Patent Publication Number: US-2003224491-A1

Title: Continuous fermentation process

Description:
FIELD OF THE INVENTION  
       [0001] The present invention relates to a continuous process for the manufacture of proteins. In particular, the invention relates to fermentation assemblies and processes for manufacturing proteins.  
       SUMMARY OF THE INVENTION  
       [0002] In accordance with the present invention it has been found that splitting of cultivation media used in a continuous fermentation process allows one to study the influence on growth and metabolite-production of microorganisms, and thus to determine optimal conditions for the fermentation process. A continuously delivered fermentation medium can generally be split into as many fractions as it contains ingredients. Examples of such ingredients are carbon, nitrogen, phosphorus, and sulfur sources as well as vitamins and complex substrates such as corn steep, yeast extract, and other natural products. Furthermore, every required mineral, micro- or trace element can be provided separately as a solution of a water-soluble salt, such as a chloride, sulfate or nitrate. In this manner, a fermentation medium of any desired composition can be obtained, provided that the desired amounts of the ingredients are (water)-soluble and no disturbing interactions (e.g., precipitation, reaction) occur in the individual feed solutions or in the fermentation medium.  
       [0003] One embodiment of the invention is a fermentation assembly containing a vessel for culturing living cells, at least two storage flasks in fluid communication with the vessel for supply of liquids and a first transport means for transferring the liquids from the storage flasks to the vessel, individual appliances operably connected to the transport means for monitoring the supply of the contents of the storage flasks to the vessel, a harvest flask in fluid communication with the vessel and a second transport means for transferring the fermentation broth from the vessel to the harvest flask, and a device operably connected to the first transport means for controlling and maintaining a constant dilution rate in the vessel with varying rates of individual supply of liquid from the storage flasks to the vessel.  
       [0004] Another embodiment of the invention is a process for the manufacture of a protein. This process includes providing a continuous culture of living cells in a fermentation reactor; and individually feeding nutrients and other agents required for the growth of the cells into the reactor at a constant dilution rate to achieve optimal production of the protein.  
       [0005] A further embodiment of the invention is a fermentation assembly. This fermentation assembly includes a fermentor  1  equipped with inlet tubes  2   a  in fluid communication with a storage flask  2  for supply of liquids to the fermentor, a pump  3  operably connected to the inlet tubes for transporting liquids from the storage flask  2  to the fermentor  1 , a scale  4  in contact with each storage flask for monitoring the amount of liquid supplied to and discharged from the fermentor, a gas inlet  9  and outlet tubes  10  in communication with the fermentor for introducing and removing gas therefrom, a pump  6  operably connected to an outlet tube  5   a  which is in fluid connection with the fermentor, wherein the pump discharges fermentation broth from the fermentor to a harvest flask  5 , a main controlling unit  7  operably connected to the fermentation assembly for overall process monitoring and steering, a controlling unit  11  operably connected to individual control systems  17  for monitoring and steering temperature, pH, gas pressure, fermentor content, and antifoam agents, a circuit  12  for monitoring gas supply and taking samples including an outlet tube from the fermentor, which circuit is operably connected to the outlet to be pump  13 , and gas inlet and outlet flow control devices  14  and  15  operably connected to the gas inlet and outlet tubes  9 ,  10 . 
     
    
    
     BRIEF DESCRIPTION OF DRAWINGS  
     [0006]FIG. 1 is a diagram depicting a fermentation assembly according to the present invention.  
     [0007]FIG. 2 depicts the design of a consensus phytase amino acid sequence.  
     [0008]FIG. 3 depicts the nucleotide sequence of the consensus phytase-1 gene (fcp) and of the primers used to construct it.  
     [0009]FIG. 4 depicts the alignment and consensus sequences of five Basidiomycetes phytases.  
     [0010]FIG. 5 depicts the design of the consensus phytase-10 amino acid sequence.  
     [0011]FIG. 6 depicts the nucleotide and amino acid sequences of consensus phytase-10.  
     [0012]FIG. 7 depicts the amino acid alignment for the design of consensus phytase-11.  
     [0013]FIG. 8 depicts the nucleotide and amino acid sequence of phytase-1-thermo[8]-Q50T-K91A.  
     [0014]FIG. 9 depicts the nucleotide and amino acid sequence of consensus phytase-10-thermo[3]-Q50T-K91A.  
     [0015]FIG. 10 depicts the nucleotide and amino acid sequence of  A. fumigatus  ATCC 13073 phytase α-mutant.  
     [0016]FIG. 11 depicts the nucleotide and amino acid sequence of consensus phytase-7.  
     [0017]FIG. 12 depicts a differential scanning calorimetry (DSC) of consensus phytase-10 ( 12 A) and consensus phytase-1 ( 12 B).  
     [0018]FIG. 13 depicts DSC of consensus phytase-10-thermo[3]-Q50T ( 13 A) and consensus phytase-10-thermo-[3]-Q50T-K91A ( 13 B).  
     [0019]FIG. 14 is a graph comparing the temperature optima between consensus phytase-1, consensus phytase-10, and consensus phytase-10-thermo[3]-Q50TF.  
     [0020]FIG. 15 is a graph depicting the pH 1-dependent activity profile ( 15 A) and substrate specificity ( 15 B) of consensus phytase 10, consensus phytase-10-thermo[3]-Q50T, and consensus phytase-10-thermo[3]-Q50T-K91A.  
     [0021]FIG. 16 depicts the, pH-dependent activity profile ( 16 A) and substrate specificity ( 16 B) of consensus phytase-1-thermo[g]-Q50T and consensus phytase-1-thermo[8]-Q50T-K91A.  
     [0022]FIG. 17 depicts DSC of consensus phytase-1-thermo[8]-Q50T ( 17 A) and consensus phytase-1-thermo[8]-Q50T-K91A ( 17 B).  
     [0023]FIG. 18 is a graph comparing the temperature optima between consensus phytase-1, consensus phytase-1-thermo[3], and consensus phytase-1-thermo[8].  
     [0024]FIG. 19 depicts the pH-dependent activity profile ( 19 A) and substrate specificity ( 19 B) of consensus phytase-1, consensus phytase-7, and the phytase from  A. niger  NRRL 3135.  
     [0025]FIG. 20 depicts DSC of the phytase from  A. fumigatus  ATCC 13073 ( 20 B) and of its stabilized α-mutant ( 20 A).  
     [0026]FIG. 21 is a graph depicting the temperature optima of  A. fumigatus  ATCC 13073 wild type phytase, its α-mutant, and a further stabilized α-mutant.  
     [0027]FIG. 22 depicts the amino acid sequence of consensus phytase-12 (consphy 12). 
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
     [0028] In one aspect, the present invention is a continuous process for the manufacture of proteins by means of a protein-producing microorganism.  
     [0029] More particularly, the invention is a continuous process for the manufacture of a protein using a protein-producing microorganism that may be immobilized on a solid carrier and/or the nutrients and other agents required for growth of the microorganism and optimal production of protein therefrom are fed into a reactor individually at a constant dilution rate.  
     [0030] In a preferred aspect, the invention is a process for the manufacture of a protein using a fermentation assembly that includes (a) a vessel suitable for carrying out reactions with living or inactivated cells; (b) at least two storage flasks connected to the vessel for supply of liquids and means to transport the liquids from the storage flasks to the vessel; (c) individual appliances monitoring the supply of the contents of the storage flasks to the vessel; (d) a harvest flask connected to the vessel and means to transport fermentation broth from the vessel to the harvest flask; and (e) a device for controlling and maintaining a constant dilution rate in the vessel with varying rates of supply of individual liquids from the storage flasks to the vessel.  
     [0031] Any conventional fermentation vessel can be used as the vessel in step (a) above for the purpose of this invention. The vessel may be made of materials such as stainless steel, glass or ceramics, and may have a volume of from e.g., 100 ml to 2500 m 3  although these figures are not critical to the invention. For continuous operation, the inside of the vessel is optionally equipped with a receptacle or sieve plate for uptake of immobilized living cells. These cells may be present in the vessel as a bed of immobilized cells, such as one selected from the group consisting of a fixed bed, an expanded bed, a moving bed, and combinations thereof.  
     [0032] Further, the fermentation vessel is connected to a series of storage flasks that contain nutrient solutions and solutions for maintaining and controlling a desired pH and other parameters, such as foam formation, redox potential, etc., in the fermentation broth. Depending on the particular needs of the fermentation, there may be separate storage flasks for individual supply of substrates to the vessel, which substrates serve as the carbon, nitrogen or mineral source for the living cells in the vessel. In a particular embodiment, at least one of the at least two storage flasks contains a controlling agent. Said controlling agent has the purpose of regulating the pH of the contents of the flask containing it, and may be an acid, e.g., hydrochloric acid, or a base, e.g., sodium hydroxide.  
     [0033] The present process is advantageously carried out at a constant dilution rate in the fermentation vessel. As used herein, the term “dilution rate” means the total volume of liquids supplied to the fermentation vessel per volume of the fermentation vessel per hour (h −1 ).  
     [0034] Accordingly, it is a particular feature of the present invention to carry out the fermentation process at a constant dilution rate in the fermentation vessel while varying the supply of individual nutrient components or other additives during the fermentation process. To facilitate this task, a storage flask containing an inert component, e.g., water is optionally provided in the fermentation vessel to complement the supply of liquids added to the fermentation vessel, thus keeping the total supply of liquid constant in the fermentation vessel.  
     [0035] The assembly that is preferably used to carry out the process of this invention further includes means to transport the individual components of the fermentation medium from the storage flasks to the fermentation vessel, and appliances for monitoring the amount of liquid supplied to the fermentation vessel. Every combination of measuring instruments (e.g., volumetric or mass flow rate by either gravimetric, anemometric, magnetic, ultrasonic, Venturi, J. cross-relation, thermal, Coriolis, or radiometric) and transfer units (e.g., pumps or pressure difference) can be used for this purpose. Additionally, every transfer unit can be applied as a dosing unit (e.g., gear, peristaltic, piston, membrane or excenter pump). For operation on a small scale, the supply is suitably monitored by weighing the storage flasks that contain nutrient or additive solutions in a predetermined concentration.  
     [0036] As used herein, the “means to transport” individual components of the fermentation medium from the storage flasks to the fermentation vessel includes tubing, piping, and other types of conventional liquid transfer apparatus.  
     [0037] The device for controlling and maintaining a constant dilution rate in the fermentation vessel is suitably a system containing a measuring instrument that monitors the flow from the storage flasks and a controlling unit, e.g., a computer-software control that calculates the actual mass flow rates, compares them to the desired value, and adjusts the pump setting accordingly. An appropriate system is, e.g., the Process Automation System, National Instruments, Bridge View, USA, for Windows NT 4.0 (represented by National Instruments, Sonnenbergstrasse 53, 5408 Ennetbaden, Switzerland) that is connected to the various operating units (scales, pumps) through a serial-interface box (Rocket Port, Comtrol Europe Ltd, Great Britain, represented by Technosoftware AG Rothackerstrasse 13, 5702 Niederlenz, Switzerland).  
     [0038] An assembly that can be used in the process of this invention is depicted in FIG. 1. As FIG. 1 shows, the fermentation vessel  1  (Fermentor) is equipped with inlet tubes  2   a  from storage flasks  2  (e.g., suitably equipped with a stirrer) for supply of salt solution (Salts), nutrient solution (Nutrients), particular substrates (e.g., Substrate  1  and Substrate  2 ) for supply of, e.g., distinct carbon sources, agents for controlling the pH (Base), water for controlling a constant dilution rate, and antifoam agents. Pumps  3  transport liquids from the storage flasks  2  to the fermentor  1 . Scales  4  monitor the amount of liquids supplied to and discharged from the fermentor. Further, the fermentor has inlet tubes  9  for oxygen supply and outlet tubes  10  for exhaust controlled by units  14  and  15 , respectively. Pump  6  discharges fermentation broth via outlet tubes  5   a  to a harvest flask  5 . A main controlling unit  7  monitors and steers the overall process. Controlling unit  11  monitors and steers individual control systems  17  for temperature, pH, gas pressure, fermentor content, and supply of antifoam agents. Circuit  12  including pump  13  is used for taking samples from the fermentation broth and for providing a controlled gas flow for moving the fermentation broth. Inlet and outlet gas flow is controlled by flow control  14  and  15 . Sterile filters  16  are provided optionally. Optionally, the fermentation vessel  1  is equipped with a thermostating unit  8 .  
     [0039] In the process of the present invention, any protein-producing microorganism either of natural origin, e.g. naturally occurring fungal or bacterial microorganisms or microorganisms which have been transformed by protein encoding DNA whereby such transformed microorganisms can be bacteria, fungi or yeast, preferably from the genus Peniophora, Aspergillus, Hansenula or Pichia, especially  Aspergillus niger, Aspergillus awanari, Aspergillus sojae, Aspergillus oryzae, Hansenula polymorpha  or  Pichia pastoris.    
     [0040] In this context, the skilled person in the art selects such a protein-producing microorganism which is known to be useful for the production of a desired protein.  
     [0041] In a preferred embodiment of the present invention the protein has the activity of an enzyme such as catalase, lactase, phenoloxidase, oxidase; oxidoreductase, glucanase cellulase, xylanase and other polysaccharides, peroxidase, lipase, hydrolase, esterase, cutinase, protease and other proteolytic enzymes, aminopeptidase, carboxypeptidase, phytase, lyase, pectinase and other pectinolytic enzymes, amylase, glucosidase, mannosidase, isomerase, invertase, transferase, ribonuclease, chitinase, and desoxyribonuclease. In another preferred embodiment of the present invention, the protein is a therapeutic protein such as an antibody, a vaccine, an antigen, or an antibacterial and/or a health-beneficial protein such as lactoternin, lactoperoxidase or lysozyme.  
     [0042] It will be understood by those skilled in the art that the term “activity” includes not only native activities referring to naturally occurring enzymes or therapeutic functions, but also those activities or functions which have been modified by amino acid substitutions, deletions, additions or other modifications which may be made to enhance or modify the desired activity, the thermostability, pH tolerance, and/or further properties.  
     [0043] In a most preferred embodiment of the invention, the selected protein is a protein having the activity of a phytase. Examples of proteins having the activity of a phytase are described in EP 684 313, EP 897 010, EP 897 985 or in Examples 6 to 16 and FIGS.  2 - 22  of the present invention.  
     [0044]FIG. 2: Design of the consensus phytase sequence. The letters represent the amino acid residues in the one-letter code. The following sequences were used for the alignment: phyA from  Aspergillus terreus  9A-1 (Mitchell et al, 1997; from amino acid (aa) 27), phyA from  A. terreus  cbs116.46; (van Loon et al., 1998; from aa 27), phyA from  Aspergillus niger  var.  awamori  (Piddington et al, 1993; from aa 27), phyA from  A. niger  T213; Mitchell et al. 1997 from aa 27), phyA from  A. niger  strain NRRL3135 (van Hartingsveldt et al, 1993; from aa 27), phyA from  Aspergillus fumigatus  ATCC 13073 (Pasamontes et al, 1997; from aa 25), phyA from  A. fumigatus  ATCC 32722 (EP 897 985; FIG. 1; from aa 27), phyA from  A. fuimigatus  ATCC 58128 (EP 897 985; FIG. 1; from aa 27), phyA from  A. fumigatus  ATCC 26906 (EP 897 985, FIG. 1; from aa 27), phyA from  A. fumigatus  ATCC 32239 (EP 897 985; FIG. 1; from aa 30), phyA from  Emericella nidulans  (Pasamontes et al, 1997a; from aa 25), phyA from  Talaromyces thermophilus  (Pasamontes et al, 1997a; from aa 24), and phyA from  Myceliophthora thermophila  (Mitchell et al, 1997; from aa 19). As used herein, the phrase “from aaX” means that the sequence used to generate a consensus sequence began at amino acid number X counting from the start codon. The alignment was calculated using the program PILEUP (program menu for the Wisconsin Package, version 8, September 1994, Genetics Computer Group, 575 Science Drive, Madison, Wis., USA 53711; see also Devereux et al., 1984). The location of the gaps was refined by hand. Capitalized amino acid residues in the alignment at a given position belong to the amino acid coalition that establish the consensus residue. In bold, beneath the calculated consensus sequence, the amino acid sequence of the finally constructed consensus phytase (Fcp) is shown. The gaps in the calculated consensus sequence were filled by hand according to principals stated in Example 6.  
     [0045]FIG. 3: DNA sequence of the consensus phytase-1 gene (fcp) and of the primers used for the gene construction. The calculated amino acid sequence (FIG. 2) was converted into a DNA sequence using the program BACKTRANSLATE (Devereux et al., 1984) and the codon frequency table of highly expressed yeast genes (GCG program package, 9.0). The signal peptide of the phytase from  A. terreus  cbs.116.46 was fused to the N-terminus. The bold bases represent the sequences of the oligonucleotides used to generate the gene. The names of the respective oligonucleotides are alternately noted above or below the sequence. The underlined bases represent the start and stop codon of the gene. The bases written in italics show the two introduced Eco RI sites.  
     [0046]FIG. 4: Alignment and consensus sequence of five Basidiomycetes phytases. The letters represent the amino acid residues in the one-letter code. The amino acid sequences of the phytases from  Paxillus involutus , phyA1 (from aa 21) and phyA2 (from aa 21, WO 98/28409),  Trametes pubescens  (from aa 24, WO 98/28409),  Agrocybe pediades  (from aa 19, WO 98/28409), and  Peniophora lycii  (from aa 21, WO 98/28409) starting with the amino acid residues mentioned in parentheses, were used for the alignment and the calculation of the corresponding consensus sequence called “Basidio” (Example 7). The alignment was performed by the program PILEUP. The location of the gaps was refined by hand. The consensus sequence was calculated by the program PRETTY from the Sequence Analysis Package Release 9.0 (Devereux et al., 1984). PRETTY prints sequences with their columns alligned and can display a consensus sequence for the alignment. While a vote weight of 0.5 was assigned to the two  P. involutus  phytases, all other genes were used with a vote weight of 1.0 for the consensus sequence calculation. At positions where the program was not able to determine a consensus residue, the Basidio sequence contains a dash. Capitalized amino acid residues in the alignment at a given position belong to the amino acid coalition that establish the consensus residue.  
     [0047]FIG. 5: Design of consensus phytase-10 amino acid sequence. Adding the phytase sequence of  Thermomyces lanuginosus  (Berka et al., 1998) and the consensus sequence of the phytases from five Basidiomycetes (FIG. 4) to the alignment of FIG. 2, an improved consensus sequence was calculated by the program PRETTY. Additionally, the amino acid sequence of  A. niger  T213 was omitted; therefore, a vote weight of 0.5 was used for the remaining  A. niger  phytase sequences. For further information see Example 8.  
     [0048]FIG. 6: DNA and amino acid sequence of consensus phytase-10. The amino acid sequence is written above the corresponding DNA sequences using the one-letter code. The sequence of the oligonucleotides which were used to assemble the gene are in bold letters. The labels of oligonucleotides and the amino acids which were changed compared to those for consensus phytase-1 are underlined. The fcp10 gene was assembled from the following oligonucleotides: CP-1, CP-2, CP-3.10, CP-4.10, CP-5.10, CP-6, CP-7.10, CP-8.10, CP-9.10, CP-10.10, CP-11.10, CP-12.10, CP-13.10, CP-14.10, CP-15.10, CP-16.10, CP-17.10, CP18.10, CP-19.10, CP-20.10, CP-21.10, and CP-22.10. The newly synthesized oligonucleotides are additionally marked by the number 10. The phytase contains the following 32 exchanges relative to consensus phytase-1: Y54F, E58A, D69K, D70G, A94K, N134Q, 1158V, S187A, Q188N, D197N, S204A, T214L, D220E, L234V, A238P, D246H, T251N, Y259N, E267D, E277Q, A283D, R291I, A320V, R329H, S364T, 1366V, A379K, S396A, G404A, Q415E, A437G, A463E. The mutations accentuated in bold letters revealed a stabilizing effect on consensus phytase-1 when tested as single mutations in consensus phytase-1.  
     [0049]FIG. 7: Alignment for the design of consensus phytase-11. In contrast to the design of consensus phytase-10, for the design of the amino acid sequence of consensus phytase-11, all Basidiomycete phytases (FIG. 4) were used as independent sequences using an assigned vote weight of 0.2 for each Basidiomycete sequence. Additionally, the amino acid sequence of  A. niger  T213 phytase was used in that alignment, again.  
     [0050]FIG. 8: DNA and amino acid sequence of consensus phytase-1-thermo[8]-Q50T-K91A. The amino acid sequence is written above the corresponding DNA sequence using the one-letter code. The replaced amino acid residues are underlined. The stop codon of the gene is marked by a star (*).  
     [0051]FIG. 9: DNA and amino acid sequence of consensus phytase-10-thermo[3]-Q50T-K91A. The amino acid sequence is written above the corresponding DNA sequence using the one-letter code. The replaced amino acid residues are underlined. The stop codon of the gene is marked by a star (*).  
     [0052]FIG. 10: DNA and amino acid sequence of  A. fumigatus  ATCC 13073 phytase α-mutant. The amino acid sequence is written above the corresponding DNA sequence using the one-letter code. The replaced amino acid residues are underlined. The stop codon of the gene is marked by a star (*).  
     [0053]FIG. 11: DNA and amino acid sequence of consensus phytase-7. The amino acids are written above the corresponding DNA sequence using the one-letter code. The sequences of the oligonucleotides used to assemble the gene are in bold letters. Oligonucleotides and amino acids that were exchanged are underlined and their corresponding triplets are highlighted in small cases. The fcp7 gene was assembled from the following oligonucleotides: CP-1, CP-2, CP-3, CP-4.7, CP-5.7, CP-6, CP-7, CP-8.7, CP-9, CP-10.7, CP-11.7, CP-12.7, CP-13.7, CP-14.7, CP-15.7, CP-16, CP-17.7, CP-18.7, CP-19.7, CP-20, CP-21, and CP-22. The newly synthesized oligonucleotides are additionally marked by number 7. The phytase contains the following 24 exchanges in comparison to the original consensus phytase-1: S89D, S92G, A94K, D164S, P201S, G203A, G205S, H212P, G224A, D226T, E255T, D256E, V258T, P265S, Q292H, G300K, Y305H, A314T, S364G, M365I, A397S, S398A, G404A, and A405S.  
     [0054]FIG. 12: Differential scanning calorimetry (DSC) of consensus phytase-1 and consensus phytase-10. The protein samples were concentrated to about 50-60 mg/ml and extensively dialyzed against 10 mM sodium acetate, pH 5.0. A constant heating rate of 10° C./min was applied up to 95° C. DSC of consensus phytase-10 (12A) yielded a melting temperature of 85.4° C., which is 7.3° C. higher than the melting point of consensus phytase-1 (78.1° C., 12B).  
     [0055]FIG. 13: Differential scanning calorimetry (DSC) of consensus phytase-1-thermo[3]-Q50T and consensus phytase-10-thermo[3]-Q50T-K91A. The protein samples were concentrated to about 50-60 mg/ml and extensively dialyzed against 10 mM sodium acetate, pH 5.0. A constant heating rate of 10° C./min was applied up to 95° C. DSC of consensus phytase-10-thermo-[3]-Q50T ( 13 A) yielded a melting temperature of 88.6° C., while the melting point of consensus phytase-10-thermo[3]-Q50T-K91A was found at: 89.25° C. ( 13 B).  
     [0056]FIG. 14: Comparison of the temperature optimum between consensus phytase-1, consensus phytase-10 and consensus phytase-10-thermo[3]-Q50T. For the determination of the temperature optimum, the phytase standard assay was performed at a series of temperatures between 37° C. and 86° C. (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). The diluted supernatant of transformed  S. cerevisiae  strains was used for the determination. The other components of the supernatant showed no influence on the determination of the temperature optimum: Δ, consensus phytase-1; ⋄, consensus phytase-10; ▪, consensus phytase 10-thermo[3]-Q50T.  
     [0057]FIG. 15: pH-dependent activity profile and substrate specificity of consensus phytase-10 and its variants thermo[3]-Q50T and thermo[3]-Q50T-K91A. FIG. 15A) shows the pH-dependent activity profile of consensus phytase-10 (□), consensus phytase-10-thermo[3]-Q50T (Δ), and consensus phytase-10-thermo[3]-Q50T-K91A (Δ).The phytase activity was determined using a standard assay in appropriate buffers (see Example 15) at different pH-values (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). FIG. 15B) shows the corresponding substrate specificity tested by replacement of phytate by the indicated compounds in a standard assay; open bars, consensus phytase-10 (white bars, consensus phytase-10-thermo-Q50T; dark bars, consensus phytase-10-thermo-Q50T-K91A). The numbers correspond to the following compounds: 1, phytate; 2, p-nitrophenyl phosphate; 3, phenyl phosphate; 4, fructose-1,6-bisphosphate; 5, fructose-6-phosphate; 6, glucose-6-phosphate; 7, ribose-5-phosphate; 8, DL-glycerol-3-phosphate; 9, glycerol-2-phosphate; 10, 3-phosphoglycerate; 11, phosphoenolpyruvate; 12, AMP; 13, ADP; 14, ATP.  
     [0058]FIG. 16: pH-dependent activity profile and substrate specificity of consensus phytase-1-thermo[8]-Q50T and of consensus phytase-1-thermo[8]-Q50T-K91A. FIG. 16A) shows the pH-dependent activity profile of the Q50T-(▪) and the Q50T-K91A-variant (Δ). The phytase activity was determined using the standard assay in appropriate buffers (see Example 15) at different pH-values (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). FIG. 16B) shows the corresponding substrate specificities tested by replacement of phytate by the indicated compounds in the standard assay (open bars, consensus phytase-1-thermo[8]-Q50T; filled bars, consensus phytase-1-thermo[8]-Q50T-K91A). The substrates are listed in the legend of FIG. 15.  
     [0059]FIG. 17: Differential scanning calorimetry (DSC) of consensus phytase-1-thermo[8]-Q50T (FIG. 17A) and consensus phytase-1-thermo[8]-Q50T-K91A (FIG. 17B). The protein samples were concentrated to about 50-60 mg/mT. and extensively dialyzed against 10 mM sodium acetate, pH 5.0. A constant heating rate of 10° C./min was applied up to 95° C. DSC of consensus phytase-1-thermo[8]-Q50T (FIG. 17A) showed a melting temperature of 84.72° C., while the melting point of consensus phytase-1-thermo[8]-Q50T-K91A (FIG. 17B) was found at 85.70° C.  
     [0060]FIG. 18: Comparison of the temperature optimum between consensus phytase-1, consensus phytase-1-thermo[3] and consensus phytase-1-thermo[8]. For the determination of the temperature optimum, the phytase standard assay was performed at a series of temperatures between 37° C. and 86° C. (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). Purified protein from the supernatant of transformed  S. cerevisiae  strains was used for the determination. O, consensus phytase-1; □, consensus phytase-1-thermo[3]; Δ, consensus phytase-1-thermo[8].  
     [0061]FIG. 19: Comparison of the pH-dependent activity profile (FIG. 19A) and substrate specificity (FIG. 19B) of consensus phytase-1, consensus phytase-7, and of the phytase from  A. niger  NRRL 3135. FIG. 19A) shows the pH-dependent activity profile of consensus phytase-1 (▪), the phytase, from  A. niger  NRRL 3135 (O), and of consensus phytase-7 (Δ).The phytase activity was determined using the standard assay in appropriate buffers (see Example 15) at different pH-values (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). FIG. 19B) shows the corresponding substrate specificity tested by replacement of phytate by the indicated compounds in the standard assay (black bars,  A. niger  NRRL 3135 phytase; grey bars, consensus phytase-1, dashed bars, consensus phytase-7). The substrates are listed in the legend of FIG. 15.  
     [0062]FIG. 20: Differential scanning calorimetry (DSC) of the phytase from  A. fumigatus  ATCC 13073 (FIG. 20B) and of its stabilized α-mutant (FIG. 20A), which contains the following amino acid exchanges: F55Y, V100I, F114Y, A243L, S265P, N294D.  
     [0063] The protein samples were concentrated to about 50-60 mg/ml and extensively dialyzed against 10 mM sodium acetate, pH 5.0. A constant heating rate of 10° C./min was applied up to 95° C. DSC of consensus  A. fumigatus  13073 phytase (FIG. 20B) revealed a melting temperature of 62.50° C., while the melting point of the α-mutant (FIG. 20A) was found at 67.02° C.  
     [0064]FIG. 21: Comparison of the temperature optimum of  A. fumigatus  13073 wild-type phytase, its α-mutant, and a further stabilized α-mutant (E59A-S126N-R329H-S364T-G404A). For the determination of the temperature optimum, the phytase standard assay was performed at a series of temperatures between 37° C. and 75° C. (see Mitchell et al., 1997, Lehmann et al., 2000 and further references mentioned therein). The diluted supernatants of transformed  S. cerevisiae  strains were used for the determination. The other components of the supernatant showed no influence on the determination of the temperature optimum. O,  A. fumigatus  ATCC 13073 phytase; Δ,  A. fumigatus  ATCC 13073 α-mutant; □,  A. fumigatus  ATCC 13073 alpha-mutant-59A-S126N-R29H-S364T-G404A)-Q27T; ▪,  A. fumigatus  ATCC 13073 α-mutant-(E59A-S126N-R329H-S364T-G404A)-Q27T-K68A. The mutations Q51T and K92A in the  A. fumigatus  a-mutants correspond to −1 Q50T and K91A in consensus phytase, respectively.  
     [0065]FIG. 22: Amino acid sequence of consensus phytase-12 (consphy12), which contains a number of active site residues transferred from the “basidio” consensus sequence (FIG. 4) to consensus phytase-10-thermo[3]-Q50T-K91A.  
     [0066] The culture medium used in the fermentation process in accordance with the present invention includes nutrients for the cells or microorganisms such as digestible nitrogen sources and inorganic substances, vitamins, micro- and trace elements and other growth-promoting factors. In addition, the culture medium contains a carbon source. Various organic or inorganic substances may be used as nitrogen sources in the fermentation process, such as nitrates, ammonium salts, yeast extract, meat extract, peptone, casein, cornsteep liquor, amino acids and urea. Typical inorganic substances that can be used in the fermentation are calcium, iron, zinc, nickel, manganese, cobalt, copper, molybdenum, and alkali salts such as chlorides, sulfates and phosphates as well as boric acid. As a carbon source, glycerol or sugar-like mono-, di-, oligo- or polysaccharides, e.g., glucose, fructose, sucrose, maltose, starch, glycogen, cellulose or substrates containing such substances, e.g., molasses, glucose syrups and fructose syrups can be used. The concentration of glucose and/or methanol in the total feed stream may vary from about 10 to about 500 g/l for each component, and is preferably from about 200 to about 300 g/l. While the fermentation medium is principally an aqueous medium, such medium may contain organic solvents such as alcohols, e.g. methanol, ethanol or isopropanol. Further, the fermentation medium may also be a dispersion or suspension, in which case the fermentation is suitably carried out with stirring.  
     [0067] For continuous operation, the cells are optionally immobilized on a solid porous carrier. Any solid porous carrier with any porosity, size and geometry conventionally used in fermentation processes and exerting no toxic effects on the particular cell or microorganism which is to be immobilized can be used for the purpose of this invention. Examples of such carriers are those made from inorganic material and having a pore diameter of from about 0.5 to about 100 μm, preferably from about 10 to about 30 μm diameter. Examples of inorganic materials are ceramics and natural minerals such as steatite, zeolite, bentonite, silicates (glasses), aluminum silicates, aluminum oxide, magnesium aluminum silicates and magnesium aluminum oxides. Such carriers are commercially available, e.g., from Ceramtec, Marktredwitz, Germany, Schott Engineering GmbH, Mainz, Germany and others. Preferably, the carriers are spherical with a mean diameter of from about 0.2 to about 20 mm diameter. The carriers can be loaded with the living cells in a manner known per se by contacting the carrier particles with an appropriate cell culture. If desired, the carrier particles loaded with the cells can be further processed by applying a membrane-type coating layer, such as described in German Offenlegungsschrift DE 3421049. Suitably, the carrier is present in the fermentation vessel on a fixed bed. Further, the culture medium, its components and their containments, respectively are suitably sterilized prior to use if autosterilization (e.g., by methanol, ethanol, ammonia) cannot be guaranteed. Heat sterilization with steam (e.g., at 121° C. and 1 bar pressure for 20 minutes) and filtration (0.2 μm) for sensitive components are preferred. Alternative sterilization methods may be applied. Media components need not necessarily be sterilized when running the process in continuous mode.  
     [0068] Depending on the particular cell or organism used, the fermentation may be carried out at a pH between about 2 and about 11. In a preferred aspect of the invention, the fermentation process for the manufacture of phytase is carried out using the microorganism,  Hansenula polymorpha  transformed by a phytase encoding DNA sequence as described in EP 897 010, EP 897 985, or Example 11 of the present case. According to that particular aspect of the invention, the preferred carbon source is a mixture of glucose and methanol. Further, in accordance with that particular aspect of the invention, the fermentation may be carried out at a pH between about 4 and 5, preferably at about pH 4.6. A preferred temperature range for carrying out such fermentation process is between about 10° C. and 50° C., more preferably the fermentation temperature is about 30° C. The aeration rate is preferably adjusted to between about 0.01 and about 1.5 volume of gas per volume of liquid with a dissolved oxygen concentration (DO) of between 0.01% and about 500%. A DO of 100% denotes oxygen saturation of the solution at atmospheric pressure (1 bar) and reactor temperature. The fermentation can be carried out at a pressure of from about 0.1 to about 100 bar, preferably, the fermentation is carried out at atmospheric pressure, i.e., at about 1 bar. The dilution rate can vary from about 0.001 to about 0.5 per hour.  
     [0069] The following examples are provided to further illustrate the process of the present invention. These examples are illustrative only and are not intended to limit the scope of the invention in any way.  
     EXAMPLES  
     Example 1  
     [0070] Storage solutions for feed medium were prepared as follows:  
                                               1.1 CaCl 2 /H 3 BO 3  Solution                                                    CaCl 2 .2H 2 O    18.75 g/l           H 3 BO 3     0.0125 g/l                      
 
