasmid DNA designed using proprietary software. Plasmid designs were specific to each of the host organisms engineered in this work. The plasmid DNA was physically constructed by a standard DNA assembly method. This plasmid DNA was then used to integrate metabolic pathway inserts by one of two host-specific methods, each described below. 
       C. glutamicum  Pathway Integration 
     A “loop-in, single-crossover” genomic integration strategy has been developed to engineer  C. glutamicum  strains.  FIG. 14  illustrates genomic integration of loop-in only and loop-in/loop-out constructs and verification of correct integration via colony PCR. Loop-in only constructs (shown under the heading “Loop-in”) contained a single 2-kb homology arm (denoted as “integration locus”), a positive selection marker (denoted as “Marker”)), and gene(s) of interest (denoted as “promoter-gene-terminator”). A single crossover event integrated the plasmid into the  C. glutamicum  chromosome. Integration events are stably maintained in the genome by growth in the presence of antibiotic (25p g/ml kanamycin). Correct genomic integration in colonies derived from loop-in integration were confirmed by colony PCR with UF/IR and DR/IF PCR primers. 
     Loop-in, loop-out constructs (shown under the heading “Loop-in, loop-out) contained two 2-kb homology arms (5′ and 3′ arms), gene(s) of interest (arrows), a positive selection marker (denoted “Marker”), and a counter-selection marker. Similar to “loop-in” only constructs, a single crossover event integrated the plasmid into the chromosome of  C. glutamicum . Note: only one of two possible integrations is shown here. Correct genomic integration was confirmed by colony PCR and counter-selection was applied so that the plasmid backbone and counter-selection marker could be excised. This results in one of two possibilities: reversion to wild-type (lower left box) or the desired pathway integration (lower right box). Again, correct genomic loop-out is confirmed by colony PCR. (Abbreviations: Primers: UF=upstream forward, DR=downstream reverse, IR=internal reverse, IF=internal forward.) 
       S. cerevisiae  Pathway Integration 
     A “split-marker, double-crossover” genomic integration strategy has been developed to engineer  S. cerevisiae  strains.  FIG. 11  illustrates genomic integration of complementary, split-marker plasmids and verification of correct genomic integration via colony PCR in  S. cerevisiae . Two plasmids with complementary 5′ and 3′ homology arms and overlapping halves of a URA3 selectable marker (direct repeats shown by the hashed bars) were digested with meganucleases and transformed as linear fragments. A triple-crossover event integrated the desired heterologous genes into the targeted locus and re-constituted the full URA3 gene. Colonies derived from this integration event were assayed using two 3-primer reactions to confirm both the 5′ and 3′ junctions (UF/IF/wt-R and DR/IF/wt-F). For strains in which further engineering is desired, the strains can be plated on 5-FOA plates to select for the removal of URA3, leaving behind a small single copy of the original direct repeat. This genomic integration strategy can be used for gene knock-out, gene knock-in, and promoter titration in the same workflow. 
     Cell Culture 
     The workflow established for  S. cerevisiae  involved a hit-picking step that consolidated successfully built strains using an automated workflow that randomized strains across the plate. For each strain that was successfully built, up to four replicates were tested from distinct colonies to test colony-to-colony variation and other process variation. If fewer than four colonies were obtained, the existing colonies were replicated so that at least four wells were tested from each desired genotype. 
     The colonies were consolidated into 96-well plates with selective medium (SD-ura for  S. cerevisiae ) and cultivated for two days until saturation and then frozen with 16.6% glycerol at −80° C. for storage. The frozen glycerol stocks were then used to inoculate a seed stage in minimal media with a low level of amino acids to help with growth and recovery from freezing. The seed plates were grown at 30° C. for 1-2 days. The seed plates were then used to inoculate a main cultivation plate with minimal medium and grown for 48-88 hours. Plates were removed at the desired time points and tested for cell density (OD600), viability and glucose, supernatant samples stored for LC-MS analysis for product of interest. 
     Cell Density 
     Cell density was measured using a spectrophotometric assay detecting absorbance of each well at 600 nm. Robotics were used to transfer fixed amounts of culture from each cultivation plate into an assay plate, followed by mixing with 175 mM sodium phosphate (pH 7.0) to generate a 10-fold dilution. The assay plates were measured using a Tecan M1000 spectrophotometer and assay data uploaded to a LIMS database. A non-inoculated control was used to subtract background absorbance. Cell growth was monitored by inoculating multiple plates at each stage, and then sacrificing an entire plate at each time point. 
     To minimize settling of cells while handling large number of plates (which could result in a non-representative sample during measurement) each plate was shaken for 10-15 seconds before each read. Wide variations in cell density within a plate may also lead to absorbance measurements outside of the linear range of detection, resulting in underestimate of higher OD cultures. In general, the tested strains so far have not varied significantly enough for this be a concern. 
     Liquid-Solid Separation 
     To harvest extracellular samples for analysis by LC-MS, liquid and solid phases were separated via centrifugation. Cultivation plates were centrifuged at 2000 rpm for 4 minutes, and the supernatant was transferred to destination plates using robotics. 75p L of supernatant was transferred to each plate, with one stored at 4° C., and the second stored at 80° C. for long-term storage. 
     First-Round Genetic Engineering Results in  Corynebacteria glutamicum  and  Saccharomyces cerevisiae    
     A library approach was taken to screen heterologous pathway enzymes to establish the histamine pathway. For histidine decarboxylase, 18 heterologous sequences were tested from Bacteria, Archaea, Viridiplantae, Vertebrata, Metazoa, and Arthropoda sources listed in Table 1. The histidine decarboxylases were codon-optimized and expressed in both  Saccharomyces cerevisiae  and  Corynebacteria glutamicum  hosts. 
     Histidine biosynthesis is subject to feedback inhibition, therefore a feedback deregulated ATP phosphoribosyltransferase was tested with the histidine decarboxylases to improve production of histidine, the substrate for histidine decarboxylase. The ATP phosphoribosyltransferases tested were from  Salmonella typhimurium  and  Corynebacteria glutamicum , harboring known deletions and point mutations that render them resistant to feedback inhibition. 
     First-round genetic engineering results are shown in Table 1 and  FIGS. 2  ( C. glutamicum ) and  3  ( S. cerevisiae ). 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 First-round genetic engineering results in  Corynebacteria glutamicum  and  Saccharomyces cerevisiae   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 E1  
                   
                   
                   
                   
                 E2  
               
               
                   
                   
                   
                 Enzyme  
                 Enzyme  
                 Codon 
                   
                 Enzyme  
                   
                 Enzyme  
                 Codon 
               
               
                   
                   
                 E1 
                 1— 
                 1— 
                 Optimi- 
                 E2 
                 2— 
                 E2 
                 2— 
                 Optimi- 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity  
                 source 
                 zation 
                 Uniprot 
                 activity  
                 Modifi- 
                 source 
                 zation 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 organism 
                 Abbrev. 
                 ID 
                 name 
                 cations 
                 organism 
                 Abbrev. 
               
               
                   
               
            
           
           
               
            
               
                 
                   Corynebacterium 
                   glutamicum 
                 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 CgHISM 
                 985.0 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                   
                   
                   
                   
                   
               
               
                 N_06 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 695.7 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                   
                   
                   
                   
                   
               
               
                 N_07 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 385.0 
                 P00862 
                 histidine 
                   Lactobacillus  sp. 
                 Cg 
                   
                   
                   
                   
                   
               
               
                 N_08 
                   
                   
                 decarboxylase 
                 (strain 30a) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 14370.2 
                 B71459 
                 histidine 
                 
                   Acinetobacter 
                 
                 Cg 
                   
                   
                   
                   
                   
               
               
                 N_11 
                   
                   
                 decarboxylase 
                 
                   baumannii 
                 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 401.1 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                   
                   
                   
                   
                   
               
               
                 N_13 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 7529.8 
                 P00862 
                 histidine 
                 
                   Lactobacillus 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_15 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 4.0 
                 P00862 
                 histidine 
                 
                   Lactobacillus 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_16 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 3.9 
                 P23738 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_17 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 75.0 
                 Q05733 
                 histidine 
                 
                   Drosophila 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_19 
                   
                   
                 decarboxylase 
                 
                   melanogaster 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 3.8 
                 J6KM89 
                 histidine 
                 
                   Chromo- 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_24 
                   
                   
                 decarboxylase 
                 
                   bacterium 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 1.7 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_25 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 458.5 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_26 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 462.1 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_27 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 1258.2 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_28 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISM 
                 2126.4 
                 P00862 
                 histidine 
                   Lactobacillus  sp. 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_30 
                   
                   
                 decarboxylase 
                 (strain 30a) 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 234.7 
                 Q05733 
                 histidine 
                 
                   Drosophila 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_33 
                   
                   
                 decarboxylase 
                 
                   melanogaster 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 11905.3 
                 B71459 
                 histidine 
                 
                   Acinetobacter 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_35 
                   
                   
                 decarboxylase 
                   baumannii  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 CgHISM 
                 615.0 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_39 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                   
                   
                   
               
            
           
           
               
            
               
                 
                   Saccharomyces 
                   cerevisiae 
                 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 ScHISM 
                 36145.0 
                 P00862 
                 histidine 
                 
                   Lactobacillus  
                 
                 Sc 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_16 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 L231F,  
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 2369.9 
                 P54772 
                 histidine 
                 
                   Solanum 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_17 
                   
                   
                 decarboxylase 
                 
                   lycopersicum 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 1747.7 
                 P23738 
                 histidine 
                 
                   Mus musculus 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_18 
                   
                   
                 decarboxylase 
                   
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 2432.4 
                 Q05733 
                 histidine 
                 
                   Drosophila 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_20 
                   
                   
                 decarboxylase 
                 
                   melanogaster 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 43606.9 
                 J6KM89 
                 histidine 
                 
                   Chromo- 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_21 
                   
                   
                 decarboxylase 
                 
                   bacterium 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 43021.9 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Sc 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_22 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 L231F,  
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                 T235A 
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 36145.8 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_23 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 ScHISM 
                 47208.0 
                 P00862 
                 histidine 
                 
                   Lactobacillus 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_24 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 3130.1 
                 P23738 
                 histidine 
                 
                   Mus musculus 
                 
                 Cg 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_25 
                   
                   
                 decarboxylase 
                   
                   
                   
                 phosphoribosyl- 
                 L231F,  
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                 T235A 
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 3262.5 
                 Q05733 
                 histidine 
                 
                   Drosophila 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_26 
                   
                   
                 decarboxylase 
                 
                   melanogaster 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 90811.0 
                 J6KM89 
                 histidine 
                 
                   Chromo- 
                 
                 Cg 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_28 
                   
                   
                 decarboxylase 
                 
                   bacterium 
                 
                   
                   
                 phosphoribosyl- 
                 L231F, T235A 
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 42708.8 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_29 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 27660.1 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_30 
                   
                   
                 decarboxylase 
                 
                   barkeri  
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 Fusaro / 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 ScHISM 
                 33356.6 
                 P00862 
                 histidine 
                 
                   Lactobacillus  
                 
                 Sc 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_31 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 L231F,  
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 711.5 
                 P54772 
                 histidine 
                 
                   Solanum 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_32 
                   
                   
                 decarboxylase 
                 
                   lycopersicum 
                 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 1523.1 
                 P23738 
                 histidine 
                 
                   Mus musculus 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_33 
                   
                   
                 decarboxylase 
                   
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 transferase 
                   
                   
                   
               
               
                 ScHISM 
                 43170.7 
                 E3QMN8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Sc 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 Sc 
               
               
                 N_37 
                   
                   
                 decarboxylase 
                   barkeri  str. 
                   
                   
                 phosphoribosyl- 
                 L231F,  
                 
                   bacterium 
                 
                   
               
               
                   
                   
                   
                   
                 Wiesmoor 
                   
                   
                 transferase 
                 T235A 
                 
                   glutamicum 
                 
                   
               
               
                 ScHISM 
                 30675.5 
                 Q467R8 
                 histidine 
                 
                   Methanosarcina 
                 
                 Sc 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Sc 
               
               
                 N_38 
                   
                   
                 decarboxylase 
                   barkeri  (strain 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 Fusaro /  
                   
                   
                 transferase 
                   
                   
                   
               
               
                   
                   
                   
                   
                 DSM 804) 
                   
                   
                   
                   
                   
                   
               
               
                 ScHISM 
                 38293.2 
                 P00862 
                 histidine 
                 
                   Lactobacillus  
                 
                 Cg 
                 P00499 
                 ATP 
                 Deletion of 
                 
                   Salmonella  
                 
                 Cg 
               
               
                 N_39 
                   
                   
                 decarboxylase 
                 sp. 
                   
                   
                 phosphoribosyl- 
                 Q207-E208 
                 
                   typhimurium 
                 
                   
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
               
               
                   
               
               
                 Note: 
               
               
                 “Cg” refers to codon optimization for  Corynebacterium   glutamicum ; “Sc” refers to codon optimization for  Saccharomyces   cerevisiae . 
               
