CN114341344A - 用于改进醛脱氢酶活性的工程微生物体和方法 - Google Patents

用于改进醛脱氢酶活性的工程微生物体和方法 Download PDF

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CN114341344A
CN114341344A CN202080046801.5A CN202080046801A CN114341344A CN 114341344 A CN114341344 A CN 114341344A CN 202080046801 A CN202080046801 A CN 202080046801A CN 114341344 A CN114341344 A CN 114341344A
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阿米特·M·沙阿
哈里什·纳加拉让
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Abstract

公开了增加或提高己二胺、己酸或己内酰胺的生物合成的生物合成方法和工程微生物。该工程微生物包括选定的醛脱氢酶活性。

Description

用于改进醛脱氢酶活性的工程微生物体和方法
相关申请的交叉引用
本申请要求于2019年4月24日提交的序列号为62/837,888、2019年6月11日提交的序列号为62/860,123、和2019年6月11日提交的序列号为62/860,160的美国临时专利申请的权益，其公开内容通过引用整体并入本文。
并入序列表
本申请包含标题为“GNO0099WO Sequence Listing2.txt”的序列表，该序列表创建于2020年4月23日，大小为319千字节。该序列表通过引用并入本文。
背景技术
尼龙是可以通过二胺与二羧酸的缩聚或内酰胺的缩聚来合成的聚酰胺。尼龙6,6由己二胺(HMD)和己二酸的反应而产生，尼龙6由己内酰胺开环聚合而产生。因此，己二酸、己二胺和己内酰胺是尼龙生产中的重要中间体。
微生物体已被改造来产生一些尼龙中间体。然而，由于不希望的对途径中间体和最终产品的酶活性，工程微生物体可能产生不期望的副产物。因此，此类副产物和杂质会增加生物合成的化合物的成本和复杂性，并且可能降低所期望的产物的效率或产量。
发明内容
本文提供具有以下途径的非天然存在的微生物有机体：6-氨基己酸途径、己内酰胺途径、己二胺途径、己内酯途径、1,6-己二醇途径或这些途径中的一种或多种的组合。该微生物有机体包含编码醛脱氢酶的至少一种外源核酸，醛脱氢酶与己二酰辅酶A反应以形成己二酸-半缩醛。醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物、或琥珀酰辅酶A和乙酰辅酶A俩底物具有更大的转换数、更大的催化效率或其组合。非天然存在的微生物有机体还可以包括编码产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺、己二胺所必需的酶的另外的外源核酸，酶的量足以产生相应的产物。在一些情况下，这些外源核酸中的一种或多种对于微生物有机体可以是异源的。
还公开了产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺、己二胺的方法。该方法可以包括培养产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺和/或己二胺的非天然存在的微生物有机体，其中微生物有机体表达编码醛脱氢酶的至少一种外源核酸，该醛脱氢酶与己二酰辅酶A反应生成己二酸-半缩醛。该方法包括在产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺、己二胺的条件下培养非天然存在的微生物有机体足够的时间段。
一方面，提供非天然存在的微生物有机体，该非天然存在的微生物有机体包含编码醛脱氢酶的至少一种外源核酸，该醛脱氢酶与己二酰辅酶A反应形成己二酸-半缩醛，其中所述醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物、或琥珀酰辅酶A和乙酰辅酶A俩底物具有更大的催化效率和/或所述醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物、或琥珀酰辅酶A和乙酰辅酶A俩底物具有更高的转换数。
一方面，提供产生己二酸-半缩醛的方法，该方法包括将上述方面和实施方案中任一个的非天然存在的微生物体在足够的时间段和条件下培养以产生己二酸-半缩醛。
一方面，提供产生6-氨基己酸(6ACA)的方法，该方法包括将上述方面和实施方案中任一个的非天然存在的微生物有机体在足够的时间段和条件下培养以产生6ACA。在一些实施例中，该方法还包括从微生物有机体、发酵液或两者中回收6ACA。
一方面，提供产生己二胺的方法，该方法包括将上述方面和实施方案中任一个的非天然存在的微生物有机体在足够的时间段和条件下培养以产生己二胺。在一些实施方案中，该方法还包括从微生物有机体、发酵液或两者中回收己二胺。在一些实施方案中，非天然存在的微生物有机体包含各自编码己二胺途径酶的两个、三个、四个、五个、六个或七个外源核酸序列。
一方面，提供产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺、己二胺的方法，该方法包括将上述方面和实施方案中任一个的非天然存在的微生物有机体在足够的时间段和条件下培养以产生6-氨基己酸、1,6-己二醇、己内酯、己内酰胺和己二胺。在一些实施方案中，该方法还包括从微生物有机体、发酵液或两者中回收6-氨基己酸、1,6-己二醇、己内酯、己内酰胺和己二胺。在一些实施方案中，非天然存在的微生物有机体包含各自编码6-氨基己酸、1,6-己二醇、己内酯、己内酰胺、己二胺途径酶的两个、三个、四个、五个、六个或七个外源核酸序列。
一方面，提供使用所公开的方法合成的生物衍生的6-氨基己酸、己二胺或己内酰胺。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶不包含SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶包含与SEQ ID NOs:4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187和188中的任何一个的至少25、50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％氨基酸序列一致性的氨基酸序列。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶包含与SEQ ID NOs:4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187和188中的任何一个的至少25、50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约65％、70％、75％、80％、85％、90％、95％、99％或100％氨基酸序列一致性的氨基酸序列。