     [0071] This solution was sterilized at 121° C. for 20 minutes.  
                                               1.2 Microelements Solution                                                    (NH 4 ) 2 Fe(SO 4 ) 2 .6H 2 O    2.5 g/l           CuSO 4 .5H 2 O    0.2 g/l           ZnSO 4 .7H 2 O   0.75 g/l           MnSO 4 .5H 2 O    1.0 g/l           Na-EDTA    2.5 g/l                      
 
     [0072] This solution was sterilized at 121° C. for 20 minutes.  
                                               1.3 Trace Elements Solution                                                    NiSO 4 .6H 2 O   0.025 g/l           CoCl 2 .6H 2 O   0.025 g/l           Na 2 MoO 4 .2H 2 O   0.025 g/l           KI   0.025 g/l                      
 
     [0073] This solution was sterilized at 121° C. for 20 minutes.  
                                               1.4 Salts + Vitamin Solution                                                        KH 2 PO 4     50.0   g/l           NH 4 H 2 PO 4     100.0   g/l           MgSO 4 .7H 2 O   45.0   g/l           (NH 4 ) 2 SO 4     50.0   g/l           KCl   23.0   g/l           NaCl   5.0   g/l           vitamin solution   5.0   ml/l                      
 
     [0074] (D-biotin, 600 mg/l and thiamin.HCl 200 g/l in 50% isopropanol/water)  
     [0075] The vitamin solution was sterilized by filtration (0.2 μm), and added to the salt solution that was sterilized at 121° C. for 20 minutes.  
     [0076] 1.5 Glucose Solution  
     [0077] 770 g of D-glucose.H 2 O were dissolved in 480 g of water and sterilized (121° C., 20 minutes) to yield a 1 liter solution containing 57% (by weight) of D-glucose.  
     [0078] 1.6 Methanol  
     [0079] Pure methanol was assumed to be sterile and poured into a sterilized flask.  
     [0080] 1.7 Antifoam  
     [0081] A sterilized (121° C., 20 minutes) solution of 10 antifoam (STRUKTOL J 673, Schill &amp; Seilacher, Hamburg, Germany) was provided for supply on demand by foam-control.  
     [0082] 1.8 Base  
     [0083] A solution of about 12.5% (by weight) of ammonia in sterile water was poured into a sterilized flask.  
     EXAMPLE 2  
     [0084] A fixed bed bioreactor (1 liter) was set up following the principle illustrated in FIG. 1 with individual storage flasks provided for solutions 1.1 to 1.8 of Example 1. The fixed bed of porous steatite spheres (4 mm diameter, pore diameter 10-30 μm, 280 pores per ml, CeramTec, Marktredwitz, Germany) was contained by a sieve plate at the top. The reactor was sterilized (121° C., 20 minutes), and thereafter filled with an inoculum culture of  Hansenula polymorpha  transformed with a phytase encoding DNA as described, e.g. in EP 897 010, EP 897 985 or Example 11. EP 897010 and EP 897985 are incorporated by reference as if recited in full herein. Then the connection to the storage flasks was established. The inoculum culture was grown on a medium containing glycerol as a carbon source instead of glucose. The reactor was switched to batch operation until all glycerol was consumed, which was determined by a rise of the dissolved oxygen concentration. Then the feed stream was turned on and the fermentation was run under process conditions as given below:  
                                                              Temperature       30° C.               pH   4.6   Diluted oxygen concentration 52-62                                 % ox otal     10 5     N/m 2             P O2     10 5     N/m 2             Dilution rate   0.0067   h −1             Aeration rate   100   ml/min           V fluid     1190   ml −1             V fixed bed     950   ml −1                        
 
     [0085] Substrate composition as provided by storage flasks 1-8; (actual concentrations in feed stream given):  
                                                      D-glucose    305 g/l           Methanol    264 g/l           CaCl 2 /H 3 BO 3  Solution   12.2 g/l           Microelement Solution   20.9 g/l           Trace Element Solution   17.2 g/l           Salts + vitamin Solution   44.7 g/l                      
 
     [0086] Analytics:  
     [0087] Bio-Rad Protein Assay Kit I (Bio-Rad, Glattbrugg, Switzerland) was used to determine the total protein concentration. A factor for the calculation of phytase concentration (c phyt ) from total protein concentration (c tp ) was determined as C phyt =0.76 c tp .  
     [0088] To determine the biomass in the medium two samples of 1 ml were centrifuged, washed with 1 ml of water, centrifuged again, dried at 85° C. for two days and weighed.  
     [0089] Results:  
     [0090] Under the above process conditions, the biomass was 59 g/l. Given a dilution rate of 0.0067 per hour the productivity was 0.078 g of phytase per liter per hour.  
     [0091] In a fermentation that was run fed-batch-wise, the biomass was 125 g/l; the productivity, however, was calculated to 0.054 g phytase per liter per hour.  
     Example 3  
     [0092] A fermentation was carried out as set forth in Example 2 but omitting the steatite spheres (i.e., without immobilization of the microorganism). A nutrient and a salt and vitamin solution of the following composition were pumped into the reactor separately:  
                                               Nutrient Solution:                                                        NiSO 4 .6H 2 O   8.33   mg/l           CoCl 2 6.H 2 O   8.33   mg/l           Na 2 MoO 4 .2H 2 O   8.33   mg/l           KI   8.33   mg/l           (NH 4 ) 2 Fe(SO 4 ) 2 .6H 2 O   833.33   mg/l           CuSO   66.67   mg/l           ZnSO 4 .7H 2 O   250   mg/l           MnSO 4 .5H 2 O   333.33   mg/l           Na-EDTA   833.33   mg/l           CaCl 2 .2H 2 O   6250   mg/l           H 3 BO 3     4.17   mg/l                      
 
     [0093]                                               Salts + Vitamins Solution:                                                        KH 4 PO 4     50.0   g/l           NH 4 H 2 PO 4     100.0   g/l           MgSO 4 .7H 2 O   45.0   g/l           (NH 4 ) 2 Fe(SO 4 )   50.0   g/l           KCl   23.0   g/l           NaCl   5.0   g/l           vitamin solution   5.0   ml/l                        
     [0094] (D-biotin, 600 mg/l and thiamin.HCl 200 g/l in 50% isopropanol/water)  
     [0095] The supply of these two solutions was adjusted to provide in the feed stream a concentration of 51 g/l of the Nutrient Solution and 61 g/l of the Salts+Vitamins Solution. The dilution rate was adjusted to 0.009 h −1 . The pH was kept at 4.6 by addition of 12.5 wt % ammonium hydroxide. Glucose Solution as in Example 1 and methanol were fed into the reactor separately to maintain a glucose concentration of 275 g/l and a methanol concentration of 260 g/l in the feed stream.  
     [0096] The productivity of this fermentation was 0.088 g phytase per liter per hour. Biomass in outflow was 58 g/l.  
     Example 4  
     [0097] A fermentation process as set forth in Example 3 was carried out, but adjusting glucose concentration to 290 g/l and methanol concentration to 260 g/l, and keeping the dilution rate constant at 0.009 h −1 , the productivity was 0.092 g phytase per liter per hour. Biomass in outflow was 60.4 g/l.  
     Example 5  
     [0098] A fermentation process as set forth in Example 3 was carried out, but adjusting glucose concentration to 270 g/l and methanol concentration to 280 g/l, and keeping the dilution rate constant at 0.009 h −1 , the productivity was 0.094 g phytase per liter per hour. Biomass in outflow was 56.8 g/l.  
     Example 6  
     Design of the Amino Acid Sequence of Consensus Phytase-1  
     [0099] Alignment of the Amino Acid Sequences  
     [0100] The alignment was calculated using the program PILEUP from the GCG Sequence Analysis Package Release 9.0 (Devereux et al., 1984) with the standard parameters (gap creation penalty 12, gap extension penalty 4). The location of the gaps was refined using a text editor. Table 1 shows the sequences (see FIG. 2), without the signal sequence, that were used for the performance of the alignment starting with the amino acid (aa) as mentioned in Table 1, e.g., “aa 27” means that the alignment began at the 27 amino acid from the start codon.  
               TABLE 1                       Origin and vote weight of the phytase amino acid sequences       used for the design of consensus phytase-1                                    phyA from  Aspergillus terreus  9A-1, aa 27, vote weight 0.5 (Mitchell       et al., 1997)       phyA from  Aspergillis terreus cbs  116.46, aa 27, vote weight 0.5 (EP       897 985; HG. 1)       phyA from  Aspergillus niger  var.  awamori , aa 27, vote weight 0.33       (Piddington et al., 1993)       phyA from  Aspergillus niger  T213, aa 27, vote weight 0.33       phyA from  Aspergillus niger  strain NRRL3135, aa 27, vote weight 0.33       (van Hartingsveldt et al., 1993)       phyA from  Aspergillus fumigatus  ATCC 13073, aa 26, vote weight 0.2       (Pasamontes et al., 1997)       phyA from  Aspergillus fumigatus  ATCC 32722, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 58128, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 26906, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 32239, aa 30, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Emericella nidulans , aa 25, vote weight 1.0 (Pasamontes et al.,       1997a)       phyA from  Talaromyces thermophilus  ATCC 20186, aa 24, vote weight       1.0 (Pasamontes et al., 1997a)       phyA from  Myceliophthora thermophila , aa 19, vote weight 1.0 (Mitchell       et al., 1997)                  
 
     [0101] Calculation of the Amino Acid Sequence of Consensus Phytase-1  
     [0102] Using the refined alignment as input, the consensus sequence was calculated by the program PRETTY from the GCG Sequence Analysis Package Release 9.0 (Devereux et al., 1984). PRETTY prints sequences with their columns aligned and can display a consensus sequence for an alignment. A vote weight that pays regard to the similarity between the amino acid sequences of the aligned phytases was assigned to all sequences. The vote weight was set in such a way that the combined impact of all phytases from one sequence subgroup (same species, but from different strains), e.g. the amino acid sequences of all phytases from  A. fumigatus , on the election was set one, that means that each sequence contributes a value of 1 divided by the number of strain sequences (see Table 1). By this means, it was possible to prevent very similar amino acid sequences, e.g. of the phytases from different  A. fumigatus  strains, from dominating the calculated consensus sequence.  
     [0103] The program PRETTY was started with the following parameters: The plurality defining the number of votes below which there is no consensus was set on 2.0. The threshold, which determines the scoring matrix value below which an amino acid residue may not vote for a coalition of residues, was set on 2. PRETTY used the PrettyPep.Cmp consensus scoring matrix for peptides.  
     [0104] Ten positions of the alignment (positions 46, 66, 82, 138, 162, 236, 276, 279, 280, 308; FIG. 2), for which the program was not able to determine a consensus residue, were filled by hand according to the following rules: if a most frequent residue existed, this residue was chosen (positions 138, 236, 280); if a prevalent group of similar equivalent residues occurred, the most frequent or, if not available, one residue of this group was selected (positions 46, 66, 82, 162, 276, 308). If there was neither a prevalent residue nor a prevalent group, one of the occurring residues was chosen according to common assumptions on their influence on the protein stability (position 279). Eight other positions (positions 132, 170, 204, 211, 275, 317, 384, 447; FIG. 2) were not filled with the amino acid residue selected by the program but normally with amino acids that occur with the same frequency as the residues that were chosen by the program. In most cases, the slight underrating of the three  A. niger  sequences (sum of the vote weights: 0.99) was eliminated by this correction.  
     [0105] Conversion of the Consensus Phytase-1 Amino Acid Sequence to a DNA Sequence  
     [0106] The first 26 amino acid residues of the  A. terreus  cbs116.46 phytase were used as a signal peptide and, therefore, fused to the N-terminus of all consensus phytases. For this stretch, we used a special method to calculate the corresponding DNA sequence. Purvis et al (1987) proposed that the incorporation of rare codons in a gene has an influence on the folding efficiency of the protein. The DNA sequence for the signal sequence was calculated using the approach of Purvis et al (1987), which is hereby incorporated by reference as if recited in full herein, and optimized for expression in  S. cerevisiae . For the remaining parts of the protein, we used the codon frequency table of highly expressed  S. cerevisiae  genes, obtained from the GCG program package, to translate the calculated amino acid sequence into a DNA sequence.  
     [0107] The resulting sequence of the fcp gene is shown in FIG. 3.  
     [0108] Construction and Cloning of the Consensus Phytase-1 Gene  
     [0109] The calculated DNA sequence of consensus phytase-1 (fcp) was divided into oligonucleotides of 85 bp, alternately using the sequence of the sense and the anti-sense strand. Every oligonucleotide overlaps 20 bp with its previous and its following oligonucleotide of the opposite strand. The location of all primers, purchased from Microsynth, Balgach (Switzerland) and obtained in a PAGE-purified form, is indicated in FIG. 3.  
     [0110] PCR-Reactions  
     [0111] In three PCR reactions, the synthesized oligonucleotides were composed to the entire gene. For the PCR, the High Fidelity Kit from Boehringer Mannheim (Boehringer Mannheim, Germany) and the thermo cycler The Protokol (TM) from AMS Biotechnology (Europe) Ltd. (Lugano, Switzerland) were used.  
     [0112] Oligonucleotides CP-1 to CP-10 (Mix 1, FIG. 3) were mixed to a concentration of 0.2 pmol/μl of each oligonucleotide. A second oligonucleotide mixture (Mix 2) was prepared with CP-9 to CP-22 (0.2 pmol/μl of each oligonucleotide). Additionally, four short primers were used in the PCR reactions:  
                                                      Eco  RI               CP-a:   5′-TATAT GAATTC ATGGGCGTGTTCGTC-3′   (SEQ ID No. 1)               CP-b:   5′-TGAAAAGTTCATTGAAGGTTTC-3′   (SEQ ID No. 2)               CP-c:   5′-TCTTCGAAAGCAGTACAAGTAC-3′   (SEQ ID No. 5)                                    Eco  RI       CP-e:   5′-TATAT GAATTC TTAAGCGAAAC-3′   (SEQ ID No. 4)          
 
     [0113]                                                      PCR reaction a:   10 μl Mix 1 (2.0 pmol of each oligonucleotide)               2 μl nucleotides (10 mM each nucleotide)               2 μl primer CP-a (10 pmol/μl)               2 μl primer CP-c (10 pmo/μl)               10.0 μl PCR buffer               0.75 μl polymerase mixture (2.6 U)               73.25 μl H 2 O           PCR reaction b:   10 μl Mix 2 (2.0 pmol of each oligonucleotide)               2 μl nucleotides (10 mM each nucleotide)               2 μl primer CP-b (10 pmol/μl)               2 μl primer CP-e (10 pmol/μl)               10.0 μl PCR buffer               0.75 μl polymerase mixture (2.6 U)               73.25 μl H 2 O                        
     [0114] Reaction conditions for PCR reactions a and b:  
                                                      step 1    2 minutes - 45° C.           step 2   30 seconds - 72° C.           step 3   30 seconds - 94° C.           step 4   30 seconds - 52° C.           step 5    1 minute - 72° C.                      
 
     [0115] Steps 3 to 5 were repeated 40 times.  
     [0116] The PCR products (670 and 905 bp) were purified by an agarose gel electrophoresis (0.9% agarose) and subsequently subjected to gel extraction (QIAEX II Gel Extraction Kit, Qiagen, Hilden, Germany). The purified DNA fragments were used for the PCR reaction c.  
                                                          PCR reaction C:   6 μl   PCR product of reaction a (≈50 ng)               6 μl   PCR product of reaction b (≈50 ng)               2 μl   primer CP-a (10 pmol/μl)               2 μl   primer CP-e (10 pmol/μl)               10.0 μl   PCR buffer               0.75 μl   polymerase mixture (2.6 U)               73.25 μl   H 2 O                      
 
     [0117] Reaction conditions for PCR reaction c:  
                                                      step 1    2 minutes - 94° C.           step 2   30 seconds - 94° C.           step 3   30 seconds - 55° C.           step 4    1 minutes - 72° C.                      
 
     [0118] Steps 2 to 4 were repeated 31-times.  
     [0119] The resulting PCR product (1.4 kb) was purified as mentioned above, digested with Eco RI, and ligated in an Eco RI-digested and dephosphorylated pBsk(−)-vector (Stratagene, La Jolla, Calif., USA). 1 μl of the ligation mixture was used to transform  E. coli  XL-1 competent cells (Stratagene, La Jolla, Calif., USA). All standard procedures were carried out as described by Sambrook et al. (1987). The DNA sequence of the constructed consensus phytase gene (fcp, FIG. 3) was controlled by sequencing as known in the art.  
     Example 7  
     Design of an Improved Consensus Phytase (Consensus Phytase-10) Amino Acid Sequence  
     [0120] The alignments used for the design of consensus phytase-10 were calculated using the program PILEUP from the GCG Sequence Analysis Package Release 9.0 (Devereux et al., 1984) with the standard parameters (gap creation penalty 12, gap extension penalty 4). The location of the gaps was refined using a text editor.  
     [0121] The following sequences were used for the alignment of the Basiodiomycete phytases starting with the amino acid (aa) mentioned in Table 2:  
               TABLE 2                       Origin and vote weight of five  Basidiomycete phytases  used for the       calculation of the corresponding amino acid consensus sequence (basidio)                                    phyA1 from  Paxillus involutus  NN005693, aa 21, vote weight 0.5 (WO       98/28409)       phyA2 from  Paxillus involutus  NN005693, aa 21, vote weight 0.5 (WO       98/28409)       phyA from  Trametes pubescens  NN9343, aa 24, vote weight 1.0 (WO       98/28409)       phyA from  Agrocybe pediades  NN009289, aa 19, vote weight 1.0 (WO       98/28409)       phyA from  Peniophora lycii  NN006113, aa 21, vote weight 1.0 (WO       98/28409)                  
 
     [0122] The alignment is shown in FIG. 4.  
     [0123] In Table 3 the genes, which were used for the performance of the final alignment, are arranged. The first amino acid (aa) of the sequence, which is used in the alignment is mentioned behind the organism&#39;s designation.  
               TABLE 3                       Origin and vote weight of the phytase sequences used for the design of       consensus phytase 10                                    phyA from  Aspergillus terreus  9A-1, aa 27, vote weight 0.5 (Mitchell       et al., 1997)       phyA from  Aspergillus terreus  cbs116.46, aa 27, vote weight 0.5 (EP       897 985; FIG. 1)       phyA from  Aspergillus niger  var.  awamori , aa 27, vote weight 0.5       (Piddington et al., 1993)       phyA from  Aspergillus niger  strain NRRL3135, aa 27, vote weight 0.5       (van Hartingsveldt et al., 1993)       phyA from  Aspergillus fumigatus  ATCC 13073, aa 26, vote weight 0.2       (Pasamontes et al., 1997)       phyA from  Aspergillus fumigatus  ATCC 32722, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 58128, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 26906, aa 26, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Aspergillus fumigatus  ATCC 32239, aa 30, vote weight 0.2       (EP 897 985; FIG. 1)       phyA from  Emericella nidulans , aa 25, vote weight 1.0 (Pasamontes et al.,       1997a)       phyA from  Talaromyces thermophilus  ATCC 20186, aa 24, vote weight       1.0 (Pasamontes et al., 1997a)       phyA from  Myceliopthora thermophila , aa 19, vote weight 1.0 (Mitchell       et al., 1997)       phyA from  Thermomyces lanuginosa , aa 36, vote weight 1.0 (Berka et al.,       1998)       Consensus sequence of five  Basidiomycete phytases , vote weight 1.0       (Basidio, FIG. 4)                  
 
     [0124] The corresponding alignment is shown in FIG. 5.  
     [0125] Calculation of the Amino Acid Sequence of Consensus Phytase-10  
     [0126] To improve the alignment, we combined the consensus sequence of five phytases from four different Basidiomycetes, called Basidio, still containing the undefined sequence positions (see FIG. 4), nearly all phytase sequences used for calculation of the original consensus phytase, and one new phytase sequence from the  Ascomycete Thermomyces lanuginosus  to a larger alignment. We set plurality on 2.0 and threshold on 3. The vote weights used are listed in Table 3. The alignment and the corresponding consensus sequence are presented in FIG. 5. The new consensus phytase-10 sequence has 32 different amino acids in comparison to the original consensus phytase (consensus phytase-1). Positions for which the program PRETTY was not able to calculate a consensus amino acid residue were filled according to rules mentioned in Example 6. None of the residues suggested by the program was replaced.  
     [0127] We included all Basidiomycete phytases as single amino acid sequences but assigning a vote weight of 0.2 in the alignment. The corresponding alignment is shown in FIG. 7. he calculated consensus amino acid sequence (consensus phytase-11) has the following differences to the sequence of consensus phytase-10: D35X, X(K)69K, X(E)100E, A101R, Q134N, X(K)153N, X(H)190H, X(A)204S, X(E)220D, E222T, V227A, X(R)271R, H287A, X(D)288D, X(K)379K, X(I)3891, E390X, X(E)415E, X(A)416A, X(R)446L, E463A, where the numbering is as in FIG. 6.  
     [0128] As used herein, “X” means that the program was not able to calculate a consensus amino acid; the amino acid in parenthesis corresponds to the amino acid finally included in the consensus phytase-10.  
     [0129] We also checked single amino acid replacements suggested by the improved consensus phytase sequences 10 and 11 on their influence on the stability of the original consensus phytase-1. The approach is described in Example 8.  
     [0130] Conversion of Consensus Phytase-0 Amino Acid Sequence to a DNA Sequence  
     [0131] The first 26 amino acid residues of  A. terreus  cbs116.46 phytase were used as a signal peptide and, therefore, fused to the N-terminus of consensus phytase-10. The procedure used is further described in Example 6.  
     [0132] The resulting sequence of the fcp10 gene is shown in FIG. 6.  
     [0133] Construction and Cloning of the Consensus Phytase-10 Gene (fcp10)  
     [0134] The calculated DNA sequence of fcp10 was divided into oligonucleotides of 85 bp, alternately using the sequence of the sense and the anti-sense strand. Every oligonucleotide overlaps 20 bp with its previous and its following oligonucleotide of the opposite strand. The location of all primers, purchased from Microsynth, Balgach (Switzerland) and obtained in a PAGE-purified form, is indicated in FIG. 6.  
     [0135] PCR-Reactions  
     [0136] In three PCR reactions, the synthesized oligonucleotides were composed to the entire gene. For the PCR, the High Fidelity Kit from Boehringer Mannheim (Boehringer Mannheim, Mannheim; Germany) and the thermocycler The Protokol™ from AMS Biotechnology (Europe) Ltd. (Lugano, Switzerland) were used. The following oligonucleotides were used in a concentration of 0.2 pmol/ml.  
     [0137] Mix 1.10: CP-1, CP-2, CP-3.10, CP-4.10, CP-5.10, CP-6, CP-7.10, CP-8.10, CP-9.10, CP-10.10  
     [0138] Mix 2.10: CP-9.10, CP-10.10, CP-11.10, CP-12.10, CP-13.10, CP-14.10, CP-15.10, CP-16.10, CP-17.10, CP-18.10, CP-19.10, CP-20.10, CP-21.10, CP-22.10  
     [0139] The newly synthesized oligonucleotides are marked by number 10. The phytase contains the following 32 exchanges, which are underlined in FIG. 6, in comparison to the original consensus phytase-1: Y54F, E58A, D69K, D70G, A94K, N134Q, I158V, S187A, Q188N, D197N, S204A, T214L, D220E, L234V, A238P, D246H, T251N, Y259N, E267D, E277Q, A283D, R291I, A320V, R329H, S364T, 1366V, A379K, S396A, G404A, Q415E, A437G, A463E.  
     [0140] Four short PCR primers were used for the assembling of the oligonucleotides:  
                                            Eco  RI               CP-a:   5′-TATAT GAATTC ATGGGCGTGTTCGTC-3′   (SEQ ID No. 1)               CP-b:   5′-TGAAAAGTTCATTGAAGGTTTC-3′   (SEQ ID No. 2)               CP-c.10:   5′-TCTTCGAAAGCAGTACACAAAC-3′   (SEQ ID No. 5)                         Eco  RI       CP-e:   5′-TATAT GAATTC TTAAGCGAAAC-3′   (SEQ ID No. 4)          
 
     [0141]                                          PCR reaction a:   10 μl   Mix 1.10 (2.0 pmol of each oligonucleotide)           2 μl   nucleotides (10 mM each nucleotide)           2 μl   primer CP-a(10 pmol/ml)           2 μl   primer CP-c.10 (10 pmol/ml)           10.0 μl   PCR buffer           0.75 μl   polymerase mixture (2.6 U)           73.25 μl   H 2 O       PCR reaction b:   10 μl   Mix 2.10 (2.0 pmol of each oligonucleotide)           2 μl   nucleotides (10 mM each nucleotide)           2 μl   primer CP-b (10 pmol/ml)           2 μl   primer CP-e (10 pmol/ml)           10.0 μl   PCR buffer           0.75 μl   polymerase mixture (2.6 U)           73.25 μl   H 2 O                    
     [0142] Reaction conditions for PCR reactions a and b:  
                                                      step 1    2 minutes - 45° C.           step 2   30 seconds - 72° C.           step 3   30 seconds - 94° C.           step 4   30 seconds - 52° C.           step 5    1 minutes - 72° C.                      
 
     [0143] Steps 3 to 5 were repeated 40 times.  
     [0144] The PCR products (670 and 905 bp) were purified by an agarose gel electrophoresis (0.9% agarose) subsequently followed by gel extraction (QIAEX II Gel Extraction Kit, 2 Qiagen, Hilden, Germany). The purified DNA fragments were used for the PCR reaction c.  
     [0145] PCR reaction c:  
     [0146] 6 μl PCR product of reaction a (≈50 ng)  
     [0147] 6 μl PCR product of reaction b (≈50 ng)  
     [0148] 2 μl primer CP-a (10 pmol/ml)  
     [0149] 2 μl primer CP-e (10 pmol/ml).  
     [0150] 10.0 μl PCR buffer  
     [0151] 0.75 μl polymerase mixture (2.6 U)  
     [0152] 73.25 μl H 2 O  
     [0153] Reaction conditions for PCR reaction c:  
                                                      step 1    2 minutes - 94° C.           step 2   30 seconds - 94° C.           step 3   30 seconds - 55° C.           step 4    1 minute - 72° C.                      
 