            
           
         
       
     
     Second-Round Genetic Engineering Results in  Corynebacteria glutamicum  and  Saccharomyces cerevisiae    
     A library approach was taken to improve histamine production by separately expressing each upstream pathway enzyme with a constitutive promoter to screen for the rate-limiting step. The histidine pathway enzymes screened are listed in Table 2. In addition, the enzymes in Table 2, the strains contained the best enzymes from first round: the  Corynebacteria glutamicum  host contains histidine decarboxylase (UniProt ID B7I459) (SEQ ID NO: 1) and ATP phosphoribosyltransferase (UniProt ID P00499) (SEQ ID NO: 5) containing the deletion Q207-E208, and the  Saccharomyces cerevisiae  host contains histidine decarboxylase (UniProt ID J6KM89)(SEQ ID NO: 6) and ATP phosphoribosyltransferase (UniProt ID Q9Z472) (SEQ ID NO: 7) containing the amino acid substitutions N215K, L231F and T235A. 
     Second-round genetic engineering results are shown in Table 2 and  FIGS. 4  ( C. glutamicum ) and  5  ( S. cerevisiae ). 
     In  C. glutamicum , a titer of 24 mg/L was achieved after two rounds of engineering from the integration of two genes: a histidine decarboxylase gene from  Acinetobacter baumannii , and constitutive expression of an imidazoleglycerol-phosphate dehydratase from  C. glutamicum.    
     In  S. cerevisiae , a titer of 385 mg/L was achieved in two rounds of engineering from the integration of three genes: a histidine decarboxylase gene from  Chromobacterium  sp. LK1 (SEQ ID NO: 6), an ATP phosphoribosyltransferase from  C. glutamicum  containing the amino acid substitutions N215K, L231F, and T235A (SEQ ID NO: 7), and a constitutively expressed ATP phosphoribosyltransferase from  S. cerevisiae  (SEQ ID NO: 3). 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Second-round genetic engineering results in genetic engineering results in 
               
               
                 Corynebacteria glutamicum and Saccharomyces cerevisiae 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 E1 Codon 
               
               
                   
                 Titer 
                 E1 
                   
                 Enzyme 1- 
                 Optimization 
               
               
                 Strain name 
                 (μg/L) 
                 Uniprot ID 
                 Enzyme 1-activity name 
                 source organism 
                 Abbrev. 
               
               
                   
               
               
                 Corynebacteria 
                   
                   
                   
                   
                   
               
               
                 glutamicum 
                   
                   
                   
                   
                   
               
               
                 CgHISMN_41 
                 13702.1 
                 O68602 
                 1-(5-phosphoribosyl)5[(5- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphoribosylamino) 
                 glutamicum 
                   
               
               
                   
                   
                   
                 methylideneamino] 
                   
                   
               
               
                   
                   
                   
                 imidazole-4- 
                   
                   
               
               
                   
                   
                   
                 carboxamide isomerase 
                   
                   
               
               
                 CgHISMN_42 
                 12671.2 
                 Q9KJU3 
                 Imidazoleglycerol- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 glutamicum 
                   
               
               
                 CgHISMN_43 
                 11800.4 
                 Q9KJU4 
                 Histidinol-phosphate 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 aminotransferase 
                 glutamicum 
                   
               
               
                 CgHISMN_44 
                 8667.2 
                 Q8NNT5 
                 Histidinol dehydrogenase 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                   
                 glutamicum 
                   
               
               
                 CgHISMN_45 
                 12375.3 
                 Q9Z471 
                 Phosphoribosyl- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 ATP pyrophosphatase 
                 glutamicum 
                   
               
               
                 CgHISMN_46 
                 10963.6 
                 O31139 
                 Imidazole glycerol 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate synthase subunit 
                 glutamicum 
                   
               
               
                 CgHISMN_47 
                 16246.0 
                 O69043 
                 Imidazole glycerol 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate synthase subunit 
                 glutamicum 
                   
               
               
                 CgHISMN_48 
                 13038.8 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                   
                 glutamicum 
                   
               
               
                 CgHISMN_49 
                 10749.0 
                 Q8NNT9 
                 phosphoribosyl- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 AMP cyclohydrolase 
                 glutamicum 
                   
               
               
                 CgHISMN_50 
                 12960.8 
                 O68602 
                 1-(5-phosphoribosyl)5[(5- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphoribosylamino) 
                 glutamicum 
                   
               
               
                   
                   
                   
                 methylideneamino] 
                   
                   
               
               
                   
                   
                   
                 imidazole-4- 
                   
                   
               
               
                   
                   
                   
                 carboxamide isomerase 
                   
                   
               
               
                 CgHISMN_51 
                 9958.4 
                 Q9KJU3 
                 Imidazoleglycerol- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 glutamicum 
                   
               
               
                 CgHISMN_52 
                 18963.0 
                 Q9KJU4 
                 Histidinol-phosphate 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 aminotransferase 
                 glutamicum 
                   
               
               
                 CgHISMN_53 
                 20328.9 
                 Q8NNT5 
                 Histidinol dehydrogenase 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                   
                 glutamicum 
                   
               
               
                 CgHISMN_54 
                 20051.4 
                 O31139 
                 Imidazole glycerol phosphate 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 synthase subunit 
                 glutamicum 
                   
               
               
                 CgHISMN_55 
                 15070.9 
                 O69043 
                 Imidazole glycerol phosphate 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 synthase subunit 
                 glutamicum 
                   
               
               
                 CgHISMN_56 
                 12799.1 
                 O68602 
                 1-(5-phosphoribosyl)5[(5- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphoribosylamino) 
                 glutamicum 
                   
               
               
                   
                   
                   
                 methylideneamino] 
                   
                   
               
               
                   
                   
                   
                 imidazole-4- 
                   
                   
               
               
                   
                   
                   
                 carboxamide isomerase 
                   
                   
               
               
                 CgHISMN_57 
                 24773.6 
                 Q9KJU3 
                 Imidazoleglycerol- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 glutamicum 
                   
               
               
                 CgHISMN_58 
                 15268.6 
                 Q9KJU4 
                 Histidinol-phosphate 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 aminotransferase 
                 glutamicum 
                   
               
               
                 CgHISMN_59 
                 12555.0 
                 Q8NNT5 
                 Histidinol dehydrogenase 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                   
                 glutamicum 
                   
               
               
                 CgHISMN_60 
                 17725.6 
                 Q9Z471 
                 Phosphoribosyl- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 ATP pyrophosphatase 
                 glutamicum 
                   
               
               
                 CgHISMN_61 
                 18777.4 
                 O69043 
                 Imidazole glycerol 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 phosphate synthase subunit 
                 glutamicum 
                   
               
               
                 CgHISMN_62 
                 19782.8 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                   
                 glutamicum 
                   
               
               
                 CgHISMN_63 
                 15092.7 
                 Q8NNT9 
                 phosphoribosyl- 
                 Corynebacterium 
                 Native 
               
               
                   
                   
                   
                 AMP cyclohydrolase 
                 glutamicum 
                   
               
               
                 Saccharomyces 
                   
                   
                   
                   
                   
               
               
                 cerevisiae 
                   
                   
                   
                   
                   
               
               
                 ScHISMN_41 
                 385518.2 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_42 
                 70003.1 
                 P00815 
                 histidinol dehydrogenase, 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphoribosyl-AMP cyclohydrolase, 
                 cerevisiae 
                   
               
               
                   
                   
                   
                 phosphoribosyl-ATP diphosphatase 
                   
                   
               
               
                 ScHISMN_43 
                 75039.5 
                 P33734 
                 Imidazole glycerol phosphate 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 synthase subunit HisF 
                 cerevisiae 
                   
               
               
                 ScHISMN_44 
                 71402.5 
                 P07172 
                 histidinol-phosphate transaminase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_46 
                 64866.5 
                 P06633 
                 Imidazoleglycerol- 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 cerevisiae 
                   
               
               
                 ScHISMN_48 
                 113026.6 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_49 
                 79488.5 
                 P00815 
                 histidinol dehydrogenase, 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphoribosyl-AMP cyclohydrolase, 
                 cerevisiae 
                   
               
               
                   
                   
                   
                 phosphoribosyl-ATP diphosphatase 
                   
                   
               
               
                 ScHISMN_50 
                 92719.6 
                 P33734 
                 Imidazole glycerol phosphate 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 synthase subunit HisF 
                 cerevisiae 
                   
               
               
                 ScHISMN_51 
                 88847.1 
                 P07172 
                 histidinol-phosphate transaminase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_52 
                 70650.9 
                 P38635 
                 histidinol-phosphatase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_53 
                 74127.8 
                 P06633 
                 Imidazoleglycerol- 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 cerevisiae 
                   
               
               
                 ScHISMN_56 
                 73080.2 
                 P00815 
                 histidinol dehydrogenase, 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphoribosyl-AMP cyclohydrolase, 
                 cerevisiae 
                   
               
               
                   
                   
                   
                 phosphoribosyl-ATP diphosphatase 
                   
                   
               
               
                 ScHISMN_57 
                 78656.1 
                 P33734 
                 Imidazole glycerol phosphate 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 synthase subunit HisF 
                 cerevisiae 
                   
               
               
                 ScHISMN_58 
                 69769.0 
                 P07172 
                 histidinol-phosphate transaminase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_59 
                 59139.1 
                 P38635 
                 histidinol-phosphatase 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                   
                 cerevisiae 
                   
               
               
                 ScHISMN_60 
                 65506.7 
                 P06633 
                 Imidazoleglycerol- 
                 Saccharomyces 
                 Native 
               
               
                   
                   
                   
                 phosphate dehydratase 
                 cerevisiae 
               
               
                   
               
            
           
         
       
     
     Third-Round Genetic Engineering Results in  Saccharomyces cerevisiae    
     Histamine production was further pursued in  S. cerevisiae , and we designed plasmids to integrate additional copies of upstream pathway genes expressed by a strong constitutive promoter to avoid native regulation of a gene (Table 3). An expanded search was undertaken to test additional histidine decarboxylases that have similar sequences to the enzymes initially identified as active (Table 3). 
     In parallel we pursued modulating native gene expression to further improve histamine production. Our engineering approach was to take a best  S. cerevisiae  strain from the second round and test either a strong or weak constitutive promoter in place of the native promoter. Gene targets for promoter changes were selected to redirect flux supply precursors to histidine. Strain designs being tested include designs for increasing pentose phosphate pathway flux by expressing a non-native feedback deregulated glucose-6-phosphate dehydrogenase (zwf) and decreasing the “lower” pentose phosphate pathway flux thru the sugar isomerase enzymes. 
     Promoter replacements for lower expression of genes that are thought to be essential (i.e., cannot be deleted), but were expected to increase the upper glycolysis metabolite pool available for histamine production, targeted: 1) enolase (Eno2), to reduce flux through lower glycolysis, 2) pyruvate dehydrogenase (PDH, Lpd1) for lower flux through the C3/C2 node, and 3) pentose phosphate pathway sugar isomerases, which use the histamine metabolite precursor ribose-5-phosphate (Tall). An illustrative list of promoter-swap (“proswap”) and deletion (“knockout”) targets in  S. cerevisiae  includes: 
     
       
         
           
               
               
               
               
               
             
               
                   
                   
               
               
                   
                 Annotation_name 
                 Type 
                 Promoter_name 
                 Gene_name 
               
               
                   
                   
               
             
            
               
                   
                 YDR380W 
                 knockout 
                   
                 Aro10 
               
               
                   
                 YDL047W 
                 knockout 
                   
                 Sit4 
               
               
                   
                 YML035C 
                 knockout 
                   
                 Amd1 
               
               
                   
                 YMR020W 
                 knockout 
                   
                 Fms1 
               
               
                   
                 YNL229C 
                 knockout 
                   
                 Ure2 
               
               
                   
                 YJL052W 
                 proswap 
                 pRnr1 
                 Tdh1 
               
               
                   
                 YJR009C 
                 proswap 
                 pRnr1 
                 Tdh2 
               
               
                   
                 YGR192C 
                 proswap 
                 pRnr1 
                 Tdh3 
               
               
                   
                 YFL018C 
                 proswap 
                 pRnr1 
                 Lpd1 
               
               
                   
                 YHR174W 
                 proswap 
                 pRnr1 
                 Eno2 
               
               
                   
                 YNR001C 
                 proswap 
                 pRnr1 
                 Cit1 
               
               
                   
                 YCR012W 
                 proswap 
                 pRnr1 
                 Pgk1 
               
               
                   
                 YLR354C 
                 proswap 
                 pRnr1 
                 Tal1 
               
               
                   
                 YBR117C 
                 proswap 
                 pRnr1 
                 Tkl2 
               
               
                   
                 YPR074C 
                 proswap 
                 pRnr1 
                 Tkl1 
               
               
                   
                 YML035C 
                 proswap 
                 pRev1 
                 Amd1 
               
               
                   
                 YHR216W 
                 proswap 
                 pRev1 
                 Imd2 
               
               
                   
                 YOR155C 
                 proswap 
                 pRev1 
                 Isn1 
               
               
                   
                 YNL229C 
                 proswap 
                 pRev1 
                 Ure2 
               
               
                   
                 YER086W 
                 proswap 
                 pRnr1 
                 Ilv1 
               
               
                   
                 YDR380W 
                 proswap 
                 pRnr1 
                 Aro10 
               
               
                   
                 YEL009C 
                 proswap 
                 pRev1 
                 Gcn4 
               
               
                   
                   
               
            
           
         
       