在一些实施方案中，所述醛脱氢酶使用NADH作为辅因子。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶包括与SEQ ID NOs:7、28、60、和107中的任何一个的至少25、50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％的氨基酸序列一致性的氨基酸序列。在一些实施方案中，醛脱氢酶包括与SEQ ID NOs:7、28、60和188中的任何一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约65％、70％、75％、80％、85％、90％、95％、99％或100％的氨基酸序列一致性的氨基酸序列。在一些实施方案中，所述醛脱氢酶使用NADH作为辅因子。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶包含与SEQ ID NOs:53、77、82、94和152中的任何一个的至少25、50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％的氨基酸序列一致性的氨基酸序列。在一些实施方案中，所述醛脱氢酶使用NADH、NADPH或两者作为辅因子。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物具有更高的催化效率。在一些实施方案中，所述醛脱氢酶对己二酰辅酶A底物的催化效率是对琥珀酰辅酶A底物的催化效率的至少两倍。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶对己二酰辅酶A底物比对乙酰辅酶A底物具有更高的催化效率。在一些实施方案中，所述醛脱氢酶对己二酰辅酶A底物的催化效率是对乙酰辅酶A底物的催化效率的至少五倍。在一些实施方案中，所述醛脱氢酶对己二酰辅酶A底物比对乙酰辅酶A底物具有更高的转换数。
在一些实施方案中，非天然存在的微生物有机体的醛脱氢酶进一步与6-氨基己酰辅酶A反应以形成6-氨基己酸半缩醛。
在一些实施方案中，包含编码醛脱氢酶的至少一个外源核酸的非天然存在的微生物有机体比对照微生物有机体将更多的己二酰辅酶A转化为己二酸半缩醛，除了所述对照微生物有机体不包含编码醛脱氢酶的外源核酸之外，所述对照微生物有机体与非天然存在的微生物有机体基本相同。
在一些实施方案中，编码与己二酰辅酶A反应以形成己二酸-半缩醛的醛脱氢酶的所述至少一种外源核酸对于微生物有机体是异源的。
在一些实施方案中，非天然存在的微生物有机体包含6-氨基己酸途径。在一些实施方案中，6-氨基己酸途径包括：(i)转氨酶，(ii)6-氨基己酸脱氢酶，或(iii)转氨酶和6-氨基己酸脱氢酶两者。在一些实施方案中，非天然存在的微生物有机体还包含编码一种或多种6-氨基己酸途径酶的一种或多种另外的外源核酸。在一些实施方案中，编码一种或多种6-氨基己酸途径酶的外源核酸对于微生物有机体是异源的。
在一些实施方案中，非天然存在的微生物有机体包含己二胺途径。在一些实施方案中，己二胺途径包含：(i)6-氨基己酰辅酶A转移酶，(ii)6-氨基己酰辅酶A合酶，(iii)6-氨基己酰辅酶A还原酶，(iv)己二胺转氨酶，(v)己二胺脱氢酶，(v)或一种或多种酶(i)-(v)的组合。在一些实施方案中，微生物有机体还包含一种或多种另外的外源核酸，该一种或多种另外的外源核酸编码一种或多种己二胺途径酶例如将6-氨基己酸转化为6-氨基己酸半缩醛的羧酸还原酶(CAR)。随后，6-氨基己酸半缩醛可转化为己二胺。在一些实施方案中，编码一种或多种己二胺途径酶的外源核酸对于微生物有机体是异源的。
在一些实施方案中，非天然存在的微生物有机体包含己内酰胺途径。在一些实施方案中，己内酰胺途径包含氨基水解酶。在一些实施方案中，微生物有机体还包含编码氨基水解酶的一种或多种另外的外源核酸。在一些实施方案中，编码氨基水解酶的外源核酸对于微生物有机体是异源的。
在一些实施方案中，非天然存在的微生物有机体包含1,6-己二醇途径。在一些实施方案中，1,6-己二醇途径包含以下酶：催化6ACA转化为6-氨基己酰辅酶A的6-氨基己酰辅酶A转移酶或合成酶；催化6-氨基己酰辅酶A转化为6-氨基己酸半缩醛的6-氨基己酰辅酶A还原酶；催化6-氨基己酸半缩醛转化为6-氨基己醇的6-氨基己酸半缩醛还原酶；催化6ACA转化为6-氨基己酸半缩醛的6-氨基己酸还原酶；己二酰辅酶A还原酶，己二酰辅酶A至己二酸半缩醛；催化己二酸半缩醛转化为6-羟基己酸的己二酸半缩醛还原酶；催化6-羟基己酸转化为6-羟基己酰辅酶A的6-羟基己酰辅酶A转移酶或合成酶；催化6-羟基己酰辅酶A转化为6-羟基己醛的6-羟基己酰辅酶A还原酶；催化6-羟基己醛转化为HDO的6-羟基己醛还原酶；催化6-氨基己醇转化为6-羟基己醛的6-氨基己醇氨基转移酶或氧化还原酶；催化6-羟基己酸转化为6-羟基己醛的6-羟基己酸还原酶；催化ADA转化为己二酸半缩醛的己二酸还原酶；和催化己二酰辅酶A转化为ADA的己二酰辅酶A转移酶、水解酶或合酶。
在一些实施方案中，非天然存在的微生物有机体包含从己二酸或己二酰辅酶A到己内酯的途径。在一些实施方案中，从己二酸或己二酰辅酶A到己内酯的途径包括以下酶：己二酰辅酶A还原酶、己二酸半缩醛还原酶、6-羟基己酰辅酶A转移酶或合成酶、6-羟基己酰辅酶A环化酶或自发环化、己二酸还原酶、己二酰辅酶A转移酶、己二酰辅酶A合成酶或己二酰辅酶A水解酶、6-羟基己酸环化酶、6-羟基己酸激酶、6-羟基己酰磷酸环化酶或自发环化、磷酸反式-6-羟基己酰化酶。
在一些实施例中，非天然存在的微生物有机体的醛脱氢酶来源于原核物种。在一些实施例中，醛脱氢酶来源于Acidaminococcus、Collinsella、Peptostreptococcaceae或Romboustsia。
在一些实施方案中，非天然存在的微生物有机体包括Acinetobacter、Actinobacillus、Anaerobiospirillum、Aspergillus、Bacillus、Clostridium、Corynebacterium、Escherichia、Gluconobacter、Klebsiella、Kluyveromyces、Lactococcus、Lactobacillus、Mannheimia、Pichia、Pseudomonas、Rhizobium、Rhizopus、Saccharomyces、Schizosaccharomyces、Streptomyces和Zymomonas的物种。在一些实施方案中，非天然存在的微生物有机体是Escherichia.Coli菌株。
在一些实施方案中，培养在包含糖的发酵液中进行。
附图说明
