Abstract:
The present invention is related to a population of  Clostridium acetobutylicum  useful for the production of 1,3-propanediol (PDO), wherein said population comprises at least one strain of a  Clostridium acetobutylicum  sp. comprising mutations selected among the mutations identified in table 1, wherein relative percentages of said mutations are selected among specific genes.

Description:
[0001]    The present invention concerns a new modified microorganism for the production of 1,3-propanediol. This microorganism is adapted for growth and production of 1,3-propanediol from a culture medium with high glycerine content and specifically with a high concentration of industrial glycerine. The invention also concerns culture conditions of said adapted microorganisms and process for the production of 1,3-propanediol. The invention concerns, finally, 1,3-propanediol produced by the modified microorganism and its applications. 
       BACKGROUND OF THE INVENTION 
       [0002]    1,3-propanediol (PDO), also called trimethylene glycol or propylene glycol, is one of the oldest know fermentation products. It was originally identified as early as 1881 by August Freund in a glycerine fermented culture containing  Clostridium pasteurianum . PDO is a typical product of glycerine fermentation and has been found in anaerobic conversions of other organic substrates. Only very few organisms, all of them bacteria, are able to form it. They include enterobacteria of the genera  Klebsiella  ( K. pneumoniae ),  Enterobacter  ( E. agglomerans ) and  Citrobacter  ( C. freunddi ),  Lactobacilli  ( L. brevis  and  L. buchneri ) and  Clostridia  of the  C. butyricum  and the  C. pasteurianum  group. 
         [0003]    PDO, as a bifunctional organic compound, may potentially be used for many synthesis reactions, in particular as a monomer for polycondensations to produce polyesters, polyethers, polyurethanes, and in particular, polytrimethylene terephtalate (PTT). These structure and reactivity features lead to several applications in cosmetics, textiles (clothing fibers or flooring) or plastics (car industry and packing or coating). 
         [0004]    PDO can be produced by different chemical routes but they generate waste stream containing extremely polluting substances and the cost of production is high. Thus, chemically produced PDO can not compete with the petrochemically available diols like 1,2-ethanediol, 1,2-propanediol and 1,4-butanediol. To increase this competitiveness, in 1995, DuPont started a research program for the biological conversion of glucose to PDO. Although this process is environmentally friendly it has the disadvantage to i) use vitamin B12 a very expensive cofactor and ii) be a discontinuous process due to the instability of the producing strain. 
         [0005]    Due to the availability of large amounts of glycerine produced by the bio-diesel industry, a continuous, vitamin-B12-free process with a higher carbon yield would on the contrary, be advantageous. 
         [0006]    It is known in the art that PDO can be produced from glycerine, an unwanted by-product of the biodiesel production that contains roughly 80-85% of glycerine mixed with salts and water. 
         [0007]      C. butyricum  was previously described as being able to grow and produce PDO from industrial glycerine in batch and two-stage continuous fermentation (Papanikolaou et al., 2000). However, at the highest glycerine concentration, the maximal PDO titer obtained was 48.1 g·L −1  at a dilution rate of 0.02 h −1 , meaning a productivity of 0.9 g·L −1 ·h −1 . The cultures were conducted with a maximum glycerine concentration in the fed medium of 90 g·L −1  and in the presence of yeast extract, a costly compound containing organic nitrogen that is known by the man skilled in the art to help increase bacterial biomass production. 
         [0008]    Application WO2006/128381 discloses the use of this glycerine for the production of PDO with batch and fed-batch cultures using natural PDO producing organisms such as  Klebsiella pneumoniae, C. butyricum  or  C. pasteuricum . Furthermore, the medium used in WO2006/128381 also contains yeast extract. As described in this patent application, the maximal productivity reached was comprised between 0.8 and 1.1 g. 
         [0009]    The performance of a  C. acetobutylicum  strain modified to contain the vitamin B12-independent glycerine-dehydratase and the PDO-dehydrogenase from  C. butyricum , called “ C. acetobutylicum  DG1 pSPD5” has been described in Gonzalez-Pajuelo et al., 2005. This strain originally grows and produces PDO in a fed medium containing up to 120 g·l −1  of pure glycerine. In addition, analyses in a fed medium containing a maximum of 60 g·l −1  of pure or industrial glycerine did not point out to any differences. These results have been obtained in presence of yeast extract. Moreover, no test was performed with concentrations of industrial glycerine higher than 60 g·l −1 . When comparing a wild-type  C. butyricum  to the modified microorganism “ C. acetobutylicum  DG1 pSPD5”, a globally similar behaviour was observed. 
         [0010]    In patent application PCT/EP2010/056078 the inventors have described a process to adapt the strain of  C. acetobutylicum  DG1 pSPD5 such as described in Gonzalez-Pajuelo et al. (2005) to grow in a medium with a high concentration of industrial glycerine and without yeast extract. The resulting strain is able to produce PDO in medium containing up to 120 g·l −1  of industrial glycerine with a titer up to 53.5 g·L −1  of PDO, a yield up to 0.53 g·g −1  and a productivity up to 2.86 g·L −1 ·h −1 . 
         [0011]    In the present patent application, the inventors highlight the main genetics modifications of the adapted microorganism useful for the production of PDO, such as obtained after adaptation in presence of high concentration of industrial glycerine. 
       BRIEF DESCRIPTION OF THE INVENTION 
       [0012]    The present invention concerns a population of  Clostridium acetobutylicum  useful for the production of 1,3-propanediol (PDO), wherein said population comprises at least one strain of a  Clostridium acetobutylicum  sp. comprising mutations selected among the mutations identified in Table 1, wherein relative percentages of said mutations are selected among the following gene families: 
         [0000]    
       
         
               
               
               
             
           
               
                   
                   
               
               
                   
                 Gene family and function 
                 Minimum % 
               
               
                   
                   
               
             
             
               
                   
                 Transcription translation regulation 
                 12-15 
               
               
                   
                 Transporters 
                 10-12 
               
               
                   
                 Hypothetical proteins 
                  8-11 
               
               
                   
                 Energy metabolism 
                  7-10 
               
               
                   
                 Intergenic 
                  7-10 
               
               
                   
                 Carbohydrate metabolism 
                 5-7 
               
               
                   
                 Membrane proteins 
                 2-5 
               
               
                   
                 Nucleic acid metabolism 
                 2-5 
               
               
                   
                 Amino acid metabolism 
                 1-3 
               
               
                   
                 Cell division 
                 1-3 
               
               
                   
                 Sporulation 
                 1-3 
               
               
                   
                 Cell adhesion 
                 0-1 
               
               
                   
                 Cellulase 
                 0-1 
               
               
                   
                 Glycerol metabolism 
                 0-1 
               
               
                   
                 Lipid metabolism 
                 0-1 
               
               
                   
                 Proteases/Peptidases 
                 0-1 
               
               
                   
                 Cell motility 
                 0-1 
               
               
                   
                   
               
             
          
         
       
     
         [0013]    Particularly, the population of the invention comprises at least one strain of  Clostridium acetobutylicum  selected among the group consisting of:
       strain DG1 pSPD5 PD0001VE05c01 deposited at CNCM under accession number I-4378;   strain DG1 pSPD5 PD0001VE05c05 deposited at CNCM under accession number I-4379;   strain DG1 pSPD5 PD0001VE05c07 deposited at CNCM under accession number I-4380.
 
CNCM means “Collection Nationale de Cultures de Microorganismes” at the Pasteur Institute, Paris.
       
