CN112980810A - 转氨酶突变体及其应用 - Google Patents

转氨酶突变体及其应用 Download PDF

Info

Publication number
CN112980810A
CN112980810A CN202110451381.5A CN202110451381A CN112980810A CN 112980810 A CN112980810 A CN 112980810A CN 202110451381 A CN202110451381 A CN 202110451381A CN 112980810 A CN112980810 A CN 112980810A
Authority
CN
China
Prior art keywords
gly
ala
val
glu
leu
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202110451381.5A
Other languages
English (en)
Other versions
CN112980810B (zh
Inventor
洪浩
詹姆斯·盖吉
肖毅
张娜
焦学成
马玉磊
牟慧艳
王祖建
孙凯华
李响
赵桐
曹珊
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Kailaiying Biotechnology Co ltd
Asymchem Laboratories Tianjin Co Ltd
Original Assignee
Asymchem Laboratories Tianjin Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Asymchem Laboratories Tianjin Co Ltd filed Critical Asymchem Laboratories Tianjin Co Ltd
Publication of CN112980810A publication Critical patent/CN112980810A/zh
Priority to JP2023547522A priority Critical patent/JP2024506887A/ja
Priority to US18/264,091 priority patent/US20240124853A1/en
Priority to EP21924110.6A priority patent/EP4273237A1/en
Priority to PCT/CN2021/104841 priority patent/WO2022166103A1/zh
Priority to KR1020237030000A priority patent/KR20230141852A/ko
Application granted granted Critical
Publication of CN112980810B publication Critical patent/CN112980810B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/001—Amines; Imines
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/10—Nitrogen as only ring hetero atom
    • C12P17/12—Nitrogen as only ring hetero atom containing a six-membered hetero ring
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/14—Nitrogen or oxygen as hetero atom and at least one other diverse hetero ring atom in the same ring
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y206/00—Transferases transferring nitrogenous groups (2.6)
    • C12Y206/01—Transaminases (2.6.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y206/00—Transferases transferring nitrogenous groups (2.6)
    • C12Y206/01—Transaminases (2.6.1)
    • C12Y206/01018—Beta-alanine-pyruvate transaminase (2.6.1.18)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Developing Agents For Electrophotography (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

本发明提供了一种转氨酶突变体及其应用。该转氨酶突变体由SEQ ID NO:2发生一种或多种氨基酸突变得到或者为以野生型CvTA转氨酶的序列SEQ ID为NO：1作为参照，第54‑63位及第84‑96位的保守氨基酸发生突变的突变体。此类突变体的催化活性相比野生型转氨酶均有不同程度的提升，因而利用这些突变体进行手性胺类化合物的合成，尤其是大位阻手性胺的合成，有利于提高生产效率，降低工业生产成本。且采用生物酶进行绿色化学合成，有利于减少工业三废。