     
     Promoters were selected based on expression data from Lee et al [7]. 
     Additional genetic engineering results for  S. cerevisiae  are shown in Table 3 and  FIG. 10 . The parent strain for the strain designs shown in Table 3 (also the reference strain, ScHISMN_41) contained a histidine decarboxylase (UniProt ID J6KM89) and an ATP phosphoribosyltransferase (UniProt ID Q9Z472) harboring the amino acid substitutions N215K, L231F and T235A, and the ATP phosphoribosyltransferase from  S. cerevisiae . The reference strain had a histamine titer of 131 mg/L. 
     Improved titer was observed in strains that expressed each of the following enzymes from a strong constitutive promoter: 
     1. Transketolase (EC 2.2.1.1) (SEQ ID NO: 27), which catalyzes the interconversion of sugars in the pentose phosphate pathway and produces ribose-5-phosphate, which is a precursor to PPRP, the initial metabolite in the histidine biosynthesis pathway. 
     2. Ribose-phosphate pyrophosphokinase (EC 2.7.6.1) (SEQ ID NO: 28) (highest titer: 191 mg/L relative to control in experiment 131 mg/L). 
     3. ATP phosphoribosyltransferase (EC 2.4.2.17) (SEQ ID NO: 3). 
     4. Trifunctional histidinol dehydrogenase (EC 1.1.1.23)/phosphoribosyl-AMP cyclohydrolase (EC 3.5.4.19)/phosphoribosyl-ATP diphosphatase (EC 3.6.1.31) (SEQ ID NO: 29). 
     5. Histidinol-phosphate aminotransferase (EC 2.6.1.9) (SEQ ID NO: 14). 
     6. 5′ProFAR isomerase (EC 5.3.1.16) (SEQ ID NO: 31). 
     7. Imidazole glycerol phosphate synthase (EC 4.3.1.B2) (SEQ ID NO: 21). 
     8. Triose-phosphate isomerase (EC 5.3.1.1), harboring the amino acid substitutions harboring the amino acid substitutions I170V (SEQ ID NO: 32) or I170T [8]. 
     9. Glucose-6-phosphate 1-dehydrogenase (EC 1.1.1.49), harboring the amino acid substitution A243T (SEQ ID NO: 26). 
     10. Various histidine decarboxylases (EC 4.1.1.22):
         a. UniProt ID A0A089YPE5 (SEQ ID NO: 33)   b. UniProt ID A0A126S6G9 (SEQ ID NO: 34)   c. UniProt ID A0A0A1R6V3 (SEQ ID NO: 35)   d. UniProt ID A0A1W0CM88 (SEQ ID NO: 36)   e. UniProt ID P00862 (SEQ ID NO: 4)   f. UniProt ID A0A0K6GJ74 (SEQ ID NO: 37)   g. UniProt ID T0QL99 (SEQ ID NO: 38)   h. UniProt ID A0A1B8HLR1 (SEQ ID NO: 39)       

     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Third-round genetic engineering results in  Saccharomyces cerevisiae   
               
               
                 Built and tested strain designs: 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 Enzyme  
                 E1 
                 Enzyme  
                   
                 Enzyme  
                 Enzyme 
                   
                 Enzyme  
                 E3 
                 Enzyme  
               
               
                   
                   
                 E1 
                 1— 
                 Modi- 
                 1— 
                 E2 
                 2— 
                 2— 
                 E3 
                 3— 
                 Modi- 
                 3— 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity  
                 fica- 
                 source 
                 Uniprot 
                 activity  
                 source 
                 Uniprot 
                 activity  
                 fica- 
                 source 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 tions 
                 organism 
                 ID 
                 name 
                 organism 
                 ID 
                 name 
                 tions 
                 organism 
               
               
                   
               
               
                 Sc 
                 39059 
                 A0A0C 
                 Histidine 
                   
                 
                   Lactobacillus 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 HISM 
                   
                 1PR48 
                 decarboxy- 
                   
                 
                   fructivorans 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 N_100 
                   
                   
                 lase 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 144871 
                 T0QL99 
                 Histidine 
                   
                 
                   Aeromonas 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 decarboxy- 
                   
                 
                   salmonicida 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 101 
                   
                   
                 lase 
                   
                 subsp. 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 pectinolytica 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 34mel 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 143763 
                 A0A1B8 
                 Histidine 
                   
                 Morganella 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 HLR1 
                 decarboxy- 
                   
                 psychrotoler- 
                   
                   
                   
                   
                   
                   
                   
               
               
                 102 
                   
                   
                 lase 
                   
                 ans 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 155931 
                 Q9Z472 
                 ATP 
                   
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                 P0A717 
                 Ribose- 
                   
                 
                   Escheri- 
                 
               
               
                 MN_ 
                   
                   
                 phosphori- 
                   
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   chia 
                 
               
               
                 l03 
                   
                   
                 bosyl- 
                   
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   coli 
                 
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                 pyro- 
                   
                 (strain 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                 phos- 
                   
                 K12) 
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                 pho- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydrolase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 151846 
                 P0A717 
                 Ribose- 
                   
                 
                   Escherichia 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphate 
                   
                 
                   coli  
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 104 
                   
                   
                 pyrophos- 
                   
                 (strain 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 pho- 
                   
                 K12) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 kinase 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 191110 
                 Q12265 
                 Ribose- 
                   
                 
                   Saccharo- 
                 
                 P38620 
                 Ribose- 
                 
                   Saccharo- 
                 
                 Q680A5 
                 Ribose- 
                   
                 
                   Arabi- 
                 
               
               
                 MN_ 
                   
                   
                 phosphate 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   dopsis 
                 
               
               
                 105 
                   
                   
                 pyropho- 
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   thaliana 
                 
               
               
                   
                   
                   
                 spho- 
                   
                 (strain  
                   
                 phokinase 
                 S288c 
                   
                 pyro- 
                   
                 (Mouse- 
               
               
                   
                   
                   
                 kinase 
                   
                 ATCC 
                   
                   
                   
                   
                 phos- 
                   
                 ear 
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                 pho- 
                   
                 cress) 
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 160586 
                 P32895 
                 Ribose- 
                   
                 
                   Saccharo- 
                 
                 P38689 
                 Ribose- 
                 
                   Saccharo- 
                 
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphate 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                   
                   
                   
               
               
                 106 
                   
                   
                 pyrophos- 
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 
                   cerevisiae 
                 
                   
                   
                   
                   
               
               
                   
                   
                   
                 pho- 
                   
                 (strain  
                   
                 phokinase 
                 S288c 
                   
                   
                   
                   
               
               
                   
                   
                   
                 kinase 
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 157191 
                 P23254 
                 Trans- 
                   
                 
                   Saccharo- 
                 
                 P32895 
                 Ribose- 
                 
                   Saccharo- 
                 
                 P38689 
                 Ribose- 
                   
                 
                   Saccharo- 
                 
               
               
                 MN_ 
                   
                   
                 ketolase 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   myces 
                 
               
               
                 107 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   cerevisiae 
                 
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                 phokinase 
                 S288c 
                   
                 pyro- 
                   
                 S288c 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                 phos- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                 pho- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 168183 
                 P23254 
                 Trans- 
                   
                 
                   Saccharo- 
                 
                 Q12265 
                 Ribose- 
                 
                   Saccharo- 
                 
                 P38620 
                 Ribose- 
                   
                 
                   Saccharo- 
                 
               
               
                 MN_ 
                   
                   
                 ketolase 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   myces 
                 
               
               
                 108 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   cerevisiae 
                 
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                 phokinase 
                 S288c 
                   
                 pyro- 
                   
                 S288c 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                 phos- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                 pho- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 125249 
                 P23254 
                 Trans- 
                   
                 
                   Saccharo- 
                 
                 Q12265 
                 Ribose- 
                 
                   Saccharo- 
                 
                 Q680A5 
                 Ribose- 
                   
                 
                   Arabi- 
                 
               
               
                 MN_ 
                   
                   
                 ketolase 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   dopsis 
                 
               
               
                 109 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   thaliana 
                 
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                 phokinase 
                 S288c 
                   
                 pyro- 
                   
                 (Mouse- 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                 phos- 
                   
                 ear 
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                 pho- 
                   
                 cress) 
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 157653 
                 P23254 
                 Trans- 
                   
                 
                   Saccharo- 
                 
                 P0A717 
                 Ribose- 
                 
                   Escherichia 
                 
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 ketolase 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   coli 
                 
                   
                   
                   
                   
               
               
                 110 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                 pyrophos- 
                 (strain 
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                 phokinase 
                 K12) 
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 136093 
                 P23254 
                 Trans- 
                   
                 
                   Saccharo- 
                 
                 P15019 
                 Trans- 
                 
                   Saccharo- 
                 
                 P0A717 
                 Ribose- 
                   
                 
                   Escheri- 
                 
               
               
                 MN_ 
                   
                   
                 ketolase 
                   
                 
                   myces 
                 
                   
                 aldolase 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   chia 
                 
               
               
                 111 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                   
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   coli 
                 
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                   
                 S288c 
                   
                 pyro- 
                   
                 (strain 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                 phos- 
                   
                 K12) 
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                 pho- 
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                 kinase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                   
                 P06775 
                 Histidine 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 permease 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 112 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 160417 
                 P00815 
                 trifunctional 
                   
                 
                   Saccharo- 
                 
                 P40545 
                 5&#39;ProFAR 
                 
                   Saccharo- 
                 
                 O59667 
                 Bifunc- 
                   
                 Schizo- 
               
               
                 MN_ 
                   
                   
                 histidinol 
                   
                 
                   myces 
                 
                   
                 isomerase 
                 
                   myces 
                 
                   
                 tional 
                   
                 saccharo- 
               
               
                 113 
                   
                   
                 dehydro- 
                   
                 
                   cerevisiae 
                 
                   
                   
                 
                   cerevisiae 
                 
                   
                 phos- 
                   
                 myces 
               
               
                   
                   
                   
                 genase/ 
                   
                 (strain  
                   
                   
                 S288c 
                   
                 phoribo- 
                   
                 pombe 
               
               
                   
                   
                   
                 phos- 
                   
                 ATCC 
                   
                   
                   
                   
                 syl-AMP 
                   
                 ATCC 
               
               
                   
                   
                   
                 phoribo- 
                   
                 204508 / 
                   
                   
                   
                   
                 cyclo- 
                   
                 24843 
               
               
                   
                   
                   
                 syl-AMP 
                   
                 S288c) 
                   
                   
                   
                   
                 hydrolase  
                   
                   
               
               
                   
                   
                   
                 cyclo- 
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                 and 
                   
                   
               
               
                   
                   
                   
                 hydro- 
                   
                 yeast) 
                   
                   
                   
                   
                 phos- 
                   
                   
               
               
                   
                   
                   
                 lase/ 
                   
                   
                   
                   
                   
                   
                 phoribo- 
                   
                   
               
               
                   
                   
                   
                 phos- 
                   
                   
                   
                   
                   
                   
                 syl-ATP 
                   
                   
               
               
                   
                   
                   
                 phoribo- 
                   
                   
                   
                   
                   
                   
                 pyrophos- 
                   
                   
               
               
                   
                   
                   
                 syl-ATP 
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
               
               
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 116907 
                 P06633 
                 Imidazole- 
                   
                 
                   Saccharo- 
                 
                 P07172 
                 Histidinol- 
                 
                   Saccharo- 
                 
                 P38635 
                 Histidi- 
                   
                 
                   Saccharo- 
                 
               
               
                 MN_ 
                   
                   
                 glycerol- 
                   
                 
                   myces 
                 
                   
                 phosphate 
                 
                   myces 
                 
                   
                 nol- 
                   
                 
                   myces 
                 
               
               
                 114 
                   
                   
                 phosphate 
                   
                 
                   cerevisiae 
                 
                   
                 amino- 
                 
                   cerevisiae 
                 
                   
                 phos- 
                   
                 
                   cerevisiae 
                 
               
               
                   
                   
                   
                 dehydratase 
                   
                 (strain  
                   
                 trans- 
                 S288c 
                   
                 phatase 
                   
                 S288c 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 ferase 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 131308 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 41 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 123614 
                 P00815 
                 trifunctional 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 histidinol 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 73 
                   
                   
                 dehydro- 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 genase/ 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phos- 
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phoribo- 
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 syl-AMP 
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 cyclohydro- 
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 lase/ 
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phos- 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phoribo- 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 syl-ATP 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 129393 
                 P06633 
                 Imidazole- 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 glycerol- 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 74 
                   
                   
                 phosphate 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 dehydratase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 151455 
                 P07172 
                 Histidinol- 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphate 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 75 
                   
                   
                 aminotrans- 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 ferase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 138833 
                 P00498 
                 ATP 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphori- 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 76 
                   
                   
                 bosyl- 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 164217 
                 P40545 
                 5′ProFAR 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 isomerase 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 77 
                   
                   
                   
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 159871 
                 P33734 
                 Imidazole 
                   
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 glycerol 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 78 
                   
                   
                 phosphate 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 synthase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 145179 
                 P00942 
                 Triosephos- 
                 I170V 
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phate 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 79 
                   
                   
                 isomerase 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 137192 
                 P00942 
                 Triosephos- 
                 I170T 
                 
                   Saccharo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phate 
                   
                 
                   myces 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 80 
                   
                   
                 isomerase 
                   
                 
                   cerevisiae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 204508 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 S288c) 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 (Baker&#39;s 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 yeast) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 139699 
                 Q9Z472 
                 ATP 
                   
                 
                   Coryne- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphori- 
                   
                 
                   bacterium 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 81 
                   
                   
                 bosyl- 
                   
                 
                   glutamicum 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 / 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 148665 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphori- 
                 L231F, 
                 
                   bacterium 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 82 
                   
                   
                 bosyl- 
                 T235A 
                 
                   glutamicum 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /LMG 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 109350 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 phosphori- 
                 L231F, 
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                   
                   
                   
               