 
         [0017]    In a particular embodiment of the invention, the population comprises the above strains further mutated with at least one of the following point mutations:
       C is replaced with T at locus CA_C0175, position 198989 in the  Clostridium acetobutylicum  genome, coding for a predicted sugar phosphate isomerase, homolog of an eukaryotic glucokinase regulator (carbohydrate metabolism)   G is replaced with A at locus CA_C1300, position 1444099 in the  Clostridium acetobutylicum  genome, coding for an RNA polymerase sigma factor RPOD transcription and translation regulation)   C is replaced with T at locus CA_C2670, position 2787387 in the  Clostridium acetobutylicum  genome, coding for a Glu-tRNAGln amidotransferase subunit A (transcription and translation regulation)   C is replaced with T at locus CA_C3339, position 3512658 in the  Clostridium acetobutylicum  genome, coding for an ATPase component of an ABC transporter (two ATPase domains)   C is replaced with T at locus CA_C1610, position 1752341 in the  Clostridium acetobutylicum  genome, coding for a branched-chain amino acid permease (transporter).       
 
         [0023]    The present invention also concerns a method for the production of 1,3-propanediol, comprising culturing a population of  Clostridium acetobutylicum  useful for the production of 1,3-propanediol (PDO) according to the invention in a culture medium comprising glycerine as sole source of carbon and recovering the 1,3-propanediol produced from the culture medium. 
       DETAILED DESCRIPTION OF THE INVENTION 
       [0024]    Population of  Clostridium acetobutylicum  Useful for the Production of 1,3-propanediol (PDO) 
         [0025]    A population of  Clostridium acetobutylicum  useful for the production of 1,3-propanediol means one or more strains of  Clostridium acetobutylicum  genetically modified for the production of 1,3-propanediol from glycerine as sole source of carbon. Such strains are known in the art and disclosed, particularly, in applications WO200104324 and WO2008052595. The population according to the invention may be a combination of several strains, the majority of which comprising the mutations according to the invention, as well as a single strain, and particularly strain DG1 pSPD5 PD0001VE05c01, DG1 pSPD5 PD0001VE05c05 or DG1 pSPD5 PD0001VE05c07 deposited at CNCM under accession numbers I-4378, I-4379, I-4380 respectively, or strain DG1 pSPD5 PD0001VE05c08. 
         [0026]    Mutations are changes of nucleotides in the strain genome, more particularly SNPs (“Single Nucleotide Polymorphisms”), identified when compared to the mother strain DG1 pSPD5 PD0001VT. Said strain is disclosed in WO200104324 and is derived from strain ATCC824 which genome sequence has been published (Nölling et al., 2001). 
         [0027]    Mutations can occur in coding or non-coding sequences. These mutations can be synonymous wherein there is not modification of the corresponding amino acid or non-synonymous wherein the corresponding amino acid is altered. Synonymous mutations do not have any impact on the function of translated proteins, but may have an impact on the regulation of the corresponding genes or even of other genes, if the mutated sequence is located in a binding site for a regulator factor. Non-synonymous mutations may have an impact on the function of the translated protein as well as on regulation depending the nature of the mutated sequence. 
         [0028]    The population of  Clostridium acetobutylicum  useful for the production of 1,3-propanediol may preferably comprise one of those deposited strains comprising additional modifications, at least one of the following modifications:
       C replaced with T at locus CA_C0175, position 198989 in the  Clostridium acetobutylicum  genome, coding for a predicted sugar phosphate isomerase, homolog of an eukaryotic glucokinase regulator (carbohydrate metabolism)   G replaced with A at locus CA_C1300, position 1444099 in the  Clostridium acetobutylicum  genome, coding for an RNA polymerase sigma factor RPOD transcription and translation regulation)   C replaced with T at locus CA_C2670, position 2787387 in the  Clostridium acetobutylicum  genome, coding for a Glu-tRNAGln amidotransferase subunit A (transcription and translation regulation)   C replaced with T at locus CA_C3339, position 3512658 in the  Clostridium acetobutylicum  genome, coding for an ATPase component of an ABC transporter (two ATPase domains)   C replaced with T at locus CA_C1610, position 1752341 in the  Clostridium acetobutylicum  genome, coding for a branched-chain amino acid permease (transporter).       
 
         [0034]    It may preferably comprise any combinations of these mutations, comprising 1, 2, 3, 4 or 5 of these mutations. 
         [0035]    The population of strains of the invention is capable of growing on a medium comprising up to 120 g·L −1  of glycerine and more particularly of industrial glycerine. 
         [0036]    The strains of the population of the invention may be obtained using standard techniques of mutagenesis and/or gene replacement in  Clostridium , such as disclosed in application WO2008040387 which contents are incorporated herein by reference. 
         [0037]    The person skilled in the art may start from one of the strains disclosed in applications WO200104324 and WO2008052595 as well as use one of the strains c01, c05 or c07 deposited at CNCM under accession numbers I-4378, I-4379, I-4380 respectively, and introduce additional mutations. 
         [0038]    In a preferred embodiment, the population of the invention comprises strain DG1 pSPD5 PD0001VE05c08, which mutations are identified in Table 1. The person skilled in the art knows how to introduce the mutations into a  Clostridium  strain to generate a strain similar to strain DG1 pSPD5 PD0001VE05c08, starting from one of strains DG1 pSPD5 PD0001VE05c01, DG1 pSPD5 PD0001VE05c05 or DG1 pSPD5 PD0001VE05c07, deposited at CNCM under accession numbers I-4378, I-4379, I-4380 respectively and using standard gene replacement and recombination techniques. 
       Culture Medium Comprising Glycerine 
       [0039]    An “appropriate culture medium” or a “culture medium” refers to a culture medium optimized for the growth and the diol-production of the  Clostridium  strains or population. The fermentation process is generally conducted in reactors with a synthetic, particularly inorganic, culture medium of known defined composition adapted to the  Clostridium  species used and containing glycerine. 
         [0040]    The term “synthetic medium” means a culture medium comprising a chemically defined composition on which organisms are grown. In the culture medium of the present invention, glycerine is advantageously the single source of carbon. 
         [0041]    The terms “glycerine” and ‘glycerol” are synonymous and used interchangeably in this invention to refer to the same molecule. 
         [0042]    In a particular embodiment, glycerine is added to the medium in the form of glycerine composition comprising at least 50% of glycerine, preferably at least 85% of glycerine. 
         [0043]    Advantageously, the glycerine used in the culture medium of the invention is industrial glycerine. “Industrial glycerine” means a glycerine product obtained from an industrial process without substantial purification. Industrial glycerine can also be designated as “raw glycerine”. Industrial glycerine comprises more than 70% of glycerine, preferably more than 80%, water and impurities such as mineral salts and fatty acids. The maximum content of glycerine in industrial glycerine is generally 90%, more generally about 85%. 
         [0044]    Industrial processes form which industrial glycerine is obtained are, inter alia, manufacturing methods where fats and oils, particularly fats and oils of plant origin, are processed into industrial products such as detergent or lubricants. In such manufacturing methods, industrial glycerine is considered as a by-product. 
         [0045]    In a particular embodiment, the industrial glycerine is a by-product from biodiesel production and comprises known impurities of glycerine obtained from biodiesel production, comprising about 80 to 85% of glycerine with salts, water and some other organic compounds such as fatty acids. Industrial glycerine obtained from biodiesel production has not been subjected to further purification steps. 
         [0046]    Preferably, the culture medium comprises high concentrations of glycerine. 
         [0047]    The terms “high glycerine content” or “high concentration of glycerine” means more than 90 g·L −1  of glycerine in the culture medium. In preferred embodiments, the concentration is comprised between 90 and 200 g·L −1  of glycerine, more particularly between 90 and 140 g/L of glycerine, preferably about 120 g·L −1  of glycerine. 
         [0048]    Preferably, the culture medium is a synthetic medium without addition of organic nitrogen. 
         [0049]    Such culture media are disclosed in the art, particularly in PCT/EP2010/056078 filed on May 5, 2010 and PCT/EP2010/064825 filed on May 10, 2010, which contents are incorporated herein by reference. 
       Culturing the Microorganisms 
       [0050]    In the method of the invention, the production is advantageously done in a batch, fed-batch or continuous process. Culturing microorganisms at industrial scale for the production of 1,3-propanediol is known in the art, particularly disclosed in PCT/EP2010/056078 filed on May 5, 2010 and PCT/EP2010/064825 filed on May 10, 2010, which content are incorporated herein by reference. 
         [0000]    1,3-propanediol Recovery 
         [0051]    Methods for recovering and eventually purifying 1,3-propanediol from a fermentation medium are known to the skilled person. 1,3-propanediol may be isolated by distillation. In most embodiments, 1,3-propanediol is distilled from the fermentation medium with a by-product, such as acetate, and then further purified by known methods. 
         [0052]    A particular purification method is disclosed in applications WO2009/068110 and WO 2010/037843, which content is incorporated herein by reference. 
     