Description

转氨酶突变体及其应用
技术领域
本发明涉及生物酶应用领域，具体而言，涉及一种转氨酶突变体及其应用。
背景技术
大位阻手性胺化合物（本申请中指潜在手性羰基旁边的基团比甲基大的一类化合物）是一类光学活性物质和功能性分子，是药物和配体合成中广泛应用的重要的手性中间体。然而，目前合成此类手性化合物的相关报道较少，而且无法取得令人满意的结果。例如，羰基一边有大位阻苯基叔丁基氨基酯的选择性为76%（Angew. Chem. Int. Ed. 2007, 46,4367）。目前对于小位阻（本申请中指潜在手性羰基旁边的基团为H或甲基的一类化合物）的不对称加氢反应，一些金属催化剂，比如，钌，铑，钯等均有报道取得较好的立体选择性，但是随着羰基临近位阻的变大，催化效果变得较差。
不对称加氢是一类对于生产精细化学品以及药物前体的技术，不过这一技术有一些缺点，比如需要使用高压氢气，对生产的安全性造成很大挑战，需要昂贵的过渡金属催化剂，这些有毒的催化剂还需要在后处理过程中小心对待并除去。而且这个过程中需要花费大量的人力物力筛选对应的配体。很多时候这个过程还存在立体选择性不高的问题（Science, 2010, 329, 305）。
转氨酶催化的不对称转氨反应为手性胺化合物的合成提供了一种经济、绿色的合成思路。不过，转氨酶结构上天然存在大小两个口袋，因而其能够转化的底物有较大的局限性，目前尚未发现具有对于大位阻的手性胺化合物具有催化活性的转氨酶。一般来说，潜在手性羰基旁边的基团如果比甲基大，转氨酶的小口袋就无法接受。因而转氨酶催化大位阻的手性胺化合物的合成是一个长久以来存在的挑战。
发明内容
本发明的主要目的在于提供一种转氨酶突变体及其应用，以解决现有技术中转氨酶难以催化合成大位阻手性胺化合物的问题。
为了实现上述目的，根据本发明的一个方面，提供了一种转氨酶突变体，该转氨酶突变体选自：（1）由SEQ ID NO: 2发生氨基酸突变得到，氨基酸突变为如下任意一种：F22A、L59A、W60A、C61A、F88A、Y89A、K90A、T107A、N151A、Y153A、F166A、E171A、K193A、V234A、I262A、I297A、K304A、F320A、T321A、E368A、L380A、F397A、R405A、F409A、D416A、S417A、C418A、S424A；或者（2）由SEQ ID NO: 2发生氨基酸突变得到，氨基酸突变为F88A以及以下位置中的任意一种或多种突变：L59、P83、F84、Y85、N86、T87、Y89、K90、T91、F409及A417；或者（3）由野生型转氨酶的保守氨基酸突变得到，以野生型CvTA转氨酶的序列SEQ ID为NO：1作为参照，保守氨基酸位于SEQ ID NO：1的第54-63位及第84-96位，保守氨基酸至少发生如下任意一种突变：L59A、W60A、C61A、F84S/C/G/L/R/V/P/A/M/E/K/W/Q/T/H/D/Y/N/I、Y85M、N86H/Q/P/D/K/Y/S/L/M/I/T/W/F/V/R/S/K/G、T87K/C/E/N/Q/H/F/R/D/I/M/W/A/P/V/S/L/G、F88A、Y89D/A/、K90A/F/G/D/C/S/A/E/L/V/I/R/T/Y/M/H/N/P/Q/W、T91M/W/V/G/C/R/A/F/I/M/E/L/S/K/H/Q/D/N/P/Y，/表示或 。
进一步地，（3）中，野生型转氨酶为与SEQ ID NO：1具有69%以上、优选75%以上，更优选80%以上，进一步优选85%以上同源性，且具有保守氨基酸的野生型转氨酶；优选地，野生型转氨酶来源于表9和表11所示的酶编号为1至54所示的来源中的任意一种。
进一步地，（3）中，保守氨基酸发生如下任一种组合突变：（a）L59A+F88A；（b）L59A+F88A+F89D；（c）L59A+F88A+F89D+N86H；（d）L59A+F88A+F89D+N86H +Y85M。
进一步地，（1）或（2）中，氨基酸突变为如下任意一种：
Figure DEST_PATH_IMAGE001
Figure 154402DEST_PATH_IMAGE002
Figure DEST_PATH_IMAGE003
Figure 192765DEST_PATH_IMAGE004
Figure DEST_PATH_IMAGE005
Figure 530206DEST_PATH_IMAGE006
Figure DEST_PATH_IMAGE007
为了实现上述目的，根据本发明的第二个方面，提供了一种DNA分子，DNA分子编码上述任一种转氨酶突变体。
为了实现上述目的，根据本发明的第三个方面，提供了一种重组质粒，重组质粒连接有上述DNA分子。
为了实现上述目的，根据本发明的第四个方面，提供了一种手性胺化合物的合成方法，采用上述任一种转氨酶突变体在氨基供体的作用下对式I所示酮类底物进行转氨基反应，得到手性胺化合物；
Figure 489457DEST_PATH_IMAGE008
式I，
Ar1表示取代或未取代的芳基或亚芳基，或者取代或未取代的杂亚芳基；A r2表示取代或未取代的芳基，或者环烷基；以及可选的R，R表示C原子数为1~5的取代或未取代的亚烷基，R与Ar1或A r2连接成环；其中，取代的芳基或亚芳基、取代的杂亚芳基或取代的亚烷基的取代基选自卤素、羟基或氨基，取代的杂亚芳基中的杂原子选自N、O或S。
进一步地，取代的芳基或亚芳基中的取代基为卤素或-S-CH3，卤素或-S-CH3位于芳基或亚芳基的邻位、间位或对位中的任意一个或多个位置，优选地，卤素为F、Cl或Br。
进一步地，Ar1表示取代或未取代的芳基或亚芳基，Ar2表示未取代的芳基，R表示C原子数为1~5的未取代的亚烷基，且R与Ar1连接成环。
进一步地，Ar1表示未取代的杂亚芳基，Ar2表示卤素取代的芳基，R表示羟基取代的C原子数为1~5的亚烷基，且R与Ar1连接成环。
进一步地，Ar1表示卤素和氨基取代的亚芳基，Ar2为碳原子数为3~8的环烷基。
进一步地，Ar1表示未取代的环烷基或芳基，Ar2为取代或未取代的环烷基或芳基。
进一步地，Ar1表示羟基、甲基、乙基或-S-CH3取代的环烷基或芳基，Ar2为未取代的芳基或烷基。
进一步地，酮类底物为：
Figure DEST_PATH_IMAGE009
进一步地，氨基供体为异丙胺、异丙胺盐酸盐、丙氨酸、正丁胺或苯胺。
应用本发明的技术方案，通过筛选出一类对特定底物均具有一定催化活性的转氨酶，并分析此类转氨酶具有保守的氨基酸序列区域，通过对保守区域的氨基酸进行定点突变及定向筛选，从而获得了一系列催化活性比野生型有所提高的转氨酶突变体。利用这些突变体进行手性胺类化合物的合成，尤其是大位阻手性胺的合成，有利于提高生产效率，降低工业生产成本。且采用生物酶进行绿色化学合成，有利于减少工业三废。
附图说明
构成本申请的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。在附图中：
图1示出了根据本发明实施例的多序列（SEQ ID NO：1及SEQ ID NO：3至SEQ IDNO：20）比对的部分结果的示意图；
图2示出了根据本发明实施例的多序列（SEQ ID NO：20至SEQ ID NO：38）比对的部分结果的示意图；以及
图3示出了根据本发明实施例的多序列（SEQ ID NO：39至SEQ ID NO：56）比对的部分结果的示意图。
具体实施方式
需要说明的是，在不冲突的情况下，本申请中的实施例及实施例中的特征可以相互组合。下面将结合实施例来详细说明本发明。
术语解释：
大位阻手性胺化合物：本申请中指潜手性羰基旁边的基团大于甲基的酮类化合物，其中，大于甲基的基团 比如，可以是乙基、丙基、叔丁基或苯基叔丁基等。
现有技术中尚未报道过具有对于大位阻的手性胺化合物具有催化活性的转氨酶，为了改善这一状况，发明人对现有已报道的转氨酶进行了大批量筛选，筛选了上千种野生型转氨酶，结果与预期类似，大多都没有催化活性（即根本检测不到目标产物）。但仍发现了一些能够催化合成此类大位阻手性胺化合物的转氨酶（表10及表12），尽管活性（催化底物的转化率）较低（低于1%），但至少提供了潜在改进的基础。
发明人将这些具有一定催化活性的与Chromobaterium violaceum的CvTA转氨酶（野生型序列为SEQ ID NO：1）同属或不同属来源的转氨酶，以CvTA转氨酶作为氨基酸序列比对的参照序列进行了多序列比对，比对结果如图1至图3所示，发现存在多处保守氨基酸序列区域。比如，第37位的T、第41的G、第43-37位的YLWDS、第49位的G、第51-52位的KI、第54-63位的DGMAGLWCVN、第66-69位的GYGR、第75位的A、第78-79位的QM、第81-82位的EL、第84-96位的FYNTFFKTTHPAV等等。
由多序列比对结果可以看出，第54位到第63位的保守氨基酸序列区域以及第84位到第96位的保守氨基酸序列区域，两个区域在Chromobacterium属的转氨酶中高度保守，且在结构上均位于底物结合附近位置，是比较重要的氨基酸。一般而言，蛋白序列中高度保守的位点突变容易造成酶活性的损失，因此一般不选择在高度保守的位置进行突变。而此类转氨酶具有一定的转氨酶活性，或许对此类保守氨基酸序列区域的氨基酸进行某种突变后，有助于提高此类转氨酶对大位阻底物的催化活性。
为进一步验证这些保守氨基酸位点的突变对此类大位阻底物的催化活性有改善，发明人首先对来源于Chromobaterium violaceum的CvTA转氨酶（野生型序列为SEQ ID NO：1）进行了测试，并在CvTA转氨酶发生如下突变后的变体作为进化母本：T7C、S47C、F89Y、M166F、Y168A、V379A、Q380L、R416D及A417S，将该母本的氨基酸序列记为SEQ ID NO：2。
SEQ ID NO：1的序列如下（459个氨基酸）：
MQKQRTTSQWRELDAAHHLHPFTDTASLNQAGARVMTRGEGVYLWDSEGNKIIDGMAGLWCVNVGYGRKDFAEAARRQMEELPFYNTFFKTTHPAVVELSSLLAEVTPAGFDRVFYTNSGSESVDTMIRMVRRYWDVQGKPEKKTLIGRWNGYHGSTIGGASLGGMKYMHEQGDLPIPGMAHIEQPWWYKHGKDMTPDEFGVVAARWLEEKILEIGADKVAAFVGEPIQGAGGVIVPPATYWPEIERICRKYDVLLVADEVICGFGRTGEWFGHQHFGFQPDLFTAAKGLSSGYLPIGAVFVGKRVAEGLIAGGDFNHGFTYSGHPVCAAVAHANVAALRDEGIVQRVKDDIGPYMQKRWRETFSRFEHVDDVRGVGMVQAFTLVKNKAKRELFPDFGEIGTLCRDIFFRNNLIMRACGDHIVSAPPLVMTRAEVDEMLAVAERCLEEFEQTLKARGLA。
SEQ ID NO：2的序列如下：
MQKQRTCSQWRELDAAHHLHPFTDTASLNQAGARVMTRGEGVYLWDCEGNKIIDGMAGLWCVNVGYGRKDFAEAARRQMEELPFYNTFYKTTHPAVVELSSLLAEVTPAGFDRVFYTNSGSESVDTMIRMVRRYWDVQGKPEKKTLIGRWNGYHGSTIGGASLGGFKAMHEQGDLPIPGMAHIEQPWWYKHGKDMTPDEFGVVAARWLEEKILEIGADKVAAFVGEPIQGAGGVIVPPATYWPEIERICRKYDVLLVADEVICGFGRTGEWFGHQHFGFQPDLFTAAKGLSSGYLPIGAVFVGKRVAEGLIAGGDFNHGFTYSGHPVCAAVAHANVAALRDEGIVQRVKDDIGPYMQKRWRETFSRFEHVDDVRGVGMALAFTLVKNKAKRELFPDFGEIGTLCRDIFFRNNLIMDSCGDHIVSAPPLVMTRAEVDEMLAVAERCLEEFEQTLKARGLA。
进一步，以该母本为基础行进行了蛋白质工程改造，得到了一系列转氨酶突变体，结果发现这些转氨酶突变体对多种酮类底物的催化效率得到极大提升，可以用于多种手性胺化合物的合成，尤其是大位阻手性胺化合物的高效合成。
进而，发明人对上述不同来源的野生型转氨酶（即SEQ ID NO：3至SEQ ID NO：56所示）均在相同的保守位点进行了相同的突变。比如，按照在SEQ ID NO：1基础上发生L59A突变，对其他种属来源的转氨酶的对应保守位点发生相同的L到A的突变，发现对大位阻底物均有明显改善的催化活性。类似地，按照在SEQ ID NO：1基础上发生L59A+F88A突变，对其他种属来源的转氨酶的相应保守氨基酸位点进行相同的突变，同样获得了对大位阻底物的催化活性。
实施例部分将以SEQ ID NO：1为参照的第54-63位及第84-96位的保守氨基酸突变为例，详细说明不同种属来源的野生型转氨酶在与CvTA转氨酶具有相同的保守氨基酸位点区域进行相同的氨基酸突变之后，对大位阻底物的催化活性的变化。
基于此，在本申请一种优选的实施例中，上述转氨酶突变体选自：
（1）由SEQ ID NO: 2发生氨基酸突变得到，氨基酸突变为如下任意一种：F22A、L59A、W60A、C61A、F88A、Y89A、K90A、T107A、N151A、Y153A、F166A、E171A、K193A、V234A、I262A、I297A、K304A、F320A、T321A、E368A、L380A、F397A、R405A、F409A、D416A、S417A、C418A、S424A；或者
（2）由SEQ ID NO: 2发生氨基酸突变得到，氨基酸突变为F88A以及以下位置中的任意一种或多种突变：L59、P83、F84、Y85、N86、T87、Y89、K90、T91、F409及A417；或者
（3）由野生型转氨酶的保守氨基酸突变得到，以其中野生型CvTA转氨酶的序列SEQID为NO：1作为参照，保守氨基酸位于SEQ ID NO：1的第54-63位及第84-96位，保守氨基酸至少发生如下任意一种突变：L59A、W60A、C61A、F84S/C/G/L/R/V/P/A/M/E/K/W/Q/T/H/D/Y/N/I、Y85M、N86H/Q/P/D/K/Y/S/L/M/I/T/W/F/V/R/S/K/G、T87K/C/E/N/Q/H/F/R/D/I/M/W/A/P/V/S/L/G F88A、Y89D/A/、K90A/F/G/D/C/S/A/E/L/V/I/R/T/Y/M/H/N/P/Q/W、T91M/W/V/G/C/R/A/F/I/M/E/L/S/K/H/Q/D/N/P/Y，/表示或 。
在本申请一种优选的实施例中，上述野生型转氨酶为与SEQ ID NO：1具有69%以上、优选75%以上，更优选80%以上，进一步优选85%以上同源性，且具有保守氨基酸的野生型转氨酶。比如可以是70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%以上同源性的野生型转氨酶。比如，还可以是99.1%、99.2%、99.3%、99.4%、99.5%、99.6%、99.7%、99.8%甚至99.9%以上同源性的野生型转氨酶。
在本申请一种优选的实施例中，野生型转氨酶来源于酶编号为1至54所示的54种来源中的任意一种。
在本申请一种优选的实施例中，保守氨基酸发生如下任一种组合突变：（a）L59A+F88A；（b）L59A+F88A+F89D；（c）L59A+F88A+F89D+N86H；（d）L59A+F88A+F89D+N86H +Y85M。
在本申请一种优选的实施例中，转氨酶突变为表1至表9所示的转氨酶突变体。
上述各转氨酶突变体对酮类底物的催化活性相比野生型均有大幅提高，实现了利用转氨酶催化合成大位阻的手性胺化合物，不需要使用重金属催化剂，实现绿色化学合成，且使用活性提高的转氨酶进行转氨反应，容易得到高活性高选择性的一系列手性胺化合物。
在本申请一种典型的实施例中，还提供了一种DNA分子，DNA分子编码权利前述任一种转氨酶突变体。该DNA分子编码的转氨酶突变体对一系列酮类底物化合物具有较高的催化活性，有利于通过生物酶催化的方法合成此类手性胺化合物。
本发明的上述DNA分子还可以以“表达盒”的形式存在。“表达盒”是指线性或环状的核酸分子，涵盖了能够指导特定核苷酸序列在恰当宿主细胞中表达的DNA和RNA序列。一般而言，包括与目标核苷酸有效连接的启动子，其任选的是与终止信号和/或其他调控元件有效连接的。表达盒还可以包括核苷酸序列正确翻译所需的序列。编码区通常编码目标蛋白，但在正义或反义方向也编码目标功能RNA，例如反义RNA或非翻译的RNA。包含目标多核苷酸序列的表达盒可以是嵌合的，意指至少一个其组分与其至少一个其他组分是异源的。表达盒还可以是天然存在的，但以用于异源表达的有效重组形成获得的。
在本申请一种典型的实施例中，还提供了一种重组质粒，重组质粒连接有前述DNA分子。上述重组质粒中的DNA分子置于重组质粒的适当位置，使得上述DNA分子能够正确地、顺利地复制、转录或表达。
虽然本发明在限定上述DNA分子时所用限定语为“含有”，但其并不意味着可以在DNA序列的两端任意加入与其功能不相关的其他序列。本领域技术人员知晓，为了满足重组操作的要求，需要在DNA序列的两端添加合适的限制性内切酶的酶切位点，或者额外增加启动密码子、终止密码子等，因此，如果用封闭式的表述来限定将不能真实地覆盖这些情形。
本发明中所使用的术语“质粒”包括双链或单链线状或环状形式的任何质粒、粘粒、噬菌体或农杆菌二元核酸分子，优选为重组表达质粒，可以是原核表达质粒也可以是真核表达质粒，但优选原核表达质粒，在某些实施方案中，重组质粒选自pET-21b（+）、pET-22b（+）、pET-3a（+）、pET-3d（+）、pET-11a（+）、pET-12a（+）、pET-14b、pET-15b（+）、pET-16b（+）、pET-17b（+）、pET-19b（+）、pET-20b（+）、pET-21a（+）、pET-23a（+）、pET-23b（+）、pET-24a（+）、pET-25b（+）、pET-26b（+）、pET-27b（+）、pET-28a（+）、pET-29a（+）、pET-30a（+）、pET-31b（+）、pET-32a（+）、pET-35b（+）、pET-38b（+）、pET-39b（+）、pET-40b（+）、pET-41a（+）、pET-41b（+）、pET-42a（+）、pET-43a（+）、pET-43b（+）、pET-44a（+）、pET-49b（+）、pQE2、pQE9、pQE30、pQE31、pQE32、pQE40、pQE70、pQE80、pRSET-A、pRSET-B、pRSET-C、pGEX-5X-1、pGEX-6p-1、pGEX-6p-2、pBV220、pBV221、pBV222、pTrc99A、pTwin1、pEZZ18、pKK232-8、pUC-18或pUC-19。更优选，上述重组质粒是pET-22b（+）。
根据本发明一种典型的实施方式，提供一种宿主细胞，宿主细胞含有上述任一种重组质粒。适用于本发明的宿主细胞包括但不仅限于原核细胞或真核细胞。优选原核细胞为大肠杆菌BL21细胞或大肠杆菌DH5α感受态细胞，真核细胞为酵母。
在本申请一种典型的实施例中，还提供了一种手性胺化合物的合成方法，该合成方法采用上述任一种转氨酶突变体在氨基供体的作用下对式I所示酮类底物进行转氨基反应，得到手性胺化合物；
Figure 621361DEST_PATH_IMAGE010
式I，
Ar1表示取代或未取代的芳基或亚芳基，或者取代或未取代的杂亚芳基；A r2表示取代或未取代的芳基，或者环烷基；以及可选的R，R表示C原子数为1~5的取代或未取代的亚烷基，R与Ar1或A r2连接成环；其中，取代的芳基或亚芳基、取代的杂亚芳基或取代的亚烷基的取代基选自卤素、羟基或氨基，取代的杂亚芳基中的杂原子选自N、O或S。
由于本发明的上述转氨酶对上述式I所示底物具有更高的酶催化活性，因而利用本发明的转氨酶突变体制备手性胺类化合物，尤其是大位阻的手性胺类化合物，不仅能够降低生产成本，而且所获得的大位阻手性胺的纯度和ee值更高。
上述取代的芳基或亚芳基中的取代基为卤素或-S-CH3，卤素或-S-CH3位于芳基或亚芳基的邻位、间位或对位中的任意一个或多个位置，优选地，卤素为F、Cl或Br。
在一种优选的实施例中，Ar1表示取代或未取代的芳基或亚芳基，Ar2表示未取代的芳基，R表示C原子数为1~5的未取代的亚烷基，且R与Ar1连接成环。
在一种优选的实施例中，Ar1表示未取代的杂亚芳基，Ar2表示卤素取代的芳基，R表示羟基取代的C原子数为1~5的亚烷基，且R与Ar1连接成环。
在一种优选的实施例中，Ar1表示卤素和氨基取代的亚芳基，Ar2为碳原子数为3~8的环烷基。
在一种优选的实施例中，Ar1表示未取代的环烷基或芳基，Ar2为取代或未取代的环烷基或芳基。
在一种优选的实施例中，Ar1表示羟基、甲基、乙基或-S-CH3取代的环烷基或芳基，Ar2为未取代的芳基或烷基。
在一种优选的实施例中，酮类底物为：
Figure DEST_PATH_IMAGE011
在一种优选的实施例中，氨基供体为异丙胺、异丙胺盐酸盐、丙氨酸、正丁胺或苯胺。
以下将结合具体实施例对上述技术方案和技术效果进行说明。以下实施例中所用到的底物如下：
Figure 147021DEST_PATH_IMAGE012
Figure DEST_PATH_IMAGE013
Figure 288152DEST_PATH_IMAGE014
（一）第一部分 以SEQ ID NO ：2 为母本进行改造
实施例1
在母本基础上进行定向突变，具体突变位点见下表，并按照如下反应条件对突变体的催化活性进行检测：
1 mL体系包括1 mg底物1，1 mg PLP，1 mg异丙胺盐酸盐，50 mg酶粉，pH8.0 100mM 磷酸盐buffer，50oC反应40 h。
检测结果见下表：
表1：
Figure DEST_PATH_IMAGE015
Figure 582867DEST_PATH_IMAGE016
。
注：上表中，0代表转化率<1%，+代表转化率大于等于1%且小于5%，++代表转化率大于等于5%且小于等于10%。
实施例2
在实施例1的基础上进行组合突变，并按照与实施例1相同的反应条件对组合突变进行活性筛选，结果见下表。
表2：
Figure DEST_PATH_IMAGE017
Figure 918296DEST_PATH_IMAGE018
。
注：上表中，0代表转化率<1%，+代表转化率大于等于1%且小于5%，++代表转化率大于等于5%且小于10%，+++代表转化率大于等于10%且小于15%，++++代表转化率大于等于15%且小于20%。
实施例3
在实施例2的基础上进行组合突变，并按照与实施例1相同的反应条件对组合突变进行活性筛选，结果见下表。
表3：
Figure 134513DEST_PATH_IMAGE019
Figure 548177DEST_PATH_IMAGE020
。
注：0代表转化率<1%，+代表转化率大于等于1%且小于5%，++代表转化率大于等于5%且小于10%，+++代表转化率大于等于10%且小于15%，++++代表转化率大于等于15%且小于20%，+++++代表转化率大于等于20%且小于等于40%。
实施例4
在实施例3的基础上进行组合突变，并按照与实施例1相同的反应条件对组合突变进行活性筛选，结果见下表。
表4：
Figure DEST_PATH_IMAGE021
Figure 697399DEST_PATH_IMAGE022
。
注：上表中，0代表转化率<1%，+代表转化率大于等于1%且小于5%，++代表转化率大于等于5%且小于10%，+++代表转化率大于等于10%且小于15%，++++代表转化率大于等于15%且小于20%，+++++代表转化率大于等于20%且小于40%，++++++代表转化率大于等于40%。
实施例5
在实施例4的基础上进行组合突变，具体突变位点见下表，并按照如下反应条件对突变体的催化活性进行检测：
1 mL体系包括10 mg底物1，1 mg PLP，1 mg异丙胺盐酸盐，5 mg酶粉，pH8.0 100mM 磷酸盐buffer，50oC反应40 h，结果见下表。
表5：
Figure DEST_PATH_IMAGE023
Figure 436685DEST_PATH_IMAGE024
。
注：上表中，+代表转化率大于等于0%且小于20%，++代表转化率大于等于20%且小于40%。
实施例6
在实施例5的基础上进行组合突变，并按照与实施例5相同的反应条件对组合突变进行活性筛选，结果见下表。
表6：
Figure DEST_PATH_IMAGE025
Figure 936936DEST_PATH_IMAGE026
。
注：+代表转化率大于等于0%且小于20%，++代表转化率大于等于20%且小于40%，+++代表转化率大于等于40%且小于80%。
实施例7
在实施例6的基础上进行组合突变，并按照与实施例5相同的反应条件对组合突变进行活性筛选，结果见下表。
表7：
Figure DEST_PATH_IMAGE027
Figure 655756DEST_PATH_IMAGE028
。
注：上表中，+代表转化率大于等于0%且小于20%，++代表转化率大于等于20%且小于40%，+++代表转化率大于等于40%且小于80%，++++代表转化率大于等于80%。
实施例8
在实施例7的基础上进行组合突变，并按照与实施例5相同的反应条件对组合突变进行活性筛选，结果见下表。
表8：
Figure DEST_PATH_IMAGE029
。
注：上表中，+代表转化率大于等于0%且小于20%，++代表转化率大于等于20%且小于40%，+++代表转化率大于等于40%且小于80%，++++代表转化率大于等于80%。
实施例9
在实施例8的基础上进行组合突变。反应条件如下所述：
1 mL体系包括10 mg底物1，1 mg PLP，1 mg异丙胺盐酸盐，3 mg酶粉，pH8.0 100mM 磷酸盐buffer，50oC反应40 h，结果见下表。
表9：
Figure 925063DEST_PATH_IMAGE030
Figure DEST_PATH_IMAGE031
Figure 100829DEST_PATH_IMAGE032
。
注：上表中，+代表转化率大于等于0%且小于20%，++代表转化率大于等于20%且小于40%，+++代表转化率大于等于40%且小于80%，++++代表转化率大于等于80%。
实施例 10
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），10g
Figure DEST_PATH_IMAGE033
（底物1），搅拌均匀，再加入SEQ ID NO：2基础上突变的CvTA的转氨酶突变体（F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I）的酶液10 mL(0.5wt，0.5 g/mL)调pH=8.5~9.0。升温至50℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 822798DEST_PATH_IMAGE034
。
经HPLC检测，纯度＞98%，de值＞99%，收率84%。
实施例 11
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），10g
Figure DEST_PATH_IMAGE035
（底物2），搅拌均匀，再加入SEQ ID NO：2基础上突变的CvTA的转氨酶突变体（F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I）酶液10 mL(0.5wt，0.5 g/mL)调pH=8.5~9.0。升温至50℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 843843DEST_PATH_IMAGE036
。
经HPLC检测，纯度＞98%，de值＞99%，收率82%。
实施例 12
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），10g
Figure DEST_PATH_IMAGE037
（底物3），搅拌均匀，再加入SEQ ID NO：2基础上突变的CvTA的转氨酶突变体（F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I）酶液10 mL(0.5wt，0.5 g/mL)调pH=8.5~9.0。升温至50℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 203543DEST_PATH_IMAGE038
。
经HPLC检测，纯度＞98%，ee值＞99%，收率80%。
实施例 13
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），10g
Figure DEST_PATH_IMAGE039
（底物4），搅拌均匀，再加入SEQ ID NO：2基础上突变的CvTA的转氨酶突变体（F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I）酶液10 mL(0.5wt，0.5 g/mL)调pH=8.5~9.0。升温至50℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 550211DEST_PATH_IMAGE040
。
经HPLC检测，纯度＞98%，de值＞99%，收率89%。
实施例 14
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），10g
Figure DEST_PATH_IMAGE041
（底物13），搅拌均匀，再加入SEQ ID NO：2基础上突变的CvTA的转氨酶突变体（F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I）酶液10 mL(0.5 wt，0.5 g/mL)调pH=8.5~9.0。升温至50℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 759475DEST_PATH_IMAGE042
。
经HPLC检测，纯度＞98%，ee值98%，收率81%。
实施例15
采用SEQ ID NO：2基础上突变的CvTA的转氨酶突变体(F88A+L59A+Y89D+N86H+Y85M+K90G+T91M+P83S+V234A+L380A+S417I+A168E+A379L+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+M91I)的酶液，参照实施例10至实施例14的催化合成步骤，对相应的底物进行催化反应，结果分别如下：
（1）催化底物1合成目标产物的纯度＞99%，de值＞99%，收率87%；
（2）催化底物2催化合成目标产物的纯度＞98%，de值＞99%，收率84%；
（3）催化底物3合成目标产物的纯度＞98%，ee值＞99%，收率82%；
（4）催化底物4合成目标产物的纯度＞99%，de值＞99%，收率91%；
（5）催化底物13合成目标产物的纯度＞98%，ee值＞99%，收率85%。
从以上实施例的描述中，可以看出，上述实施例实现了如下技术效果：
1）使用活性提高的转氨酶进行转氨反应，可以高活性，高选择性的得到一系列手性胺化合物。
2）使用转氨酶突变体进行生物催化，不需要使用重金属催化剂，实现绿色化学。
3）该转氨酶突变体催化效率高，反应体积少，产品收率高，大大降低了三废，节约生产成本。
（二）第二部分 进一步对不同种属来源的转氨酶突变体对此类大位阻底物的催化活性进行了筛选和验证。
实施例16对其他来源的野生型转氨酶的催化活性测试
本实施例选择了与来源于Chromobaterium violaceum的野生型转氨酶（即SEQ IDNO：1）同属但不同种，且氨基酸序列与SEQ ID NO：1相比，同源性在82%以上的野生型转氨酶（具体见表10），并测试了其对底物4的催化活性，结果与CvTA野生型转氨酶类似（1%以下），具体结果见表11。
表10：来源于Chromobacterium的其它野生型转氨酶及其与SEQ ID NO：1的序列同源性。
Figure DEST_PATH_IMAGE043
。
10* ：编号10的酶不属于Chromobacterium属的，但因同源性较高，将其列在该表中。
实施例17 对目标大位阻底物催化活性突变体筛选
在本实施例中，对上表10中的野生型转氨酶在与SEQ ID NO：1对应的第59位进行L59A突变，同时还进行Y85M，N86H，F88A及F89D的任意一个或多个突变之后，对目标大位阻底物（底物4）的催化活性均有不同程度的提高（见表11）。该结果表明，与SEQ ID NO：1同源性从82%到95%不等的转氨酶，在保守氨基酸位点发生相同的氨基酸突变后，对大位阻底物的催化活性都有提升。
表11：
Figure 584212DEST_PATH_IMAGE044
。
上表中， +代表转化率在1-20%之间，++代表转化率在20%以上。
实施例18
除了Chromobacterium来源的转氨酶，本申请还测试了与Chromobateriumviolaceum有69%到87%同源性的其他来源的转氨酶。同样地，其对底物4的催化活性低于1%。然而对这些转氨酶的保守氨基酸序列区域发生如表11相同的氨基酸突变后，均获得了不同程度的活性提升。具体的信息列见表12和表13。
表12：与Chromobaterium violaceum具有更低的同源性，但来源于不同属的其他转氨酶。
Figure DEST_PATH_IMAGE045
Figure 562532DEST_PATH_IMAGE046
。
表13：
Figure DEST_PATH_IMAGE047
Figure 310127DEST_PATH_IMAGE048
。
上表中，+代表转化率在1-20%之间，++代表转化率在20%以上。
实施例 19
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5vol），24ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure DEST_PATH_IMAGE049
（底物5），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml）调pH=7.5~8.0。升温至30℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=12，再用50ml 二氯甲烷萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜35℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 6688DEST_PATH_IMAGE050
。
经HPLC检测，纯度＞98%，ee值＞99%，收率84%。
实施例20
室温向250 mL四口瓶内加入60 mL 100 mmol/L 磷酸盐缓冲液（6vol），18ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.0~8.5。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure 838377DEST_PATH_IMAGE033
（底物1），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml），调pH=8.0~8.5。调温至20℃搅拌反应过夜。反应完全后，将体系调酸至pH=2，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=13，再用50ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 671204DEST_PATH_IMAGE034
。
经HPLC检测，纯度＞98%，de值＞99%，收率89%。
实施例 21
室温向250 mL四口瓶内加入70 mL 100 mmol/L 磷酸盐缓冲液（7vol），13ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.0~7.5。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure 562937DEST_PATH_IMAGE035
（底物2），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml）升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=13，再用50ml二氯甲烷萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜35℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 215635DEST_PATH_IMAGE036
。
经HPLC检测，纯度大于97%，de值＞99%，收率86%。
实施例 22
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5vol），38ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure DEST_PATH_IMAGE051
（底物6），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml），调pH=8.5~9.0。调温至10℃搅拌反应过夜。反应完全后，将体系调酸至pH=2，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=13，再用50ml 二氯甲烷萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜35℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 647753DEST_PATH_IMAGE052
。
经HPLC检测，纯度＞95 %，de值＞99%，收率：85%。
实施例 23
室温向250 mL四口瓶内加入70 mL 100 mmol/L 磷酸盐缓冲液（7vol），17ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure 305393DEST_PATH_IMAGE037
（底物3），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml），调pH=7.5~8.0。升温至29℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=12，再用50ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 633606DEST_PATH_IMAGE038
。
经HPLC检测，纯度＞96%，ee值＞99%，收率：77%。
实施例 24
室温向500 mL四口瓶内加入140 mL 100 mmol/L 磷酸盐缓冲液（7vol），50ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.2g 磷酸吡哆醛（1wt%），2g
Figure DEST_PATH_IMAGE053
（底物7），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液20 ml（5wt，0.5 g/ml），调pH=7.5~8.0。升温至32℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用100mL 二氯甲烷萃取。水相调pH=12，再用100ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 39180DEST_PATH_IMAGE054
。
经HPLC检测，纯度＞95%，ee值＞99%，收率：80%。
实施例 25
室温向500 mL四口瓶内加入220 mL 100 mmol/L 磷酸盐缓冲液（11vol），50ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.2g 磷酸吡哆醛（1wt%），2g
Figure DEST_PATH_IMAGE055
（底物8），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液40 ml（10wt，0.5 g/ml），调pH=7.5~8.0。升温至32℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用100mL 二氯甲烷萃取。水相调pH=12，再用100ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 9410DEST_PATH_IMAGE056
。
经HPLC检测，纯度＞95%，ee值＞99%，收率：86%。
实施例 26
室温向500 mL四口瓶内加入180 mL 100 mmol/L 磷酸盐缓冲液（12vol），36ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.15g 磷酸吡哆醛（1wt%），1.5g
Figure DEST_PATH_IMAGE057
（底物9） ，搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液30 ml（1wt，0.5 g/ml），调pH=7.5~8.0。升温至35℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用100mL 二氯甲烷萃取。水相调pH=12，再用100ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 82408DEST_PATH_IMAGE058
。
经HPLC检测，纯度＞96%，ee值＞99%，收率：85%。
实施例 27
室温向500 mL四口瓶内加入180 mL 100 mmol/L 磷酸盐缓冲液（12vol），26ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.15 g 磷酸吡哆醛（1wt%），1.5g
Figure DEST_PATH_IMAGE059
（底物10），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液30 ml（10wt，0.5 g/ml），调pH=7.5~8.0。升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=1，变性蛋白。过滤后滤液用100mL 二氯甲烷萃取。水相调pH=12，再用100ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 847102DEST_PATH_IMAGE060
。
经HPLC检测，纯度＞97%，ee值＞99%，收率：79%。
实施例 28
室温向500 mL四口瓶内加入110 mL 100 mmol/L 磷酸盐缓冲液（11vol），23ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure DEST_PATH_IMAGE061
（底物11），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml），调pH=7.5~8.0。升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=2，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=12，再用50ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 241436DEST_PATH_IMAGE062
。
经HPLC检测，纯度＞95%，收率：73%。
实施例 29
室温向500 mL四口瓶内加入110 mL 100 mmol/L 磷酸盐缓冲液（11vol），23ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.1g 磷酸吡哆醛（1wt%），1g
Figure DEST_PATH_IMAGE063
（底物12），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 ml（5wt，0.5 g/ml），调pH=7.5~8.0。升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=2，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=12，再用50ml 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 749778DEST_PATH_IMAGE064
。
经HPLC检测，纯度＞95%，ee值＞99%，收率：67%。
实施例 30
室温向500 mL四口瓶内加入110 mL 100 mmol/L 磷酸盐缓冲液（11vol），23 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=7.5~8.0。再加入0.1 g 磷酸吡哆醛（1wt%），1 g
Figure 146124DEST_PATH_IMAGE039
（底物4），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 mL（5wt，0.5 g/mL），调pH=7.5~8.0。升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=2，变性蛋白。过滤后滤液用50mL 二氯甲烷萃取。水相调pH=12，再用50mL 乙酸乙酯萃取三次。合并有机相用无水硫酸镁干燥后有机相于T＜45℃，P≤-0.08Mpa条件下浓缩至无馏分。得到目标产物
Figure 816140DEST_PATH_IMAGE040
。
经HPLC检测，纯度＞95%，de值＞99%，收率：80%。
实施例 31
室温向250 mL四口瓶内加入50 mL 100 mmol/L 磷酸盐缓冲液（5 vol），20 mL5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.5~9.0。再加入0.1 g 磷酸吡哆醛（1 wt%），1g
Figure 665147DEST_PATH_IMAGE041
（底物13），搅拌均匀，再加入SEQ ID NO：1基础上的CvTA的转氨酶突变体（L59A+Y85M+N86H+F88A+F89D）酶液10 mL(0.5 wt，0.5 g/mL)调pH=8.5~9.0。升温至40℃搅拌反应过夜。反应完全后，将体系调酸至pH=2-3，变性蛋白。过滤后滤液用50 mL甲基叔丁基醚萃取。水相调pH=12，再用50 mL甲基叔丁基醚萃取2次。合并有机相用无水硫酸镁干燥后于T＜40℃，P≤-0.06Mpa条件下浓缩至无馏分。得到目标产物
Figure 446022DEST_PATH_IMAGE042
。
经HPLC检测，纯度93%，ee值98%，收率73%。
实施例 32
以底物1为例，对反应的酶量进行优化，具体操作如下：
室温向10 mL小瓶内加入1.2 mL 100 mmol/L 磷酸盐缓冲液（6vol），0.4 ml5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.0~8.5。再加入0.002g 磷酸吡哆醛（1wt%），0.02 g
Figure 962454DEST_PATH_IMAGE033