               
                 84 
                   
                   
                 bosyl- 
                 T235A 
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                   
                   
                   
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydro- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 lase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 144154 
                 Q9Z472 
                 ATP 
                   
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                 P00942 
                 Triose- 
                 I170V 
                 
                   Saccharo- 
                 
               
               
                 MN_ 
                   
                   
                 phosphori- 
                   
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   myces 
                 
               
               
                 85 
                   
                   
                 bosyl- 
                   
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   cerevisiae 
                 
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                 iso- 
                   
                 S288c 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                 merase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydro- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 lase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 145171 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                 P00942 
                 Triose- 
                 I170V 
                 
                   Saccharo- 
                 
               
               
                 MN_ 
                   
                   
                 phosphori- 
                 L231F, 
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                 phos- 
                   
                 
                   myces 
                 
               
               
                 86 
                   
                   
                 bosyl- 
                 T235A 
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   cerevisiae 
                 
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                 iso- 
                   
                 S288c 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                 merase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydrolase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 166497 
                 Q9Z472 
                 ATP 
                   
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                 A4QEF2 
                 Glucose- 
                 A243T 
                 
                   Coryne- 
                 
               
               
                 MN_ 
                   
                   
                 phosphori- 
                   
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                 6-phos- 
                   
                 
                   bacterium 
                 
               
               
                 87 
                   
                   
                 bosyl- 
                   
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                 phate 
                   
                 
                   glutami- 
                 
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                 1- 
                   
                 
                   cum 
                 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                 dehydro- 
                   
                 (strain R) 
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                 genase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 / 
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydro- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 lase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 152555 
                 Q9Z472 
                 ATP 
                 N215K, 
                 
                   Coryne- 
                 
                 P00815 
                 tri- 
                 
                   Saccharo- 
                 
                 A4QEF2 
                 Glucose- 
                 A243T 
                 
                   Coryne- 
                 
               
               
                 MN_ 
                   
                   
                 phosphori- 
                 L231F, 
                 
                   bacterium 
                 
                   
                 functional 
                 
                   myces 
                 
                   
                 6-phos- 
                   
                 
                   bacterium 
                 
               
               
                 88 
                   
                   
                 bosyl- 
                 T235A 
                 
                   glutamicum 
                 
                   
                 histidinol 
                 
                   cerevisiae 
                 
                   
                 phate  
                   
                 
                   glutami- 
                 
               
               
                   
                   
                   
                 transferase 
                   
                 (strain  
                   
                 dehydro- 
                 S288c 
                   
                 1- 
                   
                 
                   cum 
                 
               
               
                   
                   
                   
                   
                   
                 ATCC 
                   
                 genase/ 
                   
                   
                 dehydro- 
                   
                 (strain R) 
               
               
                   
                   
                   
                   
                   
                 13032 /  
                   
                 phos- 
                   
                   
                 genase 
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20300 /  
                   
                 syl-AMP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 JCM 
                   
                 cyclo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 1318 /  
                   
                 hydro- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 LMG 
                   
                 lase/ 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 3730 /  
                   
                 phos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 NCIMB 
                   
                 phoribo- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 10025) 
                   
                 syl-ATP 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 diphos- 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 phatase 
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 143866 
                 O66000 
                 Histidine 
                   
                 
                   Oenococcus 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 decarboxy- 
                   
                 
                   oeni 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 89 
                   
                   
                 lase 
                   
                 ( Leuconostoc   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   oenos ) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 124157 
                 A0A0R 
                 Histidine 
                   
                 
                   Lactobacillus 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 1Y874 
                 decarboxy- 
                   
                 
                   aviarius 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 90 
                   
                   
                 lase 
                   
                 subsp. 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 aviarius  
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 DSM 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 20655 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 68849 
                 A0A1H 
                 Histidine 
                 S9R 
                 
                   Pseudo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 1TEB8 
                 decarboxy- 
                   
                 
                   monas 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 92 
                   
                   
                 lase 
                   
                 sp. bs2935 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 157127 
                 A0A089 
                 Histidine 
                   
                 
                   Pseudo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 YPE5 
                 decarboxy- 
                   
                 
                   monas 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 93 
                   
                   
                 lase 
                   
                 
                   rhizosphaerae 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 175497 
                 A0A126 
                 Histidine 
                   
                 
                   Pseudo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 S6G9 
                 decarboxy- 
                   
                 
                   monas putida 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 94 
                   
                   
                 lase 
                   
                 ( Arthrobacter   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 
                   siderocap- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   sulatus ) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 116642 
                 A0A0J6 
                 Histidine 
                   
                 
                   Chromo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 KM89 
                 decarboxy- 
                   
                   bacterium  sp. 
                   
                   
                   
                   
                   
                   
                   
               
               
                 95 
                   
                   
                 lase 
                   
                 LK1 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 171681 
                 A0A0A1 
                 Histidine 
                   
                 
                   Citrobacter 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 R6V3 
                 decarboxy- 
                   
                 
                   pasteurii 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 96 
                   
                   
                 lase 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 171393 
                 A0A1W 
                 Histidine 
                   
                 
                   Chromo- 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 0CM88 
                 decarboxy- 
                   
                 
                   bacterium 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 97 
                   
                   
                 lase 
                   
                 
                   haemolyticum 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 152065 
                 P00862 
                 Histidine 
                   
                 
                   Lactobacillus 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                   
                 decarboxy- 
                   
                 sp. (strain 
                   
                   
                   
                   
                   
                   
                   
               
               
                 98 
                   
                   
                 lase 
                   
                 30a) 
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHIS 
                 148362 
                 A0A0K6 
                 Histidine 
                   
                 
                   Lactobacillus 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 MN_ 
                   
                 GJ74 
                 decarboxy- 
                   
                 
                   reuteri 
                 
                   
                   
                   
                   
                   
                   
                   
               
               
                 99 
                   
                   
                 lase 
               
               
                   
               
               
                 Note: 
               
               
                 E1, E2, and E3 genes were codon-optimized according to modified codon usage for Cg and Sc 
               
            
           
         
       
     
     Example 2—Host Evaluation for Histamine Production 
     Histamine production was also tested in two additional hosts,  Bacillus subtilus  and  Yarrowia lipolytica , which were engineered to express the enzymes from the best-performing  Corynebacterium glutamicum  and  Saccharomyces cerevisiae  strains. 
     Host evaluation designs were selected to express 1-3 enzymes and, each design was tested with four different codon optimizations based on the host organisms  C. glutamicum, S. cerevisiae, B. subtilis , and  Y. lipolytica . The codon optimizations tested were based on the Kazusa codon usage tables tabulated for each host for gene codon optimization (www.kazusa.or.jp/codon/). 
     Histamine production was demonstrated in  Y. lipolytica  ( FIG. 6 ) and  B. subtilis  ( FIG. 7 ) and further improved in  C. glutamicum  ( FIG. 9 ) and  S. cerevisiae  ( FIG. 8 ). 
     In  Y. lipolytica  ( FIG. 6 , Table 4, below) the best performing strain produced 505 mg/L histamine and expressed the histidine decarboxylase from  Acinetobacter baumannii  strain AB0057 (UniProt ID B7I459), where the DNA sequence was codon-optimized for  Y. lipolytica , and the ATP phosphoribosyltransferase from  S. cerevisiae  S288c (UniProt ID P00498), where the DNA sequence was codon optimized for  Y. lipolytica . The same two genes were also tested where the DNA sequence was codon-optimized for  B. subtilis  and  S. cerevisiae  and the resulting strains produced no histamine titer. 
     The second best-performing strain in  Y. lipolytica  also expressed the histidine decarboxylase from  Acinetobacter baumannii  strain AB0057 (UniProt ID B7I459), where the DNA sequence was codon-optimized for  Y. lipolytica , and the ATP phosphoribosyltransferase from  Salmonella typhimurium  LT2 (UniProt ID P00499), where the DNA was codon optimized for  Y. lipolytica . Versions of these two genes were also tested where the DNA sequence was codon optimized for  B. subtilis  (which produced 0 titer), codon-optimized for  S. cerevisiae  (which produced 33 micrograms histamine) and codon-optimized using a combined codon table for  S. cerevisiae  and  C. glutamicum  (produced 97 mg/L histamine). 
     The third best-performing strain in  Y. lipolytica  produced 258 mg/L histamine and expressed the histidine decarboxylase from  Chromobacterium  sp. LK1 (UniProt ID A0A0J6KM89), where the DNA sequence was codon optimized for  Y. lipolytica , and the ATP phosphoribosyltransferase from  C. glutamicum  ATCC 13032 (UniProt ID Q9Z472) harboring the amino acid substitutions N215K, L231F, T235A (SEQ ID NO: 7), where the DNA sequence was codon-optimized for  Y. lipolytica  (SEQ ID NO: 64). Versions of these two genes were also tested where the DNA sequences were codon-optimized for  S. cerevisiae  (SEQ ID NO: 65, 66) or  B. subtilis  (SEQ ID NO: 67, 68), and these  Y. lipolytica  strains produced 1.8 mg/L and 0.3 mg/L, respectively. Accordingly, codon-optimization of genes affects expression in  Y. lipolytica.    
     In  B. subtilis  ( FIG. 7 , Table 5, below) the best performing strain produced 18 mg/L histamine and expressed the histamine decarboxylase from  Lactobacillus  sp. (strain 30a) (UniProt ID P00862)(SEQ ID NO: 4) with the ATP phosphoribosyltransferase from  Salmonella typhimurium  LT2 (UniProt ID P00499)(SEQ ID NO: 5) where the DNA sequence was codon optimized for  Bacillus subtilis  (SEQ ID NO: 69, 59). The same two genes were also tested where the DNA sequence was codon-optimized for  S. cerevisiae  (SEQ ID NO: 70, 60) or modified codon usage table for  C. glutamicum  and  S. cerevisiae  (SEQ ID NO: 71, 62), and these strains produced 6.7 mg/L or 0 mg/L histamine, respectively. 
     The host evaluation designs were also tested in  S. cerevisiae  and  C. glutamicum . In  S. cerevisiae  ( FIG. 8 , Table Table 6, below) the best-performing strain produced 111 mg/L histamine and expressed the histamine decarboxylase from  Chromobacterium  sp. LK1 (UniProt ID A0A0J6KM89)(SEQ ID NO: 51) and the ATP phosphoribosyltransferase from  Saccharomyces cerevisiae  S288c (UniProt ID P00498)(SEQ ID NO: 3), where the DNA sequences were codon-optimized for  Y. lipolytica  (SEQ ID NO: 63, 53). The same two genes were also tested where the DNA sequences were codon optimized for  S. cerevisiae  (SEQ ID NO: 65, 57) and  B. subtilis  (SEQ ID NO: 67, 55) produced 86 mg/L and 101 mg/L, respectively. 
     In  C. glutamicum  ( FIG. 9 , Table 7), the best-performing strain produced 68 mg/L histamine and expressed the histamine decarboxylase from  Acinetobacter baumannii  (strain AB0057) (UniProt ID B7I459) (SEQ ID NO: 1) with the ATP phosphoribosyltransferase from  Saccharomyces cerevisiae  S288c (UniProt ID P00498) (SEQ ID NO: 3) where the DNA sequences were codon-optimized using a modified codon usage table for  C. glutamicum  and  S. cerevisiae  (SEQ ID NO: 72, 73). The same two genes were also tested where the DNA sequence was codon-optimized for  Y. lipolytica  (SEQ ID NO: 52, 53) or  S. cerevisiae  (SEQ ID NO: 56, 57), and these strains produced 16 mg/L and 18 microgram/L histamine, respectively. 
     The second best-performing strain in  C. glutamicum  produced 15 mg/L histamine and also expressed a histidine decarboxylase from  Acinetobacter baumannii  strain AB0057 (UniProt ID B7I459) (SEQ ID NO: 1), where the DNA sequence was codon optimized for  Y. lipolytica  (SEQ ID NO: 52), and an ATP phosphoribosyltransferase from  Salmonella typhimurium  LT2 (UniProt ID P00499) (SEQ ID NO: 5), where the DNA was codon optimized for  Y. lipolytica  (SEQ ID NO: 58). These same two genes were also tested, where the DNA sequences were codon-optimized for  B. subtilis  (SEQ ID NO: 54, 59) (which produced 8 mg/L histamine) or codon-optimized for  S. cerevisiae  (SEQ ID NO: 56, 60)(which produced 9.3 mg/L histamine). 
     Since the best performing strain is in the host  Y. lipolytica , further strain improvements can be pursued in this host organism. Designs that can further enhance histamine production in  Y. lipolytica  include: 
     1. Transketolase (EC 2.2.1.1) (SEQ ID NO: 27), which catalyzes the interconversion of sugars in the pentose phosphate pathway and produces ribose-5-phosphate, which is a precursor to PPRP, the initial metabolite in the histidine biosynthesis pathway. 
     2. Ribose-phosphate pyrophosphokinase (EC 2.7.6.1) (SEQ ID NO: 28). 
     3. ATP phosphoribosyltransferase (EC 2.4.2.17) (SEQ ID NO: 5). 
     4. Trifunctional histidinol dehydrogenase (EC 1.1.1.23)/phosphoribosyl-AMP cyclohydrolase (EC 3.5.4.19)/phosphoribosyl-ATP diphosphatase (EC 3.6.1.31) (SEQ ID NO: 20). 
     5. Histidinol-phosphate aminotransferase (EC 2.6.1.9) (SEQ ID NO: 14). 
     6. 5′ProFAR isomerase (EC 5.3.1.16) (SEQ ID NO: 31). 
     7. Imidazole glycerol phosphate synthase (EC 4.3.1.B2) (SEQ ID NO: 21). 
     8. Triose-phosphate isomerase (EC 5.3.1.1) harboring the amino acid substitution I170V (SEQ ID NO: 32). 
     9. Glucose-6-phosphate 1-dehydrogenase (EC 1.1.1.49) harboring the amino acid substitution A243T (SEQ ID NO: 26). 
     10. Various histidine decarboxylases:
         a. UniProt ID A0A089YPE5 (SEQ ID NO: 33)   b. UniProt ID A0A126S6G9 (SEQ ID NO: 34)   c. UniProt ID A0A0A1R6V3 (SEQ ID NO: 35)   d. UniProt ID A0A1W0CM88 (SEQ ID NO: 36)   e. UniProt ID P00862 (SEQ ID NO: 4)   f. UniProt ID A0A0K6GJ74 (SEQ ID NO: 37)   g. UniProt ID TOQL99 (SEQ ID NO: 38)   h. UniProt ID A0A1B8HLR1 (SEQ ID NO: 39)       

     Example 3—Improvement of Histamine Production in  Yarrowia lipolytica  Engineered to Produce Histamine 
     Three improvement rounds of genetic engineering were carried out in  Yarrowia lipolytica.    
     First-Improvement Round Genetic Engineering in  Yarrowia lipolytica    
     Strategy: Improve flux into histidine and then histamine by overexpression of two enzymes. 
     