    
     
       FIGURES 
         [0053]      FIG. 1  describes the evolution of 1,3-propanediol production and glycerine consumption of the population and clone c08 during the chemostat cultures from inoculation up to D=0.06 h −1 . 
       
    
    
     EXAMPLES 
     Example 1 
     Isolation of Clones from the Evolved Population 
       [0054]    Clone isolation was performed on agar plates starting from a growing flask culture of the population strain  Clostridium acetobutylicum  DG1 pSPD5 PD0001VE05. The synthetic media used for flask culture contained per liter of deionized water:glycerine, 30 g; KH 2 PO 4 , 0.5 g; K 2 HPO 4 , 0.5 g; MgSO 4 , 7H 2 O, 0.2 g; CoCl 2  6H 2 O, 0.01 g; acetic acid, 99.8%, 2.2 ml; NH 4 Cl, 1.65 g; MOPS, 23.03 g, biotin, 0.16 mg; p-amino benzoic acid, 32 mg; FeSO 4 , 7H 2 O, 0.028 g; resazurin, 1 mg and cysteine, 0.5 g. The pH of the medium was adjusted to 6.5 with NH 4 OH 6N. 
         [0055]    Different media were used for isolation on agar plates:synthetic agar medium (the same as described above) with either commercial glycerine or raw glycerine and CGM (Clostridial Growth Medium) agar medium which contains per liter of deionized water:commercial or raw glycerine, 30 g; yeast extract, 5 g; KH 2 PO 4 , 0.75; K 2 HPO 4 , 0.75 g; MgSO 4 , 7H 2 O, 0.4 g; asparagine, 2 g; (NH 4 ) 2 SO 4 , 2 g; NaCl, 1 g; MnSO 4 , H2O, 10 mg; FeSO 4 , 7H 2 O, 10 mg; MOPS, 23.03 g; resasurin, 1 mg and cysteine, 15 g. The pH of the medium was adjusted to 6.6 with NH 4 OH 3N. 
         [0056]    Cells were plated from a flask culture (Table 2) in four different ways:
       on agar plates of synthetic medium with commercial glycerine;   on agar plates of synthetic medium with raw glycerine;   on agar plates of rich medium with commercial glycerine;   on agar plates of rich medium with raw glycerine.       
 
         [0061]    Isolated clones were considered pure after three subsequent subcultures on agar plates. Pure clones were then transferred into liquid rich medium which contained either commercial or raw glycerine (Table 2). Subsequently, growing liquid cultures were conserved on glycerine 20% at −80° C. until further characterization. 
         [0062]    Clones were then characterized in the following way:
       Measurement of viability after conservation: evaluation of growth rate of cells on synthetic medium;   Evaluation of growth and metabolism: measurement of OD 620 nm  during culture and PDO/glycerine yield on synthetic medium;   Genetic evaluation: PCR analysis to confirm the genotype of the strain;   Chemostat culture to compare the performances of isolated clones with those of the population (example 2);   gDNA extraction for sequence analysis of the clones (example 3).       
 
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Synthetic agar media and liquid media used for 
               
               
                 the isolation of 4 clones from the population. 
               
             
          
           
               
                 Clone 
                   
                 Liquid media for clone culture 
               
               
                 number 
                 Agar media for isolation 
                 before conservation 
               
               
                   
               
               
                 c01 
                 Synthetic medium with 
                 Rich medium with commercial 
               
               
                   
                 commercial glycerine 
                 glycerine 
               
               
                 c05 
                 Rich medium with raw 
                 Rich medium with commercial 
               
               
                   
                 glycerine 
                 glycerine 
               
               
                 c07 
                 Synthetic medium with 
                 Rich medium with raw glycerine 
               
               
                   
                 commercial glycerine 
               
               
                 c08 
                 Rich medium with 
                 Rich medium with raw glycerine 
               
               
                   
                 commercial glycerine 
               
               
                   
               
             
          
         
       
     
       Example 2 
     Performances of Clone c08 in a Chemostat Culture with High Concentrations of Raw Glycerine 
     Bacterial Strain: 
       [0068]    Isolated clone of  C. acetobutylicum  strain DG1 pSPD5 PD0001VE05 (strain was 1/cured from pSOL1 2/transformed with plasmid pSPD5 harbouring dhaB1, dhaB2 and dhaT genes, ie 1,3-propanediol operon, and 3/evolved on high concentrations of raw glycerine). The isolation protocol was described in example 1. 
       Culture Media: 
       [0069]    The synthetic media used for clostridia batch cultivations contained per liter of deionized water: glycerine, 30 g; KH 2 PO 4 , 0.5 g; K 2 HPO 4 , 0.5 g; MgSO 4 , 7H 2 O, 0.2 g; CoCl 2  6H 2 O, 0.01 g; H 2 SO 4 , 0.1 ml; NH 4 Cl, 1.5 g; biotin, 0.16 mg; p-amino benzoic acid, 32 mg and FeSO 4 , 7H 2 O, 0.028 g. The pH of the medium was adjusted to 6.3 with NH 4 OH 3N. Commercial glycerine purchased from Sigma (purity 99.5%) was used for batch cultivation. The feed medium for continuous cultures contained per liter of tap water:raw glycerine, 105 g; KH 2 PO 4 , 0.5 g; K 2 HPO 4 , 0.5 g; MgSO 4 , 7H 2 O, 0.2 g; CoCl 2  6H 2 O, 0.026 g; NH 4 Cl, 1.5 g; biotin, 0.16 mg; p-amino benzoic acid, 32 mg; FeSO 4 , 7H 2 O, 0.04 g; anti-foam, 0.05 ml; ZnSO 4 , 7H 2 O, 8 mg; CuCl 2 , 2H 2 O, 4 mg; MnSO 4 , H 2 O, 40 mg; H 3 BO 3 , 2 mg; Na 2 MoO 4 , 2H 2 O, 0.8 mg. Medium pH was not adjusted in this case. Raw glycerine, from the transesterification process for biodiesel, was supplied by Novance (Venette, France) and had the following purity: glycerine 84.8% (w/w). 
       Experimental Set-Up: 
       [0070]    Continuous cultures were performed in a 51 bioreactor Tryton (Pierre Guerin, France) with a working volume of 2000 ml. The culture volume was kept constant at 2000 ml by automatic regulation of the culture level. Cultures were stirred at 200 RPM, the temperature was set to 35° C. and pH maintained constant at 6.5 by automatic addition of NH 4 OH 5.5N. The POR measurement (mV) was controlled during the entire culture. To create anaerobic conditions, the sterilized medium in the vessel was flushed with sterile O 2 -free nitrogen for one hour at 60° C. and flushed again until 35° C. was attained (flushing during 2 hours). The bioreactor gas outlet was protected from oxygen by a pyrogallol arrangement (Vasconcelos et al, 1994). After sterilisation the feed medium was also flushed with sterile O 2 -free nitrogen until room temperature was attained and maintained under nitrogen at 200 mbar to avoid O 2  entry. 
       Batch and Continuous Cultures Process: 
       [0071]    The process used to evaluate has been described in patent application PCT/EP2010/056078 (example 2). 
         [0072]    A culture growing in a 100 ml flask on synthetic medium (the same as described above for batch culture but with addition of acetic acid, 2.2 g·L −1  and MOPS, 23.03 g·L −1 ) taken at the end of exponential growth phase was used as inoculum (5% v/v). 
         [0073]    Cultures were first grown batchwise. At the early exponential growth phase we performed a pulse of commercial glycerine: For the pulse synthetic medium (the same as described for batch culture) with 105 g·L −1  raw glycerine was added at a static flow rate during 3 hours (i.e. an addition of 18 g·L −1  of glycerine). Then the growth continued batchwise and before the end of the exponential growth phase the continuous feeding started with a dilution rate of 0.025 h −1 : The feed medium contains 105 g·L −1  of raw glycerine. 8-10 days after inoculation of the bioreactor and after 3 residence times the dilution rate was increased from 0.025 h −1  to 0.060 h −1  by different stages: Increase of 0.01 h −1  units in 48 hours—no change for 24-hours—increase of 0.01 h −1  units in 48 hours—no change for 24 hours—increase of 0.015 h −1  unit in 48 hours. After that, stabilisation of the culture was followed by 1,3-propanediol production and glycerine consumption ( FIG. 1 ) using the HPLC protocol described below. Particularly we waited until the concentration of residual glycerine was as low as possible. 
         [0074]    The overall performances of c08 clone are presented in Table 3 and compared with performances of the population under the same conditions and with performances of the strain  C. acetobutylicum  DG1 pSPD5 PD0001VT such as described in Gonzalez-Pajuelo et al. (2005). 
       Analytical Procedures: 
       [0075]    Cell concentration was measured turbidimetrically at 620 nm and correlated with cell dry weight determined directly. Glycerine, 1,3-propanediol, ethanol, butanol, acetic and butyric acids concentrations were determined by HPLC analysis. Separation was performed on a Biorad Aminex HPX-87H column and detection was achieved by refractive index. 
         [0076]    Operating conditions were as follows: mobile phase sulphuric acid 0.5 mM; flow rate 0.5 ml/min, temperature, 25° C. 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 performances of the  C. acetobutylicum  DG1 pSPD5 population PD0001VE05 (mean data 
               