（底物1），混匀，分别加入CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液 0.04mL，0.08mL，0.12mL，0.16mL，0.2mL，0.24mL，0.28mL，0.32mL，0.36mL，0.4mL（酶液浓度为0.5 g/mL，对应质量比分别为1wt，2wt，3wt，4wt，5wt，6wt，7wt，8wt，9wt，10wt，），调pH=8.0~8.5。摇床转速170rpm，升温至30℃条件下反应过夜。转化率结果如下表所示，证明5 wt底物可转化率大于99%，酶量继续增大也并无显著效果。
表14. 酶量与转化率的关系
Figure DEST_PATH_IMAGE065
。
实施例 33
以底物1为例，对反应的底物浓度进行优化。具体操作如下：
室温向10~50 mL小瓶内（根据反应体积选择合适的摇瓶）分别加入0.5mL100mmol/L 磷酸盐缓冲液，加入0.4 ml 5mol/ L 异丙胺盐酸盐溶液（2 vol），调pH=8.0~8.5。再加入0.002g 磷酸吡哆醛（1wt%），0.02g （1vol, 0.02ml）
Figure 570415DEST_PATH_IMAGE033
，混匀，加入CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液 0.2ml（1vol, 5wt，酶液浓度为0.5 g/ml），补充100 mmol/L的磷酸盐缓冲液至反应体积分别为2mL，2.2mL，2.4mL， 3mL，3.4mL， 4 mL，对应反应体积分别为100vol，110vol，120vol，150vol，170vol，200vol。调pH=8.0~8.5。摇床转速170rpm，升温至30℃条件下反应过夜。如下表所示，证明5~10 g/L浓度下，底物可转化率大于99%。继续降低底物浓度会导致反应体积增加，三废量增加。
表15. 反应体积与转化率的关系
Figure 172297DEST_PATH_IMAGE066
。
实施例 34
以底物1为例，对反应的氨基供体进行优化，具体操作如下：
室温向10 mL小瓶内加入1.2 mL 100 mmol/L 磷酸盐缓冲液（6vol），分别使用3vol氨基供体的体系（氨基供体分别为异丙胺、异丙胺盐酸盐、丙氨酸、苯胺，正丁胺），调pH=8.0~8.5。再加入0.002g 磷酸吡哆醛（1wt%），0.02g
Figure 491283DEST_PATH_IMAGE033
，混匀，加入CvTA的突变体（L59A+Y85M+N86H+F88A+F89D）酶液 0.2ml（5wt，酶液浓度为0.5 g/ml），调pH=8.0~8.5。摇床转速170rpm，升温至29℃条件下反应过夜。转化率结果如下表所示，证明异丙胺、异丙胺盐酸盐、丙氨酸、正丁胺作为氨基供体，底物可转化率大于99%。而苯胺作为氨基供体在此反应条件下转化率仅为75%。
表16. 氨基供体对反应的影响
Figure DEST_PATH_IMAGE067
。
从以上的描述中，可以看出，本发明上述的实施例实现了如下技术效果：
1）使用不同来源的转氨酶，进行保守位点突变后的转氨酶突变体，催化多种酮类底物的生物转化反应，能够一步反应得到所需目标化合物，底物类型适用广泛。
2）使用转氨酶进行该生物转化反应选择性很高，产品ee值或de值大于99%。
3）使用前述不同来源突变的转氨酶变体进行催化反应，底物浓度可达到100g/L，大大提高了生产效率。
4）使用转氨酶进行生物催化，不需要使用重金属催化剂，实现绿色化学。
5）该转氨酶催化效率高，反应体积少，合成路线短，产品收率高，大大降低了三废，节约生产成本。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
序列表
<110> 凯莱英医药集团（天津）股份有限公司
<120> 转氨酶突变体及其应用
<130> PN154608KLY
<160> 56
<170> SIPOSequenceListing 1.0
<210> 1
<211> 459
<212> PRT
<213> Chromobacterium violaceum
<400> 1
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Ala Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
Arg Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Gln Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Ala Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Val Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Lys Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Ala Val Ala Glu Arg Cys Leu Glu Glu
435 440 445
Phe Glu Gln Thr Leu Lys Ala Arg Gly Leu Ala
450 455
<210> 2
<211> 459
<212> PRT
<213> Chromobacterium violaceum
<400> 2
Met Gln Lys Gln Arg Thr Cys Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Cys Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Ala Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Tyr Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
Arg Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Phe Lys Ala Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Gln Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Ala Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Ala Leu Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Lys Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Asp
405 410 415
Ser Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Ala Val Ala Glu Arg Cys Leu Glu Glu
435 440 445
Phe Glu Gln Thr Leu Lys Ala Arg Gly Leu Ala
450 455
<210> 3
<211> 459
<212> PRT
<213> Chromobacterium vaccinii
<400> 3
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Asp Glu Lys Ile Gln Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Ala Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys His Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Gln Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Met Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Lys Arg Glu Leu Phe Pro Asp Phe Gly Glu Val
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Cys Ser Pro Pro Leu Val Met Thr Arg
420 425 430
Thr Glu Val Asp Glu Met Leu Gly Val Ala Ala Arg Cys Leu Ala Glu
435 440 445
Phe Glu Gln Ala Leu Lys Ala Arg Gly Leu Ala
450 455
<210> 4
<211> 459
<212> PRT
<213> Chromobacterium subtsugae
<400> 4
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg His Asp Phe Ala Glu Ala Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Glu
100 105 110
Arg Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys His Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Gln Leu Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Ile Gly Gln
290 295 300
Arg Val Ala Glu Gly Leu Ile Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Thr Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Ser Asp Phe Gly Glu Val
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Arg
420 425 430
Ser Glu Val Asp Glu Met Leu Ala Val Ala Ser Arg Cys Leu Glu Glu
435 440 445
Phe Glu Arg Thr Ile Lys Ala Arg Gly Leu Val
450 455
<210> 5
<211> 459
<212> PRT
<213> Chromobacterium sphagni
<400> 5
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ser Glu Val Thr Pro Ala Gly Phe Glu
100 105 110
Arg Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Gln Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Gly Leu Gly Phe Thr Pro Asp Ile Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Ile Gly Gln
290 295 300
Arg Val Ala Asp Gly Leu Ile Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Leu Ala Leu Arg Asp Glu Gly Ile Val Ala Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Ala Glu Val Asp Glu Met Leu Gly Val Ala Ala Arg Cys Leu Thr Glu
435 440 445
Phe Glu Gln Thr Ile Lys Ala Arg Gly Leu Val
450 455
<210> 6
<211> 459
<212> PRT
<213> unclassified Chromobacterium
<400> 6
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Asp Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
Arg Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Phe Gln Gln Leu Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Ala Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ser Lys Arg Glu Leu Phe Ala Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Ala Met Leu Ala Val Ala Ala Arg Cys Leu Glu Glu
435 440 445
Phe Glu Gln Thr Leu Lys Ala Arg Gly Leu Ala
450 455
<210> 7
<211> 459
<212> PRT
<213> Chromobacterium vaccinii
<400> 7
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Ile Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Glu
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Asp Glu Lys Ile Gln Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Ala Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys His Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Gln Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Val Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Val Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Lys Arg Glu Leu Phe Pro Asp Phe Gly Glu Val
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Cys Ser Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Gly Val Ala Ala Arg Cys Leu Ala Glu
435 440 445
Phe Glu Gln Ala Leu Lys Ala Arg Gly Leu Ala
450 455
<210> 8
<211> 459
<212> PRT
<213> Chromobacterium sp. 257-1
<400> 8
Met Gln Lys Gln Arg Ser Thr Ser Gln Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Met Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Ile Glu Leu Ser Ser Leu Leu Ser Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys His Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Gln Leu Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Val Ala Leu Arg Asp Glu Gly Ile Val Arg Arg Val Lys Glu Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Ala Leu Ala Ser Arg Cys Leu Glu Glu
435 440 445
Phe Glu Gln Ala Leu Lys Ala Arg Ser Leu Val
450 455
<210> 9
<211> 459
<212> PRT
<213> Chromobacterium sp. MWU14-2602
<400> 9
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Arg Lys Ile Met Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ala Leu Leu Thr Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Leu Phe Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Ala Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asp Val Gly Glu Ala
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asp Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Ala Leu Ala Ala Arg Cys Met Glu Glu
435 440 445
Phe Glu Gln Thr Leu Lys Ala Arg Gly Leu Val
450 455
<210> 10
<211> 459
<212> PRT
<213> Chromobacterium sp. ATCC 53434
<400> 10
Met Gln Lys Gln Arg Thr Thr Ser Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Met Thr His Gly Asp Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Ala Ala Arg Arg Gln Met Asp
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ser Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Arg Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Ala Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys His Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Gln Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Val Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Ser Lys Ala Thr Arg Glu Leu Phe Ala Asp Phe Gly Asp Ala
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Glu Val Asp Glu Met Leu Thr Val Ala Ala Arg Cys Leu Glu Glu
435 440 445
Phe Glu Arg Thr Leu Lys Glu Arg Gly Leu Val
450 455
<210> 11
<211> 459
<212> PRT
<213> Chromobacterium
<400> 11
Met Gln Asn Gln Arg Thr Thr Ser Gln Leu Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gln Ala Gly
20 25 30
Ala Arg Val Met Thr Arg Gly Glu Gly Val Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ala Leu Leu Cys Glu Val Thr Pro Ala Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys Phe Gly
180 185 190
Lys Asp Met Thr Pro Asp Glu Phe Gly Leu Val Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Ile
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Arg Leu Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Lys
290 295 300
Arg Val Ala Glu Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Val Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Asp Asp Ile
340 345 350
Gly Pro Tyr Leu Gln Gln Arg Trp Arg Glu Thr Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Met Leu Gly Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Lys Arg Glu Leu Phe Pro Asp Phe Gly Glu Val
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ser Pro Pro Leu Val Met Thr Arg
420 425 430
Gly Glu Val Asp Glu Met Leu Ala Val Ala Ala Arg Cys Leu Glu Glu
435 440 445
Phe Glu Gln Thr Ile Lys Ala Arg Gly Leu Val
450 455
<210> 12
<211> 459
<212> PRT
<213> Xenophilus sp. AP218F
<400> 12
Met Gln Lys Gln Arg Thr Thr Ser Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn His Gln Gly
20 25 30
Ala Arg Ile Met Thr Arg Gly Glu Gly Met Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Lys Asp Phe Ala Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ala Leu Leu Ala Glu Val Thr Pro Ala Gly Phe Glu
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Ala Gln Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Ser Arg Asp Glu Phe Gly Leu Ala Ala Ala Arg Trp Leu
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe Lys Pro Asp Met Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Met Gly Ala Val Phe Val Gly Glu
290 295 300
Arg Val Ala Lys Gly Leu Ile Ala Ala Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala Arg Ala Asn Val
325 330 335
Leu Ala Leu Arg Asp Glu Gly Ile Val Glu Gln Val Lys Arg Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Gln Phe Lys
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Glu Thr Arg Glu Leu Phe Pro Asn Phe Gly Glu Val
385 390 395 400
Gly Leu Val Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ala Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Asp Asp Ile Asp Glu Met Leu Ala Val Ala Ala Arg Cys Met Arg Glu
435 440 445
Phe Glu Gln Lys Val Glu Ser Glu Asn Leu Ser
450 455
<210> 13
<211> 460
<212> PRT
<213> Chromobacterium sp. LK11
<400> 13
Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp Arg Gln Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Gln Ser Leu Asn Gln Gln
20 25 30
Gly Ala Arg Val Met Thr Arg Gly Glu Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Val Gly Tyr Gly Arg Lys Glu Leu Ser Asp Val Ala Lys His Gln Met
65 70 75 80
Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Thr Pro Ala Gly Tyr
100 105 110
Gln His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg His Tyr Trp Asp Val Gln Gly Lys Lys Asp Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Ala Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys Tyr
180 185 190
Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp
195 200 205
Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp Ile Phe Thr Ala Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala Val Ala Arg Ala Asn
325 330 335
Ile Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys Asn Glu
340 345 350
Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe Pro Asp Phe Gly Asp
385 390 395 400
Ile Gly Met Met Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ala Ser Pro Pro Leu Val Ile Ser
420 425 430
Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala Ala Arg Cys Met Glu
435 440 445
Glu Phe Glu Arg Lys Leu Lys Asp Asp Gly Leu Val
450 455 460
<210> 14
<211> 460
<212> PRT
<213> Chromobacterium sp. LK1
<400> 14
Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp Arg Gln Leu Asp Ala
1 5 10 15
Ser His His Leu His Pro Phe Thr Asp Thr Gln Ser Leu Asn Gln Gln
20 25 30
Gly Ala Arg Val Met Thr Arg Gly Glu Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Asp Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Glu Leu Ser Asp Val Ala Lys Arg Gln Met
65 70 75 80
Asp Glu Leu Ala Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Thr Pro Ala Gly Tyr
100 105 110
Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg His Tyr Trp Asp Val Gln Gly Lys Lys Asp Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys Tyr
180 185 190
Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp
195 200 205
Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala Val Ala Arg Ala Asn
325 330 335
Val Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys His Glu
340 345 350
Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe Pro Asn Phe Gly Asp
385 390 395 400
Val Gly Leu Met Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ala Ser Pro Pro Leu Val Ile Thr
420 425 430
Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala Ala Arg Cys Met Glu
435 440 445
Thr Phe Glu Arg Lys Leu Lys Glu Ser Gly Leu Ala
450 455 460
<210> 15
<211> 460
<212> PRT
<213> Chromobacterium haemolyticum
<400> 15
Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp Arg Gln Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Gln Ser Leu Asn Gln Gln
20 25 30
Gly Ala Arg Val Met Thr Arg Ser Glu Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Glu Leu Ser Glu Val Ala Lys Asn Gln Met
65 70 75 80
Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Thr Pro Ala Gly Tyr
100 105 110
Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg His Tyr Trp Asp Val Gln Gly Lys Lys Asp Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys Tyr
180 185 190
Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp
195 200 205
Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala Val Ala Arg Val Asn
325 330 335
Ile Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys Asn Glu
340 345 350
Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe Pro Glu Phe Gly Asp
385 390 395 400
Val Gly Leu Met Cys Arg Asp Ile Phe Phe Ser Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser
420 425 430
Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala Ala Arg Cys Met Glu
435 440 445
Thr Phe Glu Arg Lys Leu Lys Asp Ser Gly Leu Val
450 455 460
<210> 16
<211> 481
<212> PRT
<213> Chromobacterium haemolyticum
<400> 16
Met Ala Lys Glu Gly Arg Gln Arg Thr Ser Thr Pro Arg Trp Arg Ile
1 5 10 15
Leu Glu Gly Ser Pro Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp
20 25 30
Arg Gln Leu Asp Ala Ala His His Leu His Pro Phe Thr Asp Thr Gln
35 40 45
Ser Leu Asn Gln Gln Gly Ala Arg Val Met Thr Arg Ser Glu Gly Ile
50 55 60
Tyr Leu Trp Asp Ser Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly
65 70 75 80
Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg Lys Glu Leu Ser Glu Val
85 90 95
Ala Lys Asn Gln Met Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys
100 105 110
Thr Thr His Pro Ala Val Val Glu Leu Ser His Leu Leu Ala Gln Val
115 120 125
Thr Pro Ala Gly Tyr Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu
130 135 140
Ala Val Asp Thr Met Ile Arg Met Val Arg His Tyr Trp Asp Val Gln
145 150 155 160
Gly Lys Lys Asp Lys Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His
165 170 175
Gly Ser Thr Ile Gly Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His
180 185 190
Glu Gln Gly Asp Leu Pro Ile Pro Gly Met Ala His Val Glu Gln Pro
195 200 205
Trp Trp Tyr Lys Tyr Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val
210 215 220
Val Ala Ala Arg Trp Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu
225 230 235 240
Lys Val Ala Ala Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val
245 250 255
Ile Ile Pro Pro Ala Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg
260 265 270
Lys Tyr Asp Val Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly
275 280 285
Arg Thr Gly Glu Trp Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp
290 295 300
Ile Phe Thr Thr Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly
305 310 315 320
Ala Val Phe Val Gly Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly
325 330 335
Asp Phe Asn His Gly Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala
340 345 350
Val Ala Arg Val Asn Ile Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp
355 360 365
Arg Val Lys Asn Glu Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu
370 375 380
Thr Phe Ser Gln Phe Glu His Val Asp Asp Val Arg Gly Val Gly Leu
385 390 395 400
Ile Gln Ala Phe Thr Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe
405 410 415
Pro Glu Phe Gly Asp Val Gly Leu Met Cys Arg Asp Ile Phe Phe Ser
420 425 430
Asn Asn Leu Ile Met Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro
435 440 445
Pro Leu Val Ile Ser Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala
450 455 460
Ala Arg Cys Met Glu Thr Phe Glu Arg Lys Leu Lys Asp Ser Gly Leu
465 470 475 480
Val
<210> 17
<211> 460
<212> PRT
<213> Chromobacterium sp. Panama
<400> 17
Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp Arg Gln Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Gln Ser Leu Asn Gln Gln
20 25 30
Gly Ala Arg Val Met Thr Arg Ser Glu Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Glu Leu Ser Asp Val Ala Lys Ser Gln Met
65 70 75 80
Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Thr Pro Ala Gly Tyr
100 105 110
Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg His Tyr Trp Asp Val Gln Gly Lys Lys Glu Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys Tyr
180 185 190
Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val Val Ala Ala Arg Trp
195 200 205
Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala Val Ala Arg Val Asn
325 330 335
Ile Glu Leu Leu Arg Asp Glu Gly Val Ile Asp Arg Val Lys Asn Glu
340 345 350
Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe Pro Glu Phe Gly Asp
385 390 395 400
Val Gly Leu Met Cys Arg Asp Ile Phe Phe Ser Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser
420 425 430
Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala Ala Arg Cys Met Glu
435 440 445
Thr Phe Glu Arg Lys Leu Lys Asp Ser Gly Leu Val
450 455 460
<210> 18
<211> 481
<212> PRT
<213> Chromobacterium haemolyticum
<400> 18
Met Ala Lys Glu Gly Arg Gln Arg Thr Ser Thr Pro Arg Trp Arg Ile
1 5 10 15
Leu Glu Gly Ser Pro Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp
20 25 30
Arg Gln Leu Asp Ala Ala His His Leu His Pro Phe Thr Asp Thr Gln
35 40 45
Ser Leu Asn Gln Gln Gly Ala Arg Val Met Thr Arg Ser Glu Gly Ile
50 55 60
Tyr Leu Trp Asp Ser Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly
65 70 75 80
Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg Lys Glu Leu Ser Asp Val
85 90 95
Ala Lys Ser Gln Met Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys
100 105 110
Thr Thr His Pro Ala Val Val Glu Leu Ser His Leu Leu Ala Gln Val
115 120 125
Thr Pro Ala Gly Tyr Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu
130 135 140
Ala Val Asp Thr Met Ile Arg Met Val Arg His Tyr Trp Asp Val Gln
145 150 155 160
Gly Lys Lys Asp Lys Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His
165 170 175
Gly Ser Thr Ile Gly Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His
180 185 190
Glu Gln Gly Asp Leu Pro Ile Pro Gly Met Ala His Val Glu Gln Pro
195 200 205
Trp Trp Tyr Lys Tyr Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val
210 215 220
Val Ala Ala Arg Trp Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu
225 230 235 240
Lys Val Ala Ala Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val
245 250 255
Ile Ile Pro Pro Ala Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg
260 265 270
Lys Tyr Asp Val Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly
275 280 285
Arg Thr Gly Glu Trp Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp
290 295 300
Ile Phe Thr Thr Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly
305 310 315 320
Ala Val Phe Val Gly Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly
325 330 335
Asp Phe Asn His Gly Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala
340 345 350
Val Ala Arg Val Asn Ile Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp
355 360 365
Arg Val Lys Asn Glu Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu
370 375 380
Thr Phe Ser Gln Phe Glu His Val Asp Asp Val Arg Gly Val Gly Leu
385 390 395 400