       
         
           
               
               
               
               
               
               
             
               
                   
               
               
                   
                 UniProt 
                   
                   
                 Codon 
                   
               
               
                 Enzyme Name 
                 ID 
                 Organism 
                 Description 
                 optmization 
                 Mutation 
               
               
                   
               
             
            
               
                 Histidine 
                 B71459 
                 Acinetobacter 
                 Last decarboxylation 
                 Yarrowia 
                 None 
               
               
                 decarboxylase 
                   
                 baumannii 
                 step of histamine 
                 lypolytica 
                   
               
               
                 (HDC) 
                   
                   
                 biosynthesis 
                   
                   
               
               
                 ATP 
                 P00498 
                 Saccharomyces 
                 Upstream step of 
                 Yarrowia 
                 None 
               
               
                 phosphoribosyltransferase 
                   
                 cerevisiae 
                 histidine biosynthesis. 
                 lypolytica 
                   
               
               
                 (ATP-PRase) 
                   
                   
                 Utilization of ATP 
                   
                   
               
               
                   
                   
                   
                 to covert PRPP to 
                   
                   
               
               
                   
                   
                   
                 PR-ATP 
               
               
                   
               
            
           
         
       
     
     Summary: ATP phosphoribosyltransferase catalyzes the first committed step of histidine biosynthesis pathway. This enzyme would be allosterically feedback-inhibited by histidine and competitively inhibited by AMP and ADP. The results did not indicate activity and/or inhibition of P00498. 
     Second-Improvement Round Genetic Engineering in  Yarrowia lipolytica    
     Strategy: Overexpression of one enzyme. The final step of histamine biosynthesis was enhanced by utilizing the best first-round histidine decarboxylase which was modified to include a solubility tag to improve protein folding. 
     
       
         
           
               
               
               
               
               
               
             
               
                   
               
               
                   
                 UniProt 
                   
                   
                 Codon 
                   
               
               
                 Enzyme Name 
                 ID 
                 Organism 
                 Description 
                 optmization 
                 Mutation 
               
               
                   
               
             
            
               
                 Histidine 
                 B71459 
                 Acinetobacter 
                 Last decarboxylation 
                 Yarrowia 
                 None 
               
               
                 decarboxylase 
                   
                 baumannii 
                 step of histamine 
                 lypolytica 
                   
               
               
                 (HDC) 
                   
                   
                 biosynthesis 
               
               
                   
               
            
           
         
       
     
     Summary: The histidine decarboxylase used for the second round of genetic engineering was the same as for the first round, although the codon optimization was different. Furthermore, an N-terminal solubility tag (MQYKLALNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDDATKTFT VT, SEQ ID NO:142) was included in the second-round enzyme. 
     Third-Improvement Round Genetic Engineering in  Yarrowia lipolytica    
     Strategy: Overexpression of two enzymes in pathways upstream of histidine biosynthesis to improve flux into phosphoribosyl pyrophosphate (PRPP). 
     
       
         
           
               
               
               
               
               
               
             
               
                   
               
               
                   
                 UniProt 
                   
                   
                 Codon 
                   
               
               
                 Enzyme Name 
                 ID 
                 Organism 
                 Description 
                 optmization 
                 Mutation 
               
               
                   
               
             
            
               
                 Ribose-phosphate 
                 E7EAU9 
                 Bacillus 
                 ATP dependent step for 
                 Yarrowia 
                 L135I 
               
               
                 pyrophosphokinase 
                   
                 amyloliquefaciens 
                 synthesis of PRPP 
                 lypolytica 
                   
               
               
                 (RPPK) 
                   
                   
                   
                   
                   
               
               
                 Glocose-6-phosphate 
                 A4QEF2 
                 Corynebacterium 
                 Upstream pathway to 
                 Yarrowia 
                 A243T 
               
               
                 1-dehydrogenase 
                   
                 glutamicum 
                 push carbon flux into 
                 lypolytica 
                   
               
               
                 (G6PDH) 
                   
                   
                 ribose-5-phosphate 
               
               
                   
               
            
           
         
       
     
     Summary: Ribose-phosphate pyrophosphokinase is competitively inhibited ADP. The L135I mutation at the ATP binding site on the enzyme relieves ADP inhibition. This strain expressed histamine at a titer of 1.68 g/L of culture medium. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 First-round results for histamine production in Yarrowia lipolytica 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 E1 
                 Enzyme 1 
                 Enzyme 1 
                 E1 Codon 
                 E2 
                 Enzyme 2 
                 E2 
                 Enzyme 2 
                 E2 Codon 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity 
                 source 
                 Optimization 
                 Uniprot 
                 activity 
                 Modifi- 
                 source 
                 Optimization 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 organism 
                 Abbrev. 
                 ID 
                 name 
                 cations 
                 organism 
                 Abbrev. 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Yarrowia 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 lipolytica 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                    0 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 01 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                    0 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 02 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 S288c 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                 505019 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Yarrowia 
               
               
                 03 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                    0 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 04 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 (strain LT2/ 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 SGSC1412/ 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATCC 700720) 
                   
               
               
                 YIHISMN_ 
                  32011 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 05 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                   
                 (strain LT2/ 
                 cerevisiae 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 SGSC1412/ 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATCC 700720) 
                   
               
               
                 YIHISMN_ 
                   833 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 06 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 (strain LT2/ 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 SGSC1412/ 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATCC 700720) 
                   
               
               
                 YIHISMN_ 
                   299 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 07 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 YIHISMN_ 
                  1778 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 08 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 YIHISMN_ 
                 257949 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 09 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 YIHISMN_ 
                    0 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 10 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                  96836 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 modified 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 modified 
               
               
                 11 
                   
                   
                 decarboxylase 
                 baumannii 
                 codon 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 
                 usage for  
                   
                 transferase 
                   
                 LT2 
                 usage for  
               
               
                   
                   
                   
                   
                 AB0057) 
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 YIHISMN_ 
                   33 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 12 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                 366139 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 13 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 YIHISMN_ 
                   23 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 14 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 YIHISMN_ 
                   26 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 modified 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 modified 
               
               
                 15 
                   
                   
                 decarboxylase 
                 sp. 
                 codon 
                   
                 phosphoribosyl 
                 L231F, 
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 usage for  
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 YIHISMN_ 
                   56 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 16 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 YIHISMN_ 
                  1406 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 17 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 YIHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 18 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                 YIHISMN_ 
                  90046 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 modified 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 modified 
               
               
                 19 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 codon 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 codon 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 usage for  
                   
                 transferase 
                   
                 S288c 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 YIHISMN_ 
                  1639 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 20 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                   
                 S288c 
                 cerevisiae 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 First-round results for production of histamine in Bacillus subtilis 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 E1 
                 Enzyme 1 
                 Enzyme 1 
                 E1 Codon 
                 E2 
                 Enzyme 2 
                 E2 
                 Enzyme 2 
                 E2 Codon 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity 
                 source 
                 Optimization 
                 Uniprot 
                 activity 
                 Modifi- 
                 source 
                 Optimization 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 organism 
                 Abbrev. 
                 ID 
                 name 
                 cations 
                 organism 
                 Abbrev. 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 BsHISMN_ 
                   
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Yarrowia 
               
               
                 01 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 919.7 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 02 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 BsHISMN_ 
                 2.4 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 modified 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 modified 
               
               
                 03 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 codon 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 usage for  
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                 9156.1 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 04 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 BsHISMN_ 
                 5057.2 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 modified 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 modified 
               
               
                 05 
                   
                   
                 decarboxylase 
                 sp. 
                 codon 
                   
                 phosphoribosyl 
                 L231F, 
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 usage for  
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                   
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 06 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 BsHISMN_ 
                 2532.4 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 07 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 13183.4 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 modified 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 modified 
               
               
                 08 
                   
                   
                 decarboxylase 
                 baumannii 
                 codon 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 
                 usage for  
                   
                 transferase 
                   
                 S288c 
                 usage for  
               
               
                   
                   
                   
                   
                 AB0057) 
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                 114.3 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 09 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 S288c 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 18336.5 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 10 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 BsHISMN_ 
                 0 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 modified 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 modified 
               
               
                 11 
                   
                   
                 decarboxylase 
                 sp. 
                 codon 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 usage for  
                   
                 transferase 
                   
                 LT2 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                 6778.2 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 12 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                 BsHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 13 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 BsHISMN_ 
                 1071.1 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 14 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 BsHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 15 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 BsHISMN_ 
                 233.4 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 16 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 16.2 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 modified 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 modified 
               
               
                 17 
                   
                   
                 decarboxylase 
                 baumannii 
                 codon 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 
                 usage for  
                   
                 transferase 
                   
                 LT2 
                 usage for  
               
               
                   
                   
                   
                   
                 AB0057) 
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                 61 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 18 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 1413.5 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 19 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 BsHISMN_ 
                 6630.6 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 20 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 BsHISMN_ 
                 43.8 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 21 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                 BsHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 modified 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 modified 
               
               
                 22 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 codon 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 codon 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 usage for  
                   
                 transferase 
                   
                 S288c 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 BsHISMN_ 
                 529 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 23 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                   
                 5288c 
                 cerevisiae 
               
               
                 BsHISMN_ 
                 15026.1 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Yarrowia 
               
               
                 24 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                 BsHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 modified 
                 Q9Z472 
                 ATP 
                   
                 Corynebacterium 
                 modified 
               
               
                 25 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 codon 
                   
                 phosphoribosyl- 
                   
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 usage for  
                   
                 transferase 
                   
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Host evaluation designs for production of histamine tested in Saccharomyces cerevisiae 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 E1 
                 Enzyme 1 
                 Enzyme 1 
                 E1 Codon 
                 E2 
                 Enzyme 2 
                 E2 
                 Enzyme 2 
                 E2 Codon 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity 
                 source 
                 Optimization 
                 Uniprot 
                 activity 
                 Modifi- 
                 source 
                 Optimization 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 organism 
                 Abbrev. 
                 ID 
                 name 
                 cations 
                 organism 
                 Abbrev. 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Saccharomyces 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 cerevisiae 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 ScHISMN_ 
                  17466 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 116 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 ScHISMN_ 
                  28646 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 117 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                 ScHISMN_ 
                  48150 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 118 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 ScHISMN_ 
                  59265 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 119 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 ScHISMN_ 
                  72566 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 modified 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 modified 
               
               
                 120 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 codon 
                   
                 phosphoribosyl 
                 L231F, 
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 usage for  
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 ScHISMN_ 
                  46418 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 121 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 ScHISMN_ 
                  64087 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 122 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 ScHISMN_ 
                  80704 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 123 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 ScHISMN_ 
                  70043 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 124 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 ScHISMN_ 
                  25331 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 125 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 ScHISMN_ 
                  33970 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 modified 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 modified 
               
               
                 126 
                   
                   
                 decarboxylase 
                 sp. 
                 codon 
                   
                 phosphoribosyl 
                 L231F, 
                 glutamicum 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 usage for  
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 usage for  
               
               
                   
                   
                   
                   
                   
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 ScHISMN_ 
                  21402 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 127 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 ScHISMN_ 
                  41854 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 128 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 ScHISMN_ 
                 101496 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 129 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                 ScHISMN_ 
                  85546 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 130 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                   
                 S288c 
                 cerevisiae 
               
               
                 ScHISMN_ 
                 111109 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Yarrowia 
               
               
                 131 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Host evaluation designs for production of histamine tested in Corynebacterium glutamicum 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 E1 
                 Enzyme 1 
                 Enzyme 1 
                 E1 Codon 
                 E2 
                 Enzyme 2 
                 E2 
                 Enzyme 2 
                 E2 Codon 
               
               
                 Strain 
                 Titer 
                 Uniprot 
                 activity 
                 source 
                 Optimization 
                 Uniprot 
                 activity 
                 Modifi- 
                 source 
                 Optimization 
               
               
                 name 
                 (μg/L) 
                 ID 
                 name 
                 organism 
                 Abbrev. 
                 ID 
                 name 
                 cations 
                 organism 
                 Abbrev. 
               