               
                 from 4 chemostats), of clone c08 PD0001VE05c08. The feed medium contained 105 g · L −1   
               
               
                 of raw glycerine, dilution rate was 0.060 h −1  and 0.025 h −1 . Values correspond to the 
               
               
                 average of samples analyzed after at least 3 residences times at dilution rate of 0.060 h −1 . 
               
             
          
           
               
                   
                   
                   
                 Average and standard 
               
               
                   
                 Average and 
                 Average and 
                 deviation of the strain 
               
               
                   
                 standard deviation 
                 standard deviation of 
                 PD0001VT 
               
               
                   
                 for clone c08 
                 the population 
                 (Gonzalez-Pajuelo et 
               
               
                   
                 PD0001VE05c08 
                 PD0001VE05 
                 al. 2005) 
               
               
                   
                   
               
             
          
           
               
                 Feed glycerol (g · l −1 ) 
                 105.49 +/− 1.07  
                 104.21 +/− 1.36  
                 58.54 
               
               
                 1,3-propanediol (g · l −1 )) 
                 51.30 +/− 0.54  
                 50.45 +/− 1.00  
                 29.76 
               
               
                 Y 1,3-PDO  (g · g −1 ) 
                 0.50 +/− 0.01 
                 0.53 +/− 0.01 
                 0.50 
               
               
                 q 1,3pdo  (q · l −1  · h −1 ) 
                 3.05 +/− 0.03 
                 3.18 +/− 0.21 
                 1.49 
               
               
                 Dilution rate (h −1 ) 
                 0.059 +/− 0.001 
                 0.063 +/− 0.004 
                 0.05 
               
               
                 Residual glycerol (g · l −1 ) 
                 3.72 +/− 1.62 
                 4.82 +/− 1.82 
                 0 
               
               
                 Biomass (g · l −1 ) 
                 1.52 +/− 0.55 
                 2.09 +/− 0.15 
                 1.64 
               
               
                 Acetic acid (g · l −1 ) 
                 2.67 +/− 0.26 
                 2.09 +/− 0.27 
                 NI 
               
               
                 Butyric acid (g · l −1 ) 
                 10.53 +/− 0.38  
                 10.98 +/− 0.37  
                 NI 
               
               
                   
               
               
                 Y 1,3-PDO : PDO yield (g/g of glycerol consumed) 
               
               
                 Q 1,3-PDO : PDO volumetric productivity 
               
               
                 NI: no information. The PD0001VT strain can not grow in a medium lacking yeast extract. 
               
             
          
         
       
     
         [0077]    These results show that the adapted population of  C. acetobutylicum  DG1 pSPD5 is able to grow on higher concentrations of industrial glycerine and thus exhibits a better titer and productivity of PDO on industrial glycerine, than the non adapted strain  C. acetobutylicum  DG1 pSPD5 PD0001VT from Gonzalez-Pajuelo et al. (2005) which can not grow in a medium lacking yeast extract. 
       Example 3 
     Genomic DNA Extraction 
       [0078]    Genomic DNA from strains PD0001VT, PD0001VE05, PD0001VE05c01, PD0001VE05c05, PD0001VE05c07 and PD0001VE05c08 was extracted using Qiagen Genomic kit 500G (Qiagen, Inc., Valencia, Calif.). Briefly, cells were grown anaerobically respectively in rich or synthetic glycerine medium (as described in example 1 and 2) in penicillin vials (70 mL) to late exponential phase (A 620  1.5 to 2.0). Strictly anaerobic conditions were maintained throughout cell lysis. Cells were collected and washed twice in SET buffer (25% sucrose, 0.05 M Tris-HCl, 0.05 M EDTA). Cell pellets were suspended in 11 mL B1 kit buffer with 44 μL RNase, 30 mg/mL lysozyme and 100 μg/mL proteinase K. The mixtures were incubated at 37° C. for 45 min, centrifuged and supernatants were used for DNA extraction according to the Qiagen DNA purification kit instructions. The DNAs were then suspended in 50 μL of 10 mM Tris-HCl (pH8.0). 
       Sequencing Analysis 
       [0079]    Genomes of the native DG1 pSPD5 PD0001VT strain and the evolved population DG1 pSPD5 PD0001VE05 were sequenced using the Roche GS FLX technology. The sequencing project was performed by Eurofins Genomics MWG/Operon (ZA de Courtabeauf-9 Avenue de la Laponie, 91978 Les Ulis Cedex) with for each strain 1 Long-Tag paired end libraries (8 Kb), generation of sequence and scaffolding of the contigs with GS FLX Titanium series chemistry using a half run (max. 600 000 reads, max 180 000-300 000 true paired end reads). 
         [0080]    Isolated clones from the evolved population were sequenced using the comparative genomic sequencing (CGS) method developed by NimbleGen (Roche NimbleGen Inc. 500 S. Rosa Rd. Madison Wis. 53719). The CGS analysis was performed in two phases: in phase 1, regions of genomic difference were identified by a comparative hybridization of DNA of the native strain and the evolved clones. In phase 2, only the identified regions of genomic differences were sequenced so as to produce a set of fully characterized single nucleotide polymorphisms (SNPs). 
       SNP Analysis 
       [0081]    Bioinformatics and SNP analysis of the evolved population were performed by Eurofins Genomics MWG/Operon. For this analysis, the read sets of both strains were separately mapped to the Genbank reference sequence ( Clostridium acetobutylicum  ATCC 824 http://www.ncbi.nlm.nih.gov/nuccore/AE001437) using the software gsMapper (Roche 454, V2.3). Three SNPs files were delivered comparing DG1 pSPD5 PD0001VT to ATCC824, DG1 pSPD5 PD0001VE05 to ATCC824 and DG1 pSPD5 PD0001VT to DG1 pSPD5 PD0001VE05. Unique SNPs between the native and the evolved strains are presented below. Low coverage (&lt;25) and low variant frequency (&lt;85%) were removed resulting in 160 unique SNPs distributed in 17 families according to the KEGG database used for the family group annotations. 
         [0082]    SNP analysis of the isolated clones was performed by NimbleGen (Roche). The SNP files were delivered comparing native DG1 pSPD5 PD0001VT to DG1 pSPD5 PD0001VE05c01, DG1 pSPD5 PD0001VE05c05, DG1 pSPD5 PD0001VE05c07 or DG1 pSPD5 PD0001VE05c08 using Genbank reference sequence ( Clostridium acetobutylicum  ATCC 824 http://www.ncbi.nlm.nih.gov/nuccore/AE001437). 
         [0083]    The sequence results are presented in Table 1 which contains the following information:
   RefStart the start position within the reference sequence, where the difference occurs   RefNuc the reference nucleotide sequence at the difference location   VarNuc the differing nucleotide sequence at the difference location   VarFreq the percentage of different reads versus total reads that fully span the difference location   Type Lists whether or not an SNP is found within an annotated gene, or between annotated genes. SNPs in genes are designated as coding. SNPs between genes are designated as intergenic   AA change categorizes coding SNPs base on whether or not they change the amino acid sequence of a protein. S indicates synonymous SNPs (no amino acid change). N indicates nonsynonymous SNPs (altered amino acid). FC (Frame-Change) indicates a modification in protein translation because of insertion or deletion of a nucleotide   ORIG_AA the amino acid associated with the reference sequence for the corresponding SNP position   SNP_AA the amino acid associated with the test sequence, for the corresponding SNP position   Locus Tag locus tag of the corresponding gene from Genbank   Function the function of the gene as described in Genbank   Family the family of the gene from KEGG   
 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Mutations between native and evolved strains. Mutations were first identified in the adapted population and then presence 
               