Ile Gln Ala Phe Thr Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe
405 410 415
Pro Glu Phe Gly Asp Val Gly Leu Met Cys Arg Asp Ile Phe Phe Ser
420 425 430
Asn Asn Leu Ile Met Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro
435 440 445
Pro Leu Val Ile Ser Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala
450 455 460
Ala Arg Cys Met Glu Thr Phe Glu Arg Lys Leu Lys Asp Ser Gly Leu
465 470 475 480
Val
<210> 19
<211> 460
<212> PRT
<213> Chromobacterium haemolyticum
<400> 19
Met Gln Asn Gln Gln Arg Thr Thr Glu Gln Trp Arg Gln Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Gln Ser Leu Asn Gln Gln
20 25 30
Gly Ala Arg Val Met Thr Arg Ser Glu Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Glu Leu Ser Asp Val Ala Lys Ser Gln Met
65 70 75 80
Asp Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Thr Pro Ala Gly Tyr
100 105 110
Glu His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg His Tyr Trp Asp Val Gln Gly Lys Lys Asp Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ser Ser Leu Gly Gly Met Lys Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Met Ala His Val Glu Gln Pro Trp Trp Tyr Lys Tyr
180 185 190
Gly Lys Asp Leu Ser Pro Asp Glu Phe Gly Val Glu Ala Ala Arg Trp
195 200 205
Val Glu Glu Lys Ile Leu Glu Ile Gly Pro Glu Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Phe Glu His Phe Gly Phe Arg Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Glu Arg Val Ala Gln Gly Leu Ile Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ser Ala Val Ala Arg Val Asn
325 330 335
Ile Glu Leu Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys Asn Glu
340 345 350
Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asp Lys Ser Lys Arg Glu Leu Phe Pro Glu Phe Gly Asp
385 390 395 400
Val Gly Leu Met Cys Arg Asp Ile Phe Phe Ser Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser
420 425 430
Lys Ala Glu Ile Asp Glu Met Leu Gly Val Ala Ala Arg Cys Met Glu
435 440 445
Thr Phe Glu Arg Lys Leu Lys Asp Ser Gly Leu Val
450 455 460
<210> 20
<211> 460
<212> PRT
<213> Vogesella oryzae
<400> 20
Met Thr Gln Gln Gln Arg Ser Thr Ala Gln Trp Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Gln
20 25 30
Gly Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Thr Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Lys Arg Gln Met
65 70 75 80
Asp Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Ala Gly Phe
100 105 110
Asp His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met
115 120 125
Ile Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Ile Ala His Ile Glu Gln Pro Trp Tyr Tyr Lys His
180 185 190
Gly Lys Asp Met Thr Pro Glu Glu Phe Gly Ile Ala Ala Ala His Trp
195 200 205
Leu Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser
225 230 235 240
Ser Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly His Gln His Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Asn Asp Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn
325 330 335
Val Lys Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Glu Glu
340 345 350
Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Gly Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu
385 390 395 400
Met Gly Thr Met Cys Arg Asp Ile Phe Phe Gly Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Thr
420 425 430
Lys Ala Glu Ile Asp Ala Thr Leu Ala Thr Ala Ala Arg Cys Ile Glu
435 440 445
Thr Phe Glu Lys Lys Leu Arg Glu Arg Gly Leu Ala
450 455 460
<210> 21
<211> 459
<212> PRT
<213> Vogesella urethralis
<400> 21
Met Thr Gln Gln Arg Asn Thr Ala Ala Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Gln Gly
20 25 30
Val Arg Met Ile Thr Arg Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Met Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Asp Val Ala Lys Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Gln Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Ala His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Val Ala Ala Ala Arg Trp Ile
195 200 205
Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Ser His Phe Gly Phe Thr Pro Asp Ile Tyr Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Asn Asp
290 295 300
Glu Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Glu Asp Ile
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Ile Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Gln Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Ala Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Asp Glu Ile Asp Gln Met Leu Ala Thr Ala Ala Arg Cys Met Glu Thr
435 440 445
Phe Glu Arg Lys Leu Lys Glu Lys Gly Leu Ala
450 455
<210> 22
<211> 460
<212> PRT
<213> Vogesella sp. LIG4
<400> 22
Met Thr Gln Gln Gln Arg Ser Thr Ala Gln Trp Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Thr Asp Thr Thr Ser Leu Asn Gln Gln
20 25 30
Gly Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Thr Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Leu Gly Tyr Gly Arg Lys Asp Leu Ser Asp Val Ala Lys Arg Gln Met
65 70 75 80
Asp Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala
85 90 95
Val Val Glu Leu Ser His Leu Leu Ala Gln Val Ala Pro Ala Gly Phe
100 105 110
Asp His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met
115 120 125
Met Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys
130 135 140
Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Ile Ala His Ile Glu Gln Pro Trp Tyr Tyr Lys His
180 185 190
Gly Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Arg Trp
195 200 205
Leu Glu Asp Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser
225 230 235 240
Thr Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu
245 250 255
Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Gln Gln His Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly
290 295 300
Asn Glu Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn
325 330 335
Val Lys Ala Leu Arg Asp Glu Gly Ile Val Lys Arg Val Lys Glu Glu
340 345 350
Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Thr Phe Gly Gln Phe
355 360 365
Glu His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu
385 390 395 400
Met Gly Thr Met Cys Arg Asp Ile Phe Phe Gly Asn Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Met Val Ser Ala Pro Pro Leu Val Ile Thr
420 425 430
Lys Ala Glu Ile Asp Thr Met Leu Gly Thr Ala Ala Arg Cys Met Glu
435 440 445
Thr Phe Glu Gln Lys Leu Arg Glu Arg Gly Leu Ala
450 455 460
<210> 23
<211> 459
<212> PRT
<213> Aquitalea
<400> 23
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Glu Gly Ile Tyr Leu Tyr Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Gln Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Asp
290 295 300
Lys Val Ala Thr Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Met Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Glu Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Lys Gln Leu Lys Glu Arg Gly Leu Val
450 455
<210> 24
<211> 459
<212> PRT
<213> Aquitalea sp. LB_tupeE
<400> 24
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Glu Gly Ile Phe Leu Phe Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Gln Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Asp
290 295 300
Lys Val Ala Thr Thr Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asp Phe Gly Glu Ile
385 390 395 400
Gly Thr Met Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Glu Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Gln Gln Leu Lys Ala Arg Gly Leu Val
450 455
<210> 25
<211> 459
<212> PRT
<213> Vogesella perlucida
<400> 25
Met Thr Gln Gln Arg Asn Thr Ala Ala Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Gln Gly
20 25 30
Val Arg Met Ile Thr Arg Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Gln Ile Met Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Asp Val Ala Lys Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Asp Val Ala Pro Gln Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Gln Glu Glu Phe Gly Val Ala Ala Ala Arg Trp Ile
195 200 205
Glu Glu Lys Ile Leu Glu Val Gly Ala Asp Lys Ile Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Gln Arg Ile Cys Arg Gln Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Thr His Phe Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Asn Asp
290 295 300
Glu Val Ala Ser Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Gln Arg Val Lys Glu Asp Ile
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Thr Phe Ser Gln Phe Glu
355 360 365
His Val Asp Asp Ile Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Ala Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Ala Glu Val Asp Gln Met Ile Asp Thr Ala Ala Arg Cys Met Ala Glu
435 440 445
Phe Glu Arg Gln Leu Lys Glu Lys Gly Leu Ala
450 455
<210> 26
<211> 459
<212> PRT
<213> Aquitalea
<400> 26
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Glu Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Ile Tyr Leu Tyr Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Asp
290 295 300
Lys Val Ala Thr Thr Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Gly Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asn Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Asp Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Lys Gln Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 27
<211> 459
<212> PRT
<213> Aquitalea denitrificans
<400> 27
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Ile Tyr Leu Tyr Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Gln Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
Gln Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Glu
290 295 300
Lys Val Ala Thr Thr Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Gly Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asn Phe Gly Glu Ile
385 390 395 400
Gly Thr Met Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Glu Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Lys Gln Leu Lys Glu Arg Gly Leu Val
450 455
<210> 28
<211> 459
<212> PRT
<213> Aquitalea
<400> 28
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Glu Gly Ile Phe Leu Tyr Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Gln Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Glu
290 295 300
Lys Val Ala Thr Thr Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Gly Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asn Phe Gly Glu Ile
385 390 395 400
Gly Thr Met Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Glu Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Gln Gln Leu Lys Glu Arg Gly Leu Val
450 455
<210> 29
<211> 459
<212> PRT
<213> Aquitalea sp. FJL05
<400> 29
Met Gln Asn Gln Arg Thr Thr Thr Glu Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asn Ser Leu Asn Glu Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Ile Tyr Leu Tyr Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Gln Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Glu Gly Phe Lys
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Lys Asp Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Lys His Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Val
195 200 205
Glu Asp Lys Val Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Ile Gln Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Val Phe Gly Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Asn Glu
290 295 300
Lys Val Ala Thr Thr Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Val Phe Gly Gln Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asn Phe Gly Glu Ile
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Lys His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Glu Glu Ile Asp Gln Met Leu Asp Thr Ala Ala Lys Cys Met Val Glu
435 440 445
Phe Glu Lys Gln Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 30
<211> 421
<212> PRT
<213> Aquitalea magnusonii
<400> 30
Met Asp Gly Ile Tyr Leu Tyr Asp Ser Glu Gly Asn Lys Ile Leu Asp
1 5 10 15
Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg Lys Asp
20 25 30
Leu Pro Glu Val Ala Lys Arg Gln Met Glu Gln Leu Ala Tyr Tyr Asn
35 40 45
Thr Phe Phe Lys Thr Thr His Pro Ala Val Val Glu Leu Ser His Leu
50 55 60
Leu Ala Glu Val Ala Pro Gln Gly Phe Lys His Val Phe Tyr Thr Asn
65 70 75 80
Ser Gly Ser Glu Ser Val Asp Thr Met Ile Arg Met Val Arg Arg Tyr
85 90 95
Trp Asp Val Lys Gly Lys Lys Asp Lys Lys Thr Leu Ile Gly Arg Trp
100 105 110
Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser Leu Gly Gly Met
115 120 125
Thr Tyr Met His Glu Gln Gly Asp Leu Pro Ile Pro Gly Ile Val His
130 135 140
Val Glu Gln Pro Trp Trp Tyr Lys His Gly Lys Asp Met Thr Pro Glu
145 150 155 160
Glu Phe Gly Leu Ala Ala Ala Lys Trp Val Glu Asp Lys Ile Leu Glu
165 170 175
Val Gly Ala Asp Lys Val Ala Ala Phe Val Gly Glu Pro Ile Gln Gly
180 185 190
Ala Gly Gly Val Ile Val Pro Pro Ser Thr Tyr Trp Pro Glu Ile Gln
195 200 205
Arg Ile Cys Gln Lys Tyr Asp Ile Leu Leu Val Ala Asp Glu Val Ile
210 215 220
Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe Gly Gln Gln Val Phe Gly
225 230 235 240
Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys Gly Leu Ser Ser Gly Tyr
245 250 255
Gln Pro Ile Gly Ala Val Phe Leu Asn Asp Lys Val Ala Thr Thr Leu
260 265 270
Ala Glu Gly Gly Asp Phe Asn His Gly Phe Thr Tyr Ser Gly His Pro
275 280 285
Val Ala Ala Ala Val Ala His Ala Asn Val Lys Ala Leu Arg Asp Glu
290 295 300
Gly Ile Val Asp Arg Val Lys Asn Asp Thr Gly Pro Tyr Met Gln Lys
305 310 315 320
Arg Trp Arg Glu Val Phe Gly Gln Phe Glu His Val Asp Asp Val Arg
325 330 335
Gly Val Gly Leu Ile Gln Ala Phe Thr Leu Val Lys Asn Lys Ala Thr
340 345 350
Arg Glu Leu Phe Pro Asn Phe Gly Glu Ile Gly Thr Leu Cys Arg Asp
355 360 365
Ile Phe Phe Lys Asn Asn Leu Ile Met Arg Ala Cys Gly Asp His Ile
370 375 380
Val Ser Ala Pro Pro Leu Val Ile Ser Lys Glu Glu Ile Asp Gln Met
385 390 395 400
Leu Glu Thr Ala Ala Lys Cys Met Val Glu Phe Glu Lys Gln Leu Lys
405 410 415
Glu Arg Gly Leu Val
420
<210> 31
<211> 459
<212> PRT
<213> Pseudogulbenkiania sp. MAI-1
<400> 31
Met Gln Asn Glu Arg Thr Thr Ala Gln Trp Arg Ala Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Glu Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Val Phe Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Lys Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Arg Leu Leu Ser Glu Val Ala Pro Gln Gly Phe Ser
100 105 110
Arg Ile Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Thr His Val Glu Gln Pro Trp Trp Tyr Arg Tyr Gly
180 185 190
Gln Gly Met Thr Pro Glu Glu Phe Gly Val Val Ala Ala Arg Trp Val
195 200 205
Glu Glu Lys Ile Leu Glu Val Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Gln Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Leu Phe Gly Phe Arg Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Lys Val Ala Gln Gly Leu Ile Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys His Asp Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Ala Ala Phe Gly Gly Phe Ala
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Thr His Glu Leu Phe Pro Asp Trp Gly Thr Val
385 390 395 400
Gly Thr Ile Cys Arg Asp Ile Phe Phe Arg His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Ala Glu Ile Asp Gln Leu Ile Glu Thr Ala Val Gln Cys Val Thr Glu
435 440 445
Cys Glu Gln Val Leu Arg Glu Arg Gly Leu Val
450 455
<210> 32
<211> 459
<212> PRT
<213> Paludibacterium paludis
<400> 32
Met Ser Gln Gln Arg Thr Thr Ala Glu Trp Arg Ala Leu Asp Ala Ala
1 5 10 15
His His Val His Pro Phe Ser Asp Ala His Ser Leu Asn Arg Gln Gly
20 25 30
Ala Arg Val Met Thr Arg Ala Asp Gly Ile Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg His Asp Met Thr Ala Ala Ala Ser Arg Gln Met Asp
65 70 75 80
Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Ala Leu Leu Ala Lys Val Ala Pro Glu Gly Phe Asn
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Ala Gly Lys Pro Ser Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Met Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Asp Tyr Met His Ala Gln Gly Gly Leu Pro
165 170 175
Ile Pro Asp Met Ala His Ile Glu Gln Pro Trp Trp Tyr Lys Phe Gly
180 185 190
Lys Asp Met Ser Arg Asp Glu Phe Gly Leu Leu Ala Ala Gly Trp Leu
195 200 205
Glu Asp Arg Ile Leu Glu Ile Gly Pro Asp Asn Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Asp Thr
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Arg Phe Gly Phe Arg Pro Asp Ile Phe Thr Ala Ala Lys
275 280 285
Gly Leu Thr Ser Gly Tyr Leu Pro Met Gly Ala Val Phe Leu Asn Glu
290 295 300
Lys Val Ala Glu Gly Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Arg Ala Asn Ile
325 330 335
Leu Ala Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Arg His Asp Thr
340 345 350
Gly Pro Tyr Met Gln Glu Arg Trp Arg Gln Val Phe Gly Gln Phe Asp
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Thr Arg Thr Leu Phe Pro Asn Trp Gly Glu Thr
385 390 395 400
Gly Leu Ile Cys Arg Asp Ile Phe Phe Arg Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Ala Asp His Met Val Ala Ser Pro Pro Leu Val Met Thr Arg
420 425 430
Ala Gln Ile Asp Glu Met Leu Gly Val Ala Glu Arg Cys Met Thr Glu
435 440 445
Phe Glu Glu Thr Leu Leu Arg Arg Gly Leu Val
450 455
<210> 33
<211> 459
<212> PRT
<213> Pseudogulbenkiania sp. NH8B
<400> 33
Met Gln Ser Glu Arg Thr Thr Glu Gln Trp Arg Ala Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Glu Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Val Phe Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Arg Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Gln Val Ala Pro Glu Gly Phe Ser
100 105 110
Arg Ile Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Arg Pro Gly Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Arg Tyr Gly
180 185 190
Gln Gly Met Ser Pro Glu Glu Phe Gly Val Val Ala Ala Arg Trp Val
195 200 205
Glu Glu Lys Ile Leu Glu Val Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Arg Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Leu Phe Gly Phe Lys Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Lys Val Ala Gln Gly Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys His Asp Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly Phe Ala
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Thr His Glu Leu Phe Pro Asn Trp Gly Glu Val
385 390 395 400
Gly Thr Ile Cys Arg Asp Ile Phe Phe Arg His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Thr Glu Ile Asp Gln Leu Ile Asp Thr Ala Val Gln Cys Val Thr Glu
435 440 445
Cys Glu Ala Val Leu Arg Gln Arg Gly Leu Val
450 455
<210> 34
<211> 459
<212> PRT
<213> Pseudogulbenkiania subflava
<400> 34
Met Gln Asn Glu Arg Thr Thr Glu Gln Trp Arg Ala Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Glu Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Val Phe Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Lys Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser Gln Leu Leu Ser Glu Val Ala Pro Lys Gly Phe Ser
100 105 110
Arg Ile Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gly Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Glu His Val Glu Gln Pro Trp Trp Tyr Arg Tyr Cys
180 185 190
Gln Gly Met Ser Pro Glu Glu Phe Gly Val Val Ala Ala Arg Trp Leu
195 200 205
Glu Ala Lys Ile Leu Glu Ile Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Gln Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Leu Phe Gly Phe His Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Lys Val Ala Gln Gly Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys Arg Asp Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly Phe Ala
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Thr His Glu Leu Phe Pro Asn Trp Gly Glu Val
385 390 395 400
Gly Thr Ile Cys Arg Asp Ile Phe Phe Arg His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Ala Glu Ile Asp Gln Leu Ile Asp Thr Ala Val Gln Cys Val Thr Glu
435 440 445
Cys Glu Glu Val Leu Arg Gln Arg Gly Leu Val
450 455
<210> 35
<211> 459
<212> PRT
<213> Pseudogulbenkiania ferrooxidans
<400> 35
Met Gln Ser Glu Arg Thr Thr Glu Gln Trp Arg Ala Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Glu Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Lys Ala Asp Gly Val Phe Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Arg Arg Gln Met Asp
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Gln Val Ala Pro Glu Gly Phe Ser
100 105 110
His Ile Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gly Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Glu Gln Pro Trp Trp Tyr Arg Tyr Gly
180 185 190
Gln Gly Met Ser Pro Glu Glu Phe Gly Val Val Ala Ala Arg Trp Val
195 200 205
Glu Glu Lys Ile Leu Glu Val Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Arg Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Leu Phe Gly Phe His Pro Asp Leu Phe Thr Ala Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Lys Val Ala Gln Gly Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Ile Asp Arg Val Lys His Asp Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly Phe Ala
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Thr His Glu Leu Phe Pro Asn Trp Gly Glu Val
385 390 395 400
Gly Thr Ile Cys Arg Asp Ile Phe Phe Arg His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Ala Glu Ile Asp Gln Leu Ile Asp Thr Ala Val Gln Cys Val Thr Glu
435 440 445
Cys Glu Ala Val Leu His Gln Arg Gly Leu Val
450 455
<210> 36
<211> 460
<212> PRT
<213> Gulbenkiania indica
<400> 36
Met Gln Thr Ala Arg Thr Thr Glu Gln Trp Arg Glu Leu Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Pro His Ser Leu Asn Gln Gln Gly
20 25 30
Ala Arg Val Ile Thr Arg Gly Glu Gly Ile His Leu Trp Asp Ser Glu