图1显示了从琥珀酰辅酶A和乙酰辅酶A到6-氨基己酸、己二胺(HMDA)、己内酰胺的示例性途径。酶命名如下：A)3-氧代己二酰辅酶A硫解酶，B)3-氧代己二酰辅酶A还原酶，C)3-羟基己二酰辅酶A脱水酶，D)5-羧基-2-戊烯酰辅酶A还原酶，E)3-氧代己二酰辅酶A/酰基辅酶A转移酶，F)3-氧代己二酰辅酶A合酶，G)3-氧代己二酰辅酶A水解酶，H)3-氧代己二酸还原酶，I)3-羟基己二酸脱水酶，J)5-羧基-2-戊烯酸还原酶，K)己二酰辅酶A/酰基辅酶A转移酶，L)己二酰辅酶A合酶，M)己二酰辅酶A水解酶，N)己二酰辅酶A还原酶(醛形成)，O)6-氨基己酸转氨酶，P)6-氨基己酸脱氢酶，Q)6-氨基己酰辅酶A/酰基辅酶A转移酶，R)6-氨基己酰辅酶A合酶，S)酰胺水解酶，T)自发环化，U)6-氨基己酰辅酶A还原酶(醛形成)，V)HMDA转氨酶，W)HMDA脱氢酶，X)己二酸还原酶，Y)己二酸激酶，Z)己二酰磷酸还原酶。
图2是醛脱氢酶裂解物数据的图示，该醛脱氢酶裂解物数据显示了对己二酰辅酶A相对于对琥珀酰辅酶A的活性。
图3A-C是动力学数据的图示，该动力学数据显示相比乙酰辅酶A和琥珀酰辅酶A，纯化的醛脱氢酶更偏好己二酰辅酶A。图3A显示由其SEQ ID No指示的各种醛脱氢酶对琥珀酰辅酶A底物、乙酰辅酶A底物和己二酰辅酶A底物的催化效率。图3B显示由其SEQ ID No指示的各种醛脱氢酶对己二酰辅酶A底物与对琥珀酰辅酶A底物的催化效率的比率。图3C显示由其SEQ ID NO指示的各种醛脱氢酶对己二酰辅酶A底物与对乙酰辅酶A底物的催化效率的比率。
图4显示使用赖氨酸作为起点合成6-氨基己酸和己二酸的示例性途径。
图5显示了使用己二酰辅酶A作为起点的示例性己内酰胺合成途径。
图6显示了从丙酮酸和琥珀半缩醛到6-氨基己酸的示例性途径。酶为：A)HODH醛缩酶，B)OHED水合酶，C)OHED还原酶，D)2-OHD脱羧酶，E)己二酸半缩醛氨基转移酶和/或己二酸半缩醛氧化还原酶(胺化)，F)OHED脱羧酶，G)6-OHE还原酶,H)2-OHD氨基转移酶和/或2-OHD氧化还原酶(胺化),I)2-AHD脱羧酶,J)OHED氨基转移酶和/或OHED氧化还原酶(胺化),K)2-AHE还原酶,L)HODH甲酸-裂解酶和/或HODH脱氢酶，M)3-羟基己二酰辅酶A脱水酶，N)2,3-脱氢己二酰辅酶A还原酶，O)己二酰辅酶A脱氢酶，P)OHED甲酸-裂解酶和/或OHED脱氢酶，Q)2-OHD甲酸裂解酶和/或2-OHD脱氢酶。缩写是：HODH＝4-羟基-2-氧代庚烷-1,7-二酸，OHED＝2-氧代庚-4-烯-1,7-二酸，2-OHD＝2-氧代庚烷-1,7-二酸，2-AHE＝2-氨基庚-4-烯-1,7-二酸，2-AHD＝2-氨基庚烷-1,7-二酸，和6-OHE＝6-氧代己-4-烯酸。
图7显示从6-氨基己酸到己二胺的示例性途径。酶为：A)6-氨基己酸激酶、B)6-AHOP氧化还原酶、C)6-氨基己酸半缩醛氨基转移酶和/或6-氨基己半缩醛氧化还原酶(胺化)、D)6-氨基己酸N-乙酰转移酶、E)6-乙酰氨基己酸激酶,F)6-AAHOP氧化还原酶,G)6-乙酰氨基己醛氨基转移酶和/或6-乙酰氨基己醛氧化还原酶(胺化),H)6-乙酰氨基己胺N-乙酰转移酶和/或6-乙酰氨基己胺水解酶(酰胺),I)6-乙酰氨基己酸辅酶A转移酶和/或6-乙酰氨基己酸辅酶A连接酶，J)6-乙酰氨基己酰辅酶A氧化还原酶，K)6-AAHOP酰基转移酶，L)6-AHOP酰基转移酶，M)6-氨基己酸辅酶A转移酶和/或6-氨基己酸辅酶A连接酶，N)6-氨基己酰辅酶A氧化还原酶。缩写是：6-AAHOP＝[(6-乙酰氨基己酰基)氧基]膦酸和6-AHOP＝[(6-氨基己酰基)氧基]膦酸。
图8显示通向1,6-己二醇的示例性生物合成途径。A)是催化6ACA转化为6-氨基己酰辅酶A的6-氨基己酰辅酶A转移酶或合成酶；B)是催化6-氨基己酰辅酶A转化为6-氨基己酸半缩醛的6-氨基己酰辅酶A还原酶；C)是催化6-氨基己酸半缩醛转化为6-氨基己醇的6-氨基己酸半缩醛还原酶；D)是催化6ACA转化为6-氨基己酸半缩醛的6-氨基己酸还原酶；E)是己二酰辅酶A还原酶，己二酰辅酶A至己二酸半缩醛；F)是催化己二酸半缩醛转化为6-羟基己酸的己二酸半缩醛还原酶；G)是催化6-羟基己酸转化为6-羟基己酰辅酶A的6-羟基己酰辅酶A转移酶或合成酶；H)是催化6-羟基己酰辅酶A转化为6-羟基己醛的6-羟基己酰辅酶A还原酶；I)是催化6-羟基己醛转化为HDO的6-羟基己醛还原酶；J)是催化6-氨基己醇转化为6-羟基己醛的6-氨基己醇氨基转移酶或氧化还原酶；K)是催化6-羟基己酸转化为6-羟基己醛的6-羟基己酸还原酶；L)是催化ADA转化为己二酸半缩醛的己二酸还原酶；以及M)是催化己二酰辅酶A转化为ADA的己二酰辅酶A转移酶、水解酶或合酶。
图9显示从己二酸或己二酰辅酶A到己内酯的示例性途径。酶为：A.己二酰辅酶A还原酶；B.己二酸半缩醛还原酶；C.6-羟基己酰辅酶A转移酶或合成酶；D.6-羟基己酰辅酶A环化酶或自发环化；E.己二酸还原酶；F.己二酰辅酶A转移酶、合成酶或水解酶；G.6-羟基己酸环化酶；H.6-羟基己酸激酶；I.6-羟基己酰磷酸环化酶或自发环化；J.磷酸反式-6-羟基己酸酶。
具体实施方式
除非另外定义，否则本文使用的所有技术和科学术语具有与本发明所属领域的普通技术人员通常理解的相同的含义。在本发明的实践中可以使用与本文描述的那些相似或等效的任何方法、装置和材料。提供以下定义以便于理解本文中经常使用的某些术语，并且并非意味着限制本公开的范围。在此提及的所有参考文献均通过引用整体并入。
本文公开了非天然存在的微生物有机体，该非天然存在的微生物有机体被改造以表达外源性醛脱氢酶(ALD)，该外源性醛脱氢酶(ALD)对己二酰辅酶A底物比对琥珀酰辅酶A底物、或乙酰辅酶A底物、或两种底物具有更大的催化效率和转换数。己二酰辅酶A是通向6-氨基己酸、己内酰胺和己二胺(本文称为尼龙中间体)生物合成生产的途径中的中间体。许多不同的途径可用于产生这些尼龙中间体。在一些实施方案中，尼龙中间体可以从如图1所示的途径产生。经由己二酰辅酶A到尼龙中间体的其他途径的细节可以在例如专利号为8,377680的美国专利中找到，并通过引用整体并入本文。
在通向尼龙中间体的各种途径中，能够将酰基辅酶A还原为其相应醛的酰基辅酶A脱氢酶可以将己二酰辅酶A转化为己二酸半缩醛(步骤N，图1)。然而，一些酰基辅酶A脱氢酶也可以与琥珀酰辅酶A和乙酰辅酶A反应。在一些实施方案中，公开了酰基辅酶A脱氢酶(产生醛)，该酰基辅酶A脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物或两种底物具有更高的催化效率、更高的转换数或两者。这提高了效率，进而提高了尼龙中间体的产量。
为了鉴定对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物或两种底物具有更大的催化效率、更大的转换数或两者的酶，使用由基因adh(SEQ ID NO:1)编码的克氏梭菌DSM555(Clostridium kluyveri DSM555)的示例性序列来鉴定其他醛脱氢酶。同源酶的鉴定如表1所示(氨基酸序列如序列表所示)。