               
                 of each mutation was verified in isolated clones (four last columns: Y for presence and N for absence of mutation). 
               
             
          
           
               
                 Ref 
                 Ref 
                 Var 
                 Var 
                   
                 AA 
                 YRIG 
                 SNP 
                 Locus 
                   
                   
                   
                   
                   
                   
               
               
                 Start 
                 Nuc 
                 Nuc 
                 Freq 
                 Type 
                 cha. 
                 AA 
                 AA 
                 Tag 
                 Function 
                 Family 
                 c01 
                 c05 
                 c07 
                 C08 
               
               
                   
               
             
          
           
               
                 15594 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 L 
                 P 
                 CA_C0009 
                 Uncharacterized 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 conserved protein, 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ortholog of YRXA 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 
                   B. subtilis 
                 
               
               
                 21339 
                 G 
                 A 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 25354 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 S 
                 CA_C0017 
                 Seryl-tRNA synthetase 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 29516 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 D 
                 CA_C0020 
                 MDR-type permease 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 31667 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C0021 
                 Seryl-tRNA synthetase 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (serine-tRNA ligase) 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 35547 
                 G 
                 A 
                 &gt;99% 
                 C 
                 S 
                 R 
                 R 
                 CA_C0024 
                 Membrane protein, 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 related to 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   Actinobacillus  protein 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (1944168) 
               
               
                 43029 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C0032 
                 Transcriptional 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator TetR/AcrR 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family 
                 regulation 
               
               
                 52503 
                 A 
                 G 
                 &gt;99% 
                 C 
                 S 
                 A 
                 A 
                 CA_C0039 
                 DNA segregation 
                 Cell division 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase FtsK/SpoIIIE 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family protein, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 contains FHA domain 
               
               
                 76769 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 F 
                 F 
                 CA_C0066 
                 ABC transporter, ATP- 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 binding protein 
               
               
                 143022 
                 G 
                 A 
                 &gt;99% 
                 C 
                 S 
                 S 
                 S 
                 CA_C0135 
                 Hypothetical protein, 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 CF-23 family 
                 proteins 
               
               
                 198989 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 T 
                 I 
                 CA_C0175 
                 Predicted sugar 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 phosphate isomerase, 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 homolog of eucaryotic 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 glucokinase regulator 
               
               
                 219199 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 M 
                 I 
                 CA_C0193 
                 Uncharacterized 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 conserved membrane 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein, affecting 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 LPS biosynthesis 
               
               
                 220222 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 N 
                 S 
                 CA_C0194 
                 Glycosyltransferase 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 involved in cell wall 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 biogenesis 
               
               
                 265152 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 S 
                 CA_C0234 
                 PTS system, fructoso- 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 specific IIBC 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 component 
               
               
                 286308 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 T 
                 CA_C0256 
                 Nitrogenase 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 molybdenum-iron 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein, alpha chain 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (nitrogenase component 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 I) gene nifD 
               
               
                 347024 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 T 
                 CA_C0291 
                 FUSION: methionine 
                 Amino acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthase I (cobalamin 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 dependent) and 5,10 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 methylenetetrahydro- 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 folate reductase 
               
               
                 364263 
                 A 
                 G 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 454074 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 E 
                 K 
                 CA_C0390 
                 Cystathionine gamma- 
                 Amino acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthase 
                 metabolism 
               
               
                 541268 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 T 
                 I 
                 CA_C0471 
                 GrpE protein HSP-70 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 cofactor 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 601197 
                 A 
                 G 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 621074 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 P 
                 L 
                 CA_C0534 
                 Phosphoenolpyruvate 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthase (gene pps) 
                 metabolism 
               
               
                 656050 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 S 
                 P 
                 CA_C0566 
                 Malate dehydrogenase 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 668495 
                 A 
                 G 
                 &gt;99% 
                 C 
                 S 
                 I 
                 I 
                 CA_C0578 
                 Cobalamine-dependent 
                 Amino acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 methionine synthase I 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (methyltransferase and 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 cobalamine-binding 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domain) 
               
               
                 723431 
                 — 
                 A 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 794841 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 T 
                 I 
                 CA_C0688 
                 1-acyl-sn-glycerine-3- 
                 Glycerine 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 phosphate acyltrans- 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ferase 
               
               
                 817479 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C0706 
                 Endo-1,4-beta 
                 Cellulase 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 glucanase (fused to 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 two ricin-B-like 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domains) 
               
               
                 819542 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 P 
                 L 
                 CA_C0707 
                 RNA polymerase sigma- 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 54 factor 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 951584 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 I 
                 V 
                 CA_C0823 
                 Predicted membrane 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 977563 
                 G 
                 A 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 978671 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 C 
                 C 
                 CA_C0850 
                 Nitroreductase family 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 metabolism 
               
               
                 991021 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 V 
                 A 
                 CA_C0861 
                 ABC-type multidrug 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transport system, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase component 
               
               
                 991449 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 R 
                 CA_C0861 
                 ABC-type multidrug 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transport system, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase component 
               
               
                 1019710 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 V 
                 . 
                 CA_C0888 
                 Phosphoglycerine 
                 Glycerine 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transferase MdoB 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 related protein, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 alkaline phosphatase 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 superfamily 
               
               
                 1068817 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 L 
                 S 
                 CA_C0925 
                 TPR-repeat-containing 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 1113238 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 N 
                 S 
                 CA_C0967 
                 Probably membrane 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 1223725 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 T 
                 CA_C1072 
                 Fe—S 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 oxidoreductase 
                 metabolism 
               
               
                 1254865 
                 T 
                 A 
                 &gt;99% 
                 C 
                 N 
                 Y 
                 N 
                 CA_C1086 
                 Transcriptional 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulators of 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 NagC/XylR family 
                 regulation 
               
               
                 1299105 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 M 
                 T 
                 CA_C1133 
                 Phage related protein, 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 YonE  B. subtilis   
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 homolog 
               
               
                 1309504 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 M 
                 I 
                 CA_C1143 
                 Exodeoxyribonuclease 
                 Cell division 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V, Alpha subunit, RecD 
               