35 40 45
Gly His Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Arg Arg Gln Met Glu
65 70 75 80
Asp Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ser Glu Val Ala Pro Glu Gly Phe Arg
100 105 110
His Ile Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Met
115 120 125
Arg Met Val Arg His Phe Trp Ala Val Gln Gly Lys Pro Glu Lys Lys
130 135 140
Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Glu His Val Glu Gln Pro Trp Trp Tyr Lys Tyr Gly
180 185 190
Lys Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Arg Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Pro Glu Arg Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Gln Thr Met Gly Phe His Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Arg Val Ala Asn Val Leu Val Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Asp Arg Val Lys Asn Glu Thr
340 345 350
Gly Pro Tyr Met Gln Glu Arg Trp Arg Ala Thr Phe Gly Asn Phe Asp
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Gly Phe Thr Leu
370 375 380
Val Arg Asn Lys Ala Thr His Glu Leu Phe Pro Glu Trp Gly Thr Ala
385 390 395 400
Gly Thr Met Cys Arg Asp Ile Phe Phe Glu Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ala Ala Pro Pro Leu Val Ile Thr Lys
420 425 430
Ala Glu Ile Asp Glu Met Leu Gly Val Ala Gly Arg Cys Leu Glu Arg
435 440 445
Phe Glu Lys Val Val Arg Glu Lys Gly Met Ala Gly
450 455 460
<210> 37
<211> 459
<212> PRT
<213> Paludibacterium sp. dN 18-1
<400> 37
Gln Lys Gln Arg Ser Thr Arg Glu Leu Arg Glu Leu Asp Ala Ala His
1 5 10 15
His Leu His Pro Phe Thr Asp Thr Thr Ser Leu Asn Gln Gln Gly Ala
20 25 30
Arg Val Ile Thr Lys Gly Glu Gly Ile Tyr Leu Trp Asp Ser Glu Gly
35 40 45
Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly
50 55 60
Tyr Gly Arg Lys Asp Leu Ser Glu Val Ala Lys Arg Gln Met Asp Asp
65 70 75 80
Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val Val
85 90 95
Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Ala Gly Phe Ser His
100 105 110
Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile Arg
115 120 125
Met Val Arg Arg Tyr Trp Glu Val Lys Gly Gln Lys Gln Lys Lys Thr
130 135 140
Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala
145 150 155 160
Ser Leu Gly Gly Met Ala Tyr Met His Gln Asp Gly Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Glu His Val Gln Gln Pro Trp Trp Tyr Gly Leu Gly
180 185 190
Gly Asp Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Lys Trp Leu
195 200 205
Glu Asp Lys Ile Leu Glu Ile Gly Pro Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys His Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln Cys Phe Gly Phe His Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Lys Val Ala Ala Met Leu Ala Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Cys Ala Asn Val
325 330 335
Arg Ala Leu Arg Asp Glu Gly Ile Ile Glu Arg Val Lys His Glu Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Val Phe Ser Arg Phe Glu
355 360 365
His Val Gly Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Pro Asn Thr Gly Glu Ile
385 390 395 400
Gly Leu Ile Cys Arg Asp Ile Phe Phe Arg His Asp Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ser Ala Pro Pro Leu Val Ile Ser Lys
420 425 430
Gln Glu Ile Asp Asp Met Leu Ala Leu Ala Ala Gln Cys Met Glu Glu
435 440 445
Phe Glu Ala Glu Leu Lys Ala Arg Asn Leu Val
450 455
<210> 38
<211> 459
<212> PRT
<213> Vogesella fluminis
<400> 38
Met Lys His Gln Arg Ser Thr Ala Gln Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Ala Gln Gly
20 25 30
Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Ile Ala Lys Arg Gln Met Glu
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Lys Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Gln Gln Pro Trp His Tyr Lys Tyr Gly
180 185 190
Lys Gly Met Ser Ala Asp Glu Phe Gly Leu Leu Ala Ala Ser Trp Ile
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly His Trp Phe
260 265 270
Gly Gln Gln Gln Phe Gly Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asn
290 295 300
Glu Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Ala Ala Val His Glu Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Ala Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Ser Gln Thr Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Arg His Gly Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Ile Ile Ser Lys
420 425 430
Asp Gln Ile Asp Glu Met Ile Ala Thr Ala Val Glu Cys Met Glu Ile
435 440 445
Phe Glu Gln Lys Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 39
<211> 459
<212> PRT
<213> Vogesella alkaliphila
<400> 39
Met Lys His Gln Arg Ser Thr Ala Gln Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Gln Gly
20 25 30
Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Lys Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Gln Gln Pro Trp His Tyr Lys Tyr Gly
180 185 190
Lys Gly Met Ser Ala Asp Glu Phe Gly Lys Leu Ala Ala Ser Trp Val
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly His Trp Phe
260 265 270
Gly His Gln Gln Phe Gly Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Met Pro Ile Gly Ala Val Phe Val Asn Asn
290 295 300
Glu Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Ala Ala Val His Glu Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Ala Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Ser Gln Thr Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Lys His Gly Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Ile Ile Ser Lys
420 425 430
Asp Gln Ile Asp Glu Leu Ile Ala Thr Ala Val Lys Cys Met Glu Ile
435 440 445
Phe Glu Gln Gln Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 40
<211> 458
<212> PRT
<213> Crenobacter sp. HX-7-9
<400> 40
Met Lys Gln Asn Thr Glu Ala Trp Arg Ala Leu Asp Ala Ala His His
1 5 10 15
Leu His Pro Phe Ser Asp Thr Lys Gly Leu Asn Ala Gln Gly Ala Arg
20 25 30
Val Ile Thr Arg Ala Glu Gly Ile Tyr Leu Trp Asp Ser Glu Gly Asn
35 40 45
Gln Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr
50 55 60
Gly Arg Lys Asp Leu Ala Glu Val Ala Lys Arg Gln Met Glu Glu Leu
65 70 75 80
Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr Asn Pro Gln Val Ala Glu
85 90 95
Leu Ser Lys Leu Leu Ala Glu Val Ala Pro Glu Gly Phe Glu His Val
100 105 110
Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile Arg Met
115 120 125
Val Arg His Tyr Trp Ser Ser Val Gly Lys Pro Glu Lys Lys Val Leu
130 135 140
Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser
145 150 155 160
Met Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro Ile Pro
165 170 175
Gly Ile Val His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly Lys Asp
180 185 190
Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Arg Trp Ile Glu Asp
195 200 205
Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val Gly Glu
210 215 220
Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr Tyr Trp
225 230 235 240
Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Met Val Cys
245 250 255
Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Gln Trp Phe Gly His
260 265 270
Glu Leu Tyr Gly Val Lys Pro Asp Leu Phe Thr Thr Ala Lys Gly Leu
275 280 285
Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Glu Arg Val
290 295 300
Ala Gln Gly Leu Ile Glu His Gly Gly Asp Phe Asn His Gly Phe Thr
305 310 315 320
Tyr Ser Gly His Pro Val Cys Cys Ala Val Ala Arg Ala Asn Val Thr
325 330 335
Ala Leu Arg Asp Glu Gly Ile Val Ala Arg Val Arg Glu Glu Thr Gly
340 345 350
Pro Tyr Met Gln Ala Lys Trp Arg Glu Thr Phe Gly Gln Phe Asp His
355 360 365
Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu Val
370 375 380
Lys Asp Lys Gln Lys Arg Glu Leu Phe Asp Asn Trp Gly Thr Met Gly
385 390 395 400
Leu Ala Cys Arg Asp Ile Phe Phe Arg Glu Asn Leu Ile Met Arg Ala
405 410 415
Cys Gly Asp His Ile Val Ala Ala Pro Pro Leu Val Ile Ser Lys Arg
420 425 430
Glu Ile Asp Glu Met Leu Gly Arg Ala Ala Arg Cys Val Glu Glu Phe
435 440 445
Glu Arg Val Val Val Arg Gly Gln Pro Gly
450 455
<210> 41
<211> 459
<212> PRT
<213> Vogesella indigofera
<400> 41
Met Lys His Gln Arg Ser Thr Ala Gln Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Glu Gln Gly
20 25 30
Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Lys Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Glu Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Gln Gln Pro Trp His Tyr Lys Tyr Gly
180 185 190
Lys Gly Met Ser Ala Asp Glu Phe Gly Lys Leu Ala Ala Ser Trp Ile
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly His Trp Phe
260 265 270
Gly Gln Gln Gln Phe Gly Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Met Pro Ile Gly Ala Val Phe Val Asn Asn
290 295 300
Glu Val Ala Thr Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Ala Ala Val His Glu Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Ala Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Ser Gln Thr Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Lys His Gly Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Ile Ile Ser Lys
420 425 430
Asp Gln Ile Asp Glu Leu Ile Thr Thr Ala Val Lys Cys Met Glu Ile
435 440 445
Phe Glu Gln Gln Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 42
<211> 458
<212> PRT
<213> Crenobacter sp. GY 70310
<400> 42
Met Lys Gln Asn Thr Glu Ala Trp Arg Ala Leu Asp Ala Ala His His
1 5 10 15
Leu His Pro Phe Ser Asp Thr Lys Gly Leu Asn Ala Gln Gly Ala Arg
20 25 30
Val Ile Thr Arg Ala Glu Gly Ile Tyr Leu Trp Asp Ser Glu Gly Asn
35 40 45
Gln Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr
50 55 60
Gly Arg Lys Asp Leu Ala Glu Val Ala Lys Arg Gln Met Glu Glu Leu
65 70 75 80
Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr Asn Pro Gln Val Ala Glu
85 90 95
Leu Ser Lys Leu Leu Ala Glu Val Ala Pro Asp Gly Phe Glu His Val
100 105 110
Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile Arg Met
115 120 125
Val Arg His Tyr Trp Ser Ser Val Gly Lys Pro Glu Lys Lys Val Leu
130 135 140
Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser
145 150 155 160
Met Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu Pro Ile Pro
165 170 175
Gly Ile Val His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly Lys Asp
180 185 190
Met Thr Pro Glu Glu Phe Gly Leu Ala Ala Ala Arg Trp Ile Glu Asp
195 200 205
Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val Gly Glu
210 215 220
Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Thr Tyr Trp
225 230 235 240
Pro Glu Val Glu Arg Ile Cys Arg Lys His Asp Ile Leu Met Val Cys
245 250 255
Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Gln Trp Phe Gly His
260 265 270
Glu Leu Tyr Gly Val Lys Pro Asp Leu Phe Thr Thr Ala Lys Gly Leu
275 280 285
Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Glu Arg Val
290 295 300
Ala Gln Gly Leu Ile Glu His Gly Gly Asp Phe Asn His Gly Phe Thr
305 310 315 320
Tyr Ser Gly His Pro Val Cys Cys Ala Val Ala Arg Ala Asn Val Thr
325 330 335
Ala Leu Arg Asp Glu Gly Ile Val Ala Arg Val Arg Glu Glu Thr Gly
340 345 350
Pro Tyr Met Gln Ala Lys Trp Arg Glu Thr Phe Gly Gln Phe Glu His
355 360 365
Ile Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu Val
370 375 380
Lys Asp Lys Gln Lys Arg Ala Leu Phe Asp Asn Trp Gly Thr Met Gly
385 390 395 400
Leu Ala Cys Arg Asp Ile Phe Phe Arg Glu Asn Leu Ile Met Arg Ala
405 410 415
Cys Gly Asp His Ile Val Ala Ala Pro Pro Leu Val Ile Ser Lys Pro
420 425 430
Glu Ile Asp Glu Met Leu Gly Arg Ala Ala Arg Cys Val Glu Glu Phe
435 440 445
Glu Arg Val Val Val Arg Gly Gln Pro Gly
450 455
<210> 43
<211> 459
<212> PRT
<213> Vogesella sp. EB
<400> 43
Met Lys His Gln Arg Ser Thr Ala Gln Trp Arg Glu Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Ala Ser Leu Asn Gly Gln Gly
20 25 30
Val Arg Met Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro Ala Val
85 90 95
Val Glu Leu Ser His Leu Leu Ala Glu Val Ala Pro Lys Gly Phe Asp
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Lys Gly Lys Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Val His Val Gln Gln Pro Trp His Tyr Lys Tyr Gly
180 185 190
Lys Gly Met Ser Ala Asp Glu Phe Gly Lys Leu Ala Ala Ser Trp Ile
195 200 205
Glu Asp Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Ile Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly His Trp Phe
260 265 270
Gly Gln Gln Gln Leu Gly Phe Thr Pro Asp Ile Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Met Pro Ile Gly Ala Val Phe Val Asn Asn
290 295 300
Glu Val Ala Ala Thr Leu Ala Ala Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala His Ala Asn Val
325 330 335
Lys Ala Leu Arg Asp Glu Gly Ile Val Ala Ala Val His Glu Asp Thr
340 345 350
Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Ala Phe Ser Arg Phe Glu
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asp Lys Ala Ser Gln Thr Leu Phe Asp Asn Trp Gly Glu Ile
385 390 395 400
Gly Leu Met Cys Arg Asp Ile Phe Phe Lys His Gly Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Ser Ala Pro Pro Leu Ile Ile Ser Lys
420 425 430
Asp Gln Ile Asp Glu Leu Ile Thr Thr Ala Val Lys Cys Met Glu Ile
435 440 445
Phe Glu Gln Gln Leu Lys Glu Arg Gly Leu Ala
450 455
<210> 44
<211> 457
<212> PRT
<213> Rhodobacteraceae bacterium CH30
<400> 44
Ser Gln Gln His Thr Thr Gln Asp Trp Arg Ala Met Asp Ala Ala His
1 5 10 15
His Leu His Pro Phe Ser Asp Thr Lys Gly Leu Asn Ala Gln Gly Ala
20 25 30
Arg Val Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu Gly
35 40 45
Asn Lys Ile Thr Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly
50 55 60
Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu Glu
65 70 75 80
Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Lys Val Ala
85 90 95
Glu Leu Ser Ala Leu Leu Ala Ser Val Ala Pro Asp Gly Phe Glu His
100 105 110
Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile Arg
115 120 125
Met Val Arg His Tyr Trp Ser Ser Val Gly Lys Pro Glu Lys Lys Val
130 135 140
Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala
145 150 155 160
Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro Ile
165 170 175
Pro Gly Ile Ala His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly Lys
180 185 190
Asp Leu Thr Pro Glu Glu Phe Gly Ile Gln Ala Ala Gln Trp Ile Glu
195 200 205
Ala Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val Gly
210 215 220
Glu Pro Val Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr Tyr
225 230 235 240
Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu Val
245 250 255
Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe Gly
260 265 270
Gln Thr His Phe Gly Phe Lys Pro Asp Met Phe Thr Thr Ala Lys Gly
275 280 285
Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Glu Arg
290 295 300
Val Ala Arg Gly Leu Ile Glu Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn Val
325 330 335
Ala Ala Leu Arg Asp Glu Gly Ile Val Glu Arg Val Lys Asn Glu Thr
340 345 350
Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Val Phe Gly Gln Phe Ala
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr Leu
370 375 380
Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu Ala
385 390 395 400
Gly Thr Gln Cys Arg Asp Ile Phe Phe Lys His Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Ile Val Cys Ala Pro Pro Leu Val Ile Thr Lys
420 425 430
Ala Glu Ile Asp Ala Met Leu Glu Thr Ala Val Ala Cys Leu Arg Glu
435 440 445
Phe Glu Gln Leu Val Ala Ala Arg Ser
450 455
<210> 45
<211> 458
<212> PRT
<213> Neisseriaceae bacterium B2N2-7
<400> 45
Met Ser Gln Gln His Thr Thr Gln Asp Trp Arg Ala Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Ser Asp Thr Lys Gly Leu Asn Ala Gln Gly
20 25 30
Ala Arg Val Ile Thr His Ala Asp Gly Ile Tyr Leu Trp Asp Ser Glu
35 40 45
Gly Asn Lys Ile Thr Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Pro Glu Val Ala Lys Arg Gln Met Glu
65 70 75 80
Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Lys Val
85 90 95
Ala Glu Leu Ser Ala Leu Leu Ala Ser Val Ala Pro Asp Gly Phe Glu
100 105 110
His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg His Tyr Trp Ser Ser Val Gly Lys Pro Glu Lys Lys
130 135 140
Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Ser Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Ala His Val Glu Gln Pro Trp Tyr Tyr Lys His Gly
180 185 190
Lys Asp Leu Thr Pro Glu Glu Phe Gly Ile Gln Ala Ala Gln Trp Ile
195 200 205
Glu Ala Lys Ile Leu Glu Val Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Val Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr
225 230 235 240
Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Gln Thr His Phe Gly Phe Lys Pro Asp Met Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Glu
290 295 300
Arg Val Ala Arg Gly Leu Ile Glu Glu Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Cys Ala Ala Val Ala His Ala Asn
325 330 335
Val Ala Ala Leu Arg Asp Glu Gly Ile Val Glu Arg Val Lys Asn Ala
340 345 350
Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Val Phe Gly Gln Phe
355 360 365
Ala His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe Thr
370 375 380
Leu Val Lys Asn Lys Ala Thr Arg Glu Leu Phe Asp Asn Trp Gly Glu
385 390 395 400
Ala Gly Thr Gln Cys Arg Asp Ile Phe Phe Lys His Asn Leu Ile Met
405 410 415
Arg Ala Cys Gly Asp His Ile Val Cys Ala Pro Pro Leu Val Ile Thr
420 425 430
Lys Ala Glu Ile Asp Ala Met Leu Glu Thr Ala Val Ala Cys Leu Arg
435 440 445
Glu Phe Glu Gln Leu Val Ala Ala Arg Ser
450 455
<210> 46
<211> 461
<212> PRT
<213> Microvirgula
<400> 46
Met Ser Thr Thr Ala Arg Ser Thr Asn Glu Tyr Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Ser Asp Met Gly Ser Leu Asn Arg Gln
20 25 30
Gly Ser Arg Val Ile Val Lys Gly Asp Gly Ile Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Thr Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Ile Gly Tyr Gly Arg Lys Asp Leu Ala Glu Val Ala Arg Gln Gln Met
65 70 75 80
Glu Asp Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Arg
85 90 95
Val Val Glu Leu Ser Thr Leu Leu Ala Glu Val Thr Gln Pro Gly Tyr
100 105 110
Gln His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ser Val Asp Thr Val
115 120 125
Ile Arg Met Val Arg Gln Tyr Trp Ser Ile Lys Gly Lys Pro Gly Lys
130 135 140
Lys Thr Leu Ile Ser Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Thr Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Ile Ala His Ile Gly Gln Pro Trp Tyr Leu Lys Glu
180 185 190
Gly Arg Asp Gly Glu Ser Glu Ala Asp Phe Gly Leu Arg Ala Ala Ser
195 200 205
Trp Leu Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala
210 215 220
Phe Val Gly Glu Pro Ile Leu Gly Ala Gly Gly Val Ile Ile Pro Pro
225 230 235 240
Ala Ser Tyr Trp Pro Glu Ile Glu Arg Ile Cys Asn Lys Tyr Asp Ile
245 250 255
Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asp
260 265 270
Trp Phe Gly His Glu His Phe Gly Phe Arg Pro His Leu Phe Thr Ile
275 280 285
Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Gly Val Ile Ile
290 295 300
Gly Asp Glu Val Ala Arg Thr Leu Ile Glu Gly Gly Asp Phe Asn His
305 310 315 320
Gly Phe Thr Tyr Ser Gly His Pro Val Ser Ala Ala Val Ala His Ala
325 330 335
Asn Val Arg Ala Leu Arg Asp Glu Gly Ile Val Glu Gln Val Lys Lys
340 345 350
Asp Thr Gly Pro Tyr Met Gln Lys Arg Trp Arg Glu Ala Phe Gly Gly
355 360 365
Phe Glu His Val Thr Asp Val Arg Gly Val Gly Leu Ile Gln Ala Phe
370 375 380
Thr Trp Val Lys Asn Lys Ala Thr Arg Glu Met Phe Ala Gly Ser Asp
385 390 395 400
Asp Val Gly Thr Leu Cys Arg Asp Ile Phe Phe Lys His Asn Leu Ile
405 410 415
Met Arg Ala Cys Gly Asp Ser Met Val Ser Ala Pro Pro Leu Ile Ile
420 425 430
Thr Arg Thr Glu Ile Asp Ala Met Leu Ser Thr Ala Val Glu Cys Val
435 440 445
Glu Gln Thr Glu Lys Leu Leu Gln Glu Arg Gly Leu Val
450 455 460
<210> 47
<211> 469
<212> PRT
<213> Neisseria shayeganii
<400> 47
Met Ser Lys Gln Asp Tyr Ser Arg Gln Asp Gly Lys His His Leu His
1 5 10 15
Pro Phe Ser Asp Asn Ala Ala Leu Ala Arg Glu Gly Val Arg Val Met
20 25 30
Val Lys Ala Lys Gly Ile Tyr Val Tyr Asp Asp Glu Gly Asn Glu Ile
35 40 45
Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg
50 55 60
Lys Glu Leu Ala Lys Val Ala Lys Lys Gln Met Lys Glu Leu Pro Phe
65 70 75 80
Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val Val Glu Leu Ser
85 90 95
Ser Thr Ile Thr Asp Ile Ala Pro Ser Gly Phe Asn His Val Phe Tyr
100 105 110
Thr Gly Ser Gly Ser Glu Ser Val Asp Thr Met Met Arg Met Val Arg
115 120 125
His Tyr Trp Ala Ser Val Gly Lys Pro Gln Lys Lys Val Ile Ile Gly
130 135 140
Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser Leu Gly
145 150 155 160
Gly Met Lys Gly Met His Asn Gln Gly Asp Leu Pro Ile Pro Asn Val
165 170 175
Val His Ile Glu Gln Pro Trp Tyr Tyr Gly Leu Ala Lys Asp Gly Glu
180 185 190
Thr Pro Glu Glu Phe Gly Leu Arg Ala Ala Asn Trp Leu Glu Glu Lys
195 200 205
Ile Glu Glu Ile Gly Ala Asp Lys Ile Ala Ala Phe Val Gly Glu Pro
210 215 220
Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Lys Thr Tyr Trp Pro
225 230 235 240
Arg Ile Glu Glu Ile Cys Arg Lys His Asp Ile Leu Leu Val Ala Asp
245 250 255
Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Trp Phe Gly His Glu
260 265 270
Thr Met Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys Gly Leu Ser
275 280 285
Ser Gly Tyr Leu Pro Ile Gly Ala Val Leu Val Asn Asp Lys Val Ala
290 295 300
Glu Gly Leu Lys Ala Gly Gly Glu Phe Asn His Gly Phe Thr Tyr Ser
305 310 315 320
Gly His Pro Val Ala Ala Ala Val Ala Lys Lys Asn Leu Glu Ile Leu
325 330 335
Gln Asp Glu Gly Ile Val Asp Gln Val Lys Asn Glu Thr Gly Pro Tyr
340 345 350
Met Lys Lys Arg Trp Glu Glu Thr Phe Ser Gln Phe Lys Tyr Val Asp
355 360 365