在一些实施方案中，通过BLAST鉴定醛脱氢酶或序列。在一些实施方案中，醛脱氢酶与表1的ALD的氨基酸序列的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、96％、97％、98％、99％或100％序列一致性。
对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物或两种底物具有更大的催化效率、更大的转换数或两者的这些醛脱氢酶来源于遗传上非常多样化的有机体。通常，序列之间的简单氨基酸序列一致性并不表示它们的共同功能。例如，表1中公开的一些示例性醛脱氢酶的成对序列比对结果如下所示。
表1序列一致性％
SEQ ID NO:7 SEQ ID NO:28 SEQ ID NO:60 SEQ ID NO:107
SEQ ID NO:7 50％ 56％ 60％
SEQ ID NO:28 50％ 53％ 57％
SEQ ID NO:60 56％ 53％ 60％
SEQ ID NO:107 60％ 67％ 60％
这些醛脱氢酶具有多个保守结构域，例如N-末端结构域、C-末端结构域和在其活性位点的半胱氨酸残基。醛脱氢酶包含具有罗斯曼折叠型核苷酸结合结构的辅因子结合结构域。罗斯曼折叠，也称为βαβ折叠，是以β链-α螺旋-β链二级结构的交替基序为特征的超二级结构。β链参与β-片层(β-sheet)的形成。βαβ折叠结构通常在具有二核苷酸辅酶的酶(例如FAD、NAD和NADP)中观察到。βαβ折叠结构与在第一个β链和α螺旋之间的紧密环区域处的特定的富含Gly的序列(GxGxxG)相关。此外，辅因子结合结构域也是与底物辅酶A结合的同一结构域。这是Ald的典型特征，在Ald中该底物辅酶A首先结合，形成中间体，然后辅因子结合且完成化学反应并进行氢化物转移。
基于多序列比对和隐马尔可夫模型(HMM)，将醛脱氢酶分组成来自欧洲生物信息学研究所的Pfam数据库(pfam.xfam.org)的Pfam PF00171，Clan CL0099。根据酶委员会的命名法，这些酶被归类为EC 1.2.1。
在一些实施方案中，与琥珀酰辅酶A、乙酰Co-A或两者相比，当己二酰辅酶A作为底物时，ALD酶具有更大的催化效率和/或转换率。在一些实施方案中，醛脱氢酶包含与SEQ IDNOs:47、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187或188中的任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％氨基酸序列一致性的氨基酸序列。在一些实施方案中，与己二酰辅酶A反应形成己二酸-半缩醛的醛脱氢酶的氨基酸序列选自SEQ ID NOs:1-4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187和188的氨基酸序列。
在一些实施方案中，对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物或两种底物具有更大的催化效率、更大的转换率或其组合的醛脱氢酶的氨基酸序列与SEQ IDNO:7、28、60和107中任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸至少约60％氨基酸序列一致。在一些实施方案中，对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物或两种底物具有更大的催化效率、更大的转换率或其组合的醛脱氢酶的氨基酸序列与SEQ ID NO:7、28、60和107中任一个的氨基酸序列的至少50、75、100、150、200、250、300个或更多个连续氨基酸至少约65％、70％、75％、80％、85％、90％、95％、96％、97％、98％、99％或100％序列一致。
在一些实施方案中，ALD酶对己二酰辅酶A底物的催化效率至少是对琥珀酰辅酶A、乙酰辅酶A或两者作为底物的至少5×、至少10×、至少25×或5-25×。
在一些实施方案中，，该酶在已知标准条件下使指定底物(例如己二酰辅酶A)向指定产物(例如己二酸半缩醛)的酶促转化相比仅对己二酰辅酶A没有特异性的酶的酶活性高至少10％、至少20％、至少30％％、至少40％、至少50％、至少60％、至少70％、至少80％或至少90％。
在一些实施方案中，所述醛脱氢酶进一步与6-氨基己酰辅酶A反应以形成6-氨基己酸半缩醛。
可以使用本领域已知的任何方法来鉴定具有降低的酶活性的细胞。例如，酶活性测试可用于鉴定具有降低的酶活性的细胞，参见例如Enzyme Nomenclature,AcademicPress,Inc.,New York 2007。可用于确定ADH降低的其他测试包括GC/MS分析。在其他示例中，可以监测NADH/NADPH的水平。例如，可以使用NADP/NADPH测试试剂盒(例如，可从ABCAMTM获得的ab65349)通过比色法或光谱法来监测NADH/NADPH。
所公开的ALD酶可用于产生尼龙中间体的途径。在一些实施方案中，非天然存在的微生物体可用于产生己二酸半缩醛或使用己二酸半缩醛作为中间体而产生的其他尼龙中间体。
在一些实施方案中，经遗传修饰的细胞(例如非天然存在的微生物体)能够产生尼龙中间体，例如6-氨基己酸、己内酰胺和己二胺。
在一些实施方案中，尼龙中间体是使用图1中描述的途径而生物合成的。在一些实施方案中，图1途径在本文描述的经遗传修饰的细胞(例如，非天然存在的微生物体)中提供，其中该途径包括编码途径酶的至少一种外源核酸，该途径酶以足以产生6-氨基己酸、己内酰胺和己二胺的量表达。
在一些实施方案中，该途径是如图1所示的HMD途径。HMD途径在本文描述的经遗传修饰的细胞(例如，非天然存在的微生物体)中提供，其中HMD途径包括编码HMD途径酶的至少一种外源核酸，HMD途径酶以足以产生HMD的量表达。所述酶为：1A是3-氧代己二酰辅酶A硫解酶；1B是3-氧代己二酰辅酶A还原酶；1C是3-羟基己二酰辅酶A脱水酶；1D是5-羧基-2-戊烯酰辅酶A还原酶；1E是3-氧代己二酰辅酶A/酰基辅酶A转移酶；1F是3-氧代己二酰辅酶A合酶；1G是3-氧代己二酰辅酶A水解酶；1H是3-氧代己二酸还原酶；1I是3-羟基己二酸脱水酶；1J是5-羧基-2-戊烯酸还原酶；1K是己二酰辅酶A/酰基辅酶A转移酶；1L是己二酰辅酶A合酶；1M是己二酰辅酶A水解酶；1N是己二酰辅酶A还原酶(醛形成)；1O是6-氨基己酸转氨酶；1P是6-氨基己酸脱氢酶；1Q是6-氨基己酰辅酶A/酰基辅酶A转移酶；1R是6-氨基己酰辅酶A合酶；1S是酰胺水解酶；1T是自发环化；1U是6-氨基己酰辅酶A还原酶(醛形成)；1V是HMDA转氨酶；1W是HMDA脱氢酶。
参考图1，在一些实施方案中，非天然存在的微生物体具有以下一种或多种途径：ABCDNOPQRUVW；ABCDNOPQRT；或ABCDNOPS。包括ALD酶以产生己二酸半缩醛的其他示例性途径包括专利号为8,377,680的美国专利中描述的那些，专利号为8,377,680的美国专利通过引用其全部内容并入本文。