               
                   
               
               
                 CgHISMN_ 
                   
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 70 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                 68395.9 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 modified 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 modified 
               
               
                 71 
                   
                   
                 decarboxylase 
                 baumannii 
                 codon 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 codon 
               
               
                   
                   
                   
                   
                 (strain 
                 usage for  
                   
                 transferase 
                   
                 S288c 
                 usage for  
               
               
                   
                   
                   
                   
                 AB0057) 
                 Cg and Sc 
                   
                   
                   
                   
                 Cg and Sc 
               
               
                 CgHISMN_ 
                 18 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Saccharo- 
               
               
                 72 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 S288c 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                 16325.5 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Yarrowia 
               
               
                 73 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 S288c 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                 4883.6 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 74 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 CgHISMN_ 
                   
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 75 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                 CgHISMN_ 
                   
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 76 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                 CgHISMN_ 
                 5.4 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 77 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 CgHISMN_ 
                 88.6 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 78 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 sp. LK1 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 CgHISMN_ 
                   
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 79 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 CgHISMN_ 
                 8368.2 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Bacillus 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Bacillus 
               
               
                 80 
                   
                   
                 decarboxylase 
                 baumannii 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                   
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                 9.3 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Saccharo- 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Saccharo- 
               
               
                 81 
                   
                   
                 decarboxylase 
                 baumannii 
                 myces 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 
                 cerevisiae 
                   
                 transferase 
                   
                 LT2 
                 cerevisiae 
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                 15529.4 
                 B71459 
                 Histidine 
                 Acinetobacter 
                 Yarrowia 
                 P00499 
                 ATP 
                   
                 Salmonella 
                 Yarrowia 
               
               
                 82 
                   
                   
                 decarboxylase 
                 baumannii 
                 lipolytica 
                   
                 phosphoribosyl- 
                   
                 typhimurium 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 
                   
                   
                 transferase 
                   
                 LT2 
                   
               
               
                   
                   
                   
                   
                 AB0057) 
                   
                   
                   
                   
                   
                   
               
               
                 CgHISMN_ 
                   
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Bacillus 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Bacillus 
               
               
                 83 
                   
                   
                 decarboxylase 
                 sp. 
                 subtilis 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 subtilis 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 CgHISMN_ 
                 2.6 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Saccharo- 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Saccharo- 
               
               
                 84 
                   
                   
                 decarboxylase 
                 sp. 
                 myces 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 myces 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                 cerevisiae 
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                 cerevisiae 
               
               
                 CgHISMN_ 
                 6134 
                 P00862 
                 Histidine 
                 Lactobacillus 
                 Yarrowia 
                 Q9Z472 
                 ATP 
                 N215K, 
                 Corynebacterium 
                 Yarrowia 
               
               
                 85 
                   
                   
                 decarboxylase 
                 sp. 
                 lipolytica 
                   
                 phosphoribosyl- 
                 L231F, 
                 glutamicum 
                 lipolytica 
               
               
                   
                   
                   
                   
                 (strain 30a) 
                   
                   
                 transferase 
                 T235A 
                 ATCC 13032 
                   
               
               
                 CgHISMN_ 
                 197 
                 A0A0J6 
                 Histidine 
                 Chromo- 
                 Bacillus 
                 P00498 
                 ATP 
                   
                 Saccharomyces 
                 Bacillus 
               
               
                 86 
                   
                 KM89 
                 decarboxylase 
                 bacterium 
                 subtilis 
                   
                 phosphoribosyl- 
                   
                 cerevisiae 
                 subtilis 
               
               
                   
                   
                   
                   
                 sp. LK1 
                   
                   
                 transferase 
                   
                 S288c 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 SEQ ID NO Cross-Reference Table 
               
            
           
           
               
               
               
               
               
            
               
                 SEQ 
                   
                   
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 ID 
                 Sequence Type with 
                 Uniprot 
                   
                   
                 Codon 
               
               
                 NO 
                 Modifications 
                 ID 
                 Activity name 
                 Source organism 
                 Optimization Abbrev. 
               
               
                   
               
               
                  1 
                 AA seq for 
                 B71459 
                 histidine decarboxylase 
                 Acinetobacter baumannii 
                   
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                   
               
               
                  2 
                 AA seq for 
                 Q9KJU3 
                 Imidazoleglycerol- 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q9KJU3 
                   
                 phosphate dehydratase 
                   
                   
               
               
                  3 
                 AA seq for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P00498 
                   
                   
                   
                   
               
               
                  4 
                 AA seq for 
                 P00862 
                 histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                   
               
               
                   
                 enzyme P00862 
                   
                   
                   
                   
               
               
                  5 
                 AA seq for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium 
                   
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  6 
                 AA seq for 
                 J6KM89 
                 histidine decarboxylase 
                 Chromobacterium sp. LK1 
                   
               
               
                   
                 enzyme J6KM89 
                   
                   
                   
                   
               
               
                  7 
                 AA seq for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                 with substitution 
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  8 
                 AA seq for 
                 E3QMN8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                   
               
               
                   
                 enzyme E3QMN8 
                   
                   
                 str. Wiesmoor 
                   
               
               
                  9 
                 AA seq for 
                 Q467R8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                   
               
               
                   
                 enzyme Q467R8 
                   
                   
                 (strain Fusaro/DSM 804) 
                   
               
               
                  10 
                 AA seq for 
                 Q05733 
                 histidine decarboxylase 
                 Drosophila melanogaster 
                   
               
               
                   
                 enzyme Q05733 
                   
                   
                   
                   
               
               
                  11 
                 AA seq for 
                 P54772 
                 histidine decarboxylase 
                 Solanum lycopersicum 
                   
               
               
                   
                 enzyme P54772 
                   
                   
                   
                   
               
               
                  12 
                 AA seq for 
                 P23738 
                 histidine decarboxylase 
                 Mus musculus 
                   
               
               
                   
                 enzyme P23738 
                   
                   
                   
                   
               
               
                  13 
                 AA seq for  
                 O68602 
                 1-(5-phosphoribosyl)5[(5- 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme O68602 
                   
                 phosphoribosylamino) 
                   
                   
               
               
                   
                   
                   
                 methylideneamino] 
                   
                   
               
               
                   
                   
                   
                 imidazole-4- 
                   
                   
               
               
                   
                   
                   
                 carboxamide isomerase 
                   
                   
               
               
                  14 
                 AA seq for 
                 Q9KJU4 
                 Histidinol-phosphate 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q9KJU4 
                   
                 aminotransferase 
                   
                   
               
               
                  15 
                 AA seq for 
                 Q8NNT5 
                 Histidinol dehydrogenase 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q8NNT5 
                   
                   
                   
                   
               
               
                  16 
                 AA seq for 
                 Q9Z471 
                 Phosphoribosyl- 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q9Z471 
                   
                 ATP pyrophosphatase 
                   
                   
               
               
                  17 
                 AA seq for 
                 O31139 
                 Imidazole glycerol phosphate 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme O31139 
                   
                 synthase subunit 
                   
                   
               
               
                  18 
                 AA seq for 
                 O69043 
                 Imidazole glycerol phosphate 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme O69043 
                   
                 synthase subunit 
                   
                   
               
               
                  19 
                 AA seq for 
                 Q8NNT9 
                 phosphoribosyl- 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q8NNT9 
                   
                 AMP cyclohydrolase 
                   
                   
               
               
                  22 
                 AA seq for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                   
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                   
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  20 
                 AA seq for 
                 P00815 
                 histidinol dehydrogenase, 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P00815 
                   
                 phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 AMP cyclohydrolase, 
                   
                   
               
               
                   
                   
                   
                 phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 ATP diphosphatase 
                   
                   
               
               
                  21 
                 AA seq for 
                 P33734 
                 Imidazole glycerol phosphate 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P33734 
                   
                 synthase subunit HisF 
                   
                   
               
               
                  23 
                 AA seq for 
                 P07172 
                 histidinol- 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P07172 
                   
                 phosphate transaminase 
                   
                   
               
               
                  24 
                 AA seq for 
                 P06633 
                 Imidazoleglycerol- 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P06633 
                   
                 phosphate dehydratase 
                   
                   
               
               
                  25 
                 AA seq for 
                 P38635 
                 histidinol-phosphatase 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P38635 
                   
                   
                   
                   
               
               
                  26 
                 AA seq for 
                 A4QEF2 
                 Glucose-6-phosphate  
                 Corynebacterium glutamicum 
                   
               
               
                   
                 enzyme A4QEF2 with 
                   
                 1-dehydrogenase 
                   
                   
               
               
                   
                 substitution A243T 
                   
                 (G6PD) (EC 1.1.1.49) 
                 (strain R) 
                   
               
               
                  27 
                 AA seq for 
                 P23254 
                 Transketolase 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P23254 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  28 
                 AA seq for 
                 Q12265 
                 Ribose-phosphate  
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme Q12265 
                   
                 pyrophosphokinase 5 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                 (EC 2.7.6.1) 
                 (Baker&#39;s yeast) 
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase 5) 
                   
                   
               
               
                  30 
                 DNA seq1 for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q9Z472 
                   
                   
                   
                   
               
               
                  29 
                 DNA seq1 for 
                 P00815 
                 histidinol dehydrogenase, 
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P00815 
                   
                 phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 AMP cyclohydrolase, 
                   
                   
               
               
                   
                   
                   
                 phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 ATP diphosphatase 
                   
                   
               
               
                  31 
                 AA seq for 
                 P40545 
                 1-(5-phosphoribosyl)-5-[(5- 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P40545 
                   
                 phosphoribosylamino) 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                 methylideneamino] 
                 (Baker&#39;s yeast) 
                   
               
               
                   
                   
                   
                 imidazole-4-carboxamide 
                   
                   
               
               
                   
                   
                   
                 isomerase (EC 5.3.1.16) 
                   
                   
               
               
                   
                   
                   
                 (5-proFAR isomerase) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosylformimino-5- 
                   
                   
               
               
                   
                   
                   
                 aminoimidazole carboxamide 
                   
                   
               
               
                   
                   
                   
                 ribotide isomerase) 
                   
                   
               
               
                  32 
                 AA seq for 
                 P00942 
                 Triosephosphate isomerase 
                 Saccharomyces cerevisiae 
                   
               
               
                   
                 enzyme P00942 with 
                   
                 (TIM) (EC 5.3.1.1) 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                 substitution 1170V 
                   
                 (Triose-phosphate isomerase) 
                 (Baker&#39;s yeast) 
                   
               
               
                  33 
                 AA seq for enzyme 
                 A0A089 
                 Histidine decarboxylase 
                 Pseudomonas rhizosphaerae 
                   
               
               
                   
                 A0A089YPE5 
                 YPE5 
                 (HDC) (EC 4.1.1.22) 
                   
                   
               
               
                  34 
                 AA seq for enzyme 
                 A0A126 
                 Histidine decarboxylase 
                 Pseudomonas putida 
                   
               
               
                   
                 AOA12656G9 
                 56G9 
                 (HDC) (EC 4.1.1.22) 
                 (Arthrobacter siderocapsulatus) 
                   
               
               
                  35 
                 AA seq for enzyme 
                 A0A0A1 
                 Histidine decarboxylase 
                 Citrobacter pasteurii 
                   
               
               
                   
                 A0A0A1R6V3 
                 R6V3 
                 (HDC) (EC 4.1.1.22) 
                   
                   
               
               
                  36 
                 AA seq for enzyme 
                 A0A1W0 
                 Histidine decarboxylase 
                 Chromobacterium haemolyticum 
                   
               
               
                   
                 A0A1W0CM88 
                 CM88 
                 (HDC) (EC 4.1.1.22) 
                   
                   
               
               
                  37 
                 AA seq for enzyme 
                 A0A0K6 
                 Histidine decarboxylase 
                 Lactobacillus reuteri 
                   
               
               
                   
                 A0A0K6GJ74 
                 GJ74 
                 proenzyme 
                   
                   
               
               
                  38 
                 AA seq for 
                 T0QL99 
                 Histidine decarboxylase 
                 Aeromonas salmonicida 
                   
               
               
                   
                 enzyme T0QL99 
                   
                 (EC 4.1.1.22) (Fragment) 
                 subsp. pectinolytica 34mel 
                   
               
               
                  39 
                 AA seq for enzyme 
                 A0A1B8 
                 Histidine decarboxylase 
                 Morganella psychrotolerans 
                   
               
               
                   
                 A0A1B8HLR1 
                 HLR1 
                 (HDC) (EC 4.1.1.22) 
                   
                   
               
               
                  40 
                 AA seq for enzyme 
                 A0A0C1 
                 Histidine decarboxylase 
                 Lactobacillus fructivorans 
                   
               
               
                   
                 A0A0C1PR48 
                 PR48 
                 proenzyme 
                   
                   
               
               
                  41 
                 AA seq for 
                 P0A717 
                 Ribose-phosphate 
                 Escherichia coli (strain K12) 
                   
               
               
                   
                 enzyme P0A717 
                   
                 pyrophosphokinase 
                   
                   
               
               
                   
                   
                   
                 (RPPK) (EC 2.7.6.1) 
                   
                   
               
               
                   
                   
                   
                 (5-phospho-D-ribosyl 
                   
                   
               
               
                   
                   
                   
                 alpha-1-diphosphate) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 diphosphate synthase) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase)  
                   
                   
               
               
                   
                   
                   
                 (P-Rib-PP synthase)  
                   
                   
               
               
                   
                   
                   