     [0154] Steps 2 to 4 were repeated 31 times.  
     [0155] The resulting PCR product (1.4 kb) was purified as mentioned above, digested with Eco RI, and ligated in an Eco RI-digested and dephosphorylated pBsk(−)-vector (Stratagene, La Jolla, Calif., USA). 1 μl of the ligation mixture was used to transform  E. coli  XL-1 competent cells (Stratagene, La Jolla, Calif., USA). All standard procedures were carried out as described by Sambrook et al. (1987). The DNA sequence of the constructed gene (fcp10) was checked by sequencing as known in the art.  
     Example 8  
     Increasing the Thermostability of Consensus Phytase-1 by Introduction of Single Mutations Suggested by the Amino Acid Sequence of Consensus Phytase-10 and/or Consensus Phytase-11  
     [0156] To increase the thermostability of homologous genes, it is also possible to test the stability effect of each differing amino acid residue between the protein of interest and the calculated consensus sequence, and to combine all stabilizing mutations into the protein of interest. We used the consensus phytase-1 as the protein of interest, and tested the effect on the protein stability of 34 amino acids, which differed between consensus phytase-1 on one hand and consensus phytases-10 and/or -11 on the other hand, by single mutation.  
     [0157] To construct muteins for expression in  A. niger, S. cerevisiae , or  H. polymorpha , the corresponding expression plasmid containing the consensus phytase gene was used as template for site-directed mutagenesis (see Examples 11-13). Mutations were introduced using the “quick exchange™ site-directed mutagenesis kit” from Stratagene (La Jolla, Calif., USA) following the manufacturer&#39;s protocol, and using the corresponding primers. All mutations made and their corresponding primers are summarized in Table 4. Plasmids harboring the desired mutation were identified by DNA sequence analysis as known in the art.  
                   TABLE 4                          Primers used for site-directed mutagenesis of consensus phytases           (Exchanged bases are highlighted in bold. The introduction of a       restriction site is marked above the sequence. When a restriction site       is written in parenthesis, the mentioned site was destroyed by       introduction of the mutation.)                     mutation   Primer set                                                      Kpn  I               Q50T   5′-CACTTGTGG GGTACC TACTCTCCATACTTCTC-3′   (SEQ ID No. 6)           5′-GAGAAGTATGGAGAGTA GGTACC CCACAAGTG-3′   (SEQ ID No. 7)               Y54F   5′-GGTCAATACTCTCCATTCTTCTCTTTGGAAG-3′   (SEQ ID No. 8)           5′-CTTCCAAAGAGAAGAATGGAGAGTATTGACC-3′   (SEQ ID No. 9)               E58A   5′-CATACTTCTCTTTGGCAGACGAATCTGC-3′   (SEQ ID No. 10)           5′-GCAGATTCGTCTGCCAAAGAGAAGTATG-3′   (SEQ ID No. 11)                                               Aat  II       D69K   5′-CTCCA GACGTC CCAAAGGACTGTAGAGTTAC-3′   (SEQ ID No. 12)           5′-GTAACTCTACAGTCCTTTGG GACGTC TGGAG-3′   (SEQ ID No. 13)                                               Aat  II       D70G   5′-CTCCA GACGTC CCAGACGGCTGTAGAGTTAC-3′   (SEQ ID No. 14)           5′-GTAACTCTACAGCCGTCTGG GACGTC TGGAG-3′   (SEQ ID No. 15)               K91A   5′-GATACCCAACTTCTTCTGCGTCTAAGGCTTACTCTG-3′   (SEQ ID No. 16)           5′-CAGAGTAAGCCTTAGACGCAGAAGAAGTTGGGTATC-3′   (SEQ ID No. 17)                                                   Sca  I       A94K   5′-CTTCTAAGTCTAAGA AGTACT CTGCTTTG-3′   (SEQ ID No. 18)           5′-CAAAGCAG AGTACT TCTTAGACTTAGAAG-3′   (SEQ ID No. 19)               A101R   5′-GCTTACTCTGCTTTGATTGAACGGATTCAAAAGAACGCTAC-3′   (SEQ ID No. 20)           5′-GTAGCGTTCTTTTGAATCCGTTCAATCAAAGCAGAGTAAGC-3′   (SEQ ID No. 21)               N134Q   5′-CCATTCGGTGAACAGCAAATGGTTAACTC-3′   (SEQ ID No. 22)           5′-GAGTTAACCATTTGCTGTTCACCGAATGG-3′   (SEQ ID No. 23)                                                  Nru  I       K153N   5′-GATACAAGGCTC TCGCGA GAAACATTGTTC-3′   (SEQ ID No. 24)           5′-GGAACAATGTTTC TCGCGA GAGCCTTGTATC-3′   (SEQ ID No. 25)                                                  Bss  HI       I158V   5′-GATTGTTCCATTCGT GCGCGC TTCTGGTTC-3′   (SEQ ID No. 26)           5′-GAACCAGAA GCGCGC ACGAATGGAACAATC-3′   (SEQ ID No. 27)                                                    Bcl  I       D197N   5′-CTCCAGTTATTAACG TGATCA TTCCAGAAGG-3′   (SEQ ID No. 28)           5′-CCTTCTGGAA TGATCA CGTTAATAACTGGAG-3′   (SEQ ID No. 29)                                               Apa  I       S187A   5′-GGCTGACCCAG GGGCCC AACCACACCAAGC-3′   (SEQ ID No. 30)           5′-GCTTGGTGTGGTT GGGCCC CTGGGTCAGCC-3′   (SEQ ID No. 31)                                                Nco  I       T214L   5′-CACTTTGGA CCATGG TCTTTGTACTGCTTTCG-3′   (SEQ ID No. 32)           5′-CGAAAGCAGTACAAAGA CCATGG TCCAAAGTG-3′   (SEQ ID No. 33)                                                     Avr  II       E222T   5′-GCTTTCGAAGACTCTA CCCTAG GTGACGACGTTG-3′   (SEQ ID No. 34)           5′-CAACGTCGTCA CCTAGG GTAGAGTCTTCGAAAGC-3′   (SEQ ID No. 35)               V227A   5′-GGTGACGACGCTGAAGCTAACTTCAC-3′   (SEQ ID No. 36)           5′-GTGAAGTTAGCTTCAGCGTCGTCACC-3′   (SEQ ID No. 37)                                               Sac  II       L234V   5′-CTAACTTCA CCGCGG TGTTCGCTCCAG-3′   (SEQ ID No. 38)           5′-CTGGAGCGAACACCGCGGTGAAGTTAG-3′   (SEQ ID No. 39)               A238P   5′-GCTTTGTTCGCTCCACCTATTAGAGCTAGATTGG-3′   (SEQ ID No. 40)           5′-CCAATCTAGCTCTAATAGGTGGAGCGAACAAAGC-3′   (SEQ ID No. 41)                                             Hpa  I       T251N   5′-GCCAGGT GTTAAC TTGACTGACGAAG-3′   (SEQ ID No. 42)           5′-TTCGTCAGTCAA GTTAAC ACCTGGC-3′   (SEQ ID No. 43)                                             Aat  II       Y259N   5′-GACGAA GACGTC GTTAACTTGATGGAC-3′   (SEQ ID No. 44)           5′-GTCCATCAAGTTAAC GACGTC TTCGTC-3′   (SEQ ID No. 45)                                               Asp  I       E267D   5′-GTCCATTC GACACT GTCGCTAGAACTT C-3′   (SEQ ID No. 46)           5′-GAAGTTCTAGC GACAGT GTCGAATGGAC-3′   (SEQ ID No. 47)               E277Q   5′-CTGACGCTACTCAGCTGTCTCCATTC-3′   (SEQ ID No. 48)           5′-GAATGGAGACAGCTGAGTAGCGTCAG-3′   (SEQ ID No. 49)               A283D   5′-GTCTCCATTCTGTGATTTGTTCACTCAC-3′   (SEQ ID No. 50)           5′-GTGAGTGAACAAATCACAGAATGGAGAC-3′   (SEQ ID No. 51)                                              Ksp  I       H287A   5′-GCTTTGTTCA CCGCGG ACGAATGGAG-3′   (SEQ ID No. 52)           5′-CTCCATTCGT CCGCGG TGAACAAAGC-3′   (SEQ ID No. 53)                                              Bam  HI       R291I   5′-CACGACGAAT GGATCC AATACGACTAC-3′   (SEQ ID No. 54)           5′-GTAGTCGTATT GGATCC ATTCGTCGTG-3′   (SEQ ID No. 55)                                                Bsi  WI       Q292A   5′-GACGAATGGAGAG CGTACG ACTACTTG-3′   (SEQ ID No. 56)           5′-CAAGTAGT CGTACG CTCTCCATTCGTC-3′   (SEQ ID No. 57)                                                 Hpa  I       A320V   5′-GGTGTTGGTTTC GTTAAC GAATTGATTGC-3′   (SEQ ID No. 58)           5′-GCAATCAATTC GTTAAC GAAACCAACACC-3′   (SEQ ID No. 59)                                               (Bgl II)       R329H   5′-GCTAGATTGACT CACTCT CCAGTTCAAG-3′   (SEQ ID No. 60)           5′-CTTGAACTGGAGA GTGAGT CAATCTAGC-3′   (SEQ ID No. 61)                                                    Eco  RV       S364T   5′-CTCACGACAACACTAT GATATC TATTTTCTTC-3′   (SEQ ID No. 62)           5′-GAAGAAAATA GATATC ATAGTGTTGTCGTGAG-3′   (SEQ ID No. 63)                                               Nco  I       I366V   5′-CGACAACT CCATGG TTTCTATTTTCTTCGC-3′   (SEQ ID No. 64)           5′-GCGAAGAAAATAGAAA CCATGG AGTTGTCG-3′   (SEQ ID No. 65)                                            Kpn  I       A379K   5′-GTACAAC GGTACC AAGCCATTGTCTAC-3′   (SEQ ID No. 66)           5′-GTAGACAATGGCTT GGTACC GTTGTAC-3′   (SEQ ID No. 67)               S396A   5′-CTGACGGTTACGCTGCTTCTTGGAC-3′   (SEQ ID No. 68)           5′-GTCCAAGAAGCAGCGTAACCGTCAG-3′   (SEQ ID No. 69)               G404A   5′-CTGTTCCATTCGCTGCTAGAGCTTAC-3′   (SEQ ID No. 70)           5′-GTAAGCTCTAGCAGCGAATGGAACAG-3′   (SEQ ID No. 71)               Q415E   5′-GATGCAATGTGAAGCTGAAAAGGAACC-3′   (SEQ ID No. 72)           5′-GGTTCCTTTTCAGCTTCACATTGCATC-3′   (SEQ ID No. 73)                                                Sal  I       A437G   5′-CACGGTTGTGGT GTCGAC AAGTTGGG-3′   (SEQ ID No. 74)           5′-CCCAACTT GTCGAC ACCACAACCGTG-3′   (SEQ ID No. 75)                                                 Mun  I       A463E   5′-GATCTGGTGG CAATTG GGAGGAATGTTTCG-3′   (SEQ ID No. 76)           5′-CGAAACATTCCTCC CAATTG CCACCAGATC-3′   (SEQ ID No. 77)                  
 
     [0158] and accordingly for other mutations.  
     [0159] The temperature optimum of the purified phytases, expressed in  Saccharomyces cerevisiae  (Example 14), was determined as outlined in Example 14. Table 5 shows the effect on the stability of consensus phytase-1 for each mutation introduced.  
     [0160] Table 5: Stability Effect of the Individual Amino Acid Replacements in Consensus Phytase-1  
     [0161] (+or − means a positive, respectively, negative effect on the protein stability up to 1° C., ++ and − means a positive, respectively, negative effect on the protein stability between 1° C. and 3° C.; the number 10 or 11 corresponds to the consensus phytase sequence that suggested the amino acid replacement.)  
                           TABLE 5                          stabilizing   neutral   destabilizing                                         mutation   effect   Mutation   effect   Mutation   effect               E58A (10)   +   D69A   ±   Y54F (10)   −       D69K (11)   +   D70G (10)   ±   V73I   −       D197N (10)   +   N134Q (10)   ±   A94K (10)   −       T214L (10)   ++   G186H   ±   A101R (11)   −       E222T (11)   ++   S187A (10)   ±   K153N (11)   −       E267D (10)   +   T214V   ±   I158V (10)   −−       R291I*   +   T251N (10)   ±   G203A   −−       R329H (10)   +   Y259N (10)   ±   G205S   −       S364T (10)   ++   A283D (10)   ±   A217V   −       A379K (11)   +   A320V (10)   ±   V227A (11)   −−       G404A (10)   ++   K445T   ±   L234V (10)   −               A463E (10)   ±   A238P (10)   −−                       E277Q (10)   −−                       H287A (11)   −−                       Q292A (10)   −−                       I366V (10)   −−                       S396A (10)   −−                       Q415E (11)   −−                       A437G (10)   −−                       E451R   −−                          
 
     [0162] We combined eight positive mutations (E58A, D197N, E267D, R291I, R329H, S364T, A379K, G404A) in consensus phytase-1 using the primers and the technique mentioned above in this example. Furthermore, the mutations Q50T and K91A were introduced which mainly influence the catalytical characteristics of the phytase (see EP 897 985 as well as Example 14). The DNA and amino acid sequence of the resulting phytase gene (consensus phytase-1-thermo[8]-Q50T-K91A) is shown in FIG. 8. In this way, the temperature optimum and the melting point of the consensus phytase was increased by 7° C. (FIGS. 16, 17,  18 ).  
     [0163] Using the results of Table 5, we further improved the thermostability of consensus phytase 10 by the back mutations K94A, V158I, and A396S that revealed a strong negative influence on the stability of consensus phytase-1. The resulting protein is consensus phytase-10-thermo [3]. We also introduced the mutations Q50T and K91A which mainly influence the catalytical characteristics of consensus phytase (see EP 897 485 as well as Example 14 and FIGS. 15 and 16). The resulting DNA and amino acid sequence is shown in FIG. 9. The optimized phytase showed a 4° C. higher temperature optimum and melting point than consensus phytase-10 (FIGS. 13 and 14). The phytase also had a strongly increased specific activity with phytate as substrate of 250 U/mg at pH 5.5 (FIG. 15).  
     Example 9  
     Stabilization of the Phytase of  A. fumigatus  ATCC 13073 by Replacement of Amino Acid Residues with the Corresponding Consensus Phytase-1 and Consensus Phytase-10 Residues  
     [0164] At six typical positions where the  A. fumigatus  13073 phytase is the only or nearly the only phytase in the alignment of FIG. 2 that does not contain the corresponding consensus phytase amino acid residue, the non-consensus amino acid residue was replaced by the consensus one. In a first round, the following amino acids were substituted in  A. fumigatus  13073 phytase, containing the Q51T substitution and the signal sequence of  A. terreus cbs.  116.46 phytase (see FIG. 10):  
     [0165] F55(28)Y, V100(73)I, F114(87)Y, A243(220)L, S265(242)P, N294(282)D.  
     [0166] The numbers in parentheses refer to the numbering of FIG. 2.  
     [0167] In a second round, four of the seven stabilizing amino acid exchanges (E59A, R329H, S364T, G404A) found in the consensus phytase-10 sequence and, tested as single mutations in consensus phytase-1 (Table 5), were additionally introduced into the  A. fumigatus  α-mutant. The amino acid replacement S154N, shown to reduce the protease susceptibility of the phytase, was also introduced.  
     [0168] The mutations were introduced as described in example 8 (see Table 6), and expressed as described in Examples 11 to 13. The resulting  A. fumigatus  13073 phytase variants were called α-mutant and α-mutant-E59A-S154N—R329H—S364T-G404A.  
     [0169] The temperature optimum (60° C., FIG. 21) and the melting point (67.02° C., FIG. 20) of the  A. fumigatus  13073 phytase α-mutant were increased by 5-7° C. in comparison to the values of the wild-type (temperature optimum: 55° C., T m : 60° C.). The five additional amino acid replacements further increased the temperature optimum by 3° C. (FIG. 21).  
                   TABLE 6                          Mutagenesis primers for stabilization of             A. fumigatus  phytase ATCC 13073                     Mutation   Primer                                     F55Y   5′-CACGTACTCGCCATACTTTTCGCTCGAG-3′   (SEQ ID No. 78)               5′-CTCGAGCGAAAAGTATGGCGAGTACGTG-3′   (SEQ ID No. 79)                                              ( Xho  I)       E58A   5′-CCATACTTTTCG CTCGCG GACGAGCTGTCCGTG-3′   (SEQ ID No. 80)           5′-CACGGACAGCTCGTC CGCGAG CGAAAAGTAGG-3′   (SEQ ID No. 81)               V100I   5′-GTATAAGAAGCTTATTACGGCGATCCAGGCC-3′   (SEQ ID No. 82)           5′-GGCCTGGATCGCCGTAATAAGCTTCTTATAC-3′   (SEQ ID No. 83)               F114Y   5′-CTTCAAGGGCAAGTACGCCTTTTTGAAGACG-3′   (SEQ ID No. 84)           5′-CGTCTTCAAAAAGGCGTACTTGCCCTTGAAG-3′   (SEQ ID No. 85)               A243L   5′-CATCCGAGCTCGCCTCGAGAAGCATCTTC-3′   (SEQ ID No. 86)           5′-GAAGATGCTTCTCGAGGCGAGCTCGGATG-3′   (SEQ ID No. 87)               S265P   5′-CTAATGGA TGTGTCCGTTTGATACGGTAG-3′   (SEQ ID No. 88)           5′-CTACCGTATCAAACGGACACATGTCCATTAG-3′   (SEQ ID No. 89)               N294D   5′-GTGGAAGAAGTACGACTACCTTCAGTC-3′   (SEQ ID No. 90)           5′-GACTGAAGGTAGTCGTACTTCTTCCAC-3′   (SEQ ID No. 91)                                              ( Mlu  I)       R329H   5′-GCCCGGTTG ACGCAT TCGCCAGTGCAGG-3′   (SEQ ID No. 92)           5′-CCTGCACTGGCGA ATGCGT CAACCGGGC-3′   (SEQ ID No. 93)                                                 Nco  I       S364T   5′-CACACGACAACA CCATGG TTTCCATCTTC-3′   (SEQ ID No. 94)           5′-GAAGATGGAAACCATGGTGTTGTCGTGTG-3′   (SEQ ID No. 95)                                                ( Bss  HI)       G404A   5′-GTGGTGCCTTTCG CCGCGC GAGCCTACTTC-3′   (SEQ ID No. 96)           5′-GAAGTAGGCTC GCGCGG CGAAAGGCACCAC-3′   (SEQ ID No. 97)                  
 
     Example 10  
     Introduction of the Active Site Amino Acid Residues of the  A. niger  NRRL 3135 Phytase into the Consensus Phytase-1  
     [0170] We used the crystal structure of the  Aspergillus niger  NRRL 3135 phytase to define all active site amino acid residues (see Reference Example and EP 897 010, which is hereby incorporated by reference as if recited in full herein). Using the alignment of FIG. 2, we replaced the following active site residues and additionally the non-identical adjacent residues of the consensus phytase-1 by those of the  A. niger  phytase:  
     [0171] S89D, S92G, A94K, D164S, P201S, G203A, G205S, H212P, G224A, D226T, E255T, D256E, V258T, P265S, Q292H, G300K, Y305H, A314T, S364G, M365I, A397S, S398A, G404A, and A405S  
     [0172] The new protein sequence, consensus phytase-7, was backtranslated into a DNA sequence (FIG. 11) as described in Example 6. The corresponding gene (fcp7) was generated as described in Example 6 using the following oligonucleotide mixes:  
     [0173] Mix 1.7: CP-1, CP-2, CP-3, CP-4.7, CP-5.7, CP-6, CP-7, CP-8.7, CP-9, CP-10.7  
     [0174] Mix 2.7: CP-9, CP-10.7, CP-11.7, CP-12.7, CP-13.7, CP-14.7, CP-15.7, CP-16, CP-17.7, CP-18.7, CP-19.7, CP-20, CP-21, CP-22.  
     [0175] The DNA sequences of the oligonucleotides are indicated in FIG. 11. The newly synthesized oligonucleotides are additionally marked by the number “7.” After assembling of the oligonucleotides using the same PCR primers as set forth in Example 6, the gene was cloned into an expression vector as described in Examples 11-13.  
     [0176] The pH-profile of consensus phytase-7, purified after expression in  Hansenula polymorpha , was very similar to that of  A. niger  NRRL 3135 phytase (see FIG. 19).  
     Example  11   
     Expression of the Consensus Phytase Genes in  Hansenula olymorpha    
     [0177] The phytase expression vectors, used to transform  H. polymorpha  RB11 (Gellissen et al., 1994), were constructed by inserting the Eco RI fragment of pBsk − fcp or variants thereof into the multiple cloning site of the  H. polymorpha  expression vector pFPMT121, which is based on an ura3 selection marker from  S. cerevisiae , a formate dehydrogenase (FMD) promoter element and a methanol oxidase (MO) terminator element from  H. polymorpha . The 5′ end of the fcp gene is fused to the FMD promoter, and the 3end to the MOX terminator (Gellissen et al., 1996;, EP 0299 108 B). The resulting expression vectors were designated pFPMTfcp, pFPMTfcp10, pFPMTfcp7.  
     [0178] The constructed plasmids were propagated in  E. coli . Plasmid DNA was purified using standard state of the art procedures (see, for example, EP 897 985, Example 5). The expression plasmids were transformed into&#39;the  H. polymorpha  strain RP11 deficient in orotidine-5′-phosphate decarboxylase (ura3) using the procedure for preparation of competent cells and for transformation of yeast as described in Gelissen et al. (1996). Each transformation mixture was plated on YNB (0.14% w/v Difco YNB and 0.5% ammonium sulfate),containing 2% glucose and 10.8% agar and incubated at 37° C. After 4 to 5 days, individual transformant colonies were picked and grown in the liquid medium described above for 2 days at 37° C. Subsequently, an aliquot of this culture was used to inoculate fresh vials With YNB-medium containing 2% glucose. After seven further passages in selective medium, the expression vector is integrated into the yeast genome in multimeric form. Subsequently, mitotically stable transformants were obtained by two additional cultivation steps in 3 ml non-selective liquid medium (YPD, 2% glucose, 10 g yeast extract, and 20 g peptone). To obtain genetically homogeneous recombinant strains, an aliquot from the last stabilization culture was plated on a selective plate. Single colonies were isolated for analysis of phytase expression in YNB containing 2% glycerol instead of glucose to derepress the fmd promoter. Purification of the consensus phytases was done as described in Example 12.  
     Example 12  
     Expression of the Consensus Phytase Genes in  Saccharomyces cerevisiae  and Purification of the Phytases from Culture Supernatant  
     [0179] The consensus phytase genes were isolated from the corresponding Bluescript-plasmid (pBsk − fcp, pBSK − fcp10, pBsk − fcp7), and ligated into the Eco RI sites of the expression cassette of the  Saccharomyces cerevisiae  expression vector pYES2 (Invitrogen, San Diego, Calif., USA) or subcloned between the shortened GAPFL (glyceraldhyde-3-phosphate dehydrogenase) promoter and the phos terminator as described by Janes et al. (1990). The correct orientation of the gene was checked by PCR. Transformation of  S. cerevisiae  strains e.g. INVSc1 (Invitrogen, San Diego, Calif., USA) was done according to Hinnen et al. (1978). Single colonies harboring the phytase gene under the control of the GAPFL promoter were picked and cultivated in 5 ml selection medium (SD-uracil, Sherman et al., 1986) at 30° C. under vigorous shaking (250 rpm) for one day. The preculture was then added to 500 ml YPD medium (Sherman et al., 1986) and grown under the same conditions. Induction of the gall promoter was done according to the manufacturer&#39;s (Invitrogen) instructions. After four days of incubation, cell broth was centrifuged (7000 rpm, GS3 rotor, 15 min, 5° C.) to remove the cells and the supernatant was concentrated by way of ultrafiltration in Amicon 8400 cells (PM30 membranes) and ultrafree-15 centrifugal filter devices (Biomax-30K, Millipore, Bedford, Mass., USA). The concentrate (10 ml) was desalted on a 40 ml Sephadex G25 Superfine column (Pharmacia Biotech, Freiburg, Germany), with 10 mM sodium acetate, pH 5.0, serving as elution buffer. The desalted sample was brought to 2 M (NH 4 ) 2 SO 4  and directly loaded onto a 1 ml Butyl Sepharose 4 Fast Flow hydrophobic interaction chromatography column (Pharmacia Biotech, Freiburg, Germany) which was eluted with a linear gradient from 2 M to 0 M (NH 4 ) 2 SO 4  in 10 mM sodium acetate, pH 5.0. Phytase was eluted in the break-through, concentrated and loaded on a 120 ml Sephacryl S-300 gel permeation chromatography column (Pharmacia Biotech, Freiburg, Germany). Consensus phytase-1 and consensus phytase-7 eluted as a homogeneous symmetrical peak and was shown by SDS-PAGE to be approximately 95% pure.  
     Example 13  
     Expression of the Consensus Phytase Genes in  Aspergillus niger    
     [0180] The Bluescript-plasmids pBsk − fcp, pBSK − fcp10, and pBsk − fcp7 were used as template for the introduction of a Bsp HI-site upstream of the start codon of the genes and an Eco RV-site downstream of the stop codon. The Expand™ High Fidelity PCR Kit (Boehringer Mannheim, Mannheim, Germany) was used with the following primers:  
                                      Primer Asp-1:                             (SEQ ID No. 98)                                             Bsp  HI               5′-TATA TCATGA GCGTGTTCGTCGTGCTACTGTTC-3′                       Primer Asp-2 used for cloning of  fcp  and  fcp7 :                         (SEQ ID No. 99)                                                                Eco  RV               3′-ACCCGACTTACAAAGCGAATT CTATAG ATATAT-5′                       Primer Asp-3 used for cloning of  fcp10 :                         (SEQ ID No. 100)                                                                Eco  RV               3′-ACCCTTCTTACAAAGCGAATT CTATAG ATATAT-5′          
 
     [0181] The reaction was performed as described by the supplier. The PCR-amplified fcp-genes had a new Bsp HI site at the start codon, introduced by primer Asp-1, which resulted in a replacement of the second amino acid residue glycine by serine. Subsequently, the DNA-fragment was digested with Bsp HI and Eco RV and ligated into the Nco I site downstream of the glucoamylase promoter of  Aspergillus niger  (glaA) and the Eco RV site upstream of the Aspergillus nidulans tryptophan C terminator (trpC) (Mullaney et al., 1985). After this cloning step, the genes were sequenced to detect possible failures introduced by PCR. The resulting expression plasmids which basically correspond to the pGLAC vector as described in Example 9 of EP 684 313 contained the orotidine-5′-phosphate decarboxylase gene (pyr4) of  Neurospora crassa  as a selection marker. Transformation of  Aspergillus niger  and expression of the consensus phytase genes was done as described in EP 684 313. The consensus phytases were purified as described in Example 12.  
     Example 14  
     Determination of Phytase Activity and of Temperature Optimum  
     [0182] Phytase activity was determined basically as described by Mitchell et al. (1997). The activity was measured in an assay mixture containing 0.5% phytic acid (≈5 mM) in 200 mM sodium acetate, pH 5.0. After 15 minutes of incubation at 37° C., the reaction was stopped by addition of an equal volume of 15% trichloroacetic acid. The liberated phosphate was quantified by mixing 100 μl of the assay mixture with 900 μl H 2 O and 1 ml of 0.6 M H 2 SO 4 , 2% ascorbic acid and 0.5% ammonium molybdate. Standard solutions of potassium phosphate were used as reference. One unit of enzyme activity was defined as the amount of enzyme that releases 1 μmol phosphate per minute at 37° C. The protein concentration was determined using the enzyme extinction coefficient at 280 nm calculated according to Pace et al. (1995): consensus phytase-1.101; consensus phytase-7, 1.068; consensus phytase-1 10, 1.039.  
     [0183] In case of pH-optimum curves, purified enzymes were diluted in 10 mM sodium acetate, pH 5.0. Incubations were started by mixing aliquots of the diluted protein with an equal volume of 1% phytic acid (≈10 mM) in a series of different buffers: 0.4 M glycine/HCl, pH 2.5; 0.4 M acetate/NaOH, pH 3.0, 3.5, 4.0, 4.5, 5.0, 5.5; 0.4 M imidazole/HCl, pH 6.0, 6.5; 0.4 M Tris/HCl pH 7.0, 7.5, 8.0, 8.5, 9.0. Control experiments showed that the pH was only slightly affected by the mixing step. Incubations were performed for 15 minutes at 37° C. as described above.  
     [0184] For determining the substrate specificities of the phytases, phytic acid in the assay mixture was replaced by 5 mM concentrations of the respective phosphate compounds. The activity tests were performed as described above.  
     [0185] For determination of the temperature optimum, enzyme (100 μl) and substrate solution (100 μl) were pre-incubated for 5 minutes at the given temperature. The reaction was started by addition of the substrate solution to the enzyme. After 15 minutes of incubation, the reaction was stopped with trichloroacetic acid and the amount of phosphate released was determined.  
     [0186] The pH-optimum of the original consensus phytase was about pH 6.0-6.5 (80 U/mg). By introduction of the QSOT mutation, the pH-optimum shifted to pH 6.0 (130 U/mg). After introduction of K91A, the pH optimum shifted one pH-unit into the acidic pH-range showing a higher specific activity between pH 2.5 and pH 6.0. That was shown for the stabilized mutants and for consensus phytase-10, too (FIGS. 15 and 16).  
     [0187] Consensus phytase-7, which was constructed to transfer the catalytic characteristics of the  A. niger  NRRL 3135 phytase into consensus phytase-1, had a pH-profile very similar to that of  A. niger  NRRL 3135 phytase (see FIG. 19). The substrate specificity of consensus phytase-7 also resembled  A. niger  NRRL 3135 phytase more than it resembled consensus phytase-1.  
     [0188] The temperature optimum of consensus phytase-1 (71° C.) was 16-26° C. higher than the temperature optimum of the wild-type phytases (45-55° C., Table 7) which were used to calculate the consensus sequence. The improved consensus phytase-10 showed a further increase of its temperature optimum to 80° C. (FIG. 12). The temperature optimum of the consensus phytase-1-thermo[8] phytase was found in the same range (78° C.) when using the supernatant of an overproducing  S. cerevisiae  strain. The highest temperature optimum reached of 82° C. was determined for consensus phytase-10-thermo[3]-Q50T-K91A.  
               TABLE 7                          Temperature optimum and T m -value of consensus phytase and of the       phytases from  A. fumigatus ,  A. niger ,  E. nidulans , and  M. thermophila .       The determination of the temperature optimum was performed as       described in Example 14. The T m -values were determined by       differential scanning calorimetry as described in Example 15                             Temperature   Tm       Phytase   optimum [° C.]   [° C.]               Consensus phytase-10-thermo[3]-Q50T-K91A   82   89.3       Consensus phytase-10-thermo[3]-Q50T   82   88.6       Consensus phytase-10   80   85.4       Consensus phytase-1-thermo[8]-Q50T   78   84.7       Consensus phytase-1-thermo[8]-Q50T-K91A   78   85.7       Consensus phytase-1   71   78.1         A. niger  NRRL3135   55   63.3         A. fumigatus  13073   55   62.5         A. fumigatus  13073   60   67.0       α-mutant         A. fumigatus  13073   63   —       α-mutant (optimized)         A. terreus  9A-1   49   57.5         A. terreus  cbs.116.46   45   58.5         E. nidulans     45   55.7         M. thermophila     55   n.d.         T. thermophilus     45   n.d.                  
 