图1还显示了通过转移酶或合酶从6-氨基己酸到6-氨基己酰辅酶A(图1，步骤Q或R)，随后6-氨基己酰辅酶A自发环化形成己内酰胺(图1，步骤T)的途径。在其他实施方案中，6-氨基己酸被活化为6-氨基己酰辅酶A(图1，步骤Q或R)，然后还原(图1，步骤U)和胺化(图1，步骤V或W)以形成HMDA。6-氨基己酸也可以被活化为6-氨基己酰基-磷酸而不是6-氨基己酰辅酶A。6-氨基己酰基-磷酸可自发环化以形成己内酰胺。在一些实施方案中，6-氨基己酰基-磷酸可被还原为6-氨基己酸半缩醛，然后6-氨基己酸半缩醛可转化为HMDA，如图1所示。在一些实施方案中，6-氨基己酸被氨基己酸还原酶(CAR)转化为6-氨基己酸半缩醛。虽然图1未显示，氨基己酸还原酶可以催化如图1所示的氨基己酸到6-氨基己酸半缩醛的转化。
在一些实施方案中，非天然存在的微生物有机体具有己二胺途径，该途径包括：(i)6-氨基己酰辅酶A转移酶，(ii)6-氨基己酰辅酶A合酶，(iii)6-氨基己酰辅酶A还原酶，(iv)己二胺转氨酶，(v)己二胺脱氢酶，(v)或一种或多种酶(i)-(v)的组合。在其他实施方案中，非天然存在的微生物有机体具有己二胺途径，该己二胺途径包括3-氧代己二酰辅酶A硫解酶(Thl)、3-氧代己二酰辅酶A脱氢酶(Hbd)和3-氧代己二酰辅酶A脱水酶(“巴豆酸酶”或Crt)、5-羧基-2-戊烯酰辅酶A还原酶(Ter)、转氨酶(HMD TA)和羧酸还原酶(CAR)。
如本文所用，术语“非天然存在的”在用于提及微生物有机体或微生物体时旨在表示该微生物有机体具有在所提及的物种的天然存在的菌株(包括所提及的物种的野生型菌株)中通常不存在的至少一种遗传改变。遗传改变包括，例如，引入编码代谢多肽的可表达核酸的修饰、其他核酸添加、核酸缺失和/或微生物遗传材料的其他功能破坏。此类修饰包括，例如，用于所提及的物种的异源多肽、同源多肽、或异源和同源多肽的编码区及其功能片段。另外的修饰包括，例如，其中修饰改变了基因或操纵子的表达的非编码调控区。示例性代谢多肽包括6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径内的酶。
代谢修饰是指改变其天然存在状态的生化反应。因此，非天然存在的微生物体可以对编码代谢多肽或其功能片段的核酸进行遗传修饰。本文中公开了示例性代谢修饰。
如本文所用，术语“微生物”、“微生物有机体”或“微生物体”已可互换使用，旨在表示作为包括在古细菌、细菌或真核生物领域内的微观细胞存在的任何有机体。因此，该术语旨在涵盖原核或真核细胞或有机体，该原核或真核细胞或有机体具有微观尺寸，并且包括所有物种的细菌、古细菌和真细菌以及真核微生物体例如酵母和真菌。该术语还包括任何物种的细胞培养物，这些细胞培养物可以培养以产生生化物质。
如本文所用，术语“CoA”或“辅酶A”旨在表示这样的有机辅因子或辅基(酶的非蛋白质部分)：其存在是许多酶(脱辅酶)的活性所需要的以形成活性酶系统。辅酶A在某些缩合酶中起作用，在乙酰基或其他酰基转移以及脂肪酸合成和氧化、丙酮酸氧化和其他乙酰化中起作用。
如本文所用，具有化学式-OOC-(CH2)4-COO-(见图1)(IUPAC名称己二酸酯)的“己二酸根”是己二酸(IUPAC名称己二酸)的离子化形式，并且应理解，己二酸根和己二酸自始至终可互换使用以指代其任何中性或离子化形式，包括其任何盐形式的化合物。技术人员理解，具体形式将取决于pH。
如本文所用，具有化学式-OOC-(CH2)5-NH2(参见图1，缩写为6-ACA)的“6-氨基己酸根”是6-氨基己酸(IUPAC名称为6-氨基己酸)的离子化形式，并且应理解，6-氨基己酸根和6-氨基己酸自始至终可互换使用，以指代其任何中性或离子化形式，包括其任何盐形式的化合物。技术人员理解，具体形式将取决于pH。
如本文所用，“己内酰胺”(IUPAC名称氮杂环庚烷-2-酮(azepan-2-one))是6-氨基己酸的内酰胺(参见图1，缩写为CPO)。
如本文所用，“己二胺”，也称为1,6-二氨基己烷或1,6-己二胺，具有化学式H2N(CH2)6NH2(参见图1,并缩写为HMD)。
如本文所用，术语“基本厌氧”在用于提及培养或生长条件时旨在表示氧的含量小于液体培养基中溶解氧的约10％的饱和度。该术语还旨在包括维持在低于约1％氧气的气氛中的液体或固体介质的密封室。
如本文所用，术语“渗透保护剂”在用于提及培养或生长条件时旨在表示充当渗透剂并帮助如本文所述的微生物有机体在渗透胁迫中存活的化合物。渗透保护剂包括例如甜菜碱、氨基酸和糖海藻糖。其非限制性实例是甘氨酸甜菜碱、脯氨酸甜菜碱、二甲基噻亭、二甲基磺酰丙酸酯、3-二甲基磺酰-2-甲基丙酸酯、哌可酸、二甲基磺酰乙酸、胆碱、L-肉碱和四氢嘧啶。
如本文所用，术语“生长耦合的”在用于提及生化物质的生产时，旨在表示所提及的生化物质的生物合成是在微生物的生长阶段中产生的。在一个特定的实施方案中，生长耦合的生产可以是强制性的，这意味着所提及的生化物质的生物合成是在微生物的生长阶段中产生的强制性产品。
如本文所用，“代谢修饰”旨在指改变其天然存在状态的生化反应。代谢修饰可以包括，例如，通过编码参与反应的酶的一种或多种基因的功能破坏来消除生化反应活性。
如本文所用，术语“基因破坏”或其语法等价物旨在表示使编码的基因产物失活的遗传改变。遗传改变可以是，例如，整个基因的缺失、转录或翻译所需的调控序列的缺失、导致截短基因产物的基因部分缺失、或通过使编码的基因产物失活的各种突变策略中的任一种。一种特别有用的基因破坏方法是基因完全缺失，因为它减少或消除了非天然存在的微生物中遗传逆转的发生。
如本文所用，“外源的”旨在表示将所提及的分子或所提及的活性引入宿主微生物有机体中。例如，可以通过将编码核酸引入宿主遗传物质中，例如通过整合到宿主染色体中、或作为非染色体遗传物质(例如质粒)，来引入分子。因此，当用于提及编码核酸的表达时，该术语是指将编码核酸以可表达的形式引入微生物有机体中。当用于提及生物合成活性时，该术语是指引入所提及的宿主参照有机体的活性。来源可以是例如同源或异源编码核酸，该同源或异源编码核酸在引入宿主微生物后表达所提及的活性。因此，术语“内源性”是指所提及的存在于宿主中的分子或活性。类似地，当用于提及编码核酸的表达时，该术语是指包含在微生物有机体内的编码核酸的表达。
术语“异源的”是指来源于所提及的物种之外的来源的分子或活性，而“同源的”是指来源于宿主微生物有机体的分子或活性。因此，编码核酸的外源表达可利用异源或同源编码核酸之一或两者。
如本文所用，术语“约”是指所述值的±10％。术语“约”可以表示四舍五入到最接近的有效数字。因此，约5％意味着4.5％到5.5％。此外提及特定数字的约也包括该确切数字。例如，约5％还包括精确的5％。
本文中使用的术语转换数(也称为kcat)定义为对于给定的酶浓度[ET]，单个催化位点每秒将执行的底物分子的最大化学转化数。可由最大反应速率Vmax和催化剂位点浓度[ET]计算如下：
Kcat＝Vmax/[ET]。单位是s-1。
如本文所用，术语“催化效率”是酶将底物转化为产物的效率的量度。催化效率的比较也可以用作酶对不同底物的偏好(即底物特异性)的量度。催化效率越高，酶越“偏好”该底物。可由下式计算：kcat/KM，其中kcat为转换数，KM为米氏常数，KM为反应速率为Vmax一半时的底物浓度。催化效率的单位可以表示为s-1M-1。