                 (PRPP synthase) (PRPPase) 
                   
                   
               
               
                  42 
                 AA seq for 
                 Q680A5 
                 Ribose-phosphate 
                 Arabidopsis thaliana 
                   
               
               
                   
                 enzyme Q680A5 
                   
                 pyrophosphokinase 4 
                 (Mouse-ear cress) 
                   
               
               
                   
                   
                   
                 (EC 2.7.6.1) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase 4) 
                   
                   
               
               
                  43 
                 AA seq for 
                 P38620 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae  
                   
               
               
                   
                 enzyme P38620 
                   
                 pyrophosphokinase 2 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                 (EC 2.7.6.1) 
                 (Baker&#39;s yeast) 
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase 2) 
                   
                   
               
               
                  44 
                 AA seq for 
                 P32895 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae  
                   
               
               
                   
                 enzyme P32895 
                   
                 pyrophosphokinase 1 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                 (EC 2.7.6.1) 
                 (Baker&#39;s yeast) 
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase 1) 
                   
                   
               
               
                  45 
                 AA seq for 
                 P38689 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae  
                   
               
               
                   
                 enzyme P38689 
                   
                 pyrophosphokinase 3 
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                 (EC 2.7.6.1) 
                 (Baker&#39;s yeast) 
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase 3) 
                   
                   
               
               
                  46 
                 AA seq for 
                 P15019 
                 Transaldolase (EC 2.2.1.2) 
                 Saccharomyces cerevisiae  
                   
               
               
                   
                 enzyme P15019 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  47 
                 AA seq for 
                 P06775 
                 Histidine permease 
                 Saccharomyces cerevisiae  
                   
               
               
                   
                 enzyme P06775 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  48 
                 AA seq for 
                 O59667 
                 Histidine biosynthesis 
                 Schizosaccharomyces pombe 
                   
               
               
                   
                 enzyme O59667 
                   
                 bifunctional protein 
                 (strain 972/ATCC 24843) 
                   
               
               
                   
                   
                   
                 his7 [Includes: 
                 (Fission yeast) 
                   
               
               
                   
                   
                   
                 Phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 AMP cyclohydrolase 
                   
                   
               
               
                   
                   
                   
                 (EC 3.5.4.19); 
                   
                   
               
               
                   
                   
                   
                 Phosphoribosyl- 
                   
                   
               
               
                   
                   
                   
                 ATP pyrophosphatase 
                   
                   
               
               
                   
                   
                   
                 (EC 3.6.1.31)] 
                   
                   
               
               
                  49 
                 AA seq for 
                 O66000 
                 Histidine decarboxylase 
                 Oenococcus oeni 
                   
               
               
                   
                 enzyme O66000 
                   
                 proenzyme 
                 (Leuconostoc oenos) 
                   
               
               
                  50 
                 AA seq for enzyme 
                 A0A0R1 
                 Pyruvoyl family 
                 Lactobacillus aviarius subsp. 
                   
               
               
                   
                 A0A0R1Y874 
                 Y874 
                 histidine decarboxylase 
                 aviarius DSM 20655 
                   
               
               
                  51 
                 AA seq for enzyme 
                 A0A0J6K 
                 Histidine decarboxylase  
                 Chromobacterium sp. LK1 
                   
               
               
                   
                 A0A0J6KM89 
                 M89 
                 (HDC) (EC 4.1.1.22) 
                   
                   
               
               
                  52 
                 DNA seq1 for 
                 B71459 
                 Histidine decarboxylase 
                 Acinetobacter baumannii  
                 Yarrowia lipolytica 
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                   
               
               
                  53 
                 DNA seq1 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae  
                 Yarrowia lipolytica 
               
               
                   
                 enzyme P00498 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  54 
                 DNA seq2 for 
                 B71459 
                 Histidine decarboxylase 
                 Acinetobacter baumannii 
                 Bacillus subtillus 
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                   
               
               
                  55 
                 DNA seq2 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae  
                 Bacillus subtillus 
               
               
                   
                 enzyme P00498 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  56 
                 DNA seq3 for 
                 B71459 
                 Histidine decarboxylase 
                 Acinetobacter baumannii 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                   
               
               
                  57 
                 DNA seq3 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae  
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P00498 
                   
                   
                 (strain ATCC 204508/S288c) 
                   
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                   
               
               
                  58 
                 DNA seq1 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium  
                 Yarrowia lipolytica 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  59 
                 DNA seq2 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium  
                 Bacillus subtillus 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  60 
                 DNA seq3 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium  
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  61 
                 DNA seq4 for 
                 B71459 
                 Histidine decarboxylase 
                 Acinetobacter baumannii 
                 modified codon usage for 
               
               
                   
                 enzyme B71459 
                   
                 (HDC) (EC 4.1.1.22) 
                 (strain AB0057) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                  62 
                 DNA seq4 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium 
                 modified codon usage for 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                 Corynebacterium glutamicum and 
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                 Saccharomyces cerevisiae 
               
               
                  63 
                 DNA seq1 for enzyme 
                 A0A0J6K 
                 Histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 Yarrowia lipolytica 
               
               
                   
                 A0A0J6KM89 
                 M89 
                   
                   
                   
               
               
                  64 
                 DNA seq1 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 Yarrowia lipolytica 
               
               
                   
                 for enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                 with substitution 
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  65 
                 DNA seq2 for enzyme 
                 A0A0J6K 
                 Histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 Saccharomyces cerevisiae 
               
               
                   
                 A0A0J6KM89 
                 M89 
                   
                   
                   
               
               
                  66 
                 DNA seq2 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 Saccharomyces cerevisiae 
               
               
                   
                 for enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                 with substitution 
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  67 
                 DNA seq3 
                 A0A0J6K 
                 Histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 Bacillus subtillus 
               
               
                   
                 for enzyme 
                 M89 
                   
                   
                   
               
               
                   
                 A0A0J6KM89 
                   
                   
                   
                   
               
               
                  68 
                 DNA seq3 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 Bacillus subtillus 
               
               
                   
                 for enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                 with substitution 
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  69 
                 DNA seq1 for 
                 P00862 
                 Histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 Bacillus subtillus 
               
               
                   
                 enzyme P00862 
                   
                 proenzyme 
                   
                   
               
               
                  70 
                 DNA seq2 for 
                 P00862 
                 Histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P00862 
                   
                 proenzyme 
                   
                   
               
               
                  71 
                 DNA seq3 for 
                 P00862 
                 Histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 modified codon usage for 
               
               
                   
                 enzyme P00862 
                   
                 proenzyme 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                  72 
                 DNA seq5 for 
                 B71459 
                 Histidine decarboxylase 
                 Acinetobacter baumannii 
                 modified codon usage for 
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                  73 
                 DNA seq4 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae  
                 modified codon usage for 
               
               
                   
                 enzyme P00498 
                   
                   
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                  74 
                 AA seq for enzyme 
                 A0A1H1 
                 Histidine decarboxylase 
                 Pseudomonas sp. bs2935 
                 modified codon usage for 
               
               
                   
                 A0A1H1TEB8 
                 TEB8 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                  75 
                 DNA seq1 for 
                 E3QMN8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme E3QMN8 
                   
                   
                 str. Wiesmoor 
                   
               
               
                  76 
                 DNA seq1 for 
                 Q467R8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme Q467R8 
                   
                   
                 (strain Fusaro/DSM 804) 
                   
               
               
                  77 
                 DNA seq4 for 
                 P00862 
                 histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme P00862 
                   
                   
                   
                   
               
               
                  78 
                 DNA seq6 for 
                 B71459 
                 histidine decarboxylase 
                 Acinetobacter baumannii 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme B71459 
                   
                   
                 (strain AB0057) 
                   
               
               
                  79 
                 DNA seq1 for 
                 Q05733 
                 histidine decarboxylase 
                 Drosophila melanogaster 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme Q05733 
                   
                   
                   
                   
               
               
                  80 
                 DNA seq1 for 
                 J6KM89 
                 histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme J6KM89 
                   
                   
                   
                   
               
               
                  81 
                 DNA seq5 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium  
                 Corynebacterium glutamicum 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  82 
                 DNA seq5 for 
                 P00862 
                 histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P00862 
                   
                   
                   
                   
               
               
                  83 
                 DNA seq for 
                 P54772 
                 histidine decarboxylase 
                 Solanum lycopersicum 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P54772 
                   
                   
                   
                   
               
               
                  84 
                 DNA seq for 
                 P23738 
                 histidine decarboxylase 
                 Mus musculus 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P23738 
                   
                   
                   
                   
               
               
                  85 
                 DNA seq2 for 
                 Q05733 
                 histidine decarboxylase 
                 Drosophila melanogaster 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme Q05733 
                   
                   
                   
                   
               
               
                  86 
                 DNA seq2 for 
                 J6KM89 
                 histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme J6KM89 
                   
                   
                   
                   
               
               
                  87 
                 DNA seq2 for 
                 E3QMN8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme E3QMN8 
                   
                   
                 str. Wiesmoor 
                   
               
               
                  88 
                 DNA seq2 for 
                 Q467R8 
                 histidine decarboxylase 
                 Methanosarcina barkeri 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme Q467R8 
                   
                   
                 (strain Fusaro/DSM 804) 
                   
               
               
                  89 
                 DNA seq4 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 Saccharomyces cerevisiae 
               
               
                   
                 for enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                 with substitution 
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                  90 
                 DNA seq6 for 
                 P00499 
                 ATP phosphoribosyltransferase 
                 Salmonella typhimurium  
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme P00499 with 
                   
                   
                 (strain LT2/SGSC1412/ 
                   
               
               
                   
                 deletion of Q207-E208 
                   
                   
                 ATCC 700720) 
                   
               
               
                  91 
                 DNA seq for 
                 O68602 
                 1-(5-phosphoribosyl)5[(5- 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme O68602 
                   
                 phosphoribosylamino) 
                   
                   
               
               
                   
                   
                   
                 methylideneamino]imidazole- 
                   
                   
               
               
                   
                   
                   
                 4-carboxamide isomerase 
                   
                   
               
               
                  92 
                 DNA seq for 
                 Q9KJU3 
                 Imidazoleglycerol- 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q9KJU3 
                   
                 phosphate dehydratase 
                   
                   
               
               
                  93 
                 DNA seq for 
                 Q9KJU4 
                 Histidinol-phosphate  
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q9KJU4 
                   
                 aminotransferase 
                   
                   
               
               
                  94 
                 DNA seq for 
                 Q8NNT5 
                 Histidinol dehydrogenase 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q8NNT5 
                   
                   
                   
                   
               
               
                  95 
                 DNA seq for 
                 Q9Z471 
                 Phosphoribosyl-ATP 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q9Z471 
                   
                 pyrophosphatase 
                   
                   
               
               
                  96 
                 DNA seq for 
                 O31139 
                 Imidazole glycerol phosphate 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme O31139 
                   
                 synthase subunit 
                   
                   
               
               
                  97 
                 DNA seq for 
                 O69043 
                 Imidazole glycerol phosphate 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme O69043 
                   
                 synthase subunit 
                   
                   
               
               
                  98 
                 DNA seq for 
                 Q8NNT9 
                 phosphoribosyl- 
                 Corynebacterium glutamicum 
                 native 
               
               
                   
                 enzyme Q8NNT9 
                   
                 AMP cyclohydrolase 
                   
                   
               
               
                  99 
                 DNA seq5 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P00498 
                   
                 Imidazole glycerol phosphate 
                   
                   
               
               
                 100 
                 DNA seq1 for 
                 P33734 
                 synthase subunit HisF 
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P33734 
                   
                   
                   
                   
               
               
                 101 
                 DNA seq1 for 
                 P07172 
                 histidinol-phosphate  
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P07172 
                   
                 transaminase 
                   
                   
               
               
                 102 
                 DNA seq for 
                 P06633 
                 Imidazoleglycerol- 
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P06633 
                   
                 phosphate dehydratase 
                   
                   
               
               
                 103 
                 DNA seq1 for 
                 P38635 
                 histidinol-phosphatase 
                 Saccharomyces cerevisiae 
                 native 
               
               
                   
                 enzyme P38635 
                   
                   
                   
                   
               
               
                 104 
                 DNA seq 
                 A0A0C1 
                 Histidine decarboxylase 
                 Lactobacillus fructivorans 
                 modified codon usage for 
               
               
                   
                 for enzyme 
                 PR48 
                 proenzyme 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                 A0A0C1PR48 
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 105 
                 DNA seq for 
                 T0QL99 
                 Histidine decarboxylase 
                 Aeromonas salmonicida 
                 modified codon usage for 
               
               
                   
                 enzyme T0QL99 
                   
                 (EC 4.1.1.22) (Fragment) 
                 subsp. pectinolytica 34mel 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 106 
                 DNA seq 
                 A0A1B8 
                 Histidine decarboxylase 
                 Morganella psychrotolerans 
                 modified codon usage for 
               
               
                   
                 for enzyme 
                 HLR1 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                 A0A1B8HLR1 
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 107 
                 DNA seq2 for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 modified codon usage for 
               
               
                   
                 enzyme Q9Z472 
                   
                 (ATP-PRT) (ATP-PRTase) 
                 (strain ATCC 13032/ 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 DSM 20300/JCM 1318/ 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                 108 
                 DNA seq for 
                 P0A717 
                 Ribose-phosphate 
                 Escherichia coli (strain K12) 
                 modified codon usage for 
               