     Example 15  
     Determination of the Melting Point by Differential Scanning Calorimetry (DSC)  
     [0189] To determine the unfolding temperature of the phytases, differential scanning calorimetry was applied as previously published by Lehmann et al. (2000). Solutions of 50-60 mg/ml homogeneous phytase were used for the tests. A constant heating rate of 10° C./min was applied up to 90-95° C.  
     [0190] The determined melting points reflect the results obtained for the temperature optima (Table 7). The most stable consensus phytase designed is consensus phytase-10-thermo[3]-Q50T-K91A showing a melting temperature under the chosen conditions of 89.3° C. This is 26 to 33.6° C. higher than the melting points of the wild-type phytases used.  
     Example 16  
     Transfer of Basidiomycete Phytase Active Site into Consensus Phytase-10-Thermo[3]-Q50T-K91A  
     [0191] As described previously (Example 8), mutations derived from the basidiomycete phytase active site were introduced into the consensus phytase-10. The following five constructs a) to e) were prepared:  
     [0192] (a) This construct is called consensus phytase-12, and it contains a selected number of active site residues of the basidio consensus sequence. Its amino acid sequence (consphy 12) is shown in FIG. 22 (the first 26 amino acids form the signal peptide, amended positions are underlined);  
     [0193] (b) a cluster of mutations (Cluster II) was transferred to the consensus phytase-10 sequence, viz.: S80Q, Y86F, S90G, K91A, S92A, K93T, A94R, Y951;  
     [0194] (c) another cluster of mutations (Cluster III) was transferred to the consensus phytase-10 sequence, viz.: T129V, E133A, Q143N, M136S, V137S, N138Q, S139A;  
     [0195] (d) a further cluster of mutations (Cluster IV) was transferred to the consensus phytase-10 sequence, viz.: A168D, E171T, K172N, F173W;  
     [0196] (e) and finally, a further cluster of mutations (Cluster V) was transferred to the consensus phytase-10 sequence, viz.: Q297G, S298D, G300D, Y305T.  
     [0197] These constructs were expressed as described in Examples 11-13.  
     [0198] The following references are incorporated herein by reference as if recited in full herein:  
     [0199] Akanuma, S., Yamagishi, A., Tanaka, N. &amp; Oshima, T. (1998). Serial increase in the thermal stability of 3-isopropylmalate dehydrogenase from  Bacillus subtilis  by experimental evolution.  Prot. Sci.  7, 698-705.  
     [0200] Arase, A., Yomo, T., Urabe, I., Hata, Y., Katsube, Y. &amp; Okada, H. (1993). Stabilization of xylanase by random mutagenesis.  FEBS Lett.  316, 123-127.  
     [0201] Berka, R. M., Rey, M. W., Brown, K. M., Byun, T. &amp; Klotz, A. V. (1998). Molecular characterization and expression of a phytase gene from the thermophilic fungus  Thermomyces lanuginosus. Appl. Environ. Microbiol.  64, 4423-4427.  
     [0202] Blaber, M., Lindstrom, J. D., Gassner, N., Xu, J., Heinz, D. W. &amp; Matthews, B. W. (1993). Energetic cost and structural consequences of burying a hydroxyl group within the core of a protein determined from Ala‘Ser and Val‘Thr substitutions in T4 lysozyme.  Biochemistry  32, 113,63-11373.  
     [0203] Cosgrove, D. J. (1980) Inositol phosphates—their chemistry, biochemistry and physiology: studies in organic chemistry, chapter 4. Elsevier Scientific Publishing-Company, Amsterdam, Oxford, New York.  
     [0204] Devereux, J., Haeberli, P.&amp; Smithies, O. (1984) A comprehensive set of sequence analysis programs for the VAX.  Nucleic Acids Res.  12, 387-395.  
     [0205] Gellissen, G., Hollenberg, C. P., Janowicz, Z. A. (1994) Gene expression in methylotrophic yeasts. In: Smith, A. (ed.) Gene expression in recombinant microorganisms. Dekker, New York, 395-439.  
     [0206] Gellissen; G., Piontek, M., Dahlems, U., Jenzelewski, V., Gavagan, J. E., DiCosimo, R., Anton, D. I. &amp; Janowicz, Z. A. (1996) Recombinant  Hansenula polymorpha  as a biocatalyst: coexpression of the spinach glycolate oxidase (GO) and the  S. cerevisiae  catalase T (CITI) gene.  Appl. Microbiol. Biotechnol.  46, 46-54.  
     [0207] Gerber, P. and Muiller, K. (1995) Moloc molecular modeling software.  J. Comput. Aided Mol. Des.  9, 251-268  
     [0208] Hinnen, A., Hicks, J. B. &amp; Fink, G, R. (1978) Transformation of yeast.  Proc. Natl. Acad. Sci. USA  75, 1929-1933.  
     [0209] Imanaka, T., Shibazaki, M. &amp; Takagi, M. (1986). A new way of enhancing the thermostability of proteases.  Nature  324, 695-697.  
     [0210] Janes, M., Meyhack, B., Zimmermann, W. &amp; Hinnen, A. (1990) The influence of GAP promoter variants on hirudine production, average plasmid copy number and cell growth in  Saccharomyces cerevisiae. Curr. Genet.  18, 97-103.  
     [0211] Karpusas, M., Baase, W. A., Matsumura, M. &amp; Matthews, B. W. (1989). Hydrophobic packing in T4 lysozyme probed by cavity-filling mutants.  Proc. Natl. Acad. Sci . ( USA ) 86, 8237-8241.  
     [0212] Lehmann, L., Kostrewa, D., Wyss, M., Brugger, R., D&#39;Arcy, A., Pasamontes, L., van Loon, A. (2000), From DNA sequence to improved functionality: using protein sequence comparisons to rapidly design a thermostable consensus phytase,  Protein Engineering  13, 49-57.  
     [0213] Margarit, I., Campagnoli, S., Frigerio, F., Grandi, G., Fillipis, V. D. &amp; Fontana, A. (1992). Cumulative stabilizing effects of glycine to alanine substitutions in  Bacillus subtilis  neutral protease.  Prot. Eng.  5, 543-550.  
     [0214] Matthews, B. W. (1987a). Genetic and structural analysis of the protein stability problem.  Biochemistry  26, 6885-6888.  
     [0215] Matthews, B. W. (1993). Structural and genetic analysis of protein stability.  Annu. Rev. Biochem.  62, 139-160.  
     [0216] Matthews, B. W., Nicholson, H. &amp; Becktel, W. (1987). Enhanced protein thermostability from site-directed mutations that decrease the entropy of unfolding.  Proc. Natl. Acad. Sci. (USA)  84, 6663-6667.  
     [0217] Mitchell, D. B., Vogel, K., Weimann, B. J., Pasamontes, L. &amp; van Loon, A. P. G. M. (1997) The phytase subfamily of histidine acid phosphatases: isolation of genes for two novel phytases from the fungi  Aspergillus terreus  and  Myceliophthora thermophila, Microbiology  143, 245-252.  
     [0218] Mullaney, E. J., Hamer, J. E., Roberti, K. A., Yelton, M. M. &amp; Timberlake, W. E. (1985) Primary structure of the trpC gene from  Aspergillus nidulans. Mol. Gen. Genet.  199, 37-46.  
     [0219] Munoz, V. &amp; Serrano, L. (1995). Helix design, prediction and stability.  Curr. Opin. Biotechnol.  6, 382-386.  
     [0220] Pace, N. C., Vajdos, F., Fee, L., Grimsley, G. &amp; Gray, T. (1995). How to measure and predict the molar absorption coefficient of a protein.  Prot. Sci.  4, 2411-2423.  
     [0221] Pantoliano, M. W., Landner, R. C., Brian, P. N., Rollence, M. L., Wood, J. F. &amp; Poulos, T. L. (1987). Protein engineering of subtilisin BPN′: enhanced stabilization through the introduction of two cysteines to form a disulfide bond.  Biochemistry  26, 2077-2082.  
     [0222] Pasamontes, L., Haiker, M., Henriquez-Huecas, M., Mitchell, D. B. &amp; van Loon, A. P. G. M. (1997a). Cloning of the phytases from  Emericella nidulans  and the thermophilic fungus  Talaromyces thermophilus. Biochim. Biophys. Acta  1353, 217-223.  
     [0223] Pasamontes, L., Haiker, M., Wyss, M., Tessier, M. &amp; van Loon, A. P. G. M. (1997) Cloning, purification and characterization of a heat stable phytase from the fungus  Aspergillus fumigatus, Appl. Environ. Microbiol.  63, 1696-1700.  
     [0224] Piddington, C. S., Houston, C. S., Paloheimo, M., Cantrell, M., Miettinen-Oinonen, A. Nevalainen, H., &amp; Rambosek, J. (1993) The cloning and sequencing of the genes encoding phytase (phy) and pH 2.5-optimum acid phosphatase (aph) from  Aspergillus niger  var.  awamori. Gene  133, 55-62.  
     [0225] Purvis, I. J., Bettany, A. J. E., Santiago, T. C., Coggins, J. R., Duncan, K., Eason, R. &amp; Brown, A. J. P. (1987). The efficiency of folding of some proteins is increased by controlled rates of translation in vivo.  J. Mol. Biol.  193, 413-417.  
     [0226] Risse, B., Stempfer, G., Rudolph, R., Schumacher, G. &amp; Jaenicke, R. (1992). Characterization of the stability effect of point mutations of pyruvate oxidase from  Lactobacillus plantarum : protection of the native state by modulating coenzyme binding and subunit interaction.  Prot. Sci.  1, 1710-1718.  
     [0227] Sambrook, J., Fritsch, E. F. &amp; Maniatis, T. (1989)  Molecular Cloning: A Laboratory Manual,  2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.  
     [0228] Sauer, R., Hehir, K., Stearman, R., Weiss, M., Jeitler-Nilsson, A., Suchanek, E. &amp; Pabo, C. (1986). An engineered, intersubunit disulfide enhances the stability and DNA binding of the N-terminal domain of 1-repressor.  Biochemistry  25, 5992-5999.  
     [0229] Serrano, L., Day, A. G. &amp; Fersht, A. R. (1993). Step-wise mutation of bamase to binase. A procedure for engineering increased stability of proteins and an experimental analysis of the evolution of protein stability.  J. Mol. Biol.  233, 305-312.  
     [0230] Sheman, J. P., Finck, G. R. &amp; Hicks, J. B. (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor University.  
     [0231] Steipe, B., Schiller, B., Plueckthun, A. &amp; Steinbach, S. (1994). Sequence statistics reliably predict stabilizing mutations in a protein domain.  J. Mol. Biol.  240, 188-192.  
     [0232] van den Burg, B., Vriend, G., Veltman, O. R., Venema &amp; G., Eijsink, V. G. H. (1998). Engineering an enzyme to resist boiling.  Proc. Natl. Acad. Sci. (USA)  95, 2056-2060.  
     [0233] Van Etten, R. L. (1982) Human prostatic acid phosphatase: a histidine phosphatase.  Ann. NY Acad. Sci.  390, 27-50.  
     [0234] van Hartingsveldt, W., van Zeijl, C. M. F., Harteveld, G. M., Gouka, R. J., Suykerbuyk, M. E. G., Luiten, R. G. M., van Paridon, P. A., Selten, G. C. M., Veenstra, A. E., van Gorcom, R. F. M., &amp; van den Hondel, C. A. M. J. J. (1993) Cloning, characterization and overexpression of the phytase-encoding gene (phyA) of  Aspergillus niger. Gene  127, 87-94.  
     [0235] The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention and all such modifications are intended to be included within the scope of the following claims.  
    
     
       
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tatatgaatt catgggcgtg ttcgtc                                          26 

 
           
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             22  
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tgaaaagttc attgaaggtt tc                                              22 

 
           
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tcttcgaaag cagtacaagt ac                                              22 

 
           
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tatatgaatt cttaagcgaa ac                                              22 

 
           
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tcttcgaaag cagtacacaa ac                                              22 

 
           
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cacttgtggg gtacctactc tccatacttc tc                                   32 

 
           
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gagaagtatg gagagtaggt accccacaag tg                                   32 

 
           
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ggtcaatact ctccattctt ctctttggaa g                                    31 

 
           
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cttccaaaga gaagaatgga gagtattgac c                                    31 

 
           
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catacttctc tttggcagac gaatctgc                                        28 

 
           
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gcagattcgt ctgccaaaga gaagtatg                                        28 

 
           
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ctccagacgt cccaaaggac tgtagagtta c                                    31 

 
           
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gtaactctac agtcctttgg gacgtctgga g                                    31 

 
           
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ctccagacgt cccagacggc tgtagagtta c                                    31 

 
           
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gtaactctac agccgtctgg gacgtctgga g                                    31 

 
           
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gatacccaac ttcttctgcg tctaaggctt actctg                               36 

 
           
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cagagtaagc cttagacgca gaagaagttg ggtatc                               36 

 
           
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cttctaagtc taagaagtac tctgctttg                                       29 

 
           
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caaagcagag tacttcttag acttagaag                                       29 

 
           
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gcttactctg ctttgattga acggattcaa aagaacgcta c                         41 

 
           
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gtagcgttct tttgaatccg ttcaatcaaa gcagagtaag c                         41 

 
           
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ccattcggtg aacagcaaat ggttaactc                                       29 

 
           
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gagttaacca tttgctgttc accgaatgg                                       29 

 
           
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gatacaaggc tctcgcgaga aacattgttc                                      30 

 
           
             25  
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            25 

ggaacaatgt ttctcgcgag agccttgtat c                                    31 

 
           
             26  
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gattgttcca ttcgtgcgcg cttctggttc                                      30 

 
           
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gaaccagaag cgcgcacgaa tggaacaatc                                      30 

 
           
             28  
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ctccagttat taacgtgatc attccagaag g                                    31 

 
           
             29  
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ccttctggaa tgatcacgtt aataactgga g                                    31 

 
           
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ggctgaccca ggggcccaac cacaccaagc                                      30 

 
           
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gcttggtgtg gttgggcccc tgggtcagcc                                      30 

 
           
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cactttggac catggtcttt gtactgcttt cg                                   32 

 
           
             33  
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               Description of Artificial SequencePrimer  
             
           
            33 

cgaaagcagt acaaagacca tggtccaaag tg                                   32 

 
           
             34  
             34  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            34 

gctttcgaag actctaccct aggtgacgac gttg                                 34 

 
           
             35  
             34  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            35 

caacgtcgtc acctagggta gagtcttcga aagc                                 34 

 
           
             36  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            36 

ggtgacgacg ctgaagctaa cttcac                                          26 

 
           
             37  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            37 

gtgaagttag cttcagcgtc gtcacc                                          26 

 
           
             38  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            38 

ctaacttcac cgcggtgttc gctccag                                         27 

 
           
             39  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            39 

ctggagcgaa caccgcggtg aagttag                                         27 

 
           
             40  
             34  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            40 

gctttgttcg ctccacctat tagagctaga ttgg                                 34 

 
           
             41  
             34  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            41 

ccaatctagc tctaataggt ggagcgaaca aagc                                 34 

 
           
             42  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            42 

gccaggtgtt aacttgactg acgaag                                          26 

 
           
             43  
             25  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            43 

ttcgtcagtc aagttaacac ctggc                                           25 

 
           
             44  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            44 

gacgaagacg tcgttaactt gatggac                                         27 

 
           
             45  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            45 

gtccatcaag ttaacgacgt cttcgtc                                         27 

 
           
             46  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            46 

gtccattcga cactgtcgct agaacttc                                        28 

 
           
             47  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            47 

gaagttctag cgacagtgtc gaatggac                                        28 

 
           
             48  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            48 

ctgacgctac tcagctgtct ccattc                                          26 

 
           
             49  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            49 

gaatggagac agctgagtag cgtcag                                          26 

 
           
             50  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            50 

gtctccattc tgtgatttgt tcactcac                                        28 

 
           
             51  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            51 

gtgagtgaac aaatcacaga atggagac                                        28 

 
           
             52  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            52 

gctttgttca ccgcggacga atggag                                          26 

 
           
             53  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            53 

ctccattcgt ccgcggtgaa caaagc                                          26 

 
           
             54  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            54 

cacgacgaat ggatccaata cgactac                                         27 

 
           
             55  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            55 

gtagtcgtat tggatccatt cgtcgtg                                         27 

 
           
             56  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            56 

gacgaatgga gagcgtacga ctacttg                                         27 

 
           
             57  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            57 

caagtagtcg tacgctctcc attcgtc                                         27 

 
           
             58  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            58 

ggtgttggtt tcgttaacga attgattgc                                       29 

 
           
             59  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            59 

gcaatcaatt cgttaacgaa accaacacc                                       29 

 
           
             60  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            60 

gctagattga ctcactctcc agttcaag                                        28 

 
           
             61  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            61 

cttgaactgg agagtgagtc aatctagc                                        28 

 
           
             62  
             32  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            62 

ctcacgacaa cactatgata tctattttct tc                                   32 

 
           
             63  
             32  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            63 

gaagaaaata gatatcatag tgttgtcgtg ag                                   32 

 
           
             64  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            64 

cgacaactcc atggtttcta ttttcttcgc                                      30 

 
           
             65  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            65 

gcgaagaaaa tagaaaccat ggagttgtcg                                      30 

 
           
             66  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            66 

gtacaacggt accaagccat tgtctac                                         27 

 
           
             67  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            67 

gtagacaatg gcttggtacc gttgtac                                         27 

 
           
             68  
             25  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            68 

ctgacggtta cgctgcttct tggac                                           25 

 
           
             69  
             25  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            69 

gtccaagaag cagcgtaacc gtcag                                           25 

 
           
             70  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            70 

ctgttccatt cgctgctaga gcttac                                          26 

 
           
             71  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            71 

gtaagctcta gcagcgaatg gaacag                                          26 

 
           
             72  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            72 

gatgcaatgt gaagctgaaa aggaacc                                         27 

 
           
             73  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            73 

ggttcctttt cagcttcaca ttgcatc                                         27 

 
           
             74  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            74 

cacggttgtg gtgtcgacaa gttggg                                          26 

 
           
             75  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            75 

cccaacttgt cgacaccaca accgtg                                          26 

 
           
             76  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            76 

gatctggtgg caattgggag gaatgtttcg                                      30 

 
           
             77  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            77 

cgaaacattc ctcccaattg ccaccagatc                                      30 

 
           
             78  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            78 

cacgtactcg ccatactttt cgctcgag                                        28 

 
           
             79  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            79 

ctcgagcgaa aagtatggcg agtacgtg                                        28 

 
           
             80  
             33  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            80 

ccatactttt cgctcgcgga cgagctgtcc gtg                                  33 

 
           
             81  
             32  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            81 

cacggacagc tcgtccgcga gcgaaaagta gg                                   32 

 
           
             82  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            82 

gtataagaag cttattacgg cgatccaggc c                                    31 

 
           
             83  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            83 

ggcctggatc gccgtaataa gcttcttata c                                    31 

 
           
             84  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            84 

cttcaagggc aagtacgcct ttttgaagac g                                    31 

 
           
             85  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            85 

cgtcttcaaa aaggcgtact tgcccttgaa g                                    31 

 
           
             86  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            86 

catccgagct cgcctcgaga agcatcttc                                       29 

 
           
             87  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            87 

gaagatgctt ctcgaggcga gctcggatg                                       29 

 
           
             88  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            88 

ctaatggatg tgtccgtttg atacggtag                                       29 

 
           
             89  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            89 

ctaccgtatc aaacggacac atgtccatta g                                    31 

 
           
             90  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            90 

gtggaagaag tacgactacc ttcagtc                                         27 

 
           
             91  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            91 

gcccggttga cgcattcgcc agtgcagg                                        28 

 
           
             92  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            92 

gactgaaggt agtcgtactt cttccac                                         27 

 
           
             93  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            93 

cctgcactgg cgaatgcgtc aaccgggc                                        28 

 
           
             94  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            94 

cacacgacaa caccatggtt tccatcttc                                       29 

 
           
             95  
             29  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            95 

gaagatggaa accatggtgt tgtcgtgtg                                       29 

 
           
             96  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            96 

gtggtgcctt tcgccgcgcg agcctacttc                                      30 

 
           
             97  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            97 

gaagtaggct cgcgcggcga aaggcaccac                                      30 

 
           
             98  
             33  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            98 

tatatcatga gcgtgttcgt cgtgctactg ttc                                  33 

 
           
             99  
             33  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            99 

acccgactta caaagcgaat tctatagata tat                                  33 

 
           
             100  
             33  
             DNA  
             Artificial Sequence  
             
               Description of Artificial SequencePrimer  
             
           
            100 

acccttctta caaagcgaat tctatagata tat                                  33 

 
           
             101  
             440  
             PRT  
             A. terreus 9A-1  
           
            101 

Lys His Ser Asp Cys Asn Ser Val Asp His Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Glu Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr His Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Ser Ala 
 65                  70                  75                  80 

Thr Ala Phe Pro Gly Lys Tyr Ala Phe Leu Gln Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Asp Ser Glu Glu Leu Thr Pro Phe Gly Arg Asn Gln Leu Arg Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Glu Arg Tyr Asn Ala Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Thr Asp Ala Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Thr Ala Arg Gln Asp Asp His His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Ala Ile Pro Glu 
                165                 170                 175 

Gly Ser Ala Tyr Asn Asn Thr Leu Glu His Ser Leu Cys Thr Ala Phe 
            180                 185                 190 

Glu Ser Ser Thr Val Gly Asp Asp Ala Val Ala Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Gln Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Thr Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Thr Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Met Ala Arg Leu Thr Arg Ala 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Ala Pro Leu Ser Gln Thr Ser Val Glu Ser Val Ser Gln Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Pro Thr Asp Lys Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Ala Phe Val Ala Gly Leu Ser Phe Ala Gln Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Asp Cys Phe 
        435                 440 

 
           
             102  
             440  
             PRT  
             A. terreus cbs  
           
            102 

Asn His Ser Asp Cys Thr Ser Val Asp Arg Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Asp Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr Asp Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ala Leu Pro Gly Lys Tyr Ala Phe Leu Lys Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Met Gly Ser Glu Asn Leu Thr Pro Phe Gly Arg Asn Gln Leu Gln Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Arg Arg Tyr Asp Thr Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Ala Asp Ser Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Asn Ala Arg Gln Gly Asp Pro His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Val Ile Pro Glu 
                165                 170                 175 

Gly Thr Ala Tyr Asn Asn Thr Leu Glu His Ser Ile Cys Thr Ala Phe 
            180                 185                 190 

Glu Ala Ser Thr Val Gly Asp Ala Ala Ala Asp Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Lys Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Ala Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Ala Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Gln Thr Thr Val Glu Asp Ile Thr Arg Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Ile Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Gln Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Ala Val Asp Asn Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Glu Cys Phe 
        435                 440 

 
           
             103  
             441  
             PRT  
             A. niger var. awamori  
           
            103 

Asn Gln Ser Thr Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Ala Ile Ser Pro Asp Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Val 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Ile Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Ala Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile His Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Ile Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Ser Ala 
        435                 440 

 
           
             104  
             441  
             PRT  
             A. niger T213  
           
            104 

Asn Gln Ser Ser Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Val Ile Ser Pro Asp Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Val 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Ile Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Ala Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile His Tyr Asp Tyr Leu Arg Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Ile Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             105  
             441  
             PRT  
             A. niger NRRL3135  
           
            105 

Asn Gln Ser Ser Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Val Ile Ser Pro Glu Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Asp Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Ala 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Lys Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Val Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile Asn Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Val Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             106  
             440  
             PRT  
             A. fumigatus 13073  
           
            106 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             107  
             440  
             PRT  
             A. fumigatus 32722  
           
            107 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Gly 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             108  
             440  
             PRT  
             A. fumigatus 58128  
           
            108 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Ser Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             109  
             440  
             PRT  
             A. fumigatus 26906  
           
            109 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ala Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Lys Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             110  
             440  
             PRT  
             A. fumigatus 32239  
           
            110 

Gly Ser Lys Ala Cys Asp Thr Val Glu Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Gly Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Asp Leu Pro Lys Asp Cys Arg Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ala Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Glu Phe Lys Gly Lys Phe Ala Phe Leu Glu Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Met Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Lys Tyr Lys Ala Leu Ala Gly Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Asn Val Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Val Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Tyr Asn Asn Thr Leu Asp His Ser Val Cys Thr Asn Phe Glu 
            180                 185                 190 

Ala Ser Glu Leu Gly Asp Glu Val Glu Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Ala Ile Arg Ala Arg Ile Glu Lys His Leu Pro Gly Val Gln 
    210                 215                 220 

Leu Thr Asp Asp Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ala Asp Ala Ser Glu Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Ile Phe Thr His Asn Glu Trp Lys Lys Tyr Asp Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Asn Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Asp Ser Asp Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Ile Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Gly Met Ile Pro Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Gln Thr Ser Glu Glu Ser Thr Lys Glu Ser Asn Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Ala Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Lys Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Ser Glu Gln Ser Phe Ser 
        435                 440 

 
           
             111  
             439  
             PRT  
             E. nidulans  
           
            111 

Gln Asn His Ser Cys Asn Thr Ala Asp Gly Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Asn Val Ser His Val Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Ile Glu 
             20                  25                  30 

Gln Glu Ser Ala Ile Ser Glu Asp Val Pro His Gly Cys Glu Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Ser Lys Ala Tyr Ser Gly Leu Ile Glu Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ser Phe Trp Gly Gln Tyr Ala Phe Leu Glu Ser Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ile Phe Gly Glu Asn Gln Met Val Asp 
            100                 105                 110 

Ser Gly Ala Lys Phe Tyr Arg Arg Tyr Lys Asn Leu Ala Arg Lys Asn 
        115                 120                 125 

Thr Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Val Ala Ser Ala 
    130                 135                 140 

Glu Lys Phe Ile Asn Gly Phe Arg Lys Ala Gln Leu His Asp His Gly 
145                 150                 155                 160 

Ser Gly Gln Ala Thr Pro Val Val Asn Val Ile Ile Pro Glu Ile Asp 
                165                 170                 175 

Gly Phe Asn Asn Thr Leu Asp His Ser Thr Cys Val Ser Phe Glu Asn 
            180                 185                 190 

Asp Glu Arg Ala Asp Glu Ile Glu Ala Asn Phe Thr Ala Ile Met Gly 
        195                 200                 205 

Pro Pro Ile Arg Lys Arg Leu Glu Asn Asp Leu Pro Gly Ile Lys Leu 
    210                 215                 220 

Thr Asn Glu Asn Val Ile Tyr Leu Met Asp Met Cys Ser Phe Asp Thr 
225                 230                 235                 240 

Met Ala Arg Thr Ala His Gly Thr Glu Leu Ser Pro Phe Cys Ala Ile 
                245                 250                 255 

Phe Thr Glu Lys Glu Trp Leu Gln Tyr Asp Tyr Leu Gln Ser Leu Ser 
            260                 265                 270 

Lys Tyr Tyr Gly Tyr Gly Ala Gly Ser Pro Leu Gly Pro Ala Gln Gly 
        275                 280                 285 

Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Gln Ser Pro Val 
    290                 295                 300 

Gln Asp Asn Thr Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr 
305                 310                 315                 320 

Phe Pro Leu Asp Arg Lys Leu Tyr Ala Asp Phe Ser His Asp Asn Ser 
                325                 330                 335 

Met Ile Ser Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Gln Pro 
            340                 345                 350 

Leu Ser Met Asp Ser Val Glu Ser Ile Gln Glu Met Asp Gly Tyr Ala 
        355                 360                 365 

Ala Ser Trp Thr Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Leu Met 
    370                 375                 380 

Gln Cys Glu Lys Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg 
385                 390                 395                 400 

Val Val Pro Leu His Gly Cys Ala Val Asp Lys Phe Gly Arg Cys Thr 
                405                 410                 415 

Leu Asp Asp Trp Val Glu Gly Leu Asn Phe Ala Arg Ser Gly Gly Asn 
            420                 425                 430 

Trp Lys Thr Cys Phe Thr Leu 
        435 

 
           
             112  
             443  
             PRT  
             T. thermophilus  
           
            112 

Asp Ser His Ser Cys Asn Thr Val Glu Gly Gly Tyr Gln Cys Arg Pro 
  1               5                  10                  15 

Glu Ile Ser His Ser Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asp Gln Ser Glu Ile Ser Pro Asp Val Pro Gln Asn Cys Lys Ile Thr 
         35                  40                  45 

Phe Val Gln Leu Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Thr Glu Leu Tyr Ser Gln Leu Ile Ser Arg Ile Gln Lys Thr Ala 
 65                  70                  75                  80 

Thr Ala Tyr Lys Gly Tyr Tyr Ala Phe Leu Lys Asp Tyr Arg Tyr Gln 
                 85                  90                  95 

Leu Gly Ala Asn Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Ile Gln 
            100                 105                 110 

Leu Gly Ile Lys Phe Tyr Asn His Tyr Lys Ser Leu Ala Arg Asn Ala 
        115                 120                 125 

Val Pro Phe Val Arg Cys Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Arg Leu Phe Ile Glu Gly Phe Gln Ser Ala Lys Val Leu Asp Pro His 
145                 150                 155                 160 

Ser Asp Lys His Asp Ala Pro Pro Thr Ile Asn Val Ile Ile Glu Glu 
                165                 170                 175 

Gly Pro Ser Tyr Asn Asn Thr Leu Asp Thr Gly Ser Cys Pro Val Phe 
            180                 185                 190 

Glu Asp Ser Ser Gly Gly His Asp Ala Gln Glu Lys Phe Ala Lys Gln 
        195                 200                 205 

Phe Ala Pro Ala Ile Leu Glu Lys Ile Lys Asp His Leu Pro Gly Val 
    210                 215                 220 

Asp Leu Ala Val Ser Asp Val Pro Tyr Leu Met Asp Leu Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Leu Ala Arg Asn His Thr Asp Thr Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Leu Ser Thr Gln Glu Glu Trp Gln Ala Tyr Asp Tyr Tyr Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Asn Gly Gly Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Val Gly Phe Val Asn Glu Leu Ile Ala Arg Met Thr His Ser Pro 
    290                 295                 300 

Val Gln Asp Tyr Thr Thr Val Asn His Thr Leu Asp Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn 
                325                 330                 335 

Thr Met Thr Ser Ile Phe Ala Ala Leu Gly Leu Tyr Asn Gly Thr Ala 
            340                 345                 350 

Lys Leu Ser Thr Thr Glu Ile Lys Ser Ile Glu Glu Thr Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ala Trp Thr Val Pro Phe Gly Gly Arg Ala Tyr Ile Glu Met 
    370                 375                 380 

Met Gln Cys Asp Asp Ser Asp Glu Pro Val Val Arg Val Leu Val Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Glu Val Asp Ser Leu Gly Arg 
                405                 410                 415 

Cys Lys Arg Asp Asp Phe Val Arg Gly Leu Ser Phe Ala Arg Gln Gly 
            420                 425                 430 

Gly Asn Trp Glu Gly Cys Tyr Ala Ala Ser Glu 
        435                 440 

 
           
             113  
             466  
             PRT  
             M. thermophila  
           
            113 

Glu Ser Arg Pro Cys Asp Thr Pro Asp Leu Gly Phe Gln Cys Gly Thr 
  1               5                  10                  15 

Ala Ile Ser His Phe Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Val Pro 
             20                  25                  30 

Ser Glu Leu Asp Ala Ser Ile Pro Asp Asp Cys Glu Val Thr Phe Ala 
         35                  40                  45 

Gln Val Leu Ser Arg His Gly Ala Arg Ala Pro Thr Leu Lys Arg Ala 
     50                  55                  60 

Ala Ser Tyr Val Asp Leu Ile Asp Arg Ile His His Gly Ala Ile Ser 
 65                  70                  75                  80 

Tyr Gly Pro Gly Tyr Glu Phe Leu Arg Thr Tyr Asp Tyr Thr Leu Gly 
                 85                  90                  95 

Ala Asp Glu Leu Thr Arg Thr Gly Gln Gln Gln Met Val Asn Ser Gly 
            100                 105                 110 

Ile Lys Phe Tyr Arg Arg Tyr Arg Ala Leu Ala Arg Lys Ser Ile Pro 
        115                 120                 125 

Phe Val Arg Thr Ala Gly Gln Asp Arg Val Val His Ser Ala Glu Asn 
    130                 135                 140 

Phe Thr Gln Gly Phe His Ser Ala Leu Leu Ala Asp Arg Gly Ser Thr 
145                 150                 155                 160 

Val Arg Pro Thr Leu Pro Tyr Asp Met Val Val Ile Pro Glu Thr Ala 
                165                 170                 175 

Gly Ala Asn Asn Thr Leu His Asn Asp Leu Cys Thr Ala Phe Glu Glu 
            180                 185                 190 

Gly Pro Tyr Ser Thr Ile Gly Asp Asp Ala Gln Asp Thr Tyr Leu Ser 
        195                 200                 205 

Thr Phe Ala Gly Pro Ile Thr Ala Arg Val Asn Ala Asn Leu Pro Gly 
    210                 215                 220 

Ala Asn Leu Thr Asp Ala Asp Thr Val Ala Leu Met Asp Leu Cys Pro 
225                 230                 235                 240 

Phe Glu Thr Val Ala Ser Ser Ser Ser Asp Pro Ala Thr Ala Asp Ala 
                245                 250                 255 

Gly Gly Gly Asn Gly Arg Pro Leu Ser Pro Phe Cys Arg Leu Phe Ser 
            260                 265                 270 

Glu Ser Glu Trp Arg Ala Tyr Asp Tyr Leu Gln Ser Val Gly Lys Trp 
        275                 280                 285 

Tyr Gly Tyr Gly Pro Gly Asn Pro Leu Gly Pro Thr Gln Gly Val Gly 
    290                 295                 300 

Phe Val Asn Glu Leu Leu Ala Arg Leu Ala Gly Val Pro Val Arg Asp 
305                 310                 315                 320 

Gly Thr Ser Thr Asn Arg Thr Leu Asp Gly Asp Pro Arg Thr Phe Pro 
                325                 330                 335 

Leu Gly Arg Pro Leu Tyr Ala Asp Phe Ser His Asp Asn Asp Met Met 
            340                 345                 350 

Gly Val Leu Gly Ala Leu Gly Ala Tyr Asp Gly Val Pro Pro Leu Asp 
        355                 360                 365 

Lys Thr Ala Arg Arg Asp Pro Glu Glu Leu Gly Gly Tyr Ala Ala Ser 
    370                 375                 380 

Trp Ala Val Pro Phe Ala Ala Arg Ile Tyr Val Glu Lys Met Arg Cys 
385                 390                 395                 400 

Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Gly Arg Gln Glu Lys 
                405                 410                 415 

Asp Glu Glu Met Val Arg Val Leu Val Asn Asp Arg Val Met Thr Leu 
            420                 425                 430 

Lys Gly Cys Gly Ala Asp Glu Arg Gly Met Cys Thr Leu Glu Arg Phe 
        435                 440                 445 

Ile Glu Ser Met Ala Phe Ala Arg Gly Asn Gly Lys Trp Asp Leu Cys 
    450                 455                 460 

Phe Ala 
465 

 
           
             114  
             431  
             PRT  
             Consensus  
           
            114 

Asn Ser His Ser Cys Asp Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Ile Ser His Leu Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Ser Ala Ile Ser Pro Asp Val Pro Asp Asp Cys Val Thr Phe 
         35                  40                  45 

Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys 
     50                  55                  60 

Lys Ala Tyr Ser Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Phe 
 65                  70                  75                  80 

Lys Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala 
                 85                  90                  95 

Asp Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Val Asn Ser Gly Ile 
            100                 105                 110 

Lys Phe Tyr Arg Arg Tyr Lys Ala Leu Ala Arg Lys Val Pro Phe Val 
        115                 120                 125 

Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile 
    130                 135                 140 

Glu Gly Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ser Pro His Gln 
145                 150                 155                 160 

Ala Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ser Gly Tyr Asn 
                165                 170                 175 

Asn Thr Leu Asp His Gly Thr Cys Thr Ala Phe Glu Asp Ser Glu Leu 
            180                 185                 190 

Gly Asp Asp Ala Glu Ala Asn Phe Thr Ala Thr Phe Ala Pro Ala Ile 
        195                 200                 205 

Arg Ala Arg Leu Glu Ala Asp Leu Pro Gly Val Thr Leu Thr Asp Glu 
    210                 215                 220 

Asp Val Val Leu Met Asp Met Cys Pro Phe Glu Thr Val Ala Arg Thr 
225                 230                 235                 240 

Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys Ala Leu Phe Thr Glu Glu 
                245                 250                 255 