如本文所用，在6-氨基己酸、1，6-己二醇、己内酯、己内酰胺或己二胺的上下文中的术语“生物衍生的”是指这些化合物在微生物有机体中合成。
应理解，当微生物有机体中包括一种以上外源核酸时，外源核酸是指所提及的编码核酸或生物合成活性，如上文所讨论。还应理解，如本文所公开的，此类外源核酸可以在单独的核酸分子、多顺反子核酸分子或其组合上被引入宿主微生物有机体中，并且仍被认为是多于一种的外源核酸。例如，如本文所公开的，可以将微生物有机体改造成表达编码所需途径酶或蛋白质的两种或更多种外源核酸。在将编码所需活性的两种外源核酸引入宿主微生物有机体的情况下，应理解这两种外源核酸可以作为单个核酸(例如，在单个质粒上、在单独的质粒上)被引入，可以在单个位点或多个位点处整合到宿主染色体中，且仍然被认为是两种外源核酸。类似地，应当理解，可以以任何期望的组合将多于两种的外源核酸引入宿主有机体中，例如，在单个质粒上，在未整合到宿主染色体中的单独质粒上，并且质粒保持为染色体外元件，且仍被视为两种或更多种外源核酸。所提及的外源核酸或生物合成活性的数量是指编码核酸的数量或生物合成活性的数量，而不是引入宿主有机体的单独核酸的数量。
非天然存在的微生物有机体可以包含稳定的遗传改变，是指可以培养超过五代而不丧失改变的微生物。通常，稳定的遗传改变包括持续超过10代的修饰，特别地稳定的修饰将持续超过约25代，更特别地，稳定的遗传修饰将超过50代，包括无限期。
在基因破坏的情况下，特别有用的稳定的遗传改变是基因缺失。使用基因缺失来引入稳定的遗传改变对于降低回复到遗传改变之前的表型的可能性特别有用。例如，可以在一组代谢修饰内例如通过删除编码催化一个或多个反应的酶的基因来实现生化物质的稳定的生长耦合的生产。通过多次缺失，可以进一步增强生化物质的生长耦合的生产的稳定性，从而显着降低每个被破坏的活性发生多次补偿性回复的可能性。
本领域技术人员将理解，参照合适的宿主有机体例如大肠杆菌及其相应的代谢反应或合适的源有机体针对所需的遗传材料(例如所需的代谢途径的基因)来描述遗传改变，包括本文中例示的代谢修饰。然而，考虑到多种有机体的完整基因组测序和基因组学领域的高水平技术，本领域技术人员将能够容易地将本文提供的教导和指导应用于基本上所有其他有机体。例如，本文举例说明的大肠杆菌代谢改变可通过掺入来自所提及物种之外的物种的相同或类似的编码核酸而容易地应用于其他物种。此类遗传改变包括例如物种同源物的遗传改变，一般而言，特别是直系同源、旁系同源或非直系同源基因置换。
直系同源物是通过直系亲缘相关并且在不同有机体中负责基本相同或完全相同的功能的一种或多种基因，。例如，小鼠环氧化物水解酶和人环氧化物水解酶可以被认为是环氧化物水解生物学功能的直系同源物。例如，当基因共享足量的序列相似性以表明它们是同源的或通过从共同祖先进化而相关时，基因通过直系亲缘相关联。如果基因共享三维结构但不一定具有序列相似性，而共享的三维结构的数量足以表明它们已从共同祖先进化到无法识别一级序列相似性的程度，则基因也可以被认为是直系同源物。直系同源基因可以编码序列相似性为约25％至100％氨基酸序列一致性的蛋白质。如果具有小于25％的氨基酸相似性的编码蛋白质的基因的三维结构也显示出相似性，则编码蛋白质的基因也可以被认为通过直系亲缘产生。丝氨酸蛋白酶家族的成员，包括组织纤溶酶原激活剂和弹性蛋白酶被认为从共同祖先通过直系亲缘产生。
直系同源物包括通过例如进化在结构或总体活性上分化的基因或其编码的基因产物。例如，当一个物种编码表现出两种功能的基因产物并且这种功能在第二个物种中被分开到不同的基因时，这三个基因及其相应的产物被认为是直系同源物。对于生化产物的生产，本领域技术人员将理解，选择具有代谢活性的待引入或破坏的直系同源基因来构建非天然存在的微生物体。表现出可分离的活性的直系同源物的一个实例是在两个或更多个物种之间或单个物种内将不同的活性分开到不同的基因产物中。一个具体的实例是将弹性蛋白酶蛋白水解和纤溶酶原蛋白水解(两种丝氨酸蛋白酶活性)分开到不同的分子，如纤溶酶原激活剂和弹性蛋白酶中。第二个实例是支原体5'-3'核酸外切酶和果蝇DNA聚合酶III活性的分开。来自第一物种的DNA聚合酶可以被认为是来自第二物种的核酸外切酶或聚合酶之一或两者的直系同源物，反之亦然。
相反，旁系同源物是通过例如复制随后通过进化分歧而相关的同源物，并且具有相似或共同但不完全相同的功能。旁系同源物可以起源于或来源于例如相同的物种或不同的物种。例如，微粒体环氧化物水解酶(环氧化物水解酶I)和可溶性环氧化物水解酶(环氧化物水解酶II)可以被认为是旁系同源物，因为它们代表从共同的祖先共同进化而来的、催化不同的反应并在相同的物种中具有不同的功能的不同的两种酶。旁系同源物是来自相同物种的蛋白质，彼此之间具有显著的序列相似性，表明它们是同源的，或通过从共同祖先的共同进化而相关。旁系同源蛋白家族组包括HipA同源物、萤光素酶基因、肽酶等。
非直系同源基因置换是来自一个物种的非直系同源基因，其可以替代不同物种中的所提及的基因功能。取代包括，例如，与不同物种中的所提及的功能相比，能够在来源物种中执行基本相同或相似的功能。尽管一般而言，非直系同源基因置换将被鉴定为与编码所提及的功能的已知基因在结构上相关，然而结构相关性较低但功能相似的基因及其相应的基因产物仍将落入本文所用术语的含义内。例如，功能相似性要求非直系同源基因产物的活性位点或结合区与编码寻求被取代的功能的基因相比至少具有某种结构相似性。因此，非直系同源基因包括例如旁系同源基因或不相关基因。
因此，在鉴定和构建具有6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成能力的非天然存在的微生物有机体时，本领域技术人员将理解将本文提供的教导和指导应用于特定物种，代谢修饰的鉴定可包括直系同源物的鉴定和包含或失活。就编码催化相似或基本相似的代谢反应的酶的所提及的微生物中存在旁系同源物和/或非直系同源基因置换而言，本领域技术人员也可以利用这些进化相关基因。在基因破坏策略中，也可以在宿主微生物有机体、旁系同源物或直系同源物中被破坏或缺失进化相关基因，以降低或消除活性，从而确保针对破坏的酶活性的任何功能冗余不会使设计的代谢修饰短路。
直系同源物、旁系同源物和非直系同源基因置换可以通过本领域技术人员熟知的方法确定。例如，检查两个多肽的核酸或氨基酸序列将揭示比较序列之间的序列一致性和相似性。基于这样的相似性，本领域技术人员可以确定相似性是否足够高以表明蛋白质是通过从共同祖先进化而相关。本领域技术人员熟知的算法，例如Align、BLAST、Clustal W等，比较和确定原始序列的相似性或一致性，还确定序列中可被分配权重或得分的空位的存在或显著性。此类算法也是本领域已知的并且类似地可应用于确定核苷酸序列相似性或一致性。基于用于计算统计相似性或在随机多肽中发现相似匹配的可能性以及确定匹配的显著性的熟知的方法来计算用以确定相关性的足够相似性的参数。如果需要，本领域技术人员也可以在视觉上优化两个或更多个序列的计算机比较。可预期相关基因产物或蛋白质具有高相似性，例如25％至100％的序列一致性。如果扫描足够大小的数据库(大约5％)，则不相关的蛋白质的一致性可以与预期偶然发生的基本相同。5％和24％之间的序列可能代表也可能不代表足以推断出比较序列相关的同源性。在给定数据集大小的情况下，可以进行另外的确定此类匹配的显著性的统计分析，以确定这些序列的相关性。