Asp Ile Arg Ser Glu Gly Leu Ile Cys Gly Phe Thr Leu Val Lys Asp
370 375 380
Lys Ala Thr Arg Glu Leu Phe Pro Asn Thr Gly Glu Thr Gly Leu Met
385 390 395 400
Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg Ala Cys Gly
405 410 415
Asp His Met Val Ala Ala Pro Pro Leu Val Met Thr Lys Lys Glu Ile
420 425 430
Asp Lys Met Leu Ala Val Ala Glu Gln Cys Met Arg Glu Phe Glu Gln
435 440 445
Met Met Asp Lys Lys Leu Ala Lys Glu Glu Lys Ala Gly Lys Lys Glu
450 455 460
Glu Lys Ser Asp Gln
465
<210> 48
<211> 469
<212> PRT
<213> Neisseria shayeganii
<400> 48
Met Ser Lys Gln Asp Tyr Ser Arg Gln Asp Gly Lys His His Leu His
1 5 10 15
Pro Phe Ser Asp Asn Ala Ala Leu Ala Arg Glu Gly Val Arg Val Met
20 25 30
Val Lys Ala Lys Gly Ile Tyr Val Tyr Asp Asp Glu Gly Asn Glu Ile
35 40 45
Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg
50 55 60
Lys Glu Leu Ala Lys Val Ala Lys Lys Gln Met Lys Glu Leu Pro Phe
65 70 75 80
Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val Val Glu Leu Ser
85 90 95
Ala Thr Ile Thr Asp Ile Ala Pro Ser Gly Phe Asn His Val Phe Tyr
100 105 110
Thr Gly Ser Gly Ser Glu Ser Val Asp Thr Met Met Arg Met Val Arg
115 120 125
His Tyr Trp Ala Ser Val Gly Lys Pro Gln Lys Lys Val Ile Ile Gly
130 135 140
Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser Leu Gly
145 150 155 160
Gly Met Lys Gly Met His Ser Gln Gly Asp Leu Pro Ile Pro Asn Val
165 170 175
Val His Ile Glu Gln Pro Trp Tyr Tyr Gly Leu Ala Lys Asp Gly Glu
180 185 190
Ser Pro Glu Glu Phe Gly Leu Arg Ala Ala Asn Trp Leu Glu Glu Lys
195 200 205
Ile Glu Glu Ile Gly Ala Asp Lys Ile Ala Ala Phe Val Gly Glu Pro
210 215 220
Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Lys Thr Tyr Trp Pro
225 230 235 240
Arg Ile Glu Glu Ile Cys Arg Lys His Asp Ile Leu Leu Val Ala Asp
245 250 255
Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Trp Phe Gly His Glu
260 265 270
Thr Met Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys Gly Leu Ser
275 280 285
Ser Gly Tyr Leu Pro Ile Gly Ala Val Leu Val Asn Asp Lys Val Ala
290 295 300
Glu Gly Leu Lys Ala Gly Gly Glu Phe Asn His Gly Phe Thr Tyr Ser
305 310 315 320
Gly His Pro Val Ala Ala Ala Val Ala Lys Lys Asn Leu Glu Ile Leu
325 330 335
Gln Asp Glu Gly Ile Val Asp Gln Val Lys Asn Glu Thr Gly Pro Tyr
340 345 350
Met Lys Lys Arg Trp Glu Glu Thr Phe Ser Gln Phe Lys Tyr Val Asp
355 360 365
Asp Ile Arg Ser Glu Gly Leu Ile Cys Gly Phe Thr Leu Val Lys Asp
370 375 380
Lys Ala Arg Arg Glu Leu Phe Pro Asn Thr Gly Glu Thr Gly Leu Met
385 390 395 400
Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg Ala Cys Gly
405 410 415
Asp His Met Val Ala Ala Pro Pro Leu Val Met Thr Lys Lys Glu Ile
420 425 430
Asp Lys Met Leu Ala Val Ala Glu Gln Cys Met Arg Glu Phe Glu Gln
435 440 445
Met Met Asp Lys Lys Leu Ala Lys Glu Glu Lys Ala Gly Lys Lys Glu
450 455 460
Glu Lys Ser Asp Gln
465
<210> 49
<211> 478
<212> PRT
<213> Vitreoscilla sp. C1
<400> 49
Met Tyr Lys Lys Ser Thr Ile Arg Ile Phe Ala Thr Thr Ser His Lys
1 5 10 15
Met Asn Ile Asp Lys Glu Ile Asn Met Thr Asn Thr Thr Leu Asn Ala
20 25 30
Thr Ala Asn His Leu His Pro Phe Ser Asp Asn Gln Gln Leu Lys Glu
35 40 45
Lys Gly Val Arg Val Ile Thr Lys Ala Asp Gly Ile Tyr Ile Tyr Asp
50 55 60
Asp Ala Gly Asn Lys Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val
65 70 75 80
Asn Ile Gly Tyr Gly Arg Lys Glu Leu Ala Asp Ile Ala Arg Asp Gln
85 90 95
Met Asn Glu Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Ser His Pro
100 105 110
Ala Val Ile Asn Leu Ser Ala Lys Leu Ala Ser Ile Ala Pro Ala Gly
115 120 125
Phe Asn His Val Phe Tyr Thr Gly Ser Gly Ser Glu Ser Val Asp Ser
130 135 140
Met Ile Arg Met Val Arg His Tyr Trp Asp Ser Val Gly Lys Pro Ser
145 150 155 160
Lys Lys Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile
165 170 175
Gly Gly Val Ser Met Gly Gly Met Lys Gly Met His Lys Gln Gly Gly
180 185 190
Phe Pro Ile Glu Gly Ile Val His Ile Glu Gln Pro Trp Phe Tyr Gly
195 200 205
Leu His Glu Glu Gly Glu Thr Ala Glu Gln Phe Gly Ile Arg Ala Ala
210 215 220
Asn Trp Leu Glu Glu Lys Ile Leu Glu Ile Gly Ala Asp Asn Val Ala
225 230 235 240
Ala Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro
245 250 255
Pro Ser Thr Tyr Trp Pro Arg Ile Gln Glu Ile Cys Gln Lys Tyr Asp
260 265 270
Ile Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly
275 280 285
Glu Trp Phe Gly His Glu Thr Phe Gly Phe Gln Pro Asp Ile Phe Thr
290 295 300
Thr Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Ala Val Phe
305 310 315 320
Val Gly Asp Lys Val Val Glu Gly Ile Leu Lys Gly Gly Asp Phe Asn
325 330 335
His Gly Phe Thr Tyr Ala Gly His Pro Val Ala Ala Ala Val Ala Leu
340 345 350
Lys Asn Val Glu Ile Leu Asp Asp Glu Gly Ile Ile Lys Asn Val His
355 360 365
Asp Lys Thr Gly Pro Tyr Met Gln Ala Lys Trp Arg Glu Met Leu Ser
370 375 380
Gln Phe Lys Tyr Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Gly
385 390 395 400
Phe Thr Leu Val Lys Asn Lys Ala Thr Arg Glu Met Phe Pro Asn Glu
405 410 415
Gly Glu Thr Gly Thr Met Leu Arg Asp Ile Phe Phe Ser Asn Asn Leu
420 425 430
Ile Met Arg Ala Cys Gly Asp His Ile Val Ala Ser Pro Pro Leu Thr
435 440 445
Ile Thr Lys Glu Glu Ile Asp Leu Met Leu Ala Thr Ala Glu Lys Cys
450 455 460
Leu Gln Glu Phe Glu Thr Ala Met Asp Ser Lys Leu Gly Ala
465 470 475
<210> 50
<211> 454
<212> PRT
<213> Vitreoscilla
<400> 50
Met Thr Asn Thr Thr Leu Asn Ala Thr Ala Asn His Leu His Pro Phe
1 5 10 15
Ser Asp Asn Gln Gln Leu Lys Glu Lys Gly Val Arg Val Ile Thr Lys
20 25 30
Ala Asp Gly Ile Tyr Ile Tyr Asp Asp Ala Gly Asn Lys Ile Ile Asp
35 40 45
Gly Met Ala Gly Leu Trp Cys Val Asn Ile Gly Tyr Gly Arg Lys Glu
50 55 60
Leu Ala Asp Ile Ala Arg Asp Gln Met Asn Glu Leu Ala Tyr Tyr Asn
65 70 75 80
Thr Phe Phe Lys Thr Ser His Pro Ala Val Ile Asn Leu Ser Ala Lys
85 90 95
Leu Ala Ser Ile Ala Pro Ala Gly Phe Asn His Val Phe Tyr Thr Gly
100 105 110
Ser Gly Ser Glu Ser Val Asp Ser Met Ile Arg Met Val Arg His Tyr
115 120 125
Trp Asp Ser Val Gly Lys Pro Ser Lys Lys Thr Leu Ile Gly Arg Trp
130 135 140
Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Val Ser Met Gly Gly Met
145 150 155 160
Lys Gly Met His Lys Gln Gly Gly Phe Pro Ile Glu Gly Ile Val His
165 170 175
Ile Glu Gln Pro Trp Phe Tyr Gly Leu His Glu Glu Gly Glu Thr Ala
180 185 190
Glu Gln Phe Gly Ile Arg Ala Ala Asn Trp Leu Glu Glu Lys Ile Leu
195 200 205
Glu Ile Gly Ala Asp Asn Val Ala Ala Phe Val Gly Glu Pro Ile Gln
210 215 220
Gly Ala Gly Gly Val Ile Ile Pro Pro Ser Thr Tyr Trp Pro Arg Ile
225 230 235 240
Gln Glu Ile Cys Gln Lys Tyr Asp Ile Leu Leu Val Ala Asp Glu Val
245 250 255
Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe Gly His Glu Thr Phe
260 265 270
Gly Phe Gln Pro Asp Ile Phe Thr Thr Ala Lys Gly Leu Ser Ser Gly
275 280 285
Tyr Leu Pro Ile Gly Ala Val Phe Val Gly Asp Lys Val Val Glu Gly
290 295 300
Ile Leu Lys Gly Gly Asp Phe Asn His Gly Phe Thr Tyr Ala Gly His
305 310 315 320
Pro Val Ala Ala Ala Val Ala Leu Lys Asn Val Glu Ile Leu Asp Asp
325 330 335
Glu Gly Ile Ile Lys Asn Val His Asp Lys Thr Gly Pro Tyr Met Gln
340 345 350
Ala Lys Trp Arg Glu Met Leu Ser Gln Phe Lys Tyr Val Asp Asp Val
355 360 365
Arg Gly Val Gly Leu Ile Gln Gly Phe Thr Leu Val Lys Asn Lys Ala
370 375 380
Thr Arg Glu Met Phe Pro Asn Glu Gly Glu Thr Gly Thr Met Leu Arg
385 390 395 400
Asp Ile Phe Phe Ser Asn Asn Leu Ile Met Arg Ala Cys Gly Asp His
405 410 415
Ile Val Ala Ser Pro Pro Leu Thr Ile Thr Lys Glu Glu Ile Asp Leu
420 425 430
Met Leu Ala Thr Ala Glu Lys Cys Leu Gln Glu Phe Glu Thr Ala Met
435 440 445
Asp Ser Lys Leu Gly Ala
450
<210> 51
<211> 462
<212> PRT
<213> Laribacter hongkongensis
<400> 51
Met Thr Ala Asp Pro Arg Arg Thr Glu His Tyr Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Ser Asp Met Glu Ala Leu Asn Arg Gln
20 25 30
Gly Ser Arg Val Met Val Arg Gly Glu Gly Ile His Val Trp Asp Ser
35 40 45
Asp Gly Val Arg Leu Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Leu Gly Tyr Gly Arg Arg Asp Leu Ala Glu Ala Ala Tyr Ala Gln Leu
65 70 75 80
Cys Glu Leu Pro Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Arg
85 90 95
Val Ala Glu Leu Ser Ala Leu Val Ala Glu Val Ala Pro Pro Gly Phe
100 105 110
Ser His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Met Arg Met Val Arg His Tyr Trp Ala Leu Lys Gly Gln Pro Gln Lys
130 135 140
Lys Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Leu Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Pro Ala Met His Ala Gln Gly Asp Leu
165 170 175
Pro Val Ala Asp Val Val His Val Glu Gln Pro Trp Phe Leu Lys His
180 185 190
Gly Leu Asp Gly Glu Thr Glu Glu Ala Phe Gly Leu Arg Ala Ala Arg
195 200 205
Trp Val Glu Glu Lys Ile Leu Glu Thr Gly Ala Asp Arg Val Ala Ala
210 215 220
Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro
225 230 235 240
Ala Ser Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Arg His Asp Val
245 250 255
Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu
260 265 270
Trp Phe Gly His Gln Leu Tyr Gly Ile Arg Pro Asp Leu Phe Thr Val
275 280 285
Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Gly Val Val Val
290 295 300
Gly Asp Glu Val Ala Gln Val Ile Lys Ala Gly Gly Asp Phe Asn His
305 310 315 320
Gly Phe Thr Tyr Ser Gly His Pro Ala Cys Ala Ala Val Ala Ala Ala
325 330 335
Asn Val Arg Ala Leu Arg Asp Glu Gly Val Val Glu Arg Val Arg Gln
340 345 350
Asp Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly
355 360 365
Phe Ala His Leu Ala Asp Ile Arg Gly Val Gly Leu Ile Gln Gly Phe
370 375 380
Thr Trp Val Gln Asp Lys Glu Leu Arg Arg Leu Phe Pro Asp Trp Gly
385 390 395 400
Glu Val Gly Leu Leu Cys Arg Asp Val Phe Phe Arg His Asn Leu Ile
405 410 415
Met Arg Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile
420 425 430
Asn Arg Asp Glu Val Asp Ala Met Leu Glu Val Ala Val Gln Cys Val
435 440 445
Ala Glu Thr Glu Ser Val Leu Arg Glu Arg Gly Leu Ile Asp
450 455 460
<210> 52
<211> 462
<212> PRT
<213> Laribacter hongkongensis
<400> 52
Met Thr Ala Asp Pro Arg Arg Thr Glu His Tyr Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Ser Asp Met Glu Ala Leu Asn Arg Gln
20 25 30
Gly Ser Arg Val Met Val Arg Gly Glu Gly Ile His Val Trp Asp Ser
35 40 45
Asp Gly Val Arg Leu Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Leu Gly Tyr Gly Arg Arg Asp Leu Ala Glu Ala Ala Tyr Ala Gln Leu
65 70 75 80
Cys Glu Leu Pro Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Arg
85 90 95
Val Ala Glu Leu Ser Ala Leu Val Ala Glu Val Ala Pro Pro Gly Phe
100 105 110
His His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Met Arg Met Val Arg His Tyr Trp Ala Leu Lys Gly Gln Pro Gln Lys
130 135 140
Lys Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Leu Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Pro Ala Met His Ala Gln Gly Asp Leu
165 170 175
Pro Val Ala Asp Val Val His Val Glu Gln Pro Trp Phe Leu Lys His
180 185 190
Gly Leu Asp Gly Glu Thr Glu Glu Ala Phe Gly Leu Arg Ala Ala Arg
195 200 205
Trp Val Glu Glu Lys Ile Leu Glu Thr Gly Ala Asp Arg Val Ala Ala
210 215 220
Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro
225 230 235 240
Ala Ser Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Arg His Asp Val
245 250 255
Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu
260 265 270
Trp Phe Gly His Gln Leu Tyr Gly Ile Arg Pro Asp Leu Phe Thr Val
275 280 285
Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Gly Val Val Val
290 295 300
Gly Asp Ala Val Ala Gln Val Ile Lys Ala Gly Gly Asp Phe Asn His
305 310 315 320
Gly Phe Thr Tyr Ser Gly His Pro Ala Cys Ala Ala Val Ala Ala Ala
325 330 335
Asn Val Arg Ala Leu Arg Asp Glu Gly Val Val Glu Arg Val Arg Gln
340 345 350
Asp Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly
355 360 365
Phe Ala His Leu Ala Asp Ile Arg Gly Val Gly Leu Ile Gln Gly Phe
370 375 380
Thr Trp Val Gln Asp Lys Glu Leu Arg Arg Leu Phe Pro Asp Trp Gly
385 390 395 400
Glu Val Gly Leu Leu Cys Arg Asp Val Phe Phe Arg His Asn Leu Ile
405 410 415
Met Arg Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile
420 425 430
Asn Arg Asp Glu Val Asp Ala Met Leu Glu Val Ala Val Gln Cys Val
435 440 445
Ala Glu Thr Glu Ser Val Leu Arg Glu Arg Gly Leu Ile Asp
450 455 460
<210> 53
<211> 462
<212> PRT
<213> Rivicola pingtungensis
<400> 53
Met Thr Ala Asp Pro Arg Arg Thr Glu His Tyr Arg Glu Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Ser Asp Met Glu Ala Leu Asn Arg Gln
20 25 30
Gly Ser Arg Val Met Val Arg Gly Glu Gly Ile His Val Trp Asp Ser
35 40 45
Asp Gly Val Arg Leu Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Leu Gly Tyr Gly Arg Arg Asp Leu Ala Glu Ala Ala Tyr Ala Gln Leu
65 70 75 80
Cys Glu Leu Pro Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Arg
85 90 95
Val Ala Glu Leu Ser Ala Leu Val Ala Glu Val Ala Pro Pro Gly Phe
100 105 110
His His Val Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met
115 120 125
Met Arg Met Val Arg His Tyr Trp Ala Leu Lys Gly Gln Pro Gln Lys
130 135 140
Lys Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Leu Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Pro Ala Met His Ala Gln Gly Asp Leu
165 170 175
Pro Val Ala Asp Val Val His Val Glu Gln Pro Trp Phe Leu Lys His
180 185 190
Gly Leu Asp Gly Glu Thr Glu Glu Ala Phe Gly Leu Arg Ala Ala Arg
195 200 205
Trp Val Glu Glu Lys Ile Leu Glu Thr Gly Ala Asp Arg Val Ala Ala
210 215 220
Phe Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro
225 230 235 240
Ala Ser Tyr Trp Pro Glu Val Glu Arg Ile Cys Arg Arg His Asp Val
245 250 255
Leu Leu Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu
260 265 270
Trp Phe Gly His Gln Leu Tyr Gly Ile Arg Pro Asp Leu Phe Thr Val
275 280 285
Ala Lys Gly Leu Ser Ser Gly Tyr Leu Pro Ile Gly Gly Val Val Val
290 295 300
Gly Asp Ala Val Ala Gln Val Ile Lys Ala Gly Gly Asp Phe Asn His
305 310 315 320
Gly Phe Thr Tyr Ser Gly His Pro Ala Cys Ala Ala Val Ala Ala Ala
325 330 335
Asn Val Arg Ala Leu Arg Asp Glu Gly Val Val Glu Arg Val Arg Gln
340 345 350
Asp Thr Gly Pro Tyr Met Gln Gln Arg Trp Arg Glu Ala Phe Gly Gly
355 360 365
Phe Ala His Leu Ala Asp Ile Arg Gly Val Gly Leu Ile Gln Gly Phe
370 375 380
Thr Trp Val Gln Asp Lys Glu Leu Arg Arg Leu Phe Pro Asp Trp Gly
385 390 395 400
Glu Val Gly Leu Leu Cys Arg Asp Val Phe Phe Arg His Asn Leu Ile
405 410 415
Met Arg Ala Cys Gly Asp His Met Val Ala Ala Pro Pro Leu Val Ile
420 425 430
Asn Arg Asp Glu Val Asp Ala Met Leu Glu Val Ala Val Gln Cys Val
435 440 445
Ala Glu Thr Glu Ser Val Leu Arg Glu Arg Gly Leu Ile Asp
450 455 460
<210> 54
<211> 460
<212> PRT
<213> Stenoxybacter acetivorans
<400> 54
Met Ser Lys Lys Asn Tyr Ser Lys His Asp Val Lys His His Leu His
1 5 10 15
Pro Phe Ser Asp Asn Ala Ala Leu Gln Glu Glu Gly Val Arg Val Met
20 25 30
Val Lys Ala Lys Gly Val Tyr Val Tyr Asp Asn Asp Gly Asn Glu Ile
35 40 45
Leu Asp Gly Met Ala Gly Leu Trp Cys Val Asn Val Gly Tyr Gly Arg
50 55 60
Lys Glu Leu Val Arg Ala Ala Lys Lys Gln Met Glu Ile Leu Pro Phe
65 70 75 80
Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Ala Val Glu Leu Thr
85 90 95
Asp Lys Leu Val Glu Ile Thr Pro Glu Gly Phe Asn His Val Phe Tyr
100 105 110
Thr Gly Ser Gly Ser Glu Ser Val Asp Thr Met Met Arg Met Val Arg
115 120 125
His Tyr Trp Ala Ser Ile Gly Lys Pro Glu Lys Lys Ile Ile Ile Gly
130 135 140
Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly Ala Ser Leu Gly
145 150 155 160
Gly Met Lys Gly Met His Glu Gln Gly Asp Leu Pro Ile Pro Asn Val
165 170 175
Val His Ile Glu Gln Pro Trp Tyr Tyr Gly Leu Ala Lys Asp Asn Glu
180 185 190
Thr Ala Gly Glu Phe Gly Ile Arg Ala Ala Asn Trp Leu Glu Glu Lys
195 200 205
Ile Thr Glu Leu Gly Ala Asp Lys Val Ala Ala Phe Val Gly Glu Pro
210 215 220
Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Asp Gly Tyr Trp Gln
225 230 235 240
Arg Ile Glu Glu Ile Cys Arg Lys His Asp Ile Leu Leu Val Ala Asp
245 250 255
Glu Val Ile Cys Gly Tyr Gly Arg Thr Gly Asn Trp Phe Gly His Glu
260 265 270
Thr Phe Gly Phe Lys Pro Asp Ile Phe Thr Thr Ala Lys Gly Leu Ser
275 280 285
Ser Gly Tyr Ile Pro Ile Gly Ala Val Leu Val Asn Asp Arg Val Thr
290 295 300
Glu Gly Leu Leu Ala Gly Gly Glu Phe Asn His Gly Phe Thr Tyr Ser
305 310 315 320
Gly His Pro Val Ala Ala Ala Val Ala Arg Ala Asn Leu Asp Ile Leu
325 330 335
Asp Asn Glu Glu Ile Val Asp Arg Ile His Asp Lys Ile Gly Pro Tyr
340 345 350
Met Lys Lys Arg Trp Gln Glu Thr Phe Glu Pro Phe Lys Tyr Val Gly
355 360 365
Asp Ile Arg Ser Val Gly Leu Met Cys Gly Phe Thr Leu Val Lys Asn
370 375 380
Lys Glu Thr Arg Glu Phe Phe Pro Asn Thr Gly Glu Thr Gly Leu Ile
385 390 395 400
Cys Arg Asp Ile Phe Phe Lys Asn Asn Leu Ile Met Arg Ala Cys Gly
405 410 415
Asp His Ile Val Ala Ala Pro Pro Leu Val Ile Ser Lys Lys Glu Ile
420 425 430
Asp His Met Leu Ser Ile Ala Ala Lys Cys Met Arg Glu Phe Glu Val
435 440 445
Lys Met Asp Gln Ile Leu Ala Lys Gln Asp Gln Asp
450 455 460
<210> 55
<211> 461
<212> PRT
<213> Aquaspirillum sp. LM1
<400> 55
Met Thr Leu Ala Thr Arg Ser Thr Ala Asp Trp Gln Ala Leu Asp Ala
1 5 10 15
Ala His His Leu His Pro Phe Ser Asp Thr Gln Ser Leu Asn Gln Thr
20 25 30
Gly Ser Arg Val Val Val Lys Gly Asp Gly Val Tyr Ile Trp Asp Ser
35 40 45
Glu Gly Asn Arg Ile Ile Asp Gly Met Ala Gly Leu Trp Cys Val Asn
50 55 60
Leu Gly Tyr Gly Arg Glu Glu Leu Ala Asp Ala Ala Tyr Gln Gln Met
65 70 75 80
Arg Glu Leu Pro Phe Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Pro
85 90 95
Val Ile Glu Leu Ser Ala Leu Leu Ala Glu Leu Thr Pro Pro Gln Phe
100 105 110
Asn His Val Phe Leu Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val
115 120 125
Ile Arg Met Val Arg His Phe Trp Ala Ala Gln Gly Gln Ala Ser Lys
130 135 140
Lys Val Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly
145 150 155 160
Gly Ala Ser Leu Gly Gly Met Arg Tyr Met His Glu Gln Gly Asp Leu
165 170 175
Pro Ile Pro Gly Ile Glu His Ile Glu Gln Pro Tyr Trp Tyr Val Asn
180 185 190
Gly Gly Glu Leu Ser Pro His Asp Tyr Gly Leu Lys Ala Ala Gln Ala
195 200 205
Leu Glu Ala Lys Ile Leu Ala Leu Gly Ala Asp Asn Val Ala Ala Phe
210 215 220
Val Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Val Pro Pro Asp
225 230 235 240
Ser Tyr Trp Pro Glu Ile Gln Arg Ile Cys Arg Ala Tyr Asn Ile Leu
245 250 255
Leu Val Val Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Gln Trp
260 265 270
Phe Gly Ser Gln His Phe Gly Ile Glu Pro Asp Leu Met Pro Ile Ala
275 280 285
Lys Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Gly Val Met Val Ser
290 295 300
Asp Arg Val Ala Gln Val Leu Ile Gln Glu Gly Gly Asp Phe Asn His
305 310 315 320
Gly Met Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Val Ala
325 330 335
Asn Leu Lys Ile Leu Arg Asp Glu Gly Ile Val Asp Gln Val Arg Glu
340 345 350
Lys Thr Gly Pro Tyr Leu Gln Gln Lys Trp Arg Glu Thr Phe Ala Gly
355 360 365
His Pro Tyr Leu Asp Asp Val Arg Gly Val Gly Phe Leu Ala Ala Phe
370 375 380
Thr Trp Val Lys Asp Lys Ser Arg Arg Glu Leu Phe Ala Glu Pro Gly
385 390 395 400
Thr Val Gly Thr Ile Cys Arg Asp Ile Phe Phe Lys Asn Asn Ile Ile
405 410 415
Met Arg Ala Val Gly Asp Ser Met Val Cys Ala Pro Pro Leu Val Met
420 425 430
Thr Thr Thr Glu Ile Asp Glu Met Ile Ala Leu Ala Ala Arg Cys Val
435 440 445
Asp Leu Thr Val Gln Ala Leu Glu Ala Arg Gly Leu Val
450 455 460
<210> 56
<211> 459
<212> PRT
<213> Paludibacterium yongneupense
<400> 56
Met Ala Gln Glu Arg Ser Thr Gln Gln Trp Arg Asp Met Asp Ala Ala
1 5 10 15
His His Leu His Pro Phe Thr Asp Thr Asp Ser Leu Asn Arg His Gly
20 25 30
Ala Arg Val Ile Thr His Gly Glu Gly Ile Tyr Leu Trp Asp Ser Asp
35 40 45
Gly Asn Lys Ile Thr Asp Ala Met Ala Gly Leu Trp Cys Val Asn Leu
50 55 60
Gly Tyr Gly Arg Lys Asp Leu Ala Leu Val Ala Gln Gln Gln Met Glu
65 70 75 80
Thr Leu Ala Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His Pro Ala Val
85 90 95
Thr Glu Leu Ser Ala Leu Leu Ser Glu Ile Ala Pro Pro Gly Phe Ser
100 105 110
Arg Met Phe Tyr Thr Asn Ser Gly Ser Glu Ala Val Asp Thr Met Ile
115 120 125
Arg Met Val Arg Arg Tyr Trp Asp Val Gln Gly Gln Pro Gln Lys Lys
130 135 140
Thr Leu Ile Gly Arg Trp Asn Gly Tyr His Gly Ser Thr Ile Gly Gly
145 150 155 160
Ala Ser Leu Gly Gly Met Gly Tyr Met His Glu Gln Gly Asp Leu Pro
165 170 175
Ile Pro Gly Ile Ala His Ile Glu Gln Pro Trp Trp Tyr Gly Leu Gly
180 185 190
Gly Asp Ser Ser Arg Glu Ala Phe Gly Leu His Ala Ala Gly Trp Leu
195 200 205
Glu Gln Lys Ile Leu Glu Ile Gly Ala Asp Lys Val Ala Ala Phe Val
210 215 220
Gly Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Ala Ser
225 230 235 240
Tyr Trp Pro Glu Ile Glu Arg Ile Cys Arg Lys Tyr Asp Val Leu Leu
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly His Gln His Phe Gly Phe His Pro Asp Leu Phe Thr Thr Ala Lys
275 280 285
Gly Leu Ser Ser Gly Tyr Gln Pro Ile Gly Ala Val Phe Val Gly Asp
290 295 300
Arg Val Ala Glu Gly Leu Ala Arg Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Arg Ala Thr Val
325 330 335
Arg Ala Leu Arg Asp Asp Gly Ile Val Asp Arg Val Lys Asn Asp Val
340 345 350
Gly Pro Tyr Met Gln Glu Arg Trp Arg Glu Thr Phe Glu Gly Phe Asp
355 360 365
His Val Asp Asp Val Arg Gly Val Gly Leu Ile Gln Gly Phe Thr Leu
370 375 380
Val Arg Asn Arg Asp Thr Arg Glu Leu Phe Pro Asp Phe Gly Asp Ala
385 390 395 400
Gly Thr Leu Cys Arg Asp Ile Phe Phe Gly Asn Asn Leu Ile Met Arg
405 410 415
Ala Cys Gly Asp His Met Val Ala Ser Pro Pro Leu Val Val Thr Arg
420 425 430
Lys Glu Ile Asp Gln Met Leu Gly Val Ala Gln Arg Cys Val Glu Glu
435 440 445
Phe Glu Arg Glu Ile Val Arg Arg Gly Met Val
450 455