               
                 1324897 
                 A 
                 G 
                 &gt;99% 
                 C 
                 S 
                 P 
                 P 
                 CA_C1166 
                 Hypothetical protein 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 1366058 
                 T 
                 A 
                 &gt;99% 
                 C 
                 N 
                 N 
                 K 
                 CA_C1223 
                 DNA Polymerase III 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Alpha chain (dnaE) 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 1424502 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 Y 
                 C 
                 CA_C1280 
                 Transcriptional 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator of heat 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 shock genes, HrcA 
                 regulation 
               
               
                 1444099 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 R 
                 CA_C1300 
                 RNA polymerase sigma 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 factor RPOD 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 1540446 
                 A 
                 G 
                 &gt;99% 
                 C 
                 S 
                 L 
                 L 
                 CA_C1396 
                 Phosphoribosylamine- 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 glycine ligase 
                 metabolism 
               
               
                 1554592 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 K 
                 R 
                 CA_C1408 
                 Phospho-beta-gluco- 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 sidase 
                 metabolism 
               
               
                 1644651 
                 A 
                 — 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 1770126 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 I 
                 V 
                 CA_C1628 
                 DNA gyrase A subunit 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 1778678 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 T 
                 CA_C1636 
                 Uncharacterized 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein, homolog of 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   B. firmus  (2654481) 
               
               
                 1805186 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 V 
                 I 
                 CA_C1661 
                 Predicted secreted 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 nucleic acid binding 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
               
               
                 1821699 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 T 
                 CA_C1673 
                 Large subunit of NADH- 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 dependent glutamate 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthase 
               
               
                 1916660 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 N 
                 S 
                 CA_C1771 
                 Uncharacterized 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein, ykrI 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   B. subtilis  homolog 
               
               
                 1948050 
                 C 
                 T 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 2037205 
                 G 
                 A 
                 &gt;99% 
                 C 
                 S 
                 C 
                 C 
                 CA_C1886 
                 Uncharacterized phage 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 related protein 
                 proteins 
               
               
                 2114483 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 V 
                 A 
                 CA_C2003 
                 Predicted permease 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 2123888 
                 T 
                 C 
                 &gt;99% 
                 C 
                 S 
                 L 
                 L 
                 CA_C2010 
                 Predicted Fe—S 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 oxidoreductase 
                 metabolism 
               
               
                 2171503 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 D 
                 N 
                 CA_C2068 
                 Sporulation factor 
                 Sporulation 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 spoIIM, uncharacterized 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 membrane protein 
               
               
                 2231570 
                 C 
                 — 
                 &gt;99% 
                 C 
                 FC 
                   
                   
                 CA_C2137 
                 Cation transport 
                 Transporters 
                 N 
                 N 
                 N 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 P-type ATPase 
               
               
                 2294764 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 T 
                 I 
                 CA_C2201 
                 Hypothetical protein 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 2299326 
                 C 
                 G 
                 &gt;99% 
                 C 
                 N 
                 S 
                 T 
                 CA_C2205 
                 Flagellar hook- 
                 Cell motility 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 associated protein 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 FliD 
               
               
                 2307214 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 G 
                 R 
                 CA_C2215 
                 Flagellar switch 
                 Cell motility 
                 Y 
                 N 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein FliY, contains 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 CheC-like domain 
               
               
                 2342826 
                 G 
                 C 
                 &gt;99% 
                 C 
                 N 
                 P 
                 A 
                 CA_C2247 
                 Site-specific 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 recombinase, DNA 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 invertase Pin homolog 
                 regulation 
               
               
                 2392178 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 V 
                 . 
                 CA_C2288 
                 Acyl-protein 
                 Lipid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthetase, luxE 
                 metabolism 
               
               
                 2450006 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 P 
                 P 
                 CA_C2340 
                 DNA mismatch repair 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein mutS, YSHD 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   B. subtilis  ortholog 
                 regulation 
               
               
                 2477825 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 S 
                 S 
                 CA_C2367 
                 Uncharacterized 
                 Cell adhesion 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein containing 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 predicted cell 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 adhesion domain and 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ChW-repeats 
               
               
                 2493211 
                 T 
                 C 
                 &gt;99% 
                 C 
                 S 
                 H 
                 H 
                 CA_C2385 
                 Hypothetical protein 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 2595349 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C2486 
                 Transcriptional 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator, MarR/EmrR 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family 
                 regulation 
               
               
                 2693354 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 E 
                 K 
                 CA_C2588 
                 Glycosyltransferase 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 2787387 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 M 
                 I 
                 CA_C2670 
                 Glu-tRNAGln 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 amidotransferase 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 subunit A 
                 regulation 
               
               
                 2833384 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 I 
                 V 
                 CA_C2709 
                 Electron transfer 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 flavoprotein alpha- 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 subunit 
               
               
                 2836979 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C2713 
                 AT-rich DNA-binding 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 2901642 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 V 
                 . 
                 CA_C2770 
                 Amino acid transporter 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 2969858 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 M 
                 I 
                 CA_C2838 
                 Predicted nucleotide- 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 binding protein, YjeE 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family 
                 regulation 
               
               
                 3001642 
                 G 
                 A 
                 &gt;99% 
                 C 
                 S 
                 L 
                 L 
                 CA_C2867 
                 FoF1-type ATP synthase 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 alpha subunit 
                 metabolism 
               
               
                 3032956 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 H 
                 R 
                 CA_C2898 
                 Stage II sporulation 
                 Sporulation 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein R 
               
               
                 3140918 
                 T 
                 C 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 3174743 
                 G 
                 A 
                 &gt;99% 
                 C 
                 S 
                 D 
                 D 
                 CA_C3032 
                 Galactose mutarotase 
                 Carbohydrate 
                 Y 
                 N 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 related enzyme 
                 metabolism 
               
               
                 3251276 
                 G 
                 C 
                 &gt;99% 
                 C 
                 N 
                 T 
                 S 
                 CA_C3099 
                 Pseudouridylate 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 synthase, TRUA 
                 metabolism 
               
               
                 3337937 
                 G 
                 — 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 3392124 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 G 
                 R 
                 CA_C3242 
                 Uncharacterized 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Fe—S protein, 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 PflX (pyruvate formate 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 lyase activating 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein) homolog 
               
               
                 3462380 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 N 
                 N 
                 CA_C3297 
                 D-alanyl-D-alanine 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 carboxypeptidase 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family hydrolase, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 YODJ  B. subtilis   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ortholog 
               
               
                 3509372 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 E 
                 E 
                 CA_C3335 
                 Short-chain alcohol 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 dehydrogenase family 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 enzyme 
               
               
                 3512658 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 Y 
                 Y 
                 CA_C3339 
                 ATPase component of 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ABC transporter (two 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase domains) 
               
               
                 3518240 
                 T 
                 C 
                 &gt;99% 
                 C 
                 S 
                 Y 
                 Y 
                 CA_C3345 
                 Transcriptional 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator, AcrR family 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 3541557 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 I 
                 V 
                 CA_C3363 
                 Hypothetical protein 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 3565291 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 T 
                 I 
                 CA_C3387 
                 Pectate lyase 
                 Cellulase 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 3576865 
                 T 
                 C 
                 &gt;99% 
                 C 
                 N 
                 H 
                 R 
                 CA_C3392 
                 NADH-dependent 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 butanol dehydrogenase 
                 metabolism 
               
               
                 3583724 
                 C 
                 T 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 3608511 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 S 
                 S 
                 CA_C3422 
                 Sugar: proton symporter 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (possible xylulose) 
               
               
                 3614985 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 K 
                 K 
                 CA_C3428 
                 6Fe—6S prismane 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 cluster-containing 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
               
               
                 3674358 
                 T 
                 C 
                 &gt;99% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 3707038 
                 T 
                 C 
                 &gt;99% 
                 C 
                 S 
                 L 
                 L 
                 CA_C3510 
                 Membrane associated 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 methyl-accepting 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 chemotaxis protein 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (with HAMP domain) 
               