例如，用于使用BLAST算法确定两条或更多条序列的相关性的示例性参数可以如下所述。简而言之，可以使用BLASTP版本2.2.29+(2014年1月14日)和以下参数进行氨基酸序列比对：矩阵：0BLOSUM62；空位开头：11；空位延长：1；x_dropoff：50；期望：10.0；字长：3；过滤器：开。可以使用BLASTN版本2.0.6(1998年9月16日)和以下参数进行核酸序列比对：匹配：1；不匹配：-2；空位开头：5；空位延长：2；x_dropoff：50；期望：10.0；字长：11；过滤器：关闭。本领域技术人员将知道可以对上述参数进行哪些修改以例如增加或减少比较的严格性，并确定两条或更多条序列的相关性。
应当理解，本文公开的任何途径，包括图中描述的那些，均可用于生产根据需要产生任何途径中间体或产物的非天然存在的微生物有机体。如本文所公开的，这种产生中间体的微生物有机体可以与表达下游途径酶的另一种微生物组合使用以产生所需产物。然而，应理解，可利用产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体的非天然存在的微生物有机体来产生作为所需产物的中间体。
本文一般提及代谢反应、反应物或其产物，或具体提及一种或多种核酸或基因进行描述，所述一种或多种核酸或基因编码与所提及的代谢反应、反应物或产物相关或催化所提及的代谢反应、反应物或产物的酶。除非本文另有明确说明，否则本领域技术人员将理解，提及反应也构成提及反应物和反应产物。类似地，除非本文另有明确说明，否则提及反应物或产物也提及反应，提及任何这些代谢成分也提及编码催化所提及的反应、反应物或产物的酶的一个或多个基因。同样，鉴于熟知的代谢生物化学、酶学和基因组学领域，本文中对基因或编码核酸的提及也构成对相应的被编码的酶和其催化的反应以及反应的反应物和产物的提及。
非天然存在的微生物有机体可以通过引入编码参与6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径中的一种或多种途径的一种或多种酶的可表达核酸来产生。根据选择用于生物合成的宿主微生物，可以表达特定6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径中的一些或全部的核酸。例如，如果所选宿主缺乏用于所需生物合成途径的一种或多种酶，则将所缺乏的酶的可表达核酸引入宿主中用于随后的外源表达。可替换地，如果所选宿主表现出某些途径基因的内源性表达，但其他途径基因缺乏，则需要所缺乏的酶的编码核酸以实现6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成。因此，非天然存在的微生物有机体可以通过引入外源酶活性来产生以获得所需生物合成途径，或者可以通过引入一种或多种外源酶活性与一种或多种内源酶一起产生所需产物，例如6-氨基己酸、己内酰胺、己二胺或乙酰丙酸来获得所需生物合成途径。
取决于所选宿主微生物有机体的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径成分，非天然存在的微生物有机体将包括至少一种外源表达的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径编码核酸和用于一种或多种己二酸、6-氨基己酸或己内酰胺生物合成途径的至多所有编码核酸。例如，可以通过相应编码核酸的外源表达在途径酶上缺乏的宿主中建立6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成。在缺乏6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径的所有酶的宿主中，可包括途径中所有酶的外源表达，但是应理解，即使宿主包含至少一种途径酶，也均可表达途径中的所有酶。
鉴于本文提供的教导和指导，本领域技术人员将理解以可表达形式引入的编码核酸的数量将至少与所选宿主微生物有机体的己二酸、6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径的缺乏相当。因此，非天然存在的微生物有机体可以具有编码构成6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径的上述酶的至少一种、两种、三种、四种、五种、六种、七种、八种、九种、十种、十一种或十二种、至多所有核酸。在一些实施方案中，非天然存在的微生物有机体还可以包括促进或优化6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成或赋予宿主微生物有机体其他有用功能的其他遗传修饰。一种这样的其他功能性可以包括，例如，增强6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径前体中的一种或多种的合成，所述前体，例如：在己二酸合成的情况下为琥珀酰辅酶A和/或乙酰辅酶A，或在6-氨基己酸或己内酰胺合成的情况下为己二酰辅酶A或己二酸，包括本文公开的己二酸途径酶，或在6-氨基己酸合成的情况下为丙酮酸和琥珀酸半缩醛、谷氨酸、戊二酰辅酶A、高赖氨酸或2-氨基-7-氧代辛二酸，或在己二胺合成的情况下为6-氨基己酸、谷氨酸、戊二酰辅酶A、丙酮酸和4-氨基丁醛，或2-氨基-7-氧代辛二酸。
在一些实施方案中，非天然存在的微生物有机体具有编码醛脱氢酶的至少一种外源核酸，所述醛脱氢酶与己二酰辅酶A反应以形成己二酸-半缩醛并且选自包含与SEQ IDNO：1-4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187或188中任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％氨基酸序列一致性的氨基酸序列的醛脱氢酶。在一些实施方案中，非天然存在的微生物有机体具有至少一种编码醛脱氢酶的外源核酸，所述醛脱氢酶与己二酰辅酶A反应以形成己二酸-半缩醛并且选自包含与SEQ ID NO：1-4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187或188中任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约65％、70％、75％、80％、85％、90％、95％或100％氨基酸序列一致性的氨基酸序列的醛脱氢酶。