Claims (9)

1.一种转氨酶突变体，其特征在于，所述转氨酶突变体：（1）由SEQ ID NO: 2发生氨基酸突变得到，所述氨基酸突变选自如下任意一种：
Figure 953957DEST_PATH_IMAGE001
Figure 965776DEST_PATH_IMAGE002
Figure 54955DEST_PATH_IMAGE003
Figure 432758DEST_PATH_IMAGE004
Figure 658203DEST_PATH_IMAGE005
Figure 840923DEST_PATH_IMAGE006
Figure 417398DEST_PATH_IMAGE007
；
或者
（2）由野生型转氨酶的保守氨基酸突变得到，以野生型CvTA转氨酶的序列SEQ ID NO：1作为参照，所述保守氨基酸发生如下任一种组合突变：（a）L59A+F88A；（b）L59A+F88A+F89D；（c）L59A+F88A+F89D+N86H；（d）L59A+F88A+F89D+N86H +Y85M，所述野生型转氨酶来源于下表所示的酶编号为1至54中的任意一种：
Figure 360077DEST_PATH_IMAGE008
Figure 440029DEST_PATH_IMAGE009
。
2.一种DNA分子，其特征在于，所述DNA分子编码权利要求1所述的转氨酶突变体。
3.一种重组质粒，其特征在于，所述重组质粒连接有权利要求2所述的DNA分子。
4.一种手性胺化合物的合成方法，采用权利要求1所述的转氨酶突变体在氨基供体的作用下对式I所示酮类底物进行转氨基反应，得到所述手性胺化合物；
Figure 793650DEST_PATH_IMAGE010
式I，
Ar1表示取代或未取代的芳基或亚芳基，或者取代或未取代的杂亚芳基；
A r2表示取代或未取代的芳基，或者环烷基；以及
可选的R，R表示C原子数为1~5的取代或未取代的亚烷基，所述R与所述Ar1或A r2连接成环；
其中，所述取代的芳基或亚芳基、所述取代的杂亚芳基或所述取代的亚烷基的取代基选自卤素、羟基或氨基，所述取代的杂亚芳基中的杂原子选自N、O或S。
5.根据权利要求4所述的合成方法，所述取代的芳基或亚芳基中的取代基为卤素或-S-CH3，所述卤素或-S-CH3位于所述芳基或亚芳基的邻位、间位或对位中的任意一个或多个位置。
6.根据权利要求5所述的合成方法，所述卤素为F、Cl或Br。
7.根据权利要求4所述的合成方法，式I所示酮类底物为如下任意一种：
i) 所述Ar1表示取代或未取代的芳基或亚芳基，所述Ar2表示未取代的芳基，所述R表示C原子数为1~5的未取代的亚烷基，且所述R与所述Ar1连接成环；
ii) 所述Ar1表示未取代的杂亚芳基，所述Ar2表示卤素取代的芳基，所述R表示羟基取代的C原子数为1~5的亚烷基，且所述R与所述Ar1连接成环；
iii) 所述Ar1表示卤素和氨基取代的亚芳基，所述Ar2为碳原子数为3~8的环烷基；
iv) 所述Ar1表示未取代的环烷基或芳基，所述Ar2为取代或未取代的环烷基或芳基；
v) 所述Ar1表示羟基、甲基、乙基或-S-CH3取代的环烷基或芳基，所述Ar2为未取代的芳基或烷基。
8.根据权利要求4所述的合成方法，所述酮类底物为：
Figure 857421DEST_PATH_IMAGE011
9.根据权利要求4所述的合成方法，所述氨基供体为异丙胺、异丙胺盐酸盐、丙氨酸、正丁胺或苯胺。
CN202110451381.5A 2021-02-04 2021-04-26 转氨酶突变体及其应用 Active CN112980810B (zh)