               
                 3747653 
                 G 
                 A 
                 &gt;99% 
                 C 
                 N 
                 A 
                 V 
                 CA_C3551 
                 Na+ ABC transporter 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (ATP-binding protein), 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 NATA 
               
               
                 3821135 
                 C 
                 T 
                 &gt;99% 
                 C 
                 S 
                 N 
                 N 
                 CA_C3617 
                 Uncharacterized 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 membrane protein, 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 YHAG  B. subtilis   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 homolog 
               
               
                 3850220 
                 A 
                 G 
                 &gt;99% 
                 C 
                 N 
                 I 
                 T 
                 CA_C3650 
                 HD-GYP domain 
                 Proteases/ 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 containing protein 
                 Peptidases 
               
               
                 3921509 
                 C 
                 T 
                 &gt;99% 
                 C 
                 N 
                 V 
                 I 
                 CA_C3716 
                 Lon-like ATP-dependent 
                 Proteases/ 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protease 
                 Peptidases 
               
               
                 239312 
                 G 
                 A 
                  98% 
                 C 
                 N 
                 E 
                 K 
                 CA_C0214 
                 Endoglucanase, 
                 Cellulase 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 aminopeptidase M42 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 family 
               
               
                 244251 
                 C 
                 T 
                  98% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                 2410308 
                 G 
                 A 
                  98% 
                 C 
                 S 
                 L 
                 L 
                 CA_C2306 
                 Sporulation-specific 
                 Sporulation 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 sigma factor F 
               
               
                 3656844 
                 G 
                 A 
                  98% 
                 C 
                 N 
                 A 
                 T 
                 CA_C3459 
                 Homolog of cell 
                 Cell division 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 division GTPase FtsZ, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 diverged 
               
               
                 3823060 
                 A 
                 G 
                  98% 
                 C 
                 N 
                 V 
                 A 
                 CA_C3620 
                 Amino acid (probably 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 glutamine) ABC 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transporter, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 periplasmic binding 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein component 
               
               
                 3838496 
                 C 
                 T 
                  98% 
                 C 
                 N 
                 G 
                 R 
                 CA_C3637 
                 Oligopeptide ABC 
                 Transporters 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transporter, permease 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 component 
               
               
                 914 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 G 
                 R 
                 CA_C0001 
                 DNA replication 
                 Cell division 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 initiator protein, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase 
               
               
                 27240 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 R 
                 Q 
                 CA_C0019 
                 Transcriptional 
                 Transcription 
                 N 
                 Y 
                 N 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator, AcrR family 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 36341 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 D 
                 N 
                 CA_C0025 
                 Deoxycytidine 
                 Nucleic acid 
                 N 
                 Y 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 triphosphate deaminase 
                 metabolism 
               
               
                 299266 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 D 
                 N 
                 CA_C0267 
                 L-lactate dehydrogenase 
                 Energy 
                 Y 
                 N 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 376886 
                 G 
                 A 
                  97% 
                 C 
                 S 
                 K 
                 K 
                 CA_C0322 
                 Sensory protein, 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 containing EAL-domain 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 474915 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 G 
                 D 
                 CA_C0408 
                 DNA segregation 
                 Cell division 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATP-ase FtsK/SpoIIIE 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (three ATPases), 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 contains FHA domain 
               
               
                 599925 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 R 
                 K 
                 CA_C0519 
                 Dihydroorotase 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 723433 
                 G 
                 A 
                  97% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 723434 
                 GT 
                 — 
                  97% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 723871 
                 A 
                 G 
                  97% 
                 C 
                 N 
                 I 
                 M 
                 CA_C0622 
                 Polyphosphate kinase 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 809008 
                 C 
                 T 
                  97% 
                 C 
                 S 
                 L 
                 L 
                 CA_C0699 
                 Spore photoproduct 
                 Sporulation 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 lyase, splB 
               
               
                 846466 
                 G 
                 T 
                  97% 
                 C 
                 N 
                 A 
                 S 
                 CA_C0731 
                 FUSION: Nucleoside- 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 diphosphate-sugar 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 epimerase and GAF 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domain 
               
               
                 1717948 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 V 
                 I 
                 CA_C1572 
                 Fructose-1,6-bisphos- 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 phatase (YYDE 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   B. subtils  ortholog) 
               
               
                 2004797 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 S 
                 N 
                 CA_C1852 
                 Magnesium and cobalt 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transport protein 
               
               
                 2134058 
                 G 
                 A 
                  97% 
                 C 
                 S 
                 A 
                 A 
                 CA_C2020 
                 Molybdopterin 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 bioSthesis enzyme, 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 MoeA, fused to 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 molibdopterin-binding 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domain 
               
               
                 2331746 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 G 
                 R 
                 CA_C2237 
                 ADP-glucose 
                 Lipid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 pyrophosphorylase 
                 metabolism 
               
               
                 2391588 
                 G 
                 A 
                  97% 
                 C 
                 N 
                 P 
                 L 
                 CA_C2288 
                 Acyl-protein Sthetase, 
                 Lipid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 luxE 
                 metabolism 
               
               
                 2452705 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 C 
                 Y 
                 CA_C2341 
                 Collagenase family 
                 Proteases/ 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protease 
                 Peptidases 
               
               
                 2739459 
                 T 
                 C 
                  97% 
                 C 
                 N 
                 I 
                 V 
                 CA_C2630 
                 Uncharaterized 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 conserved protein, 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 YOME  B. subtilis   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ortholog 
               
               
                 2775979 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 A 
                 T 
                 CA_C2660 
                 Pyruvate carboxylase, 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 PYKA 
                 metabolism 
               
               
                 2813985 
                 G 
                 — 
                  97% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 3082247 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 L 
                 F 
                 CA_C2948 
                 ATPase components of 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ABC transporter with 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 duplicated ATPase 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domains (second domain 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 is inactivated) 
               
               
                 3242900 
                 G 
                 C 
                  97% 
                 C 
                 N 
                 V 
                 L 
                 CA_C3088 
                 NtrC family 
                 Transcription 
                 Y 
                 N 
                 N 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transcriptional 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator, ATPase 
                 regulation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domain fused 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 to two PAS domains 
               
               
                 3442855 
                 T 
                 C 
                  97% 
                 C 
                 N 
                 M 
                 V 
                 CA_C3282 
                 ABC-type 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 multidrug/protein/ 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 lipid transport system, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase component 
               
               
                 3498584 
                 C 
                 T 
                  97% 
                 C 
                 N 
                 L 
                 F 
                 CA_C3327 
                 Amino acid ABC-type 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transporter, ATPase 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 component 
               
               
                 3643224 
                 G 
                 A 
                  97% 
                 C 
                 S 
                 L 
                 L 
                 CA_C3447 
                 Protein-disulfide 
                 Sporulation 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 isomerases DsbC/DsbG 
               
               
                 3663477 
                 — 
                 T 
                  97% 
                 C 
                 FC 
                   
                   
                 CA_C3464 
                 Uncharacterized 
                 Hypothetical 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 conserved protein 
                 proteins 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (fragment) 
               
               
                 204202 
                 G 
                 A 
                  96% 
                 C 
                 N 
                 G 
                 E 
                 CA_C0180 
                 Oligopeptide ABC 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transporter, ATP- 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 binding protein 
               
               
                 803682 
                 C 
                 T 
                  96% 
                 C 
                 N 
                 T 
                 I 
                 CA_C0695 
                 Altronate 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 oxidoreductase 
                 metabolism 
               
               
                 892875 
                 G 
                 A 
                  96% 
                 C 
                 N 
                 M 
                 I 
                 CA_C0770 
                 Glycerine uptake 
                 Glycerine 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 facilitator protein, 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 permease 
               
               
                 1009389 
                 C 
                 T 
                  96% 
                 C 
                 N 
                 P 
                 S 
                 CA_C0879 
                 ABC-type polar amino 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 acid transport system, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase component 
               