Trp Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly Tyr Gly Ala 
            260                 265                 270 

Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Asn Glu Leu Ile 
        275                 280                 285 

Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr Ser Thr Asn His 
    290                 295                 300 

Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr 
305                 310                 315                 320 

Ala Asp Phe Ser His Asp Asn Ser Met Ile Ser Ile Phe Phe Ala Leu 
                325                 330                 335 

Gly Leu Tyr Asn Gly Thr Ala Pro Leu Ser Thr Thr Ser Val Glu Ser 
            340                 345                 350 

Ile Glu Glu Thr Asp Gly Tyr Ala Ala Ser Trp Thr Val Pro Phe Gly 
        355                 360                 365 

Ala Arg Ala Tyr Val Glu Met Met Gln Cys Gln Ala Glu Lys Glu Pro 
    370                 375                 380 

Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro Leu His Gly Cys 
385                 390                 395                 400 

Ala Val Asp Lys Leu Gly Arg Cys Lys Leu Asp Asp Phe Val Glu Gly 
                405                 410                 415 

Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Ala Glu Cys Phe Ala 
            420                 425                 430 

 
           
             115  
             441  
             PRT  
             Consensus phytase  
           
            115 

Asn Ser His Ser Cys Asp Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Ile Ser His Leu Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Ser Ala Ile Ser Pro Asp Val Pro Asp Asp Cys Arg Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Ala Tyr Ser Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ala Phe Lys Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Ala 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ser Gln Pro His Gln Ala Ser Pro Val Ile Asp Val Ile Ile Pro Glu 
                165                 170                 175 

Gly Ser Gly Tyr Asn Asn Thr Leu Asp His Gly Thr Cys Thr Ala Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Gly Asp Asp Val Glu Ala Asn Phe Thr Ala Leu 
        195                 200                 205 

Phe Ala Pro Ala Ile Arg Ala Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Glu Asp Val Val Tyr Leu Met Asp Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ala Arg Thr Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys 
                245                 250                 255 

Ala Leu Phe Thr His Asp Glu Trp Arg Gln Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala 
        275                 280                 285 

Gln Gly Val Gly Phe Ala Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser 
    290                 295                 300 

Pro Val Gln Asp His Thr Ser Thr Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Ser Met Ile Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Ala Pro Leu Ser Thr Thr Ser Val Glu Ser Ile Glu Glu Thr Asp Gly 
        355                 360                 365 

Tyr Ser Ala Ser Trp Thr Val Pro Phe Gly Ala Arg Ala Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Lys Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Ala Val Asp Lys Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asn Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             116  
             467  
             PRT  
             consensus phytase-1 gene  
           
            116 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser His Leu Trp 
         35                  40                  45 

Gly Gln Tyr Ser Pro Tyr Phe Ser Leu Glu Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Asp Asp Cys Arg Val Thr Phe Val Gln Val Leu Ser 
 65                  70                  75                  80 

Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys Ser Lys Ala Tyr Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Thr Pro Phe Gly Glu Asn Gln Met Val Asn Ser Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Ile Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ser Gln Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asp Val Ile Ile Pro Glu Gly Ser Gly Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Thr Cys Thr Ala Phe Glu Asp Ser Glu Leu Gly 
    210                 215                 220 

Asp Asp Val Glu Ala Asn Phe Thr Ala Leu Phe Ala Pro Ala Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala Asp Leu Pro Gly Val Thr Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Tyr Leu Met Asp Met Cys Pro Phe Glu Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys Ala Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Arg Gln Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly 
    290                 295                 300 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Ala 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Ser Met Ile Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Ala Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ser Ala Ser Trp Thr 
385                 390                 395                 400 

Val Pro Phe Gly Ala Arg Ala Tyr Val Glu Met Met Gln Cys Gln Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Ala Glu 
    450                 455                 460 

Cys Phe Ala 
465 

 
           
             117  
             1426  
             DNA  
             consensus phytase-1 gene  
           
            117 

tatatgaatt catgggcgtg ttcgtcgtgc tactgtccat tgccaccttg ttcggttcca     60 

catccggtac cgccttgggt cctcgtggta attctcactc ttgtgacact gttgacggtg    120 

gttaccaatg tttcccagaa atttctcact tgtggggtca atactctcca tacttctctt    180 

tggaagacga atctgctatt tctccagacg ttccagacga ctgtagagtt actttcgttc    240 

aagttttgtc tagacacggt gctagatacc caacttcttc taagtctaag gcttactctg    300 

ctttgattga agctattcaa aagaacgcta ctgctttcaa gggtaagtac gctttcttga    360 

agacttacaa ctacactttg ggtgctgacg acttgactcc attcggtgaa aaccaaatgg    420 

ttaactctgg tattaagttc tacagaagat acaaggcttt ggctagaaag attgttccat    480 

tcattagagc ttctggttct gacagagtta ttgcttctgc tgaaaagttc attgaaggtt    540 

tccaatctgc taagttggct gacccaggtt ctcaaccaca ccaagcttct ccagttattg    600 

acgttattat tccagaagga tccggttaca acaacacttt ggaccacggt acttgtactg    660 

ctttcgaaga ctctgaattg ggtgacgacg ttgaagctaa cttcactgct ttgttcgctc    720 

cagctattag agctagattg gaagctgact tgccaggtgt tactttgact gacgaagacg    780 

ttgtttactt gatggacatg tgtccattcg aaactgttgc tagaacttct gacgctactg    840 

aattgtctcc attctgtgct ttgttcactc acgacgaatg gagacaatac gactacttgc    900 

aatctttggg taagtactac ggttacggtg ctggtaaccc attgggtcca gctcaaggtg    960 

ttggtttcgc taacgaattg attgctagat tgactagatc tccagttcaa gaccacactt   1020 

ctactaacca cactttggac tctaacccag ctactttccc attgaacgct actttgtacg   1080 

ctgacttctc tcacgacaac tctatgattt ctattttctt cgctttgggt ttgtacaacg   1140 

gtactgctcc attgtctact acttctgttg aatctattga agaaactgac ggttactctg   1200 

cttcttggac tgttccattc ggtgctagag cttacgttga aatgatgcaa tgtcaagctg   1260 

aaaaggaacc attggttaga gttttggtta acgacagagt tgttccattg cacggttgtg   1320 

ctgttgacaa gttgggtaga tgtaagagag acgacttcgt tgaaggtttg tctttcgcta   1380 

gatctggtgg taactgggct gaatgtttcg cttaagaatt catata                  1426 

 
           
             118  
             422  
             PRT  
             P. involutus (phyA1)  
           
            118 

Ser Val Pro Lys Asn Thr Ala Pro Thr Phe Pro Ile Pro Glu Ser Glu 
  1               5                  10                  15 

Gln Arg Asn Trp Ser Pro Tyr Ser Pro Tyr Phe Pro Leu Ala Glu Tyr 
             20                  25                  30 

Lys Ala Pro Pro Ala Gly Cys Gln Ile Asn Gln Val Asn Ile Ile Gln 
         35                  40                  45 

Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Thr Thr Arg Ile Lys 
     50                  55                  60 

Ala Gly Leu Thr Lys Leu Gln Gly Val Gln Asn Phe Thr Asp Ala Lys 
 65                  70                  75                  80 

Phe Asn Phe Ile Lys Ser Phe Lys Tyr Asp Leu Gly Asn Ser Asp Leu 
                 85                  90                  95 

Val Pro Phe Gly Ala Ala Gln Ser Phe Asp Ala Gly Gln Glu Ala Phe 
            100                 105                 110 

Ala Arg Tyr Ser Lys Leu Val Ser Lys Asn Asn Leu Pro Phe Ile Arg 
        115                 120                 125 

Ala Asp Gly Ser Asp Arg Val Val Asp Ser Ala Thr Asn Trp Thr Ala 
    130                 135                 140 

Gly Phe Ala Ser Ala Ser His Asn Thr Val Gln Pro Lys Leu Asn Leu 
145                 150                 155                 160 

Ile Leu Pro Gln Thr Gly Asn Asp Thr Leu Glu Asp Asn Met Cys Pro 
                165                 170                 175 

Ala Ala Gly Asp Ser Asp Pro Gln Val Asn Ala Trp Leu Ala Val Ala 
            180                 185                 190 

Phe Pro Ser Ile Thr Ala Arg Leu Asn Ala Ala Ala Pro Ser Val Asn 
        195                 200                 205 

Leu Thr Asp Thr Asp Ala Phe Asn Leu Val Ser Leu Cys Ala Phe Leu 
    210                 215                 220 

Thr Val Ser Lys Glu Lys Lys Ser Asp Phe Cys Thr Leu Phe Glu Gly 
225                 230                 235                 240 

Ile Pro Gly Ser Phe Glu Ala Phe Ala Tyr Gly Gly Asp Leu Asp Lys 
                245                 250                 255 

Phe Tyr Gly Thr Gly Tyr Gly Gln Glu Leu Gly Pro Val Gln Gly Val 
            260                 265                 270 

Gly Tyr Val Asn Glu Leu Ile Ala Arg Leu Thr Asn Ser Ala Val Arg 
        275                 280                 285 

Asp Asn Thr Gln Thr Asn Arg Thr Leu Asp Ala Ser Pro Val Thr Phe 
    290                 295                 300 

Pro Leu Asn Lys Thr Phe Tyr Ala Asp Phe Ser His Asp Asn Leu Met 
305                 310                 315                 320 

Val Ala Val Phe Ser Ala Met Gly Leu Phe Arg Gln Pro Ala Pro Leu 
                325                 330                 335 

Ser Thr Ser Val Pro Asn Pro Trp Arg Thr Trp Arg Thr Ser Ser Leu 
            340                 345                 350 

Val Pro Phe Ser Gly Arg Met Val Val Glu Arg Leu Ser Cys Phe Gly 
        355                 360                 365 

Thr Thr Lys Val Arg Val Leu Val Gln Asp Gln Val Gln Pro Leu Glu 
    370                 375                 380 

Phe Cys Gly Gly Asp Arg Asn Gly Leu Cys Thr Leu Ala Lys Phe Val 
385                 390                 395                 400 

Glu Ser Gln Thr Phe Ala Arg Ser Asp Gly Ala Gly Asp Phe Glu Lys 
                405                 410                 415 

Cys Phe Ala Thr Ser Ala 
            420 

 
           
             119  
             422  
             PRT  
             P. involutus (phyA2)  
           
            119 

Ser Val Pro Arg Asn Ile Ala Pro Lys Phe Ser Ile Pro Glu Ser Glu 
  1               5                  10                  15 

Gln Arg Asn Trp Ser Pro Tyr Ser Pro Tyr Phe Pro Leu Ala Glu Tyr 
             20                  25                  30 

Lys Ala Pro Pro Ala Gly Cys Glu Ile Asn Gln Val Asn Ile Ile Gln 
         35                  40                  45 

Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Ala Thr Arg Ile Lys 
     50                  55                  60 

Ala Gly Leu Ser Lys Leu Gln Ser Val Gln Asn Phe Thr Asp Pro Lys 
 65                  70                  75                  80 

Phe Asp Phe Ile Lys Ser Phe Thr Tyr Asp Leu Gly Thr Ser Asp Leu 
                 85                  90                  95 

Val Pro Phe Gly Ala Ala Gln Ser Phe Asp Ala Gly Leu Glu Val Phe 
            100                 105                 110 

Ala Arg Tyr Ser Lys Leu Val Ser Ser Asp Asn Leu Pro Phe Ile Arg 
        115                 120                 125 

Ser Asp Gly Ser Asp Arg Val Val Asp Thr Ala Thr Asn Trp Thr Ala 
    130                 135                 140 

Gly Phe Ala Ser Ala Ser Arg Asn Ala Ile Gln Pro Lys Leu Asp Leu 
145                 150                 155                 160 

Ile Leu Pro Gln Thr Gly Asn Asp Thr Leu Glu Asp Asn Met Cys Pro 
                165                 170                 175 

Ala Ala Gly Glu Ser Asp Pro Gln Val Asp Ala Trp Leu Ala Ser Ala 
            180                 185                 190 

Phe Pro Ser Val Thr Ala Gln Leu Asn Ala Ala Ala Pro Gly Ala Asn 
        195                 200                 205 

Leu Thr Asp Ala Asp Ala Phe Asn Leu Val Ser Leu Cys Pro Phe Met 
    210                 215                 220 

Thr Val Ser Lys Glu Gln Lys Ser Asp Phe Cys Thr Leu Phe Glu Gly 
225                 230                 235                 240 

Ile Pro Gly Ser Phe Glu Ala Phe Ala Tyr Ala Gly Asp Leu Asp Lys 
                245                 250                 255 

Phe Tyr Gly Thr Gly Tyr Gly Gln Ala Leu Gly Pro Val Gln Gly Val 
            260                 265                 270 

Gly Tyr Ile Asn Glu Leu Leu Ala Arg Leu Thr Asn Ser Ala Val Asn 
        275                 280                 285 

Asp Asn Thr Gln Thr Asn Arg Thr Leu Asp Ala Ala Pro Asp Thr Phe 
    290                 295                 300 

Pro Leu Asn Lys Thr Met Tyr Ala Asp Phe Ser His Asp Asn Leu Met 
305                 310                 315                 320 

Val Ala Val Phe Ser Ala Met Gly Leu Phe Arg Gln Ser Ala Pro Leu 
                325                 330                 335 

Ser Thr Ser Thr Pro Asp Pro Asn Arg Thr Trp Leu Thr Ser Ser Val 
            340                 345                 350 

Val Pro Phe Ser Ala Arg Met Ala Val Glu Arg Leu Ser Cys Ala Gly 
        355                 360                 365 

Thr Thr Lys Val Arg Val Leu Val Gln Asp Gln Val Gln Pro Leu Glu 
    370                 375                 380 

Phe Cys Gly Gly Asp Gln Asp Gly Leu Cys Ala Leu Asp Lys Phe Val 
385                 390                 395                 400 

Glu Ser Gln Ala Tyr Ala Arg Ser Gly Gly Ala Gly Asp Phe Glu Lys 
                405                 410                 415 

Cys Leu Ala Thr Thr Val 
            420 

 
           
             120  
             420  
             PRT  
             T. pubescens  
           
            120 

His Ile Pro Leu Arg Asp Thr Ser Ala Cys Leu Asp Val Thr Arg Asp 
  1               5                  10                  15 

Val Gln Gln Ser Trp Ser Met Tyr Ser Pro Tyr Phe Pro Ala Ala Thr 
             20                  25                  30 

Tyr Val Ala Pro Pro Ala Ser Cys Gln Ile Asn Gln Val His Ile Ile 
         35                  40                  45 

Gln Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Ala Lys Arg Ile 
     50                  55                  60 

Gln Thr Ala Val Ala Lys Leu Lys Ala Ala Ser Asn Tyr Thr Asp Pro 
 65                  70                  75                  80 

Leu Leu Ala Phe Val Thr Asn Tyr Thr Tyr Ser Leu Gly Gln Asp Ser 
                 85                  90                  95 

Leu Val Glu Leu Gly Ala Thr Gln Ser Ser Glu Ala Gly Gln Glu Ala 
            100                 105                 110 

Phe Thr Arg Tyr Ser Ser Leu Val Ser Ala Asp Glu Leu Pro Phe Val 
        115                 120                 125 

Arg Ala Ser Gly Ser Asp Arg Val Val Ala Thr Ala Asn Asn Trp Thr 
    130                 135                 140 

Ala Gly Phe Ala Leu Ala Ser Ser Asn Ser Ile Thr Pro Val Leu Ser 
145                 150                 155                 160 

Val Ile Ile Ser Glu Ala Gly Asn Asp Thr Leu Asp Asp Asn Met Cys 
                165                 170                 175 

Pro Ala Ala Gly Asp Ser Asp Pro Gln Val Asn Gln Trp Leu Ala Gln 
            180                 185                 190 

Phe Ala Pro Pro Met Thr Ala Arg Leu Asn Ala Gly Ala Pro Gly Ala 
        195                 200                 205 

Asn Leu Thr Asp Thr Asp Thr Tyr Asn Leu Leu Thr Leu Cys Pro Phe 
    210                 215                 220 

Glu Thr Val Ala Thr Glu Arg Arg Ser Glu Phe Cys Asp Ile Tyr Glu 
225                 230                 235                 240 

Glu Leu Gln Ala Glu Asp Ala Phe Ala Tyr Asn Ala Asp Leu Asp Lys 
                245                 250                 255 

Phe Tyr Gly Thr Gly Tyr Gly Gln Pro Leu Gly Pro Val Gln Gly Val 
            260                 265                 270 

Gly Tyr Ile Asn Glu Leu Ile Ala Arg Leu Thr Ala Gln Asn Val Ser 
        275                 280                 285 

Asp His Thr Gln Thr Asn Ser Thr Leu Asp Ser Ser Pro Glu Thr Phe 
    290                 295                 300 

Pro Leu Asn Arg Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Gln Met 
305                 310                 315                 320 

Val Ala Ile Phe Ser Ala Met Gly Leu Phe Asn Gln Ser Ala Pro Leu 
                325                 330                 335 

Asp Pro Thr Thr Pro Asp Pro Ala Arg Thr Phe Leu Val Lys Lys Ile 
            340                 345                 350 

Val Pro Phe Ser Ala Arg Met Val Val Glu Arg Leu Asp Cys Gly Gly 
        355                 360                 365 

Ala Gln Ser Val Arg Leu Leu Val Asn Asp Ala Val Gln Pro Leu Ala 
    370                 375                 380 

Phe Cys Gly Ala Asp Thr Ser Gly Val Cys Thr Leu Asp Ala Phe Val 
385                 390                 395                 400 

Glu Ser Gln Ala Tyr Ala Arg Asn Asp Gly Glu Gly Asp Phe Glu Lys 
                405                 410                 415 

Cys Phe Ala Thr 
            420 

 
           
             121  
             435  
             PRT  
             A. pediades  
           
            121 

Gly Gly Val Val Gln Ala Thr Phe Val Gln Pro Phe Phe Pro Pro Gln 
  1               5                  10                  15 

Ile Gln Asp Ser Trp Ala Ala Tyr Thr Pro Tyr Tyr Pro Val Gln Ala 
             20                  25                  30 

Tyr Thr Pro Pro Pro Lys Asp Cys Lys Ile Thr Gln Val Asn Ile Ile 
         35                  40                  45 

Gln Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Gly Thr Arg Ile 
     50                  55                  60 

Gln Ala Ala Val Lys Lys Leu Gln Ser Ala Lys Thr Tyr Thr Asp Pro 
 65                  70                  75                  80 

Arg Leu Asp Phe Leu Thr Asn Tyr Thr Tyr Thr Leu Gly His Asp Asp 
                 85                  90                  95 

Leu Val Pro Phe Gly Ala Leu Gln Ser Ser Gln Ala Gly Glu Glu Thr 
            100                 105                 110 

Phe Gln Arg Tyr Ser Phe Leu Val Ser Lys Glu Asn Leu Pro Phe Val 
        115                 120                 125 

Arg Ala Ser Ser Ser Asn Arg Val Val Asp Ser Ala Thr Asn Trp Thr 
    130                 135                 140 

Glu Gly Phe Ser Ala Ala Ser His His Val Leu Asn Pro Ile Leu Phe 
145                 150                 155                 160 

Val Ile Leu Ser Glu Ser Leu Asn Asp Thr Leu Asp Asp Ala Met Cys 
                165                 170                 175 

Pro Asn Ala Gly Ser Ser Asp Pro Gln Thr Gly Ile Trp Thr Ser Ile 
            180                 185                 190 

Tyr Gly Thr Pro Ile Ala Asn Arg Leu Asn Gln Gln Ala Pro Gly Ala 
        195                 200                 205 

Asn Ile Thr Ala Ala Asp Val Ser Asn Leu Ile Pro Leu Cys Ala Phe 
    210                 215                 220 

Glu Thr Ile Val Lys Glu Thr Pro Ser Pro Phe Cys Asn Leu Phe Thr 
225                 230                 235                 240 

Pro Glu Glu Phe Ala Gln Phe Glu Tyr Phe Gly Asp Leu Asp Lys Phe 
                245                 250                 255 

Tyr Gly Thr Gly Tyr Gly Gln Pro Leu Gly Pro Val Gln Gly Val Gly 
            260                 265                 270 

Tyr Ile Asn Glu Leu Leu Ala Arg Leu Thr Glu Met Pro Val Arg Asp 
        275                 280                 285 

Asn Thr Gln Thr Asn Arg Thr Leu Asp Ser Ser Pro Leu Thr Phe Pro 
    290                 295                 300 

Leu Asp Arg Ser Ile Tyr Ala Asp Leu Ser His Asp Asn Gln Met Ile 
305                 310                 315                 320 

Ala Ile Phe Ser Ala Met Gly Leu Phe Asn Gln Ser Ser Pro Leu Asp 
                325                 330                 335 

Pro Ser Phe Pro Asn Pro Lys Arg Thr Trp Val Thr Ser Arg Leu Thr 
            340                 345                 350 

Pro Phe Ser Ala Arg Met Val Thr Glu Arg Leu Leu Cys Gln Arg Asp 
        355                 360                 365 

Gly Thr Gly Ser Gly Gly Pro Ser Arg Ile Met Arg Asn Gly Asn Val 
    370                 375                 380 

Gln Thr Phe Val Arg Ile Leu Val Asn Asp Ala Leu Gln Pro Leu Lys 
385                 390                 395                 400 

Phe Cys Gly Gly Asp Met Asp Ser Leu Cys Thr Leu Glu Ala Phe Val 
                405                 410                 415 

Glu Ser Gln Lys Tyr Ala Arg Glu Asp Gly Gln Gly Asp Phe Glu Lys 
            420                 425                 430 

Cys Phe Asp 
        435 

 
           
             122  
             419  
             PRT  
             P. lycii  
           
            122 

Ser Thr Gln Phe Ser Phe Val Ala Ala Gln Leu Pro Ile Pro Ala Gln 
  1               5                  10                  15 

Asn Thr Ser Asn Trp Gly Pro Tyr Asp Pro Phe Phe Pro Val Glu Pro 
             20                  25                  30 

Tyr Ala Ala Pro Pro Glu Gly Cys Thr Val Thr Gln Val Asn Leu Ile 
         35                  40                  45 

Gln Arg His Gly Ala Arg Trp Pro Thr Ser Gly Ala Arg Ser Arg Gln 
     50                  55                  60 

Val Ala Ala Val Ala Lys Ile Gln Met Ala Arg Pro Phe Thr Asp Pro 
 65                  70                  75                  80 

Lys Tyr Glu Phe Leu Asn Asp Phe Val Tyr Lys Phe Gly Val Ala Asp 
                 85                  90                  95 

Leu Leu Pro Phe Gly Ala Asn Gln Ser His Gln Thr Gly Thr Asp Met 
            100                 105                 110 

Tyr Thr Arg Tyr Ser Thr Leu Phe Glu Gly Gly Asp Val Pro Phe Val 
        115                 120                 125 

Arg Ala Ala Gly Asp Gln Arg Val Val Asp Ser Ser Thr Asn Trp Thr 
    130                 135                 140 

Ala Gly Phe Gly Asp Ala Ser Gly Glu Thr Val Leu Pro Thr Leu Gln 
145                 150                 155                 160 

Val Val Leu Gln Glu Glu Gly Asn Cys Thr Leu Cys Asn Asn Met Cys 
                165                 170                 175 

Pro Asn Glu Val Asp Gly Asp Glu Ser Thr Thr Trp Leu Gly Val Phe 
            180                 185                 190 

Ala Pro Asn Ile Thr Ala Arg Leu Asn Ala Ala Ala Pro Ser Ala Asn 
        195                 200                 205 

Leu Ser Asp Ser Asp Ala Leu Thr Leu Met Asp Met Cys Pro Phe Asp 
    210                 215                 220 

Thr Leu Ser Ser Gly Asn Ala Ser Pro Phe Cys Asp Leu Phe Thr Ala 
225                 230                 235                 240 

Glu Glu Tyr Val Ser Tyr Glu Tyr Tyr Tyr Asp Leu Asp Lys Tyr Tyr 
                245                 250                 255 

Gly Thr Gly Pro Gly Asn Ala Leu Gly Pro Val Gln Gly Val Gly Tyr 
            260                 265                 270 

Val Asn Glu Leu Leu Ala Arg Leu Thr Gly Gln Ala Val Arg Asp Glu 
        275                 280                 285 

Thr Gln Thr Asn Arg Thr Leu Asp Ser Asp Pro Ala Thr Phe Pro Leu 
    290                 295                 300 

Asn Arg Thr Phe Tyr Ala Asp Phe Ser His Asp Asn Thr Met Val Pro 
305                 310                 315                 320 

Ile Phe Ala Ala Leu Gly Leu Phe Asn Ala Thr Ala Leu Asp Pro Leu 
                325                 330                 335 

Lys Pro Asp Glu Asn Arg Leu Trp Val Asp Ser Lys Leu Val Pro Phe 
            340                 345                 350 

Ser Gly His Met Thr Val Glu Lys Leu Ala Cys Ser Gly Lys Glu Ala 
        355                 360                 365 

Val Arg Val Leu Val Asn Asp Ala Val Gln Pro Leu Glu Phe Cys Gly 
    370                 375                 380 

Gly Val Asp Gly Val Cys Glu Leu Ser Ala Phe Val Glu Ser Gln Thr 
385                 390                 395                 400 

Tyr Ala Arg Glu Asn Gly Gln Gly Asp Phe Ala Lys Cys Gly Phe Val 
                405                 410                 415 

Pro Ser Glu 

 
           
             123  
             369  
             PRT  
             Basidio  
           
            123 

Ser Pro Arg Thr Ala Ala Gln Leu Pro Ile Pro Gln Gln Trp Ser Pro 
  1               5                  10                  15 

Tyr Ser Pro Tyr Phe Pro Val Ala Tyr Ala Pro Pro Ala Gly Cys Gln 
             20                  25                  30 

Ile Gln Val Asn Ile Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser 
         35                  40                  45 

Gly Ala Ala Thr Arg Ile Gln Ala Ala Val Ala Lys Leu Gln Ser Ala 
     50                  55                  60 

Thr Asp Pro Lys Leu Asp Phe Leu Asn Thr Tyr Leu Gly Asp Asp Leu 
 65                  70                  75                  80 

Val Pro Phe Gly Ala Gln Ser Ser Gln Ala Gly Gln Glu Ala Phe Thr 
                 85                  90                  95 

Arg Tyr Ser Leu Val Ser Asp Asn Leu Pro Phe Val Arg Ala Ser Gly 
            100                 105                 110 

Ser Asp Arg Val Val Asp Ser Ala Thr Asn Trp Thr Ala Gly Phe Ala 
        115                 120                 125 

Ala Ser Asn Thr Pro Leu Val Ile Leu Ser Glu Gly Asn Asp Thr Leu 
    130                 135                 140 

Asp Asp Asn Met Cys Pro Ala Gly Asp Ser Asp Pro Gln Asn Trp Leu 
145                 150                 155                 160 

Ala Val Phe Ala Pro Pro Ile Thr Ala Arg Leu Asn Ala Ala Ala Pro 
                165                 170                 175 

Gly Ala Asn Leu Thr Asp Asp Ala Asn Leu Leu Cys Pro Phe Glu Thr 
            180                 185                 190 

Val Ser Glu Ser Phe Cys Asp Leu Phe Glu Pro Glu Glu Phe Ala Phe 
        195                 200                 205 

Tyr Gly Asp Leu Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Pro Leu 
    210                 215                 220 

Gly Pro Val Gln Gly Val Gly Tyr Ile Asn Glu Leu Leu Ala Arg Leu 
225                 230                 235                 240 

Thr Gln Ala Val Arg Asp Asn Thr Gln Thr Asn Arg Thr Leu Asp Ser 
                245                 250                 255 

Ser Pro Thr Phe Pro Leu Asn Arg Thr Phe Tyr Ala Asp Phe Ser His 
            260                 265                 270 

Asp Asn Gln Met Val Ala Ile Phe Ser Ala Met Gly Leu Phe Asn Gln 
        275                 280                 285 

Ser Ala Pro Leu Asp Pro Ser Pro Asp Pro Asn Arg Thr Trp Val Thr 
    290                 295                 300 

Ser Lys Leu Val Pro Phe Ser Ala Arg Met Val Val Glu Arg Leu Cys 
305                 310                 315                 320 

Gly Thr Val Arg Val Leu Val Asn Asp Ala Val Gln Pro Leu Glu Phe 
                325                 330                 335 

Cys Gly Gly Asp Asp Gly Cys Thr Leu Asp Ala Phe Val Glu Ser Gln 
            340                 345                 350 

Tyr Ala Arg Glu Asp Gly Gln Gly Asp Phe Glu Lys Cys Phe Ala Thr 
        355                 360                 365 

Pro 

 
           
             124  
             440  
             PRT  
             A. terreus 9a1  
           
            124 

Lys His Ser Asp Cys Asn Ser Val Asp His Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Glu Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr His Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Ser Ala 
 65                  70                  75                  80 

Thr Ala Phe Pro Gly Lys Tyr Ala Phe Leu Gln Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Asp Ser Glu Glu Leu Thr Pro Phe Gly Arg Asn Gln Leu Arg Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Glu Arg Tyr Asn Ala Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Thr Asp Ala Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Thr Ala Arg Gln Asp Asp His His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Ala Ile Pro Glu 
                165                 170                 175 

Gly Ser Ala Tyr Asn Asn Thr Leu Glu His Ser Leu Cys Thr Ala Phe 
            180                 185                 190 

Glu Ser Ser Thr Val Gly Asp Asp Ala Val Ala Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Gln Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Thr Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Thr Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Met Ala Arg Leu Thr Arg Ala 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Ala Pro Leu Ser Gln Thr Ser Val Glu Ser Val Ser Gln Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Pro Thr Asp Lys Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Ala Phe Val Ala Gly Leu Ser Phe Ala Gln Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Asp Cys Phe 
        435                 440 

 
           
             125  
             440  
             PRT  
             A. terreus cbs  
           
            125 

Asn His Ser Asp Cys Thr Ser Val Asp Arg Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Asp Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr Asp Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ala Leu Pro Gly Lys Tyr Ala Phe Leu Lys Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Met Gly Ser Glu Asn Leu Thr Pro Phe Gly Arg Asn Gln Leu Gln Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Arg Arg Tyr Asp Thr Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Ala Asp Ser Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Asn Ala Arg Gln Gly Asp Pro His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Val Ile Pro Glu 
                165                 170                 175 

Gly Thr Ala Tyr Asn Asn Thr Leu Glu His Ser Ile Cys Thr Ala Phe 
            180                 185                 190 

Glu Ala Ser Thr Val Gly Asp Ala Ala Ala Asp Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Lys Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Ala Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Ala Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Gln Thr Thr Val Glu Asp Ile Thr Arg Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Ile Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Gln Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Ala Val Asp Asn Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Glu Cys Phe 
        435                 440 

 
           
             126  
             441  
             PRT  
             A. niger var. awamori  
           
            126 

Asn Gln Ser Thr Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Ala Ile Ser Pro Asp Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Val 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Ile Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Ala Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile His Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Ile Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Ser Ala 
        435                 440 

 
           
             127  
             441  
             PRT  
             A. niger NRRL3135  
           
            127 

Asn Gln Ser Ser Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Val Ile Ser Pro Glu Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Asp Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Ala 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Lys Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Val Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile Asn Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Val Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             128  
             440  
             PRT  
             A. fumigatus 13073  
           
            128 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             129  
             440  
             PRT  
             A. fumigatus 32722  
           
            129 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Gly 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             130  
             440  
             PRT  
             A. fumigatus 58128  
           
            130 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Ser Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             131  
             440  
             PRT  
             A. fumigatus 26906  
           
            131 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ala Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Lys Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             132  
             440  
             PRT  
             A. fumigatus 32239  
           
            132 

Gly Ser Lys Ala Cys Asp Thr Val Glu Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Gly Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Asp Leu Pro Lys Asp Cys Arg Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ala Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Glu Phe Lys Gly Lys Phe Ala Phe Leu Glu Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Met Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Lys Tyr Lys Ala Leu Ala Gly Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Asn Val Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Val Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Tyr Asn Asn Thr Leu Asp His Ser Val Cys Thr Asn Phe Glu 
            180                 185                 190 