Priority Applications (5)

Application Number Priority Date Filing Date Title
JP2023547522A JP2024506887A (ja) 2021-02-04 2021-07-06 トランスアミナーゼ突然変異体及びその応用
US18/264,091 US20240124853A1 (en) 2021-02-04 2021-07-06 Transaminase mutants and uses thereof
EP21924110.6A EP4273237A1 (en) 2021-02-04 2021-07-06 Transaminase mutant and application thereof
PCT/CN2021/104841 WO2022166103A1 (zh) 2021-02-04 2021-07-06 转氨酶突变体及其应用
KR1020237030000A KR20230141852A (ko) 2021-02-04 2021-07-06 트랜스아미나제 돌연변이체 및 이의 응용

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN2021101507600 2021-02-04
CN202110150760.0A CN112501145A (zh) 2021-02-04 2021-02-04 转氨酶突变体及其应用

Publications (2)

Publication Number Publication Date
CN112980810A true CN112980810A (zh) 2021-06-18
CN112980810B CN112980810B (zh) 2021-08-20

Family

ID=74952866

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202110150760.0A Withdrawn CN112501145A (zh) 2021-02-04 2021-02-04 转氨酶突变体及其应用
CN202110451381.5A Active CN112980810B (zh) 2021-02-04 2021-04-26 转氨酶突变体及其应用

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN202110150760.0A Withdrawn CN112501145A (zh) 2021-02-04 2021-02-04 转氨酶突变体及其应用

Country Status (6)

Country Link
US (1) US20240124853A1 (zh)
EP (1) EP4273237A1 (zh)
JP (1) JP2024506887A (zh)
KR (1) KR20230141852A (zh)
CN (2) CN112501145A (zh)
WO (1) WO2022166103A1 (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114875006A (zh) * 2022-06-21 2022-08-09 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及手性胺化合物的制备方法
WO2022166103A1 (zh) * 2021-02-04 2022-08-11 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
WO2023050516A1 (zh) * 2021-09-28 2023-04-06 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
CN116083385A (zh) * 2022-12-02 2023-05-09 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109370998A (zh) * 2018-11-30 2019-02-22 江南大学 一种催化效率提高的ω-转氨酶突变体I215F
WO2019095161A1 (zh) * 2017-11-15 2019-05-23 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及其应用
CN110205310A (zh) * 2019-05-30 2019-09-06 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及其应用
CN110628742A (zh) * 2019-10-25 2019-12-31 凯莱英医药化学（阜新）技术有限公司 转氨酶突变体及其应用
US20210062233A1 (en) * 2017-09-01 2021-03-04 Vito Nv Method for producing chiral amines

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011005477A1 (en) * 2009-06-22 2011-01-13 Codexis, Inc. Transaminase reactions
CN111032682B (zh) * 2017-06-14 2024-04-12 科德克希思公司 用于工业生物催化的工程化转氨酶多肽
CN107828751B (zh) * 2017-11-06 2021-02-26 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及其应用
CN109957554B (zh) * 2017-12-26 2021-05-07 宁波酶赛生物工程有限公司 工程化转氨酶多肽及其应用
CN110592042B (zh) * 2019-10-28 2021-04-23 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
CN112501145A (zh) * 2021-02-04 2021-03-16 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20210062233A1 (en) * 2017-09-01 2021-03-04 Vito Nv Method for producing chiral amines
WO2019095161A1 (zh) * 2017-11-15 2019-05-23 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及其应用
CN109370998A (zh) * 2018-11-30 2019-02-22 江南大学 一种催化效率提高的ω-转氨酶突变体I215F
CN110205310A (zh) * 2019-05-30 2019-09-06 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及其应用
CN110628742A (zh) * 2019-10-25 2019-12-31 凯莱英医药化学（阜新）技术有限公司 转氨酶突变体及其应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ANDREW等: "Transaminases for chiral amine synthesis", 《CURRENT OPINION IN CHEMICAL BIOLOGY》 *
LAND等: "Engineering the Active Site of an (S)-Selective Amine Transaminase for Acceptance of Doubly Bulky Primary Amines", 《ADV. SYNTH. CATAL》 *
刘婉颐等: "生物催化合成手性胺的研究进展", 《合成化学》 *
高新星等: "转氨酶分子改造研究进展", 《生物工程学报》 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022166103A1 (zh) * 2021-02-04 2022-08-11 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
WO2023050516A1 (zh) * 2021-09-28 2023-04-06 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
CN114875006A (zh) * 2022-06-21 2022-08-09 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及手性胺化合物的制备方法
WO2023245815A1 (zh) * 2022-06-21 2023-12-28 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及手性胺化合物的制备方法
CN114875006B (zh) * 2022-06-21 2024-03-12 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及手性胺化合物的制备方法
CN116083385A (zh) * 2022-12-02 2023-05-09 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及应用
WO2024113508A1 (zh) * 2022-12-02 2024-06-06 凯莱英生命科学技术(天津)有限公司 转氨酶突变体及应用

Also Published As

Publication number Publication date
EP4273237A1 (en) 2023-11-08
KR20230141852A (ko) 2023-10-10
JP2024506887A (ja) 2024-02-15
WO2022166103A1 (zh) 2022-08-11
CN112980810B (zh) 2021-08-20
CN112501145A (zh) 2021-03-16
US20240124853A1 (en) 2024-04-18

Similar Documents

Publication Publication Date Title
CN112980810B (zh) 转氨酶突变体及其应用
CN107828751B (zh) 转氨酶突变体及其应用
CN112980899B (zh) 手性胺的合成方法
KR101910259B1 (ko) 트랜스아미나제 및 그 응용
CN113817702B (zh) 转氨酶突变体及其应用
CN110628742B (zh) 转氨酶突变体及其应用
CN114875006B (zh) 转氨酶突变体及手性胺化合物的制备方法
CN116083385A (zh) 转氨酶突变体及应用
CN111235127B (zh) 转氨酶突变体及其应用
CN115772514A (zh) 一种腈水合酶底物通道氨基酸基序的改造用于肉桂酰胺的制备
Xie et al. Vitreoscilla hemoglobin: a natural carbene transfer catalyst for diastereo-and enantioselective synthesis of nitrile-substituted cyclopropanes
CN109647489B (zh) 一种银修饰改性石墨相氮化碳材料及其在光催化中的应用
CN109647488B (zh) 一种改性Ag/g-C3N4复合材料的制备方法及其在光催化中的应用
CN114525265B (zh) 转氨酶突变体及其应用
CN116411033A (zh) 一种来源于巨大芽孢杆菌的氨基转移酶突变体及其应用
CN117230129A (zh) 一种使用氨基转移酶或其突变体制备手性芳基胺类化合物的方法
JP2023514862A (ja) キラルジアミン化合物の合成方法
CN116555211A (zh) 一种使用氨基转移酶突变体制备手性内酰胺类化合物的方法
CN114075557B (zh) 重组转氨酶及其在合成（r）-2-（2,5-二氟苯基）吡咯烷中的应用
CN116555366A (zh) 一种使用氨基转移酶突变体制备手性内酰胺类化合物的方法
CN108300744A (zh) D-杂环氨基酸的合成方法、试剂盒及应用
JP2002088090A (ja) 有機ケイ素化合物の製造法
Catal Visible-Light Photocatalysis Enables Anti-Markovnikov Hydrofunctionalization of-Olefins
CN118388391A (zh) 一种手性异吲哚啉类化合物及其制备方法
Lo New challenges for artificial metalloenzymes based on the Biotin-(Strept) avidin technology

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20221219

Address after: 300457 No. 6, Dongting Third Street, Binhai New Area Economic and Technological Development Zone, Tianjin

Patentee after: ASYMCHEM LABORATORIES (TIANJIN) Co.,Ltd.

Patentee after: Shanghai kailaiying Biotechnology Co.,Ltd.

Address before: 300457 No. 6, Dongting Third Street, Binhai New Area Economic and Technological Development Zone, Tianjin

Patentee before: ASYMCHEM LABORATORIES (TIANJIN) Co.,Ltd.