               
                 1690355 
                 C 
                 T 
                  96% 
                 C 
                 S 
                 G 
                 G 
                 CA_C1546 
                 Pyrimidine-nucleoside 
                 Nucleic acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 phosphorylase 
                 metabolism 
               
               
                 1752341 
                 C 
                 T 
                  96% 
                 C 
                 N 
                 G 
                 R 
                 CA_C1610 
                 Branched-chain amino 
                 Transporters 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 acid permease 
               
               
                 3217481 
                 A 
                 C 
                  96% 
                 C 
                 S 
                 L 
                 L 
                 CA_C3067 
                 Predicted membrane 
                 Membrane 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 3238489 
                 T 
                 C 
                  96% 
                 C 
                 S 
                 S 
                 S 
                 CA_C3086 
                 Protein containing 
                 Cell adhesion 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 cell adhesion domain 
               
               
                 447460 
                 A 
                 — 
                  95% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 N 
                 N 
                 N 
                 N 
               
               
                 670931 
                 G 
                 A 
                  95% 
                 C 
                 S 
                 N 
                 N 
                 CA_C0578 
                 Cobalamine-dependent 
                 Amino acid 
                 N 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 methionine synthase I 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 (methyltransferase and 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 cobalamine-binding 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 domain) 
               
               
                 994575 
                 G 
                 A 
                  95% 
                 C 
                 N 
                 A 
                 T 
                 CA_C0864 
                 Histidine kinase-like 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 3657101 
                 A 
                 — 
                  95% 
                 C 
                 FC 
                   
                   
                 CA_C3459 
                 Homolog of cell 
                 Cell division 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 division GTPase FtsZ, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 diverged 
               
               
                 1142263 
                 T 
                 — 
                  94% 
                 C 
                 FC 
                   
                   
                 CA_C0995 
                 Predicted membrane 
                 Membrane 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 1823156 
                 G 
                 A 
                  94% 
                 C 
                 S 
                 E 
                 E 
                 CA_C1674 
                 Small subunit of 
                 Amino acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 NADPH-dependent 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 glutamate synthase 
               
               
                 1989117 
                 C 
                 T 
                  94% 
                 C 
                 N 
                 R 
                 K 
                 CA_C1837 
                 Mismatch repair 
                 Transcription 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein MutS, ATPase 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 3481651 
                 G 
                 A 
                  94% 
                 C 
                 S 
                 S 
                 S 
                 CA_C3311 
                 TPR-repeat domain 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 fused to glycosyltrans- 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ferase 
               
               
                 126942 
                 G 
                 A 
                  93% 
                 C 
                 N 
                 E 
                 K 
                 CA_C0116 
                 Carbone-monoxide 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 dehydrogenase, beta 
                 metabolism 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 chain 
               
               
                 302716 
                 — 
                 T 
                  93% 
                 C 
                 FC 
                   
                   
                 CA_C0270 
                 Hypothetical protein 
                 Hypothetical 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 2551103 
                 G 
                 A 
                  93% 
                 C 
                 S 
                 S 
                 S 
                 CA_C2434 
                 Membrane associate 
                 Transcription 
                 Y 
                 N 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 histidine kinase with 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 HAMP domain 
                 regulation 
               
               
                 1834077 
                 C 
                 T 
                  92% 
                 C 
                 N 
                 S 
                 L 
                 CA_C1684 
                 TYPA/BIPA type 
                 Energy 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 GTPase 
                 metabolism 
               
               
                 3927304 
                 G 
                 A 
                  92% 
                 I 
                   
                   
                   
                   
                 I 
                 I 
                 Y 
                 N 
                 Y 
                 N 
               
               
                 786649 
                 — 
                 T 
                  91% 
                 C 
                 FC 
                   
                   
                 CA_C0680 
                 Predicted membrane 
                 Membrane 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 2640439 
                 C 
                 T 
                  91% 
                 C 
                 N 
                 E 
                 K 
                 CA_C2532 
                 Protein containing ChW- 
                 Cell adhesion 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 repeats 
               
               
                 3601904 
                 A 
                 — 
                  91% 
                 C 
                 FC 
                   
                   
                 CA_C3415 
                 ABC-type 
                 Transporters 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 multidrug/protein/lipid 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 transport system, 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 ATPase component 
               
               
                 838350 
                 A 
                 — 
                  89% 
                 C 
                 FC 
                   
                   
                 CA_C0723 
                 Transcriptional 
                 Transcription 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulator, AcrR family 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                 3721023 
                 G 
                 A 
                  89% 
                 C 
                 S 
                 S 
                 S 
                 CA_C3523 
                 Hypothetical protein, 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 CF-7 family 
                 proteins 
               
               
                 803924 
                 G 
                 A 
                  88% 
                 C 
                 N 
                 A 
                 T 
                 CA_C0695 
                 Altronate 
                 Carbohydrate 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 oxidoreductase 
                 metabolism 
               
               
                 3478420 
                 C 
                 T 
                  87% 
                 C 
                 N 
                 G 
                 E 
                 CA_C3309 
                 Predicted membrane 
                 Membrane 
                 N 
                 Y 
                 N 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 protein 
                 proteins 
               
               
                 3853836 
                 T 
                 C 
                  87% 
                 C 
                 N 
                 N 
                 D 
                 CA_C3652 
                 Acetolactate synthase 
                 Amino acid 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 244464 
                 C 
                 T 
                  86% 
                 C 
                 N 
                 S 
                 L 
                 CA_C0220 
                 Hypothetical protein 
                 Hypothetical 
                 Y 
                 Y 
                 Y 
                 Y 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 proteins 
               
               
                 899104 
                 G 
                 A 
                  86% 
                 C 
                 N 
                 M 
                 I 
                 CA_C0776 
                 NCAIR mutase (PurE)- 
                 Nucleic acid 
                 Y 
                 N 
                 Y 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 related protein 
                 metabolism 
               
               
                 658665 
                 T 
                 — 
                  85% 
                 C 
                 FC 
                   
                   
                 CA_C0569 
                 SACPA operon 
                 Transcription 
                 N 
                 N 
                 N 
                 N 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 antiterminator (sacT) 
                 translation 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 regulation 
               
               
                   
               
             
          
         
       
     
       REFERENCES 
       [0000]    
       
         1. González-Pajuelo M, Meynial-Salles I, Mendes F, Andrade J C, Vasconcelos I, and Soucaille P. 2005. Metabolic engineering of  Clostridium acetobutylicum  for the industrial production of 1,3-propanediol from glycerol. Metabolic Engineering 7: 329-336. 
         2. González-Pajuelo M, Meynial-Salles I, Mendes F, Soucaille P. and Vasconcelos I. 2006. Microbial conversion of a natural producer,  Clostridium butyricum  VPI 3266, and an engineered strain,  Clostridium acetobutylicum  DG (pSPD5). Applied and Environmental Microbiology, 72: 96-101. 
         3. Nölling J, Breton G, Omelchenko M V, Makarova K S, Zeng Q, Gibson R, Lee H M, Dubois J, Qiu D, Hitti J, Wolf Y I, Tatusov R L, Sabathe F, Doucette-Stamm L, Soucaille P, Daly M J, Bennett G N, Koonin E V, Smith D R. 2001. Genome sequence and comparative analysis of the solvent-producing bacterium  Clostridium acetobutylicum . Journal of bacteriology 183(16):4823-4838 
         4. Papanikolaou S, Ruiz-Sanchez P, Pariset B, Blanchard F and Fick M. 2000. High production of 1,3-propanediol from industrial glycerol by a newly isolated  Clostridium butyricum  strain. Journal of Biotechnology. 77: 191-208. 
         5. Vasconcelos I, Girbal L, Soucaille P. 1994. Regulation of carbon and electron flow in  Clostridium acetobutylicum  grown in chemostat culture at neutral pH on mixtures of glucose and glycerol. Journal of bacteriology. 176(5): 1443-1450.