Ala Ser Glu Leu Gly Asp Glu Val Glu Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Ala Ile Arg Ala Arg Ile Glu Lys His Leu Pro Gly Val Gln 
    210                 215                 220 

Leu Thr Asp Asp Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ala Asp Ala Ser Glu Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Ile Phe Thr His Asn Glu Trp Lys Lys Tyr Asp Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Asn Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Asp Ser Asp Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Ile Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Gly Met Ile Pro Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Gln Thr Ser Glu Glu Ser Thr Lys Glu Ser Asn Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Ala Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Lys Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Ser Glu Gln Ser Phe Ser 
        435                 440 

 
           
             133  
             439  
             PRT  
             E. nidulans  
           
            133 

Gln Asn His Ser Cys Asn Thr Ala Asp Gly Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Asn Val Ser His Val Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Ile Glu 
             20                  25                  30 

Gln Glu Ser Ala Ile Ser Glu Asp Val Pro His Gly Cys Glu Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Ser Lys Ala Tyr Ser Gly Leu Ile Glu Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ser Phe Trp Gly Gln Tyr Ala Phe Leu Glu Ser Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ile Phe Gly Glu Asn Gln Met Val Asp 
            100                 105                 110 

Ser Gly Ala Lys Phe Tyr Arg Arg Tyr Lys Asn Leu Ala Arg Lys Asn 
        115                 120                 125 

Thr Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Val Ala Ser Ala 
    130                 135                 140 

Glu Lys Phe Ile Asn Gly Phe Arg Lys Ala Gln Leu His Asp His Gly 
145                 150                 155                 160 

Ser Gly Gln Ala Thr Pro Val Val Asn Val Ile Ile Pro Glu Ile Asp 
                165                 170                 175 

Gly Phe Asn Asn Thr Leu Asp His Ser Thr Cys Val Ser Phe Glu Asn 
            180                 185                 190 

Asp Glu Arg Ala Asp Glu Ile Glu Ala Asn Phe Thr Ala Ile Met Gly 
        195                 200                 205 

Pro Pro Ile Arg Lys Arg Leu Glu Asn Asp Leu Pro Gly Ile Lys Leu 
    210                 215                 220 

Thr Asn Glu Asn Val Ile Tyr Leu Met Asp Met Cys Ser Phe Asp Thr 
225                 230                 235                 240 

Met Ala Arg Thr Ala His Gly Thr Glu Leu Ser Pro Phe Cys Ala Ile 
                245                 250                 255 

Phe Thr Glu Lys Glu Trp Leu Gln Tyr Asp Tyr Leu Gln Ser Leu Ser 
            260                 265                 270 

Lys Tyr Tyr Gly Tyr Gly Ala Gly Ser Pro Leu Gly Pro Ala Gln Gly 
        275                 280                 285 

Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Gln Ser Pro Val 
    290                 295                 300 

Gln Asp Asn Thr Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr 
305                 310                 315                 320 

Phe Pro Leu Asp Arg Lys Leu Tyr Ala Asp Phe Ser His Asp Asn Ser 
                325                 330                 335 

Met Ile Ser Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Gln Pro 
            340                 345                 350 

Leu Ser Met Asp Ser Val Glu Ser Ile Gln Glu Met Asp Gly Tyr Ala 
        355                 360                 365 

Ala Ser Trp Thr Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Leu Met 
    370                 375                 380 

Gln Cys Glu Lys Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg 
385                 390                 395                 400 

Val Val Pro Leu His Gly Cys Ala Val Asp Lys Phe Gly Arg Cys Thr 
                405                 410                 415 

Leu Asp Asp Trp Val Glu Gly Leu Asn Phe Ala Arg Ser Gly Gly Asn 
            420                 425                 430 

Trp Lys Thr Cys Phe Thr Leu 
        435 

 
           
             134  
             443  
             PRT  
             T. thermophilus  
           
            134 

Asp Ser His Ser Cys Asn Thr Val Glu Gly Gly Tyr Gln Cys Arg Pro 
  1               5                  10                  15 

Glu Ile Ser His Ser Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asp Gln Ser Glu Ile Ser Pro Asp Val Pro Gln Asn Cys Lys Ile Thr 
         35                  40                  45 

Phe Val Gln Leu Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Thr Glu Leu Tyr Ser Gln Leu Ile Ser Arg Ile Gln Lys Thr Ala 
 65                  70                  75                  80 

Thr Ala Tyr Lys Gly Tyr Tyr Ala Phe Leu Lys Asp Tyr Arg Tyr Gln 
                 85                  90                  95 

Leu Gly Ala Asn Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Ile Gln 
            100                 105                 110 

Leu Gly Ile Lys Phe Tyr Asn His Tyr Lys Ser Leu Ala Arg Asn Ala 
        115                 120                 125 

Val Pro Phe Val Arg Cys Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Arg Leu Phe Ile Glu Gly Phe Gln Ser Ala Lys Val Leu Asp Pro His 
145                 150                 155                 160 

Ser Asp Lys His Asp Ala Pro Pro Thr Ile Asn Val Ile Ile Glu Glu 
                165                 170                 175 

Gly Pro Ser Tyr Asn Asn Thr Leu Asp Thr Gly Ser Cys Pro Val Phe 
            180                 185                 190 

Glu Asp Ser Ser Gly Gly His Asp Ala Gln Glu Lys Phe Ala Lys Gln 
        195                 200                 205 

Phe Ala Pro Ala Ile Leu Glu Lys Ile Lys Asp His Leu Pro Gly Val 
    210                 215                 220 

Asp Leu Ala Val Ser Asp Val Pro Tyr Leu Met Asp Leu Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Leu Ala Arg Asn His Thr Asp Thr Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Leu Ser Thr Gln Glu Glu Trp Gln Ala Tyr Asp Tyr Tyr Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Asn Gly Gly Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Val Gly Phe Val Asn Glu Leu Ile Ala Arg Met Thr His Ser Pro 
    290                 295                 300 

Val Gln Asp Tyr Thr Thr Val Asn His Thr Leu Asp Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn 
                325                 330                 335 

Thr Met Thr Ser Ile Phe Ala Ala Leu Gly Leu Tyr Asn Gly Thr Ala 
            340                 345                 350 

Lys Leu Ser Thr Thr Glu Ile Lys Ser Ile Glu Glu Thr Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ala Trp Thr Val Pro Phe Gly Gly Arg Ala Tyr Ile Glu Met 
    370                 375                 380 

Met Gln Cys Asp Asp Ser Asp Glu Pro Val Val Arg Val Leu Val Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Glu Val Asp Ser Leu Gly Arg 
                405                 410                 415 

Cys Lys Arg Asp Asp Phe Val Arg Gly Leu Ser Phe Ala Arg Gln Gly 
            420                 425                 430 

Gly Asn Trp Glu Gly Cys Tyr Ala Ala Ser Glu 
        435                 440 

 
           
             135  
             440  
             PRT  
             T. lanuginosa  
           
            135 

Asn Val Asp Ile Ala Arg His Trp Gly Gln Tyr Ser Pro Phe Phe Ser 
  1               5                  10                  15 

Leu Ala Glu Val Ser Glu Ile Ser Pro Ala Val Pro Lys Gly Cys Arg 
             20                  25                  30 

Val Glu Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr 
         35                  40                  45 

Ala His Lys Ser Glu Val Tyr Ala Glu Leu Leu Gln Arg Ile Gln Asp 
     50                  55                  60 

Thr Ala Thr Glu Phe Lys Gly Asp Phe Ala Phe Leu Arg Asp Tyr Ala 
 65                  70                  75                  80 

Tyr His Leu Gly Ala Asp Asn Leu Thr Arg Phe Gly Glu Glu Gln Met 
                 85                  90                  95 

Met Glu Ser Gly Arg Gln Phe Tyr His Arg Tyr Arg Glu Gln Ala Arg 
            100                 105                 110 

Glu Ile Val Pro Phe Val Arg Ala Ala Gly Ser Ala Arg Val Ile Ala 
        115                 120                 125 

Ser Ala Glu Phe Phe Asn Arg Gly Phe Gln Asp Ala Lys Asp Arg Asp 
    130                 135                 140 

Pro Arg Ser Asn Lys Asp Gln Ala Glu Pro Val Ile Asn Val Ile Ile 
145                 150                 155                 160 

Ser Glu Glu Thr Gly Ser Asn Asn Thr Leu Asp Gly Leu Thr Cys Pro 
                165                 170                 175 

Ala Ala Glu Glu Ala Pro Asp Pro Thr Gln Pro Ala Glu Phe Leu Gln 
            180                 185                 190 

Val Phe Gly Pro Arg Val Leu Lys Lys Ile Thr Lys His Met Pro Gly 
        195                 200                 205 

Val Asn Leu Thr Leu Glu Asp Val Pro Leu Phe Met Asp Leu Cys Pro 
    210                 215                 220 

Phe Asp Thr Val Gly Ser Asp Pro Val Leu Phe Pro Arg Gln Leu Ser 
225                 230                 235                 240 

Pro Phe Cys His Leu Phe Thr Ala Asp Asp Trp Met Ala Tyr Asp Tyr 
                245                 250                 255 

Tyr Tyr Thr Leu Asp Lys Tyr Tyr Ser His Gly Gly Gly Ser Ala Phe 
            260                 265                 270 

Gly Pro Ser Arg Gly Val Gly Phe Val Asn Glu Leu Ile Ala Arg Met 
        275                 280                 285 

Thr Gly Asn Leu Pro Val Lys Asp His Thr Thr Val Asn His Thr Leu 
    290                 295                 300 

Asp Asp Asn Pro Glu Thr Phe Pro Leu Asp Ala Val Leu Tyr Ala Asp 
305                 310                 315                 320 

Phe Ser His Asp Asn Thr Met Thr Gly Ile Phe Ser Ala Met Gly Leu 
                325                 330                 335 

Tyr Asn Gly Thr Lys Pro Leu Ser Thr Ser Lys Ile Gln Pro Pro Thr 
            340                 345                 350 

Gly Ala Ala Ala Asp Gly Tyr Ala Ala Ser Trp Thr Val Pro Phe Ala 
        355                 360                 365 

Ala Arg Ala Tyr Val Glu Leu Leu Arg Cys Glu Thr Glu Thr Ser Ser 
    370                 375                 380 

Glu Glu Glu Glu Glu Gly Glu Asp Glu Pro Phe Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Arg Val Asp Arg Trp Gly 
                405                 410                 415 

Arg Cys Arg Arg Asp Glu Trp Ile Lys Gly Leu Thr Phe Ala Arg Gln 
            420                 425                 430 

Gly Gly His Trp Asp Arg Cys Phe 
        435                 440 

 
           
             136  
             466  
             PRT  
             M. thermophila  
           
            136 

Glu Ser Arg Pro Cys Asp Thr Pro Asp Leu Gly Phe Gln Cys Gly Thr 
  1               5                  10                  15 

Ala Ile Ser His Phe Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Val Pro 
             20                  25                  30 

Ser Glu Leu Asp Ala Ser Ile Pro Asp Asp Cys Glu Val Thr Phe Ala 
         35                  40                  45 

Gln Val Leu Ser Arg His Gly Ala Arg Ala Pro Thr Leu Lys Arg Ala 
     50                  55                  60 

Ala Ser Tyr Val Asp Leu Ile Asp Arg Ile His His Gly Ala Ile Ser 
 65                  70                  75                  80 

Tyr Gly Pro Gly Tyr Glu Phe Leu Arg Thr Tyr Asp Tyr Thr Leu Gly 
                 85                  90                  95 

Ala Asp Glu Leu Thr Arg Thr Gly Gln Gln Gln Met Val Asn Ser Gly 
            100                 105                 110 

Ile Lys Phe Tyr Arg Arg Tyr Arg Ala Leu Ala Arg Lys Ser Ile Pro 
        115                 120                 125 

Phe Val Arg Thr Ala Gly Gln Asp Arg Val Val His Ser Ala Glu Asn 
    130                 135                 140 

Phe Thr Gln Gly Phe His Ser Ala Leu Leu Ala Asp Arg Gly Ser Thr 
145                 150                 155                 160 

Val Arg Pro Thr Leu Pro Tyr Asp Met Val Val Ile Pro Glu Thr Ala 
                165                 170                 175 

Gly Ala Asn Asn Thr Leu His Asn Asp Leu Cys Thr Ala Phe Glu Glu 
            180                 185                 190 

Gly Pro Tyr Ser Thr Ile Gly Asp Asp Ala Gln Asp Thr Tyr Leu Ser 
        195                 200                 205 

Thr Phe Ala Gly Pro Ile Thr Ala Arg Val Asn Ala Asn Leu Pro Gly 
    210                 215                 220 

Ala Asn Leu Thr Asp Ala Asp Thr Val Ala Leu Met Asp Leu Cys Pro 
225                 230                 235                 240 

Phe Glu Thr Val Ala Ser Ser Ser Ser Asp Pro Ala Thr Ala Asp Ala 
                245                 250                 255 

Gly Gly Gly Asn Gly Arg Pro Leu Ser Pro Phe Cys Arg Leu Phe Ser 
            260                 265                 270 

Glu Ser Glu Trp Arg Ala Tyr Asp Tyr Leu Gln Ser Val Gly Lys Trp 
        275                 280                 285 

Tyr Gly Tyr Gly Pro Gly Asn Pro Leu Gly Pro Thr Gln Gly Val Gly 
    290                 295                 300 

Phe Val Asn Glu Leu Leu Ala Arg Leu Ala Gly Val Pro Val Arg Asp 
305                 310                 315                 320 

Gly Thr Ser Thr Asn Arg Thr Leu Asp Gly Asp Pro Arg Thr Phe Pro 
                325                 330                 335 

Leu Gly Arg Pro Leu Tyr Ala Asp Phe Ser His Asp Asn Asp Met Met 
            340                 345                 350 

Gly Val Leu Gly Ala Leu Gly Ala Tyr Asp Gly Val Pro Pro Leu Asp 
        355                 360                 365 

Lys Thr Ala Arg Arg Asp Pro Glu Glu Leu Gly Gly Tyr Ala Ala Ser 
    370                 375                 380 

Trp Ala Val Pro Phe Ala Ala Arg Ile Tyr Val Glu Lys Met Arg Cys 
385                 390                 395                 400 

Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Gly Arg Gln Glu Lys 
                405                 410                 415 

Asp Glu Glu Met Val Arg Val Leu Val Asn Asp Arg Val Met Thr Leu 
            420                 425                 430 

Lys Gly Cys Gly Ala Asp Glu Arg Gly Met Cys Thr Leu Glu Arg Phe 
        435                 440                 445 

Ile Glu Ser Met Ala Phe Ala Arg Gly Asn Gly Lys Trp Asp Leu Cys 
    450                 455                 460 

Phe Ala 
465 

 
           
             137  
             442  
             PRT  
             Basidio  
             
               UNSURE  
               (1)..(442)  
               n is unknown  
             
           
            137 

Xaa Ser Xaa Pro Xaa Arg Xaa Thr Ala Ala Gln Leu Pro Ile Pro Xaa 
  1               5                  10                  15 

Gln Xaa Gln Xaa Xaa Trp Ser Pro Tyr Ser Pro Tyr Phe Pro Val Ala 
             20                  25                  30 

Xaa Tyr Xaa Ala Pro Pro Ala Gly Cys Gln Ile Xaa Gln Val Asn Ile 
         35                  40                  45 

Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Ala Thr Arg 
     50                  55                  60 

Ile Gln Ala Ala Val Ala Lys Leu Gln Ser Ala Xaa Xaa Xaa Thr Asp 
 65                  70                  75                  80 

Pro Lys Leu Asp Phe Leu Xaa Asn Xaa Thr Tyr Xaa Leu Gly Xaa Asp 
                 85                  90                  95 

Asp Leu Val Pro Phe Gly Ala Xaa Gln Ser Ser Gln Ala Gly Gln Glu 
            100                 105                 110 

Ala Phe Thr Arg Tyr Ser Xaa Leu Val Ser Xaa Asp Asn Leu Pro Phe 
        115                 120                 125 

Val Arg Ala Ser Gly Ser Asp Arg Val Val Asp Ser Ala Thr Asn Trp 
    130                 135                 140 

Thr Ala Gly Phe Ala Xaa Ala Ser Xaa Asn Thr Xaa Xaa Pro Xaa Leu 
145                 150                 155                 160 

Xaa Val Ile Leu Ser Glu Xaa Gly Asn Asp Thr Leu Asp Asp Asn Met 
                165                 170                 175 

Cys Pro Xaa Ala Gly Asp Ser Asp Pro Gln Xaa Asn Xaa Trp Leu Ala 
            180                 185                 190 

Val Phe Ala Pro Pro Ile Thr Ala Arg Leu Asn Ala Ala Ala Pro Gly 
        195                 200                 205 

Ala Asn Leu Thr Asp Xaa Asp Ala Xaa Asn Leu Xaa Xaa Leu Cys Pro 
    210                 215                 220 

Phe Glu Thr Val Ser Xaa Glu Xaa Xaa Ser Xaa Phe Cys Asp Leu Phe 
225                 230                 235                 240 

Glu Xaa Xaa Pro Glu Glu Phe Xaa Ala Phe Xaa Tyr Xaa Gly Asp Leu 
                245                 250                 255 

Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Pro Leu Gly Pro Val Gln 
            260                 265                 270 

Gly Val Gly Tyr Ile Asn Glu Leu Leu Ala Arg Leu Thr Xaa Gln Ala 
        275                 280                 285 

Val Arg Asp Asn Thr Gln Thr Asn Arg Thr Leu Asp Ser Ser Pro Xaa 
    290                 295                 300 

Thr Phe Pro Leu Asn Arg Thr Phe Tyr Ala Asp Phe Ser His Asp Asn 
305                 310                 315                 320 

Gln Met Val Ala Ile Phe Ser Ala Met Gly Leu Phe Asn Gln Ser Ala 
                325                 330                 335 

Pro Leu Asp Pro Ser Xaa Pro Asp Pro Asn Arg Thr Trp Val Thr Ser 
            340                 345                 350 

Lys Leu Val Pro Phe Ser Ala Arg Met Val Val Glu Arg Leu Xaa Cys 
        355                 360                 365 

Xaa Xaa Xaa Gly Thr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
    370                 375                 380 

Xaa Xaa Xaa Xaa Xaa Xaa Val Arg Val Leu Val Asn Asp Ala Val Gln 
385                 390                 395                 400 

Pro Leu Glu Phe Cys Gly Gly Asp Xaa Asp Gly Xaa Cys Thr Leu Asp 
                405                 410                 415 

Ala Phe Val Glu Ser Gln Xaa Tyr Ala Arg Glu Asp Gly Gln Gly Asp 
            420                 425                 430 

Phe Glu Lys Cys Phe Ala Thr Pro Xaa Xaa 
        435                 440 

 
           
             138  
             424  
             PRT  
             Consensus  
           
            138 

Asn Ser His Ser Cys Asp Thr Val Asp Gly Gly Tyr Gln Cys Pro Glu 
  1               5                  10                  15 

Ile Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala Asp 
             20                  25                  30 

Glu Ser Ala Ile Ser Pro Asp Val Pro Gly Cys Arg Val Thr Phe Val 
         35                  40                  45 

Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys Ser 
     50                  55                  60 

Lys Lys Tyr Ser Ala Leu Ile Ala Ile Gln Lys Asn Ala Thr Phe Lys 
 65                  70                  75                  80 

Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp 
                 85                  90                  95 

Asp Leu Thr Pro Phe Gly Glu Gln Gln Met Val Asn Ser Gly Ile Lys 
            100                 105                 110 

Phe Tyr Arg Arg Tyr Lys Ala Leu Ala Arg Ile Val Pro Phe Val Arg 
        115                 120                 125 

Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu 
    130                 135                 140 

Gly Phe Gln Ser Ala Lys Leu Ala Asp Pro Ala Gln Ala Ser Pro Val 
145                 150                 155                 160 

Ile Asn Val Ile Ile Pro Glu Gly Gly Tyr Asn Asn Thr Leu Asp His 
                165                 170                 175 

Gly Leu Cys Thr Ala Phe Glu Pro Ser Glu Leu Gly Asp Asp Val Glu 
            180                 185                 190 

Ala Asn Phe Thr Ala Val Phe Ala Pro Pro Ile Arg Ala Arg Leu Glu 
        195                 200                 205 

Ala Leu Pro Gly Val Asn Leu Thr Asp Glu Asp Val Val Asn Leu Met 
    210                 215                 220 

Asp Met Cys Pro Phe Asp Thr Val Ala Thr Ser Asp Ala Thr Gln Leu 
225                 230                 235                 240 

Ser Pro Phe Cys Asp Leu Phe Thr His Glu Trp Gln Tyr Asp Tyr Leu 
                245                 250                 255 

Gln Ser Leu Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro 
            260                 265                 270 

Ala Gln Gly Val Gly Phe Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
        275                 280                 285 

Pro Val Gln Asp His Thr Ser Thr Asn His Thr Leu Asp Ser Asn Pro 
    290                 295                 300 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
305                 310                 315                 320 

Asn Thr Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
                325                 330                 335 

Pro Leu Ser Thr Thr Ser Val Glu Pro Ser Glu Glu Thr Asp Gly Tyr 
            340                 345                 350 

Ala Ala Ser Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Val Glu Met 
        355                 360                 365 

Met Gln Cys Glu Glu Gly Glu Lys Glu Pro Leu Val Arg Val Leu Val 
    370                 375                 380 

Asn Asp Arg Val Val Pro Leu His Gly Cys Gly Val Asp Lys Leu Gly 
385                 390                 395                 400 

Arg Cys Lys Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly 
                405                 410                 415 

Gly Asn Trp Glu Glu Cys Phe Ala 
            420 

 
           
             139  
             467  
             PRT  
             consensus phytase-10  
           
            139 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser His Leu Trp 
         35                  40                  45 

Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Lys Gly Cys Arg Val Thr Phe Val Gln Val Leu Ser 
 65                  70                  75                  80 

Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys Ser Lys Lys Tyr Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Thr Pro Phe Gly Glu Gln Gln Met Val Asn Ser Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Val Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ala Asn Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ala Gly Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Leu Cys Thr Ala Phe Glu Glu Ser Glu Leu Gly 
    210                 215                 220 

Asp Asp Val Glu Ala Asn Phe Thr Ala Val Phe Ala Pro Pro Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala His Leu Pro Gly Val Asn Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Asn Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Gln Leu Ser Pro Phe Cys Asp Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Ile Gln Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly 
    290                 295                 300 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Val 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Thr Met Val Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Lys Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ala Ala Ser Trp Thr 
385                 390                 395                 400 

Val Pro Phe Ala Ala Arg Ala Tyr Val Glu Met Met Gln Cys Glu Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Gly Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Glu Glu 
    450                 455                 460 

Cys Phe Ala 
465 

 
           
             140  
             1426  
             DNA  
             consensus phytase-10  
           
            140 

tatatgaatt catgggcgtg ttcgtcgtgc tactgtccat tgccaccttg ttcggttcca     60 

catccggtac cgccttgggt cctcgtggta attctcactc ttgtgacact gttgacggtg    120 

gttaccaatg tttcccagaa atttctcact tgtggggtca atactctcca ttcttctctt    180 

tggctgacga atctgctatt tctccagacg ttccaaaggg ttgtagagtt actttcgttc    240 

aagttttgtc tagacacggt gctagatacc caacttcttc taagtctaag aagtactctg    300 

ctttgattga agctattcaa aagaacgcta ctgctttcaa gggtaagtac gctttcttga    360 

agacttacaa ctacactttg ggtgctgacg acttgactcc attcggtgaa caacaaatgg    420 

ttaactctgg tattaagttc tacagaagat acaaggcttt ggctagaaag attgttccat    480 

tcgttagagc ttctggttct gacagagtta ttgcttctgc tgaaaagttc attgaaggtt    540 

tccaatctgc taagttggct gacccaggtg ctaacccaca ccaagcttct ccagttatta    600 

acgttattat tccagaaggt gctggttaca acaacacttt ggaccacggt ttgtgtactg    660 

ctttcgaaga atctgaattg ggtgacgacg ttgaagctaa cttcactgct gttttcgctc    720 

cacctattag agctagattg gaagctcact tgccaggtgt taacttgact gacgaagacg    780 

ttgttaactt gatggacatg tgtccattcg acactgttgc tagaacttct gacgctactc    840 

aattgtctcc attctgtgac ttgttcactc acgacgaatg gattcaatac gactacttgc    900 

aatctttggg taagtactac ggttacggtg ctggtaaccc attgggtcca gctcaaggtg    960 

ttggtttcgt taacgaattg attgctagat tgactcactc tccagttcaa gaccacactt   1020 

ctactaacca cactttggac tctaacccag ctactttccc attgaacgct actttgtacg   1080 

ctgacttctc tcacgacaac actatggttt ctattttctt cgctttgggt ttgtacaacg   1140 

gtactaagcc attgtctact acttctgttg aatctattga agaaactgac ggttacgctg   1200 

cttcttggac tgttccattc gctgctagag cttacgttga aatgatgcaa tgtgaagctg   1260 

aaaaggaacc attggttaga gttttggtta acgacagagt tgttccattg cacggttgtg   1320 

gtgttgacaa gttgggtaga tgtaagagag acgacttcgt tgaaggtttg tctttcgcta   1380 

gatctggtgg taactgggaa gaatgtttcg cttaagaatt catata                  1426 

 
           
             141  
             413  
             PRT  
             P. involutus (phyA1)  
           
            141 

Phe Pro Ile Pro Glu Ser Glu Gln Arg Asn Trp Ser Pro Tyr Ser Pro 
  1               5                  10                  15 

Tyr Phe Pro Leu Ala Glu Tyr Lys Ala Pro Pro Ala Gly Cys Gln Ile 
             20                  25                  30 

Asn Gln Val Asn Ile Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser 
         35                  40                  45 

Gly Ala Thr Thr Arg Ile Lys Ala Gly Leu Thr Lys Leu Gln Gly Val 
     50                  55                  60 

Gln Asn Phe Thr Asp Ala Lys Phe Asn Phe Ile Lys Ser Phe Lys Tyr 
 65                  70                  75                  80 

Asp Leu Gly Asn Ser Asp Leu Val Pro Phe Gly Ala Ala Gln Ser Phe 
                 85                  90                  95 

Asp Ala Gly Gln Glu Ala Phe Ala Arg Tyr Ser Lys Leu Val Ser Lys 
            100                 105                 110 

Asn Asn Leu Pro Phe Ile Arg Ala Asp Gly Ser Asp Arg Val Val Asp 
        115                 120                 125 

Ser Ala Thr Asn Trp Thr Ala Gly Phe Ala Ser Ala Ser His Asn Thr 
    130                 135                 140 

Val Gln Pro Lys Leu Asn Leu Ile Leu Pro Gln Thr Gly Asn Asp Thr 
145                 150                 155                 160 

Leu Glu Asp Asn Met Cys Pro Ala Ala Gly Asp Ser Asp Pro Gln Val 
                165                 170                 175 

Asn Ala Trp Leu Ala Val Ala Phe Pro Ser Ile Thr Ala Arg Leu Asn 
            180                 185                 190 

Ala Ala Ala Pro Ser Val Asn Leu Thr Asp Thr Asp Ala Phe Asn Leu 
        195                 200                 205 

Val Ser Leu Cys Ala Phe Leu Thr Val Ser Lys Glu Lys Lys Ser Asp 
    210                 215                 220 

Phe Cys Thr Leu Phe Glu Gly Ile Pro Gly Ser Phe Glu Ala Phe Ala 
225                 230                 235                 240 

Tyr Gly Gly Asp Leu Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Glu 
                245                 250                 255 

Leu Gly Pro Val Gln Gly Val Gly Tyr Val Asn Glu Leu Ile Ala Arg 
            260                 265                 270 

Leu Thr Asn Ser Ala Val Arg Asp Asn Thr Gln Thr Asn Arg Thr Leu 
        275                 280                 285 

Asp Ala Ser Pro Val Thr Phe Pro Leu Asn Lys Thr Phe Tyr Ala Asp 
    290                 295                 300 

Phe Ser His Asp Asn Leu Met Val Ala Val Phe Ser Ala Met Gly Leu 
305                 310                 315                 320 

Phe Arg Gln Pro Ala Pro Leu Ser Thr Ser Val Pro Asn Pro Trp Arg 
                325                 330                 335 

Thr Trp Arg Thr Ser Ser Leu Val Pro Phe Ser Gly Arg Met Val Val 
            340                 345                 350 

Glu Arg Leu Ser Cys Phe Gly Thr Thr Lys Val Arg Val Leu Val Gln 
        355                 360                 365 

Asp Gln Val Gln Pro Leu Glu Phe Cys Gly Gly Asp Arg Asn Gly Leu 
    370                 375                 380 

Cys Thr Leu Ala Lys Phe Val Glu Ser Gln Thr Phe Ala Arg Ser Asp 
385                 390                 395                 400 

Gly Ala Gly Asp Phe Glu Lys Cys Phe Ala Thr Ser Ala 
                405                 410 

 
           
             142  
             413  
             PRT  
             P. involutus (phyA2)  
           
            142 

Phe Ser Ile Pro Glu Ser Glu Gln Arg Asn Trp Ser Pro Tyr Ser Pro 
  1               5                  10                  15 

Tyr Phe Pro Leu Ala Glu Tyr Lys Ala Pro Pro Ala Gly Cys Glu Ile 
             20                  25                  30 

Asn Gln Val Asn Ile Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser 
         35                  40                  45 

Gly Ala Ala Thr Arg Ile Lys Ala Gly Leu Ser Lys Leu Gln Ser Val 
     50                  55                  60 

Gln Asn Phe Thr Asp Pro Lys Phe Asp Phe Ile Lys Ser Phe Thr Tyr 
 65                  70                  75                  80 

Asp Leu Gly Thr Ser Asp Leu Val Pro Phe Gly Ala Ala Gln Ser Phe 
                 85                  90                  95 

Asp Ala Gly Leu Glu Val Phe Ala Arg Tyr Ser Lys Leu Val Ser Ser 
            100                 105                 110 

Asp Asn Leu Pro Phe Ile Arg Ser Asp Gly Ser Asp Arg Val Val Asp 
        115                 120                 125 

Thr Ala Thr Asn Trp Thr Ala Gly Phe Ala Ser Ala Ser Arg Asn Ala 
    130                 135                 140 

Ile Gln Pro Lys Leu Asp Leu Ile Leu Pro Gln Thr Gly Asn Asp Thr 
145                 150                 155                 160 

Leu Glu Asp Asn Met Cys Pro Ala Ala Gly Glu Ser Asp Pro Gln Val 
                165                 170                 175 

Asp Ala Trp Leu Ala Ser Ala Phe Pro Ser Val Thr Ala Gln Leu Asn 
            180                 185                 190 

Ala Ala Ala Pro Gly Ala Asn Leu Thr Asp Ala Asp Ala Phe Asn Leu 
        195                 200                 205 

Val Ser Leu Cys Pro Phe Met Thr Val Ser Lys Glu Gln Lys Ser Asp 
    210                 215                 220 

Phe Cys Thr Leu Phe Glu Gly Ile Pro Gly Ser Phe Glu Ala Phe Ala 
225                 230                 235                 240 

Tyr Ala Gly Asp Leu Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Ala 
                245                 250                 255 

Leu Gly Pro Val Gln Gly Val Gly Tyr Ile Asn Glu Leu Leu Ala Arg 
            260                 265                 270 

Leu Thr Asn Ser Ala Val Asn Asp Asn Thr Gln Thr Asn Arg Thr Leu 
        275                 280                 285 

Asp Ala Ala Pro Asp Thr Phe Pro Leu Asn Lys Thr Met Tyr Ala Asp 
    290                 295                 300 

Phe Ser His Asp Asn Leu Met Val Ala Val Phe Ser Ala Met Gly Leu 
305                 310                 315                 320 

Phe Arg Gln Ser Ala Pro Leu Ser Thr Ser Thr Pro Asp Pro Asn Arg 
                325                 330                 335 

Thr Trp Leu Thr Ser Ser Val Val Pro Phe Ser Ala Arg Met Ala Val 
            340                 345                 350 

Glu Arg Leu Ser Cys Ala Gly Thr Thr Lys Val Arg Val Leu Val Gln 
        355                 360                 365 