               
                   
                 enzyme P0A717 
                   
                 pyrophosphokinase 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (RPPK) (EC 2.7.6.1) 
                   
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 (5-phospho-D-ribosyl 
                   
                   
               
               
                   
                   
                   
                 alpha-1-diphosphate) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 diphosphate synthase) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosyl 
                   
                   
               
               
                   
                   
                   
                 pyrophosphate synthase) 
                   
                   
               
               
                   
                   
                   
                 (P-Rib-PP synthase)  
                   
                   
               
               
                   
                   
                   
                 (PRPP synthase) (PRPPase) 
                   
                   
               
               
                 109 
                 DNA seq for 
                 Q12265 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme Q12265 
                   
                 pyrophosphokinase 5 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.7.6.1) (Phosphoribosyl 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 pyrophosphate synthase 5) 
                   
                   
               
               
                 110 
                 DNA seq for 
                 P32895 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P32895 
                   
                 pyrophosphokinase 1 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.7.6.1) (Phosphoribosyl 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 pyrophosphate synthase 1) 
                   
                   
               
               
                 111 
                 DNA seq for 
                 P23254 
                 Transketolase 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P23254 
                   
                   
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 112 
                 DNA seq for 
                 P06775 
                 Histidine permease 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P06775 
                   
                   
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 113 
                 DNA seq2 for 
                 P00815 
                 trifunctional histidinol 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P00815 
                   
                 dehydrogenase/ 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 phosphoribosyl-AMP 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 cyclohydrolase/ 
                   
                   
               
               
                   
                   
                   
                 phosphoribosyl-ATP 
                   
                   
               
               
                   
                   
                   
                 diphosphatase 
                   
                   
               
               
                 114 
                 DNA seq2 fo  
                 P07172 
                 Histidinol-phosphate 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P07172 
                   
                 aminotransferase 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 115 
                 DNA seq6 for 
                 P00498 
                 ATP phosphoribosyltransferase 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P00498 
                   
                 (ATP-PRT) (ATP-PRTase) 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 116 
                 DNA seq for 
                 P40545 
                 1-(5-phosphoribosyl)-5-[(5- 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P40545 
                   
                 phosphoribosylamino) 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 methylideneamino] 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 imidazole-4-carboxamide  
                   
                   
               
               
                   
                   
                   
                 isomerase (EC 5.3.1.16) 
                   
                   
               
               
                   
                   
                   
                 (5-proFAR isomerase) 
                   
                   
               
               
                   
                   
                   
                 (Phosphoribosylformimino- 
                   
                   
               
               
                   
                   
                   
                 5-aminoimidazole 
                   
                   
               
               
                   
                   
                   
                 carboxamide 
                   
                   
               
               
                   
                   
                   
                 ribotide isomerase) 
                   
                   
               
               
                 117 
                 DNA seq2 for 
                 P33734 
                 Imidazole 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P33734 
                   
                 glycerol phosphate 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 synthase hisHF 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 118 
                 DNA seq for 
                 P00942 
                 Triosephosphate isomerase 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P00942 with 
                   
                 (TIM) (EC 5.3.1.1) 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                 substitution 1170V 
                   
                 (Triose-phosphate isomerase) 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 119 
                 DNA seq3 for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 modified codon usage for 
               
               
                   
                 enzyme Q9Z472 
                   
                 (ATP-PRT) (ATP-PRTase) 
                 (strain ATCC 13032/ 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.4.2.17) 
                 DSM 20300/JCM 1318/ 
                 Saccharomyces cerevisiae 
               
               
                 120 
                 DNA seq5 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 LMG 3730/NCIMB 10025) 
                 modified codon usage for 
               
               
                   
                 for enzyme Q9Z472 
                   
                 (ATP-PRT) (ATP-PRTase) 
                 (strain ATCC 13032/ 
                 Corynebacterium glutamicum and 
               
               
                   
                 with substitution 
                   
                 (EC 2.4.2.17) 
                 DSM 20300/JCM 1318/ 
                 Saccharomyces cerevisiae 
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                 121 
                 DNA seq for 
                 O66000 
                 Histidine decarboxylase 
                 Oenococcus oeni 
                 modified codon usage for 
               
               
                   
                 enzyme O66000 
                   
                 proenzyme 
                 (Leuconostoc oenos) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 122 
                 DNA seq for enzyme 
                 A0A0R1 
                 Pyruvoyl family 
                 Lactobacillus aviarius 
                 modified codon usage for 
               
               
                   
                 A0A0R1Y874 
                 Y874 
                 histidine decarboxylase 
                 subsp. aviarius DSM 20655 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 123 
                 DNA seq for enzyme 
                 A0A1H1 
                 Histidine decarboxylase 
                 Pseudomonas sp. bs2935 
                 modified codon usage for 
               
               
                   
                 A0A1H1TEB8 with 
                 TEB8 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                 substitution S9R 
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 124 
                 DNA seq for enzyme 
                 A0A089 
                 Histidine decarboxylase 
                 Pseudomonas rhizosphaerae 
                 modified codon usage for 
               
               
                   
                 A0A089YPE5 
                 YPE5 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 125 
                 DNA seq for enzyme 
                 A0A126 
                 Histidine decarboxylase 
                 Pseudomonas putida 
                 modified codon usage for 
               
               
                   
                 A0A12656G9 
                 56G9 
                 (HDC) (EC 4.1.1.22) 
                 (Arthrobacter siderocapsulatus) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 126 
                 DNA seq4 for enzyme 
                 A0A0J6K 
                 Histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 modified codon usage for 
               
               
                   
                 A0A0J6KM89 
                 M89 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 127 
                 DNA seq for enzyme 
                 A0A0A1 
                 Histidine decarboxylase 
                 Citrobacter pasteurii 
                 modified codon usage for 
               
               
                   
                 A0A0A1R6V3 
                 R6V3 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 128 
                 DNA seq for enzyme 
                 A0A1W0 
                 Histidine decarboxylase 
                 Chromobacterium haemolyticum 
                 modified codon usage for 
               
               
                   
                 A0A1W0CM88 
                 CM88 
                 (HDC) (EC 4.1.1.22) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 Histidine decarboxylase 
                   
                 Saccharomyces cerevisiae 
               
               
                 129 
                 DNA seq6 for 
                 P00862 
                 proenzyme 
                 Lactobacillus sp. (strain 30a) 
                 modified codon usage for 
               
               
                   
                 enzyme P00862 
                   
                 (EC 4.1.1.22) (Pi chain) 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 [Cleaved into: Histidine 
                   
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 decarboxylase beta chain; 
                   
                   
               
               
                   
                   
                   
                 Histidine decarboxylase 
                   
                   
               
               
                   
                   
                   
                 alpha chain] 
                   
                   
               
               
                 130 
                 DNA seq for enzyme 
                 A0A0K6 
                 Histidine decarboxylase 
                 Lactobacillus reuteri 
                 modified codon usage for 
               
               
                   
                 A0A0K6GJ74 
                 GJ74 
                 proenzyme 
                   
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 131 
                 DNA seq for 
                 P38620 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P38620 
                   
                 pyrophosphokinase 2 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.7.6.1) (Phosphoribosyl 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 pyrophosphate synthase 2) 
                   
                   
               
               
                 132 
                 DNA seq for 
                 P38689 
                 Ribose-phosphate 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P38689 
                   
                 pyrophosphokinase 3 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.7.6.1) (Phosphoribosyl 
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 pyrophosphate synthase 3) 
                   
                   
               
               
                 133 
                 DNA seq for 
                 P15019 
                 Transaldolase (EC 2.2.1.2) 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P15019 
                   
                   
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 134 
                 DNA seq for 
                 Q680A5 
                 Ribose-phosphate 
                 Arabidopsis thaliana 
                 modified codon usage for 
               
               
                   
                 enzyme Q680A5 
                   
                 pyrophosphokinase 4 
                 (Mouse-ear cress) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 (EC 2.7.6.1) (Phosphoribosyl 
                   
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 pyrophosphate synthase 4) 
                   
                   
               
               
                 135 
                 DNA seq for 
                 O59667 
                 Histidine biosynthesis 
                 Schizosaccharomyces pombe 
                 modified codon usage for 
               
               
                   
                 enzyme O59667 
                   
                 bifunctional  
                 (strain 972/ATCC 24843) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                 protein his7 [Includes: 
                 (Fission yeast) 
                 Saccharomyces cerevisiae 
               
               
                   
                   
                   
                 Phosphoribosyl-AMP 
                   
                   
               
               
                   
                   
                   
                 cyclohydrolase (EC 3.5.4.19); 
                   
                   
               
               
                   
                   
                   
                 Phosphoribosyl-ATP 
                   
                   
               
               
                   
                   
                   
                 pyrophosphatase 
                   
                   
               
               
                   
                   
                   
                 (EC 3.6.1.31)] 
                   
                   
               
               
                 136 
                 DNA seq2 for 
                 P38635 
                 Histidinol-phosphatase 
                 Saccharomyces cerevisiae 
                 modified codon usage for 
               
               
                   
                 enzyme P38635 
                   
                 (HolPase) (EC 3.1.3.15) 
                 (strain ATCC 204508/S288c) 
                 Corynebacterium glutamicum and 
               
               
                   
                   
                   
                   
                 (Baker&#39;s yeast) 
                 Saccharomyces cerevisiae 
               
               
                 137 
                 DNA seq for 
                 A4QEF2 
                 Glucose-6-phosphate 
                 Corynebacterium glutamicum 
                 modified codon usage for 
               
               
                   
                 enzyme 
                   
                 1-dehydrogenase 
                 (strain R) 
                 Corynebacterium glutamicum and 
               
               
                   
                 A4QEF2 with 
                   
                 (G6PD) (EC 1.1.1.49) 
                   
                 Saccharomyces cerevisiae 
               
               
                   
                 substitution A243T 
                   
                   
                   
                   
               
               
                 138 
                 DNA seq7 for 
                 P00862 
                 Histidine decarboxylase 
                 Lactobacillus sp. (strain 30a) 
                 Yarrowia lipolytica 
               
               
                   
                 enzyme P00862 
                   
                 proenzyme 
                   
                   
               
               
                 139 
                 DNA seq5 
                 A0A0J6K 
                 Histidine decarboxylase 
                 Chromobacterium sp. LK1 
                 modified codon usage for 
               
               
                   
                 for enzyme 
                 M89 
                   
                   
                 Corynebacterium glutamicum and 
               
               
                   
                 A0A0J6KM89 
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                 140 
                 DNA seq6 
                 Q9Z472 
                 ATP phosphoribosyl transferase 
                 Corynebacterium glutamicum 
                 modified codon usage for 
               
               
                   
                 for enzyme Q9Z472 
                   
                   
                 ATCC 13032 
                 Corynebacterium glutamicum and 
               
               
                   
                 with substitution 
                   
                   
                   
                 Saccharomyces cerevisiae 
               
               
                   
                 N215K, L231F, T235A 
                   
                   
                   
                   
               
               
                 141 
                 DNA seq4 for 
                 Q9Z472 
                 ATP phosphoribosyltransferase 
                 Corynebacterium glutamicum 
                 Saccharomyces cerevisiae 
               
               
                   
                 enzyme Q9Z472 
                   
                   
                 (strain ATCC 13032/ 
                   
               
               
                   
                   
                   
                   
                 DSM 20300/JCM 1318/ 
                   
               
               
                   
                   
                   
                   
                 LMG 3730/NCIMB 10025) 
                   
               
               
                 142 
                 AA seq for 
                   
                   
                   
                   
               
               
                   
                 N-terminal 
                   
                   
                   
                   
               
               
                   
                 solubility tag 
                   
                   
                   
                   
               
               
                 143 
                 AA seq for 
                 E7EAU9 
                 Ribose-phosphate 
                 Bacillus amyloliquefaciens 
                 Yarrowia lypolytica 
               
               
                   
                 enzyme E7EAU9 
                   
                 pyrophosphokinase (RPPK) 
               
               
                   
               
            
           
         
       
     
     REFERENCES 
     
         
         1. Gezginc, Y., et al., Biogenic amines formation in  Streptococcus thermophilus  isolated from home-made natural yogurt. Food Chem, 2013. 138(1): p. 655-62. 
         2. Byun, B. Y. and J. H. Mah, Occurrence of biogenic amines in Miso, Japanese traditional fermented soybean paste. J Food Sci, 2012. 77(12): p. T216-23. 
         3. Landete, J. M., et al., Molecular methods for the detection of biogenic amine-producing bacteria on foods. Int J Food Microbiol, 2007. 117(3): p. 258-69. 
         4. Ferstl, R., et al., Histamine receptor 2 is a key influence in immune responses to intestinal histamine-secreting microbes. J Allergy Clin Immunol, 2014. 134(3): p. 744-746 e3. 
         5. Tabanelli, G., et al., Effect of chemico-physical parameters on the histidine decarboxylase (HdcA) enzymatic activity in  Streptococcus thermophilus  PRI60. J Food Sci, 2012. 77(4): p. M231-7. 
         6. Wauters, G., et al., Histidine decarboxylase in Enterobacteriaceae revisited. J Clin Microbiol, 2004. 42(12): p. 5923-4. 
         7. Lee, M. E., et al., A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth Biol, 2015. 4(9): p. 975-86. 
         8. Roland, B. P., et al., Triosephosphate isomerase I170V alters catalytic site, enhances stability and induces pathology in a  Drosophila  model of TPI deficiency. Biochim Biophys Acta, 2015. 1852(1): p. 61-9.