Asp Gln Val Gln Pro Leu Glu Phe Cys Gly Gly Asp Gln Asp Gly Leu 
    370                 375                 380 

Cys Ala Leu Asp Lys Phe Val Glu Ser Gln Ala Tyr Ala Arg Ser Gly 
385                 390                 395                 400 

Gly Ala Gly Asp Phe Glu Lys Cys Leu Ala Thr Thr Val 
                405                 410 

 
           
             143  
             410  
             PRT  
             T. pubescens  
           
            143 

Leu Asp Val Thr Arg Asp Val Gln Gln Ser Trp Ser Met Tyr Ser Pro 
  1               5                  10                  15 

Tyr Phe Pro Ala Ala Thr Tyr Val Ala Pro Pro Ala Ser Cys Gln Ile 
             20                  25                  30 

Asn Gln Val His Ile Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser 
         35                  40                  45 

Gly Ala Ala Lys Arg Ile Gln Thr Ala Val Ala Lys Leu Lys Ala Ala 
     50                  55                  60 

Ser Asn Tyr Thr Asp Pro Leu Leu Ala Phe Val Thr Asn Tyr Thr Tyr 
 65                  70                  75                  80 

Ser Leu Gly Gln Asp Ser Leu Val Glu Leu Gly Ala Thr Gln Ser Ser 
                 85                  90                  95 

Glu Ala Gly Gln Glu Ala Phe Thr Arg Tyr Ser Ser Leu Val Ser Ala 
            100                 105                 110 

Asp Glu Leu Pro Phe Val Arg Ala Ser Gly Ser Asp Arg Val Val Ala 
        115                 120                 125 

Thr Ala Asn Asn Trp Thr Ala Gly Phe Ala Leu Ala Ser Ser Asn Ser 
    130                 135                 140 

Ile Thr Pro Val Leu Ser Val Ile Ile Ser Glu Ala Gly Asn Asp Thr 
145                 150                 155                 160 

Leu Asp Asp Asn Met Cys Pro Ala Ala Gly Asp Ser Asp Pro Gln Val 
                165                 170                 175 

Asn Gln Trp Leu Ala Gln Phe Ala Pro Pro Met Thr Ala Arg Leu Asn 
            180                 185                 190 

Ala Gly Ala Pro Gly Ala Asn Leu Thr Asp Thr Asp Thr Tyr Asn Leu 
        195                 200                 205 

Leu Thr Leu Cys Pro Phe Glu Thr Val Ala Thr Glu Arg Arg Ser Glu 
    210                 215                 220 

Phe Cys Asp Ile Tyr Glu Glu Leu Gln Ala Glu Asp Ala Phe Ala Tyr 
225                 230                 235                 240 

Asn Ala Asp Leu Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Pro Leu 
                245                 250                 255 

Gly Pro Val Gln Gly Val Gly Tyr Ile Asn Glu Leu Ile Ala Arg Leu 
            260                 265                 270 

Thr Ala Gln Asn Val Ser Asp His Thr Gln Thr Asn Ser Thr Leu Asp 
        275                 280                 285 

Ser Ser Pro Glu Thr Phe Pro Leu Asn Arg Thr Leu Tyr Ala Asp Phe 
    290                 295                 300 

Ser His Asp Asn Gln Met Val Ala Ile Phe Ser Ala Met Gly Leu Phe 
305                 310                 315                 320 

Asn Gln Ser Ala Pro Leu Asp Pro Thr Thr Pro Asp Pro Ala Arg Thr 
                325                 330                 335 

Phe Leu Val Lys Lys Ile Val Pro Phe Ser Ala Arg Met Val Val Glu 
            340                 345                 350 

Arg Leu Asp Cys Gly Gly Ala Gln Ser Val Arg Leu Leu Val Asn Asp 
        355                 360                 365 

Ala Val Gln Pro Leu Ala Phe Cys Gly Ala Asp Thr Ser Gly Val Cys 
    370                 375                 380 

Thr Leu Asp Ala Phe Val Glu Ser Gln Ala Tyr Ala Arg Asn Asp Gly 
385                 390                 395                 400 

Glu Gly Asp Phe Glu Lys Cys Phe Ala Thr 
                405                 410 

 
           
             144  
             425  
             PRT  
             A. pediades  
           
            144 

Pro Phe Phe Pro Pro Gln Ile Gln Asp Ser Trp Ala Ala Tyr Thr Pro 
  1               5                  10                  15 

Tyr Tyr Pro Val Gln Ala Tyr Thr Pro Pro Pro Lys Asp Cys Lys Ile 
             20                  25                  30 

Thr Gln Val Asn Ile Ile Gln Arg His Gly Ala Arg Phe Pro Thr Ser 
         35                  40                  45 

Gly Ala Gly Thr Arg Ile Gln Ala Ala Val Lys Lys Leu Gln Ser Ala 
     50                  55                  60 

Lys Thr Tyr Thr Asp Pro Arg Leu Asp Phe Leu Thr Asn Tyr Thr Tyr 
 65                  70                  75                  80 

Thr Leu Gly His Asp Asp Leu Val Pro Phe Gly Ala Leu Gln Ser Ser 
                 85                  90                  95 

Gln Ala Gly Glu Glu Thr Phe Gln Arg Tyr Ser Phe Leu Val Ser Lys 
            100                 105                 110 

Glu Asn Leu Pro Phe Val Arg Ala Ser Ser Ser Asn Arg Val Val Asp 
        115                 120                 125 

Ser Ala Thr Asn Trp Thr Glu Gly Phe Ser Ala Ala Ser His His Val 
    130                 135                 140 

Leu Asn Pro Ile Leu Phe Val Ile Leu Ser Glu Ser Leu Asn Asp Thr 
145                 150                 155                 160 

Leu Asp Asp Ala Met Cys Pro Asn Ala Gly Ser Ser Asp Pro Gln Thr 
                165                 170                 175 

Gly Ile Trp Thr Ser Ile Tyr Gly Thr Pro Ile Ala Asn Arg Leu Asn 
            180                 185                 190 

Gln Gln Ala Pro Gly Ala Asn Ile Thr Ala Ala Asp Val Ser Asn Leu 
        195                 200                 205 

Ile Pro Leu Cys Ala Phe Glu Thr Ile Val Lys Glu Thr Pro Ser Pro 
    210                 215                 220 

Phe Cys Asn Leu Phe Thr Pro Glu Glu Phe Ala Gln Phe Glu Tyr Phe 
225                 230                 235                 240 

Gly Asp Leu Asp Lys Phe Tyr Gly Thr Gly Tyr Gly Gln Pro Leu Gly 
                245                 250                 255 

Pro Val Gln Gly Val Gly Tyr Ile Asn Glu Leu Leu Ala Arg Leu Thr 
            260                 265                 270 

Glu Met Pro Val Arg Asp Asn Thr Gln Thr Asn Arg Thr Leu Asp Ser 
        275                 280                 285 

Ser Pro Leu Thr Phe Pro Leu Asp Arg Ser Ile Tyr Ala Asp Leu Ser 
    290                 295                 300 

His Asp Asn Gln Met Ile Ala Ile Phe Ser Ala Met Gly Leu Phe Asn 
305                 310                 315                 320 

Gln Ser Ser Pro Leu Asp Pro Ser Phe Pro Asn Pro Lys Arg Thr Trp 
                325                 330                 335 

Val Thr Ser Arg Leu Thr Pro Phe Ser Ala Arg Met Val Thr Glu Arg 
            340                 345                 350 

Leu Leu Cys Gln Arg Asp Gly Thr Gly Ser Gly Gly Pro Ser Arg Ile 
        355                 360                 365 

Met Arg Asn Gly Asn Val Gln Thr Phe Val Arg Ile Leu Val Asn Asp 
    370                 375                 380 

Ala Leu Gln Pro Leu Lys Phe Cys Gly Gly Asp Met Asp Ser Leu Cys 
385                 390                 395                 400 

Thr Leu Glu Ala Phe Val Glu Ser Gln Lys Tyr Ala Arg Glu Asp Gly 
                405                 410                 415 

Gln Gly Asp Phe Glu Lys Cys Phe Asp 
            420                 425 

 
           
             145  
             409  
             PRT  
             P. lycii  
           
            145 

Leu Pro Ile Pro Ala Gln Asn Thr Ser Asn Trp Gly Pro Tyr Asp Pro 
  1               5                  10                  15 

Phe Phe Pro Val Glu Pro Tyr Ala Ala Pro Pro Glu Gly Cys Thr Val 
             20                  25                  30 

Thr Gln Val Asn Leu Ile Gln Arg His Gly Ala Arg Trp Pro Thr Ser 
         35                  40                  45 

Gly Ala Arg Ser Arg Gln Val Ala Ala Val Ala Lys Ile Gln Met Ala 
     50                  55                  60 

Arg Pro Phe Thr Asp Pro Lys Tyr Glu Phe Leu Asn Asp Phe Val Tyr 
 65                  70                  75                  80 

Lys Phe Gly Val Ala Asp Leu Leu Pro Phe Gly Ala Asn Gln Ser His 
                 85                  90                  95 

Gln Thr Gly Thr Asp Met Tyr Thr Arg Tyr Ser Thr Leu Phe Glu Gly 
            100                 105                 110 

Gly Asp Val Pro Phe Val Arg Ala Ala Gly Asp Gln Arg Val Val Asp 
        115                 120                 125 

Ser Ser Thr Asn Trp Thr Ala Gly Phe Gly Asp Ala Ser Gly Glu Thr 
    130                 135                 140 

Val Leu Pro Thr Leu Gln Val Val Leu Gln Glu Glu Gly Asn Cys Thr 
145                 150                 155                 160 

Leu Cys Asn Asn Met Cys Pro Asn Glu Val Asp Gly Asp Glu Ser Thr 
                165                 170                 175 

Thr Trp Leu Gly Val Phe Ala Pro Asn Ile Thr Ala Arg Leu Asn Ala 
            180                 185                 190 

Ala Ala Pro Ser Ala Asn Leu Ser Asp Ser Asp Ala Leu Thr Leu Met 
        195                 200                 205 

Asp Met Cys Pro Phe Asp Thr Leu Ser Ser Gly Asn Ala Ser Pro Phe 
    210                 215                 220 

Cys Asp Leu Phe Thr Ala Glu Glu Tyr Val Ser Tyr Glu Tyr Tyr Tyr 
225                 230                 235                 240 

Asp Leu Asp Lys Tyr Tyr Gly Thr Gly Pro Gly Asn Ala Leu Gly Pro 
                245                 250                 255 

Val Gln Gly Val Gly Tyr Val Asn Glu Leu Leu Ala Arg Leu Thr Gly 
            260                 265                 270 

Gln Ala Val Arg Asp Glu Thr Gln Thr Asn Arg Thr Leu Asp Ser Asp 
        275                 280                 285 

Pro Ala Thr Phe Pro Leu Asn Arg Thr Phe Tyr Ala Asp Phe Ser His 
    290                 295                 300 

Asp Asn Thr Met Val Pro Ile Phe Ala Ala Leu Gly Leu Phe Asn Ala 
305                 310                 315                 320 

Thr Ala Leu Asp Pro Leu Lys Pro Asp Glu Asn Arg Leu Trp Val Asp 
                325                 330                 335 

Ser Lys Leu Val Pro Phe Ser Gly His Met Thr Val Glu Lys Leu Ala 
            340                 345                 350 

Cys Ser Gly Lys Glu Ala Val Arg Val Leu Val Asn Asp Ala Val Gln 
        355                 360                 365 

Pro Leu Glu Phe Cys Gly Gly Val Asp Gly Val Cys Glu Leu Ser Ala 
    370                 375                 380 

Phe Val Glu Ser Gln Thr Tyr Ala Arg Glu Asn Gly Gln Gly Asp Phe 
385                 390                 395                 400 

Ala Lys Cys Gly Phe Val Pro Ser Glu 
                405 

 
           
             146  
             440  
             PRT  
             A. terreus 9a1  
           
            146 

Lys His Ser Asp Cys Asn Ser Val Asp His Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Glu Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr His Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Ser Ala 
 65                  70                  75                  80 

Thr Ala Phe Pro Gly Lys Tyr Ala Phe Leu Gln Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Asp Ser Glu Glu Leu Thr Pro Phe Gly Arg Asn Gln Leu Arg Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Glu Arg Tyr Asn Ala Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Thr Asp Ala Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Thr Ala Arg Gln Asp Asp His His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Ala Ile Pro Glu 
                165                 170                 175 

Gly Ser Ala Tyr Asn Asn Thr Leu Glu His Ser Leu Cys Thr Ala Phe 
            180                 185                 190 

Glu Ser Ser Thr Val Gly Asp Asp Ala Val Ala Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Gln Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Thr Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Thr Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Met Ala Arg Leu Thr Arg Ala 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Ala Pro Leu Ser Gln Thr Ser Val Glu Ser Val Ser Gln Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Pro Thr Asp Lys Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Ala Phe Val Ala Gly Leu Ser Phe Ala Gln Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Asp Cys Phe 
        435                 440 

 
           
             147  
             440  
             PRT  
             A. terreus cbs  
           
            147 

Asn His Ser Asp Cys Thr Ser Val Asp Arg Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Glu Leu Ser His Lys Trp Gly Leu Tyr Ala Pro Tyr Phe Ser Leu Gln 
             20                  25                  30 

Asp Glu Ser Pro Phe Pro Leu Asp Val Pro Asp Asp Cys His Ile Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ala Arg His Gly Ala Arg Ser Pro Thr Asp Ser 
     50                  55                  60 

Lys Thr Lys Ala Tyr Ala Ala Thr Ile Ala Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ala Leu Pro Gly Lys Tyr Ala Phe Leu Lys Ser Tyr Asn Tyr Ser 
                 85                  90                  95 

Met Gly Ser Glu Asn Leu Thr Pro Phe Gly Arg Asn Gln Leu Gln Asp 
            100                 105                 110 

Leu Gly Ala Gln Phe Tyr Arg Arg Tyr Asp Thr Leu Thr Arg His Ile 
        115                 120                 125 

Asn Pro Phe Val Arg Ala Ala Asp Ser Ser Arg Val His Glu Ser Ala 
    130                 135                 140 

Glu Lys Phe Val Glu Gly Phe Gln Asn Ala Arg Gln Gly Asp Pro His 
145                 150                 155                 160 

Ala Asn Pro His Gln Pro Ser Pro Arg Val Asp Val Val Ile Pro Glu 
                165                 170                 175 

Gly Thr Ala Tyr Asn Asn Thr Leu Glu His Ser Ile Cys Thr Ala Phe 
            180                 185                 190 

Glu Ala Ser Thr Val Gly Asp Ala Ala Ala Asp Asn Phe Thr Ala Val 
        195                 200                 205 

Phe Ala Pro Ala Ile Ala Lys Arg Leu Glu Ala Asp Leu Pro Gly Val 
    210                 215                 220 

Gln Leu Ser Ala Asp Asp Val Val Asn Leu Met Ala Met Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Val Ser Leu Thr Asp Asp Ala His Thr Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr Ala Ala Glu Trp Thr Gln Tyr Asn Tyr Leu Leu Ser 
            260                 265                 270 

Leu Asp Lys Tyr Tyr Gly Tyr Gly Gly Gly Asn Pro Leu Gly Pro Val 
        275                 280                 285 

Gln Gly Val Gly Trp Ala Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser 
    290                 295                 300 

Pro Val His Asp His Thr Cys Val Asn Asn Thr Leu Asp Ala Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Ser Asn Leu Val Ser Ile Phe Trp Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Gln Thr Thr Val Glu Asp Ile Thr Arg Thr Asp Gly 
        355                 360                 365 

Tyr Ala Ala Ala Trp Thr Val Pro Phe Ala Ala Arg Ala Tyr Ile Glu 
    370                 375                 380 

Met Met Gln Cys Arg Ala Glu Lys Gln Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Met Pro Leu His Gly Cys Ala Val Asp Asn Leu Gly 
                405                 410                 415 

Arg Cys Lys Arg Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ala 
            420                 425                 430 

Gly Gly Asn Trp Ala Glu Cys Phe 
        435                 440 

 
           
             148  
             441  
             PRT  
             A. niger var. awamori  
           
            148 

Asn Gln Ser Thr Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Ala Ile Ser Pro Asp Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Val 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Ile Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Ala Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile His Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Ile Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Ser Ala 
        435                 440 

 
           
             149  
             441  
             PRT  
             A. niger T213  
           
            149 

Asn Gln Ser Ser Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Val Ile Ser Pro Asp Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Val 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Ile Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Ala Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile His Tyr Asp Tyr Leu Arg Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Asn Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Ile Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             150  
             441  
             PRT  
             A. niger NRRL3135  
           
            150 

Asn Gln Ser Ser Cys Asp Thr Val Asp Gln Gly Tyr Gln Cys Phe Ser 
  1               5                  10                  15 

Glu Thr Ser His Leu Trp Gly Gln Tyr Ala Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asn Glu Ser Val Ile Ser Pro Glu Val Pro Ala Gly Cys Arg Val Thr 
         35                  40                  45 

Phe Ala Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Asp Ser 
     50                  55                  60 

Lys Gly Lys Lys Tyr Ser Ala Leu Ile Glu Glu Ile Gln Gln Asn Ala 
 65                  70                  75                  80 

Thr Thr Phe Asp Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Ser 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Glu Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Glu Ser Leu Thr Arg Asn Ile 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Ser Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Lys Lys Phe Ile Glu Gly Phe Gln Ser Thr Lys Leu Lys Asp Pro Arg 
145                 150                 155                 160 

Ala Gln Pro Gly Gln Ser Ser Pro Lys Ile Asp Val Val Ile Ser Glu 
                165                 170                 175 

Ala Ser Ser Ser Asn Asn Thr Leu Asp Pro Gly Thr Cys Thr Val Phe 
            180                 185                 190 

Glu Asp Ser Glu Leu Ala Asp Thr Val Glu Ala Asn Phe Thr Ala Thr 
        195                 200                 205 

Phe Val Pro Ser Ile Arg Gln Arg Leu Glu Asn Asp Leu Ser Gly Val 
    210                 215                 220 

Thr Leu Thr Asp Thr Glu Val Thr Tyr Leu Met Asp Met Cys Ser Phe 
225                 230                 235                 240 

Asp Thr Ile Ser Thr Ser Thr Val Asp Thr Lys Leu Ser Pro Phe Cys 
                245                 250                 255 

Asp Leu Phe Thr His Asp Glu Trp Ile Asn Tyr Asp Tyr Leu Gln Ser 
            260                 265                 270 

Leu Lys Lys Tyr Tyr Gly His Gly Ala Gly Asn Pro Leu Gly Pro Thr 
        275                 280                 285 

Gln Gly Val Gly Tyr Ala Asn Glu Leu Ile Ala Arg Leu Thr His Ser 
    290                 295                 300 

Pro Val His Asp Asp Thr Ser Ser Asn His Thr Leu Asp Ser Ser Pro 
305                 310                 315                 320 

Ala Thr Phe Pro Leu Asn Ser Thr Leu Tyr Ala Asp Phe Ser His Asp 
                325                 330                 335 

Asn Gly Ile Ile Ser Ile Leu Phe Ala Leu Gly Leu Tyr Asn Gly Thr 
            340                 345                 350 

Lys Pro Leu Ser Thr Thr Thr Val Glu Asn Ile Thr Gln Thr Asp Gly 
        355                 360                 365 

Phe Ser Ser Ala Trp Thr Val Pro Phe Ala Ser Arg Leu Tyr Val Glu 
    370                 375                 380 

Met Met Gln Cys Gln Ala Glu Gln Glu Pro Leu Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Pro Val Asp Ala Leu Gly 
                405                 410                 415 

Arg Cys Thr Arg Asp Ser Phe Val Arg Gly Leu Ser Phe Ala Arg Ser 
            420                 425                 430 

Gly Gly Asp Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             151  
             440  
             PRT  
             A. fumigatus 32722  
           
            151 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             152  
             440  
             PRT  
             Artificial Sequence  
             
               Description of Artificial SequenceA. fumigatus 
      ATCC32722  
             
           
            152 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Gly 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             153  
             440  
             PRT  
             A. fumigatus ATCC58128  
           
            153 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Glu Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Ser Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             154  
             440  
             PRT  
             A. fumigatus ATCC26906  
           
            154 

Gly Ser Lys Ser Cys Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Ala Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr 
         35                  40                  45 

Leu Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Ala Asn Ala 
 65                  70                  75                  80 

Thr Asp Phe Lys Gly Lys Phe Ala Phe Leu Lys Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ala Phe Gly Glu Gln Gln Leu Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Phe Asn Asn Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu 
            180                 185                 190 

Ala Ser Gln Leu Gly Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Asp Ile Arg Ala Arg Ala Lys Lys His Leu Pro Gly Val Thr 
    210                 215                 220 

Leu Thr Asp Glu Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln 
                245                 250                 255 

Leu Phe Thr His Asn Glu Trp Lys Lys Tyr Asn Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Ser Met Val Ser Ile Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Arg Thr Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Val Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asn Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Gly Glu Cys Phe Ser 
        435                 440 

 
           
             155  
             440  
             PRT  
             A. fumigatus ATCC32239  
           
            155 

Gly Ser Lys Ala Cys Asp Thr Val Glu Leu Gly Tyr Gln Cys Ser Pro 
  1               5                  10                  15 

Gly Thr Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Glu 
             20                  25                  30 

Asp Glu Leu Ser Val Ser Ser Asp Leu Pro Lys Asp Cys Arg Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ala Ser 
     50                  55                  60 

Lys Ser Lys Lys Tyr Lys Lys Leu Val Thr Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Glu Phe Lys Gly Lys Phe Ala Phe Leu Glu Thr Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Pro Phe Gly Glu Gln Gln Met Val Asn 
            100                 105                 110 

Ser Gly Ile Lys Phe Tyr Gln Lys Tyr Lys Ala Leu Ala Gly Ser Val 
        115                 120                 125 

Val Pro Phe Ile Arg Ser Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Glu Lys Phe Ile Glu Gly Phe Gln Gln Ala Asn Val Ala Asp Pro Gly 
145                 150                 155                 160 

Ala Thr Asn Arg Ala Ala Pro Val Ile Ser Val Ile Ile Pro Glu Ser 
                165                 170                 175 

Glu Thr Tyr Asn Asn Thr Leu Asp His Ser Val Cys Thr Asn Phe Glu 
            180                 185                 190 

Ala Ser Glu Leu Gly Asp Glu Val Glu Ala Asn Phe Thr Ala Leu Phe 
        195                 200                 205 

Ala Pro Ala Ile Arg Ala Arg Ile Glu Lys His Leu Pro Gly Val Gln 
    210                 215                 220 

Leu Thr Asp Asp Asp Val Val Ser Leu Met Asp Met Cys Ser Phe Asp 
225                 230                 235                 240 

Thr Val Ala Arg Thr Ala Asp Ala Ser Glu Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Ile Phe Thr His Asn Glu Trp Lys Lys Tyr Asp Tyr Leu Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Asn Ser Pro 
    290                 295                 300 

Val Gln Asp His Thr Ser Thr Asn Ser Thr Leu Asp Ser Asp Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Ile Tyr Val Asp Phe Ser His Asp Asn 
                325                 330                 335 

Gly Met Ile Pro Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Glu 
            340                 345                 350 

Pro Leu Ser Gln Thr Ser Glu Glu Ser Thr Lys Glu Ser Asn Gly Tyr 
        355                 360                 365 

Ser Ala Ser Trp Ala Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr 
    370                 375                 380 

Met Gln Cys Lys Ser Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Lys Asp Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Ser Glu Gln Ser Phe Ser 
        435                 440 

 
           
             156  
             439  
             PRT  
             E. nidulans  
           
            156 

Gln Asn His Ser Cys Asn Thr Ala Asp Gly Gly Tyr Gln Cys Phe Pro 
  1               5                  10                  15 

Asn Val Ser His Val Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Ile Glu 
             20                  25                  30 

Gln Glu Ser Ala Ile Ser Glu Asp Val Pro His Gly Cys Glu Val Thr 
         35                  40                  45 

Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Glu Ser 
     50                  55                  60 

Lys Ser Lys Ala Tyr Ser Gly Leu Ile Glu Ala Ile Gln Lys Asn Ala 
 65                  70                  75                  80 

Thr Ser Phe Trp Gly Gln Tyr Ala Phe Leu Glu Ser Tyr Asn Tyr Thr 
                 85                  90                  95 

Leu Gly Ala Asp Asp Leu Thr Ile Phe Gly Glu Asn Gln Met Val Asp 
            100                 105                 110 

Ser Gly Ala Lys Phe Tyr Arg Arg Tyr Lys Asn Leu Ala Arg Lys Asn 
        115                 120                 125 

Thr Pro Phe Ile Arg Ala Ser Gly Ser Asp Arg Val Val Ala Ser Ala 
    130                 135                 140 

Glu Lys Phe Ile Asn Gly Phe Arg Lys Ala Gln Leu His Asp His Gly 
145                 150                 155                 160 

Ser Gly Gln Ala Thr Pro Val Val Asn Val Ile Ile Pro Glu Ile Asp 
                165                 170                 175 

Gly Phe Asn Asn Thr Leu Asp His Ser Thr Cys Val Ser Phe Glu Asn 
            180                 185                 190 

Asp Glu Arg Ala Asp Glu Ile Glu Ala Asn Phe Thr Ala Ile Met Gly 
        195                 200                 205 

Pro Pro Ile Arg Lys Arg Leu Glu Asn Asp Leu Pro Gly Ile Lys Leu 
    210                 215                 220 

Thr Asn Glu Asn Val Ile Tyr Leu Met Asp Met Cys Ser Phe Asp Thr 
225                 230                 235                 240 

Met Ala Arg Thr Ala His Gly Thr Glu Leu Ser Pro Phe Cys Ala Ile 
                245                 250                 255 

Phe Thr Glu Lys Glu Trp Leu Gln Tyr Asp Tyr Leu Gln Ser Leu Ser 
            260                 265                 270 

Lys Tyr Tyr Gly Tyr Gly Ala Gly Ser Pro Leu Gly Pro Ala Gln Gly 
        275                 280                 285 

Ile Gly Phe Thr Asn Glu Leu Ile Ala Arg Leu Thr Gln Ser Pro Val 
    290                 295                 300 

Gln Asp Asn Thr Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr 
305                 310                 315                 320 

Phe Pro Leu Asp Arg Lys Leu Tyr Ala Asp Phe Ser His Asp Asn Ser 
                325                 330                 335 

Met Ile Ser Ile Phe Phe Ala Met Gly Leu Tyr Asn Gly Thr Gln Pro 
            340                 345                 350 

Leu Ser Met Asp Ser Val Glu Ser Ile Gln Glu Met Asp Gly Tyr Ala 
        355                 360                 365 

Ala Ser Trp Thr Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Leu Met 
    370                 375                 380 

Gln Cys Glu Lys Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg 
385                 390                 395                 400 

Val Val Pro Leu His Gly Cys Ala Val Asp Lys Phe Gly Arg Cys Thr 
                405                 410                 415 

Leu Asp Asp Trp Val Glu Gly Leu Asn Phe Ala Arg Ser Gly Gly Asn 
            420                 425                 430 

Trp Lys Thr Cys Phe Thr Leu 
        435 

 
           
             157  
             443  
             PRT  
             T. thermophilus  
           
            157 

Asp Ser His Ser Cys Asn Thr Val Glu Gly Gly Tyr Gln Cys Arg Pro 
  1               5                  10                  15 

Glu Ile Ser His Ser Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala 
             20                  25                  30 

Asp Gln Ser Glu Ile Ser Pro Asp Val Pro Gln Asn Cys Lys Ile Thr 
         35                  40                  45 

Phe Val Gln Leu Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser 
     50                  55                  60 

Lys Thr Glu Leu Tyr Ser Gln Leu Ile Ser Arg Ile Gln Lys Thr Ala 
 65                  70                  75                  80 

Thr Ala Tyr Lys Gly Tyr Tyr Ala Phe Leu Lys Asp Tyr Arg Tyr Gln 
                 85                  90                  95 

Leu Gly Ala Asn Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Ile Gln 
            100                 105                 110 

Leu Gly Ile Lys Phe Tyr Asn His Tyr Lys Ser Leu Ala Arg Asn Ala 
        115                 120                 125 

Val Pro Phe Val Arg Cys Ser Gly Ser Asp Arg Val Ile Ala Ser Gly 
    130                 135                 140 

Arg Leu Phe Ile Glu Gly Phe Gln Ser Ala Lys Val Leu Asp Pro His 
145                 150                 155                 160 

Ser Asp Lys His Asp Ala Pro Pro Thr Ile Asn Val Ile Ile Glu Glu 
                165                 170                 175 

Gly Pro Ser Tyr Asn Asn Thr Leu Asp Thr Gly Ser Cys Pro Val Phe 
            180                 185                 190 

Glu Asp Ser Ser Gly Gly His Asp Ala Gln Glu Lys Phe Ala Lys Gln 
        195                 200                 205 

Phe Ala Pro Ala Ile Leu Glu Lys Ile Lys Asp His Leu Pro Gly Val 
    210                 215                 220 

Asp Leu Ala Val Ser Asp Val Pro Tyr Leu Met Asp Leu Cys Pro Phe 
225                 230                 235                 240 

Glu Thr Leu Ala Arg Asn His Thr Asp Thr Leu Ser Pro Phe Cys Ala 
                245                 250                 255 

Leu Ser Thr Gln Glu Glu Trp Gln Ala Tyr Asp Tyr Tyr Gln Ser Leu 
            260                 265                 270 

Gly Lys Tyr Tyr Gly Asn Gly Gly Gly Asn Pro Leu Gly Pro Ala Gln 
        275                 280                 285 

Gly Val Gly Phe Val Asn Glu Leu Ile Ala Arg Met Thr His Ser Pro 
    290                 295                 300 

Val Gln Asp Tyr Thr Thr Val Asn His Thr Leu Asp Ser Asn Pro Ala 
305                 310                 315                 320 

Thr Phe Pro Leu Asn Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn 
                325                 330                 335 

Thr Met Thr Ser Ile Phe Ala Ala Leu Gly Leu Tyr Asn Gly Thr Ala 
            340                 345                 350 

Lys Leu Ser Thr Thr Glu Ile Lys Ser Ile Glu Glu Thr Asp Gly Tyr 
        355                 360                 365 

Ser Ala Ala Trp Thr Val Pro Phe Gly Gly Arg Ala Tyr Ile Glu Met 
    370                 375                 380 

Met Gln Cys Asp Asp Ser Asp Glu Pro Val Val Arg Val Leu Val Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Glu Val Asp Ser Leu Gly Arg 
                405                 410                 415 

Cys Lys Arg Asp Asp Phe Val Arg Gly Leu Ser Phe Ala Arg Gln Gly 
            420                 425                 430 

Gly Asn Trp Glu Gly Cys Tyr Ala Ala Ser Glu 
        435                 440 

 
           
             158  
             440  
             PRT  
             T. lanuginosa  
           
            158 

Asn Val Asp Ile Ala Arg His Trp Gly Gln Tyr Ser Pro Phe Phe Ser 
  1               5                  10                  15 

Leu Ala Glu Val Ser Glu Ile Ser Pro Ala Val Pro Lys Gly Cys Arg 
             20                  25                  30 

Val Glu Phe Val Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr 
         35                  40                  45 

Ala His Lys Ser Glu Val Tyr Ala Glu Leu Leu Gln Arg Ile Gln Asp 
     50                  55                  60 

Thr Ala Thr Glu Phe Lys Gly Asp Phe Ala Phe Leu Arg Asp Tyr Ala 
 65                  70                  75                  80 

Tyr His Leu Gly Ala Asp Asn Leu Thr Arg Phe Gly Glu Glu Gln Met 
                 85                  90                  95 

Met Glu Ser Gly Arg Gln Phe Tyr His Arg Tyr Arg Glu Gln Ala Arg 
            100                 105                 110 

Glu Ile Val Pro Phe Val Arg Ala Ala Gly Ser Ala Arg Val Ile Ala 
        115                 120                 125 

Ser Ala Glu Phe Phe Asn Arg Gly Phe Gln Asp Ala Lys Asp Arg Asp 
    130                 135                 140 

Pro Arg Ser Asn Lys Asp Gln Ala Glu Pro Val Ile Asn Val Ile Ile 
145                 150                 155                 160 

Ser Glu Glu Thr Gly Ser Asn Asn Thr Leu Asp Gly Leu Thr Cys Pro 
                165                 170                 175 

Ala Ala Glu Glu Ala Pro Asp Pro Thr Gln Pro Ala Glu Phe Leu Gln 
            180                 185                 190 

Val Phe Gly Pro Arg Val Leu Lys Lys Ile Thr Lys His Met Pro Gly 
        195                 200                 205 

Val Asn Leu Thr Leu Glu Asp Val Pro Leu Phe Met Asp Leu Cys Pro 
    210                 215                 220 

Phe Asp Thr Val Gly Ser Asp Pro Val Leu Phe Pro Arg Gln Leu Ser 
225                 230                 235                 240 

Pro Phe Cys His Leu Phe Thr Ala Asp Asp Trp Met Ala Tyr Asp Tyr 
                245                 250                 255 

Tyr Tyr Thr Leu Asp Lys Tyr Tyr Ser His Gly Gly Gly Ser Ala Phe 
            260                 265                 270 

Gly Pro Ser Arg Gly Val Gly Phe Val Asn Glu Leu Ile Ala Arg Met 
        275                 280                 285 

Thr Gly Asn Leu Pro Val Lys Asp His Thr Thr Val Asn His Thr Leu 
    290                 295                 300 

Asp Asp Asn Pro Glu Thr Phe Pro Leu Asp Ala Val Leu Tyr Ala Asp 
305                 310                 315                 320 

Phe Ser His Asp Asn Thr Met Thr Gly Ile Phe Ser Ala Met Gly Leu 
                325                 330                 335 

Tyr Asn Gly Thr Lys Pro Leu Ser Thr Ser Lys Ile Gln Pro Pro Thr 
            340                 345                 350 

Gly Ala Ala Ala Asp Gly Tyr Ala Ala Ser Trp Thr Val Pro Phe Ala 
        355                 360                 365 

Ala Arg Ala Tyr Val Glu Leu Leu Arg Cys Glu Thr Glu Thr Ser Ser 
    370                 375                 380 

Glu Glu Glu Glu Glu Gly Glu Asp Glu Pro Phe Val Arg Val Leu Val 
385                 390                 395                 400 

Asn Asp Arg Val Val Pro Leu His Gly Cys Arg Val Asp Arg Trp Gly 
                405                 410                 415 

Arg Cys Arg Arg Asp Glu Trp Ile Lys Gly Leu Thr Phe Ala Arg Gln 
            420                 425                 430 

Gly Gly His Trp Asp Arg Cys Phe 
        435                 440 

 
           
             159  
             466  
             PRT  
             M. thermophila  
           
            159 

Glu Ser Arg Pro Cys Asp Thr Pro Asp Leu Gly Phe Gln Cys Gly Thr 
  1               5                  10                  15 

Ala Ile Ser His Phe Trp Gly Gln Tyr Ser Pro Tyr Phe Ser Val Pro 
             20                  25                  30 

Ser Glu Leu Asp Ala Ser Ile Pro Asp Asp Cys Glu Val Thr Phe Ala 
         35                  40                  45 

Gln Val Leu Ser Arg His Gly Ala Arg Ala Pro Thr Leu Lys Arg Ala 
     50                  55                  60 

Ala Ser Tyr Val Asp Leu Ile Asp Arg Ile His His Gly Ala Ile Ser 
 65                  70                  75                  80 

Tyr Gly Pro Gly Tyr Glu Phe Leu Arg Thr Tyr Asp Tyr Thr Leu Gly 
                 85                  90                  95 

Ala Asp Glu Leu Thr Arg Thr Gly Gln Gln Gln Met Val Asn Ser Gly 
            100                 105                 110 

Ile Lys Phe Tyr Arg Arg Tyr Arg Ala Leu Ala Arg Lys Ser Ile Pro 
        115                 120                 125 

Phe Val Arg Thr Ala Gly Gln Asp Arg Val Val His Ser Ala Glu Asn 
    130                 135                 140 

Phe Thr Gln Gly Phe His Ser Ala Leu Leu Ala Asp Arg Gly Ser Thr 
145                 150                 155                 160 

Val Arg Pro Thr Leu Pro Tyr Asp Met Val Val Ile Pro Glu Thr Ala 
                165                 170                 175 

Gly Ala Asn Asn Thr Leu His Asn Asp Leu Cys Thr Ala Phe Glu Glu 
            180                 185                 190 

Gly Pro Tyr Ser Thr Ile Gly Asp Asp Ala Gln Asp Thr Tyr Leu Ser 
        195                 200                 205 

Thr Phe Ala Gly Pro Ile Thr Ala Arg Val Asn Ala Asn Leu Pro Gly 
    210                 215                 220 

Ala Asn Leu Thr Asp Ala Asp Thr Val Ala Leu Met Asp Leu Cys Pro 
225                 230                 235                 240 

Phe Glu Thr Val Ala Ser Ser Ser Ser Asp Pro Ala Thr Ala Asp Ala 
                245                 250                 255 

Gly Gly Gly Asn Gly Arg Pro Leu Ser Pro Phe Cys Arg Leu Phe Ser 
            260                 265                 270 

Glu Ser Glu Trp Arg Ala Tyr Asp Tyr Leu Gln Ser Val Gly Lys Trp 
        275                 280                 285 

Tyr Gly Tyr Gly Pro Gly Asn Pro Leu Gly Pro Thr Gln Gly Val Gly 
    290                 295                 300 

Phe Val Asn Glu Leu Leu Ala Arg Leu Ala Gly Val Pro Val Arg Asp 
305                 310                 315                 320 

Gly Thr Ser Thr Asn Arg Thr Leu Asp Gly Asp Pro Arg Thr Phe Pro 
                325                 330                 335 

Leu Gly Arg Pro Leu Tyr Ala Asp Phe Ser His Asp Asn Asp Met Met 
            340                 345                 350 

Gly Val Leu Gly Ala Leu Gly Ala Tyr Asp Gly Val Pro Pro Leu Asp 
        355                 360                 365 

Lys Thr Ala Arg Arg Asp Pro Glu Glu Leu Gly Gly Tyr Ala Ala Ser 
    370                 375                 380 

Trp Ala Val Pro Phe Ala Ala Arg Ile Tyr Val Glu Lys Met Arg Cys 
385                 390                 395                 400 

Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Gly Arg Gln Glu Lys 
                405                 410                 415 

Asp Glu Glu Met Val Arg Val Leu Val Asn Asp Arg Val Met Thr Leu 
            420                 425                 430 

Lys Gly Cys Gly Ala Asp Glu Arg Gly Met Cys Thr Leu Glu Arg Phe 
        435                 440                 445 

Ile Glu Ser Met Ala Phe Ala Arg Gly Asn Gly Lys Trp Asp Leu Cys 
    450                 455                 460 

Phe Ala 
465 

 
           
             160  
             440  
             PRT  
             Consensus Seq. 11  
           
            160 

Asn Ser His Ser Cys Asp Thr Val Asp Gly Tyr Gln Cys Pro Glu Ile 
  1               5                  10                  15 

Ser His Leu Trp Gly Gln Tyr Ser Pro Phe Phe Ser Leu Ala Asp Glu 
             20                  25                  30 

Ser Ala Ile Ser Pro Asp Val Pro Lys Gly Cys Arg Val Thr Phe Val 
         35                  40                  45 

Gln Val Leu Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys Ser 
     50                  55                  60 

Lys Lys Tyr Ser Ala Leu Ile Glu Arg Ile Gln Lys Asn Ala Thr Phe 
 65                  70                  75                  80 

Lys Gly Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala 
                 85                  90                  95 

Asp Asp Leu Thr Pro Phe Gly Glu Asn Gln Met Val Asn Ser Gly Ile 
            100                 105                 110 

Lys Phe Tyr Arg Arg Tyr Lys Ala Leu Ala Arg Asn Ile Val Pro Phe 
        115                 120                 125 

Val Arg Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe 
    130                 135                 140 

Ile Glu Gly Phe Gln Ser Ala Lys Leu Ala Asp Pro Ala His Gln Ala 
145                 150                 155                 160 

Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ser Gly Tyr Asn Asn 
                165                 170                 175 

Thr Leu Asp His Gly Leu Cys Thr Ala Phe Glu Asp Ser Thr Ser Glu 
            180                 185                 190 

Gln Leu Gly Asp Asp Ala Glu Ala Asn Phe Thr Ala Val Phe Ala Pro 
        195                 200                 205 

Pro Ile Arg Ala Arg Leu Glu Ala Leu Pro Gly Val Asn Leu Thr Asp 
    210                 215                 220 

Glu Asp Val Val Asn Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala 
225                 230                 235                 240 

Arg Thr Ser Asp Ala Thr Gln Leu Ser Pro Phe Cys Asp Leu Phe Thr 
                245                 250                 255 

Ala Asp Glu Trp Gln Tyr Asp Tyr Leu Gln Ser Leu Lys Tyr Tyr Gly 
            260                 265                 270 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Asn 
        275                 280                 285 

Glu Leu Ile Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr Ser 
    290                 295                 300 

Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn Ala 
305                 310                 315                 320 

Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Thr Met Val Ser Ile Phe 
                325                 330                 335 

Phe Ala Leu Gly Leu Tyr Asn Gly Thr Lys Pro Leu Ser Thr Thr Ser 
            340                 345                 350 

Val Glu Ser Ile Glu Thr Asp Gly Tyr Ala Ala Ser Trp Thr Val Pro 
        355                 360                 365 

Phe Ala Ala Arg Ala Tyr Val Glu Met Met Gln Cys Glu Ala Gly Gly 
    370                 375                 380 

Gly Gly Gly Glu Gly Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn 
385                 390                 395                 400 

Asp Arg Val Val Pro Leu His Gly Cys Gly Val Asp Lys Leu Gly Arg 
                405                 410                 415 

Cys Lys Leu Asp Asp Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly 
            420                 425                 430 

Gly Asn Trp Ala Glu Cys Phe Ala 
        435                 440 

 
           
             161  
             467  
             PRT  
             Artificial Sequence  
             
               Description of Artificial 
      Sequencephytase-1-thermo[8]-Q50T-K91A  
             
           
            161 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser His Leu Trp 
         35                  40                  45 

Gly Thr Tyr Ser Pro Tyr Phe Ser Leu Ala Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Asp Asp Cys Arg Val Thr Phe Val Gln Val Leu Ser 
 65                  70                  75                  80 

Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Ala Ser Lys Ala Tyr Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Thr Pro Phe Gly Glu Asn Gln Met Val Asn Ser Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Ile Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ser Gln Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ser Gly Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Thr Cys Thr Ala Phe Glu Asp Ser Glu Leu Gly 
    210                 215                 220 

Asp Asp Val Glu Ala Asn Phe Thr Ala Leu Phe Ala Pro Ala Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala Asp Leu Pro Gly Val Thr Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Tyr Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys Ala Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Ile Gln Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly 
    290                 295                 300 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Ala 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Thr Met Ile Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Lys Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ser Ala Ser Trp Thr 
385                 390                 395                 400 

Val Pro Phe Ala Ala Arg Ala Tyr Val Glu Met Met Gln Cys Gln Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Ala Glu 
    450                 455                 460 

Cys Phe Ala 
465 

 
           
             162  
             1404  
             DNA  
             Artificial Sequence  
             
               Description of Artificial 
      Sequencephytase-1-thermo[8]-Q50T-K91A  
             
           
            162 

atgggcgtgt tcgtcgtgct actgtccatt gccaccttgt tcggttccac atccggtacc     60 

gccttgggtc ctcgtggtaa ttctcactct tgtgacactg ttgacggtgg ttaccaatgt    120 

ttcccagaaa tttctcactt gtggggtacc tactctccat acttctcttt ggcagacgaa    180 

tctgctattt ctccagacgt tccagacgac tgtagagtta ctttcgttca agttttgtct    240 

agacacggtg ctagataccc aacttcttct gcgtctaagg cttactctgc tttgattgaa    300 

gctattcaaa agaacgctac tgctttcaag ggtaagtacg ctttcttgaa gacttacaac    360 

tacactttgg gtgctgacga cttgactcca ttcggtgaaa accaaatggt taactctggt    420 

attaagttct acagaagata caaggctttg gctagaaaga ttgttccatt cattagagct    480 

tctggttctg acagagttat tgcttctgct gaaaagttca ttgaaggttt ccaatctgct    540 

aagttggctg acccaggttc tcaaccacac caagcttctc cagttattaa cgtgatcatt    600 

ccagaaggat ccggttacaa caacactttg gaccacggta cttgtactgc tttcgaagac    660 

tctgaattag gtgacgacgt tgaagctaac ttcactgctt tgttcgctcc agctattaga    720 

gctagattgg aagctgactt gccaggtgtt actttgactg acgaagacgt tgtttacttg    780 

atggacatgt gtccattcga cactgtcgct agaacttctg acgctactga attgtctcca    840 

ttctgtgctt tgttcactca cgacgaatgg atccaatacg actacttgca aagcttgggt    900 

aagtactacg gttacggtgc tggtaaccca ttgggtccag ctcaaggtgt tggtttcgct    960 

aacgaattga ttgctagatt gactcactct ccagttcaag accacacttc tactaaccac   1020 

actttggact ctaacccagc tactttccca ttgaacgcta ctttgtacgc tgacttctct   1080 

cacgacaaca ctatgatatc tattttcttc gctttgggtt tgtacaacgg taccaagcca   1140 

ttgtctacta cttctgttga atctattgaa gaaactgacg gttactctgc ttcttggact   1200 

gttccattcg ctgctagagc ttacgttgaa atgatgcaat gtcaagctga aaaggaacca   1260 

ttggttagag ttttggttaa cgacagagtt gttccattgc acggttgtgc tgttgacaag   1320 

ttgggtagat gtaagagaga cgacttcgtt gaaggtttgt ctttcgctag atctggtggt   1380 

aactgggctg aatgtttcgc ttaa                                          1404 

 
           
             163  
             467  
             PRT  
             Artificial Sequence  
             
               Description of Artificial Sequenceconsensus 
      phytase-10-thermo[3]-Q50T-K91A  
             
           
            163 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser His Leu Trp 
         35                  40                  45 

Gly Thr Tyr Ser Pro Phe Phe Ser Leu Ala Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Lys Gly Cys Arg Val Thr Phe Val Gln Val Leu Ser 
 65                  70                  75                  80 

Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Ala Ser Lys Ala Tyr Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Thr Pro Phe Gly Glu Gln Gln Met Val Asn Ser Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Ile Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Asp Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ala Asn Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ala Gly Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Leu Cys Thr Ala Phe Glu Glu Ser Glu Leu Gly 
    210                 215                 220 

Asp Asp Val Glu Ala Asn Phe Thr Ala Val Phe Ala Pro Pro Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala His Leu Pro Gly Val Asn Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Asn Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Gln Leu Ser Pro Phe Cys Asp Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Ile Gln Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly 
    290                 295                 300 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Val 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Thr Met Val Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Lys Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ser Ala Ser Trp Thr 
385                 390                 395                 400 

Val Pro Phe Ala Ala Arg Ala Tyr Val Glu Met Met Gln Cys Glu Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Gly Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Glu Glu 
    450                 455                 460 

Cys Phe Ala 
465 

 
           
             164  
             1404  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequenceconsensus 
      phytase-10-thermo[3]-Q50T-K91A  
             
           
            164 

atgggcgtgt tcgtcgtgct actgtccatt gccaccttgt tcggttccac atccggtacc     60 

gccttgggtc ctcgtggtaa ctctcactct tgtgacactg ttgacggtgg ttaccaatgt    120 

ttcccagaaa tttctcactt gtggggtaca tactctccat tcttctcttt ggctgacgaa    180 

tctgctattt ctccagacgt tccaaagggt tgtagagtta ctttcgttca agttttgtct    240 

agacacggtg ctagataccc aacttcttct gcgtctaagg cgtactctgc tttgattgaa    300 

gctattcaaa agaacgctac tgctttcaag ggtaagtacg ctttcttgaa gacttacaac    360 

tacactttgg gtgctgacga cttgactcca ttcggtgaac aacaaatggt taactctggt    420 

attaagttct acagaagata caaggctttg gctagaaaga ttgttccatt cattagagct    480 

tctggttctg acagagttat tgcttctgct gaaaagttca ttgaaggttt ccaatctgct    540 

aagttggctg acccaggtgc taacccacac caagcttctc cagttattaa cgttattatt    600 

ccagaaggtg ctggttacaa caacactttg gaccacggtt tgtgtactgc tttcgaagaa    660 

tctgaattgg gtgacgacgt tgaagctaac ttcactgctg ttttcgctcc accaattaga    720 

gctagattgg aagctcactt gccaggtgtt aacttgactg acgaagacgt tgttaacttg    780 

atggacatgt gtccattcga cactgttgct agaacttctg acgctactca attgtctcca    840 

ttctgtgact tgttcactca cgacgaatgg attcaatacg actacttgca atctttgggt    900 

aagtactacg gttacggtgc tggtaaccca ttgggtccag ctcaaggtgt tggtttcgtt    960 

aacgaattga ttgctagatt gactcactct ccagttcaag accacacttc tactaaccac   1020 

actttggact ctaacccagc tactttccca ttgaacgcta ctttgtacgc tgacttctct   1080 

cacgacaaca ctatggtttc tattttcttc gctttgggtt tgtacaacgg tactaagcca   1140 

ttgtctacta cttctgttga atctattgaa gaaactgacg gttactctgc ttcttggact   1200 

gttccattcg ctgctagagc ttacgttgaa atgatgcaat gtgaagctga aaaggaacca   1260 

ttggttagag ttttggttaa cgacagagtt gttccattgc acggttgtgg tgttgacaag   1320 

ttgggtagat gtaagagaga cgacttcgtt gaaggtttgt ctttcgctag atctggtggt   1380 

aactgggaag aatgtttcgc ttaa                                          1404 

 
           
             165  
             467  
             PRT  
             A. fumigatus ATCC13073  
           
            165 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn His Ser Lys Ser Cys 
             20                  25                  30 

Asp Thr Val Asp Leu Gly Tyr Gln Cys Ser Pro Ala Thr Ser His Leu 
         35                  40                  45 

Trp Gly Thr Tyr Ser Pro Tyr Phe Ser Leu Glu Asp Glu Leu Ser Val 
     50                  55                  60 

Ser Ser Lys Leu Pro Lys Asp Cys Arg Ile Thr Leu Val Gln Val Leu 
 65                  70                  75                  80 

Ser Arg His Gly Ala Arg Tyr Pro Thr Ser Ser Lys Ser Lys Lys Tyr 
                 85                  90                  95 

Lys Lys Leu Ile Thr Ala Ile Gln Ala Asn Ala Thr Asp Phe Lys Gly 
            100                 105                 110 

Lys Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp 
        115                 120                 125 

Leu Thr Pro Phe Gly Glu Gln Gln Leu Val Asn Ser Gly Ile Lys Phe 
    130                 135                 140 

Tyr Gln Arg Tyr Lys Ala Leu Ala Arg Ser Val Val Pro Phe Ile Arg 
145                 150                 155                 160 

Ala Ser Gly Ser Asp Arg Val Ile Ala Ser Gly Glu Lys Phe Ile Glu 
                165                 170                 175 

Gly Phe Gln Gln Ala Lys Leu Ala Asp Pro Gly Ala Thr Asn Arg Ala 
            180                 185                 190 

Ala Pro Ala Ile Ser Val Ile Ile Pro Glu Ser Glu Thr Phe Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Val Cys Thr Lys Phe Glu Ala Ser Gln Leu Gly 
    210                 215                 220 

Asp Glu Val Ala Ala Asn Phe Thr Ala Leu Phe Ala Pro Asp Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Lys His Leu Pro Gly Val Thr Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Ser Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Ser Gln Leu Ser Pro Phe Cys Gln Leu Phe Thr His Asn 
        275                 280                 285 

Glu Trp Lys Lys Tyr Asp Tyr Leu Gln Ser Leu Gly Lys Tyr Tyr Gly 
    290                 295                 300 

Tyr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Ile Gly Phe Thr 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn Ser Thr Leu Val Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Met Tyr Val Asp Phe Ser His Asp Asn Ser Met Val Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Glu Pro Leu Ser Arg Thr 
    370                 375                 380 

Ser Val Glu Ser Ala Lys Glu Leu Asp Gly Tyr Ser Ala Ser Trp Val 
385                 390                 395                 400 

Val Pro Phe Gly Ala Arg Ala Tyr Phe Glu Thr Met Gln Cys Lys Ser 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Ala Leu Ile Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Asp Val Asp Lys Leu Gly Arg Cys Lys Leu Asn Asp 
        435                 440                 445 

Phe Val Lys Gly Leu Ser Trp Ala Arg Ser Gly Gly Asn Trp Gly Glu 
    450                 455                 460 

Cys Phe Ser 
465 

 
           
             166  
             1404  
             DNA  
             A. fumigatus ATCC13073  
           
            166 

atgggggttt tcgtcgttct attatctatc gcgactctgt tcggcagcac atcgggcact     60 

gcgctgggcc cccgtggaaa tcactccaag tcctgcgata cggtagacct agggtaccag    120 

tgctcccctg cgacttctca tctatggggc acgtactcgc catacttttc gctcgaggac    180 

gagctgtccg tgtcgagtaa gcttcccaag gattgccgga tcaccttggt acaggtgcta    240 

tcgcgccatg gagcgcggta cccaaccagc tccaagagca aaaagtataa gaagcttatt    300 

acggcgatcc aggccaatgc caccgacttc aagggcaagt acgccttttt gaagacgtac    360 

aactatactc tgggtgcgga tgacctcact ccctttgggg agcagcagct ggtgaactcg    420 

ggcatcaagt tctaccagag gtacaaggct ctggcgcgca gtgtggtgcc gtttattcgc    480 

gcctcaggct cggaccgggt tattgcttcg ggagagaagt tcatcgaggg gttccagcag    540 

gcgaagctgg ctgatcctgg cgcgacgaac cgcgccgctc cggcgattag tgtgattatt    600 

ccggagagcg agacgttcaa caatacgctg gaccacggtg tgtgcacgaa gtttgaggcg    660 

agtcagctgg gagatgaggt tgcggccaat ttcactgcgc tctttgcacc cgacatccga    720 

gctcgcctcg agaagcatct tcctggcgtg acgctgacag acgaggacgt tgtcagtcta    780 

atggacatgt gtccgtttga tacggtagcg cgcaccagcg acgcaagtca gctgtcaccg    840 

ttctgtcaac tcttcactca caatgagtgg aagaagtacg actaccttca gtccttgggc    900 

aagtactacg gctacggcgc aggcaaccct ctgggaccgg ctcaggggat agggttcacc    960 

aacgagctga ttgcccggtt gacgcgttcg ccagtgcagg accacaccag cactaactcg   1020 

actctagtct ccaacccggc caccttcccg ttgaacgcta ccatgtacgt cgacttttca   1080 

cacgacaaca gcatggtttc catcttcttt gcattgggcc tgtacaacgg cactgaaccc   1140 

ttgtcccgga cctcggtgga aagcgccaag gaattggatg ggtattctgc atcctgggtg   1200 

gtgcctttcg gcgcgcgagc ctacttcgag acgatgcaat gcaagtcgga aaaggagcct   1260 

cttgttcgcg ctttgattaa tgaccgggtt gtgccactgc atggctgcga tgtggacaag   1320 

ctggggcgat gcaagctgaa tgactttgtc aagggattga gttgggccag atctgggggc   1380 

aactggggag agtgctttag ttga                                          1404 

 
           
             167  
             467  
             PRT  
             consensus phytase-7  
           
            167 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser His Leu Trp 
         35                  40                  45 

Gly Gln Tyr Ser Pro Tyr Phe Ser Leu Glu Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Asp Asp Cys Arg Val Thr Phe Val Gln Val Leu Ser 
 65                  70                  75                  80 

Arg His Gly Ala Arg Tyr Pro Thr Asp Ser Lys Gly Lys Lys Tyr Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Thr Pro Phe Gly Glu Asn Gln Met Val Asn Ser Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Ile Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Ser Arg Val Ile Ala Ser Ala Glu Lys Phe Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ser Gln Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asp Val Ile Ile Ser Glu Ala Ser Ser Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp Pro Gly Thr Cys Thr Ala Phe Glu Asp Ser Glu Leu Ala 
    210                 215                 220 

Asp Thr Val Glu Ala Asn Phe Thr Ala Leu Phe Ala Pro Ala Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala Asp Leu Pro Gly Val Thr Leu Thr Asp Thr Glu 
                245                 250                 255 

Val Thr Tyr Leu Met Asp Met Cys Ser Phe Glu Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys Ala Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Arg His Tyr Asp Tyr Leu Gln Ser Leu Lys Lys Tyr Tyr Gly 
    290                 295                 300 

His Gly Ala Gly Asn Pro Leu Gly Pro Thr Gln Gly Val Gly Phe Ala 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr Arg Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Gly Ile Ile Ser Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Ala Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ser Ser Ala Trp Thr 
385                 390                 395                 400 

Val Pro Phe Ala Ser Arg Ala Tyr Val Glu Met Met Gln Cys Gln Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Ala Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Ala Glu 
    450                 455                 460 

Cys Phe Ala 
465 

 
           
             168  
             1426  
             DNA  
             consensus phytase-7  
           
            168 

tatatgaatt catgggcgtg ttcgtcgtgc tactgtccat tgccaccttg ttcggttcca     60 

catccggtac cgccttgggt cctcgtggta attctcactc ttgtgacact gttgacggtg    120 

gttaccaatg tttcccagaa atttctcact tgtggggtca atactctcca tacttctctt    180 

tggaagacga atctgctatt tctccagacg ttccagacga ctgtagagtt actttcgttc    240 

aagttttgtc tagacacggt gctagatacc caactgactc taagggtaag aagtactctg    300 

ctttgattga agctattcaa aagaacgcta ctgctttcaa gggtaagtac gctttcttga    360 

agacttacaa ctacactttg ggtgctgacg acttgactcc attcggtgaa aaccaaatgg    420 

ttaactctgg tattaagttc tacagaagat acaaggcttt ggctagaaag attgttccat    480 

tcattagagc ttctggttct tctagagtta ttgcttctgc tgaaaagttc attgaaggtt    540 

tccaatctgc taagttggct gacccaggtt ctcaaccaca ccaagcttct ccagttattg    600 

acgttattat ttctgacgct tcttcttaca acaacacttt ggacccaggt acttgtactg    660 

ctttcgaaga ctctgaattg gctgacactg ttgaagctaa cttcactgct ttgttcgctc    720 

cagctattag agctagattg gaagctgact tgccaggtgt tactttgact gacactgaag    780 

ttacttactt gatggacatg tgttctttcg aaactgttgc tagaacttct gacgctactg    840 

aattgtctcc attctgtgct ttgttcactc acgacgaatg gagacactac gactacttgc    900 

aatctttgaa gaagtactac ggtcacggtg ctggtaaccc attgggtcca actcaaggtg    960 

ttggtttcgc taacgaattg attgctagat tgactagatc tccagttcaa gaccacactt   1020 

ctactaacca cactttggac tctaacccag ctactttccc attgaacgct actttgtacg   1080 

ctgacttctc tcacgacaac ggtattattt ctattttctt cgctttgggt ttgtacaacg   1140 

gtactgctcc attgtctact acttctgttg aatctattga agaaactgac ggttactctt   1200 

ctgcttggac tgttccattc gcttctagag cttacgttga aatgatgcaa tgtcaagctg   1260 

aaaaggaacc attggttaga gttttggtta acgacagagt tgttccattg cacggttgtg   1320 

ctgttgacaa gttgggtaga tgtaagagag acgacttcgt tgaaggtttg tctttcgcta   1380 

gatctggtgg taactgggct gaatgtttcg cttaagaatt catata                  1426 

 
           
             169  
             467  
             PRT  
             Artificial Sequence  
             
               Description of Artificial Sequenceconsensus 
      phytase-12  
             
           
            169 

Met Gly Val Phe Val Val Leu Leu Ser Ile Ala Thr Leu Phe Gly Ser 
  1               5                  10                  15 

Thr Ser Gly Thr Ala Leu Gly Pro Arg Gly Asn Ser His Ser Cys Asp 
             20                  25                  30 

Thr Val Asp Gly Gly Tyr Gln Cys Phe Pro Glu Ile Ser Ser Asn Trp 
         35                  40                  45 

Ser Pro Tyr Ser Pro Tyr Phe Ser Leu Ala Asp Glu Ser Ala Ile Ser 
     50                  55                  60 

Pro Asp Val Pro Lys Gly Cys Arg Val Thr Phe Val Gln Val Leu Gln 
 65                  70                  75                  80 

Arg His Gly Ala Arg Phe Pro Thr Ser Gly Ala Ala Thr Arg Ile Ser 
                 85                  90                  95 

Ala Leu Ile Glu Ala Ile Gln Lys Asn Ala Thr Ala Phe Lys Gly Lys 
            100                 105                 110 

Tyr Ala Phe Leu Lys Thr Tyr Asn Tyr Thr Leu Gly Ala Asp Asp Leu 
        115                 120                 125 

Val Pro Phe Gly Ala Asn Gln Ser Ser Gln Ala Gly Ile Lys Phe Tyr 
    130                 135                 140 

Arg Arg Tyr Lys Ala Leu Ala Arg Lys Ile Val Pro Phe Ile Arg Ala 
145                 150                 155                 160 

Ser Gly Ser Asp Arg Val Ile Asp Ser Ala Thr Asn Trp Ile Glu Gly 
                165                 170                 175 

Phe Gln Ser Ala Lys Leu Ala Asp Pro Gly Ala Asn Pro His Gln Ala 
            180                 185                 190 

Ser Pro Val Ile Asn Val Ile Ile Pro Glu Gly Ala Gly Tyr Asn Asn 
        195                 200                 205 

Thr Leu Asp His Gly Leu Cys Thr Ala Phe Glu Glu Ser Glu Leu Gly 
    210                 215                 220 

Asp Asp Val Glu Ala Asn Phe Thr Ala Val Phe Ala Pro Pro Ile Arg 
225                 230                 235                 240 

Ala Arg Leu Glu Ala His Leu Pro Gly Val Asn Leu Thr Asp Glu Asp 
                245                 250                 255 

Val Val Asn Leu Met Asp Met Cys Pro Phe Asp Thr Val Ala Arg Thr 
            260                 265                 270 

Ser Asp Ala Thr Glu Leu Ser Pro Phe Cys Asp Leu Phe Thr His Asp 
        275                 280                 285 

Glu Trp Ile Gln Tyr Asp Tyr Leu Gly Asp Leu Asp Lys Tyr Tyr Gly 
    290                 295                 300 

Thr Gly Ala Gly Asn Pro Leu Gly Pro Ala Gln Gly Val Gly Phe Val 
305                 310                 315                 320 

Asn Glu Leu Ile Ala Arg Leu Thr His Ser Pro Val Gln Asp His Thr 
                325                 330                 335 

Ser Thr Asn His Thr Leu Asp Ser Asn Pro Ala Thr Phe Pro Leu Asn 
            340                 345                 350 

Ala Thr Leu Tyr Ala Asp Phe Ser His Asp Asn Thr Met Val Ala Ile 
        355                 360                 365 

Phe Phe Ala Leu Gly Leu Tyr Asn Gly Thr Lys Pro Leu Ser Thr Thr 
    370                 375                 380 

Ser Val Glu Ser Ile Glu Glu Thr Asp Gly Tyr Ser Ala Ser Trp Leu 
385                 390                 395                 400 

Val Pro Phe Ser Ala Arg Met Tyr Val Glu Met Met Gln Cys Glu Ala 
                405                 410                 415 

Glu Lys Glu Pro Leu Val Arg Val Leu Val Asn Asp Arg Val Val Pro 
            420                 425                 430 

Leu His Gly Cys Gly Val Asp Lys Leu Gly Arg Cys Lys Arg Asp Asp 
        435                 440                 445 

Phe Val Glu Gly Leu Ser Phe Ala Arg Ser Gly Gly Asn Trp Glu Glu 
    450                 455                 460 

Cys Phe Ala 
465