在其他实施方案中，非天然存在的微生物有机体具有编码醛脱氢酶的至少一种外源核酸，所述醛脱氢酶与己二酰辅酶A反应以形成己二酸-半缩醛，包含与SEQ ID NO：7、28、60或107的氨基酸序列中任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约60％氨基酸序列一致性的氨基酸序列。在其他实施方案中，非天然存在的微生物有机体具有编码醛脱氢酶的至少一种外源核酸，所述醛脱氢酶与己二酰辅酶A反应以形成己二酸-半缩醛，包含与SEQ ID NO：7、28、60或107的氨基酸序列中任一个的至少50、75、100、150、200、250、300个或更多个连续氨基酸具有至少约65％、70％、75％、80％、85％、90％、95％或100％氨基酸序列一致性的氨基酸序列。
通常，选择宿主微生物有机体使其产生6-氨基己酸、己内酰胺或己二胺途径的前体，作为天然产生的分子或作为工程产品，这提供了所需前体的从头生产或增加由宿主微生物有机体的自然产生的前体的生产。如本文所公开的，宿主有机体可以被改造以增加前体的生产。此外，已被改造以产生所需前体的微生物可用作宿主有机体并进一步改造以表达6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径的酶或蛋白质。
在一些实施方案中，非天然存在的微生物有机体由宿主产生，该宿主包含合成6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的酶促能力。在该特定实施方案中，增加6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径产物的合成或积累以例如驱动6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径反应向6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产。增加的合成或积累可以通过例如编码上述6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶中的一种或多种的核酸的过表达来实现。例如，通过一个内源基因或多个内源基因的外源表达，或通过一个异源基因或多个异源基因的外源表达可以发生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶的过表达。因此，通过使编码6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径酶的至少一种核酸、两种核酸、三种核酸、四种核酸、五种核酸、六种核酸、七种核酸、八种核酸、九种核酸、十种核酸、十一种核酸、十二种核酸、十三种核酸、十四种核酸，即，一直到所有核酸过表达，可以很容易地使天然存在的有机体生成为例如产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的非天然存在的微生物有机体。此外，非天然存在的有机体可以通过导致6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径中的酶活性增加的内源基因的诱变而产生。
在特别有用的实施方案中，使用编码核酸的外源表达。外源表达赋予宿主和应用定制表达和/或调控元件的能力，以实现由使用者控制的所需表达水平。然而，内源表达也可用于其他实施方案，例如通过去除负调节效应子或当与诱导型启动子或其他调节元件连接时诱导基因启动子。因此，可以通过提供合适的诱导剂来上调具有天然存在的诱导型启动子的内源基因，或者可以改造内源基因的调控区以掺入诱导型调控元件，从而允许调节内源基因在所需的时间的增加表达。类似地，可包括诱导型启动子作为引入非天然存在的微生物有机体中的外源基因的调控元件。
在一些实施方案中，非天然存在的微生物有机体包括一种或多种基因破坏，其中该有机体产生6-ACA、己二酸和/或HMDA。破坏发生在编码酶的基因中，当基因破坏降低酶的活性时，该酶将己二酸、6-ACA和/或HMDA的产生与有机体的生长相结合，从而基因破坏使得在非天然存在的有机体上增加己二酸、6-ACA和/或HMDA的生产。因此，在一些实施方案中，提供非天然存在的微生物有机体，该非天然存在的微生物有机体包含一种或多种基因破坏，所述一种或多种基因破坏发生在编码蛋白质或酶的基因中，其中所述一种或多种基因破坏使得在有机体中增加己二酸、6-ACA和/或HMDA的生产。如本文所公开，此类有机体包含己二酸、6-ACA和/或HMDA的生产途径。
应当理解，在方法中，可以将一种或多种外源核酸中的任一种引入微生物有机体中以产生非天然存在的微生物有机体。可以引入核酸以赋予微生物例如6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径。可替换地，可以引入编码核酸以产生具有生物合成能力的中间微生物有机体以催化一些所需反应，从而赋予6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成能力。例如，具有6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径的非天然存在的微生物有机体可以包含编码所需酶的至少两种外源核酸。在己二酸生产的情况下，至少两种外源核酸可以编码以下酶的组合：例如，琥珀酰辅酶A：乙酰辅酶A酰基转移酶和3-羟酰基辅酶A脱氢酶、或琥珀酰辅酶A：乙酰辅酶A酰基转移酶和3-羟基己二酰辅酶A脱水酶、或3-羟基己二酰辅酶A和5-羧基-2-戊烯酰辅酶A还原酶、或3-羟酰基辅酶A和己二酰辅酶A合成酶等。在己内酰胺生产的情况下，至少两种外源核酸可以编码以下酶的组合：例如，辅酶A依赖性醛脱氢酶和转氨酶、或辅酶A依赖性醛脱氢酶和酰胺水解酶、或转氨酶和酰胺水解酶。在6-氨基己酸生产的情况下，至少两种外源核酸可以编码以下酶的组合：例如，4-羟基-2-氧代庚烷-1,7-二酸(HODH)醛缩酶和2-氧代庚-4-烯-1,7-二酸(OHED)水合酶、或2-氧代庚-4-烯-1,7-二酸(OHED)水合酶和2-氨基庚烷-1,7-二酸(2-AHD)脱羧酶、3-羟基己二酰辅酶A脱水酶和己二酰辅酶A脱氢酶、谷氨酰辅酶A转移酶和6-氨基庚二酰辅酶A水解酶，或戊二酰辅酶Aβ-酮硫解酶和3-氨基庚二酸2,3-氨基变位酶。在己二胺生产的情况下，至少两种外源核酸可以编码以下酶的组合：例如，6-氨基己酸激酶和[(6-氨基己酰基)氧基]膦酸(6-AHOP)氧化还原酶、或6-乙酰氨基己酸激酶和[(6-乙酰氨基己酰基)氧基]膦酸(6-AAHOP)氧化还原酶、6-氨基己酸N-乙酰转移酶和6-乙酰氨基己酰辅酶A氧化还原酶、3-羟基-6-氨基庚二酰辅酶A脱水酶和2-氨基-7-氧代庚酸氨基转移酶，或3-氧代庚二酰辅酶A连接酶和高赖氨酸脱羧酶。因此，应当理解，在非天然存在的微生物有机体中可以包括生物合成途径的两种或更多种酶的任何组合。
类似地，应当理解，在非天然存在的微生物有机体中可以包括生物合成途径的三种或更多种酶的任何组合，例如，在己二酸生产的情况下，以下酶的组合：根据需要，琥珀酰辅酶A：乙酰辅酶A酰基转移酶、3-羟酰基辅酶A脱氢酶和3-羟基己二酰辅酶A脱水酶；或琥珀酰辅酶A：乙酰辅酶A酰基转移酶、3-羟酰基辅酶A脱氢酶和5-羧基-2-戊烯酰辅酶A还原酶；或琥珀酰辅酶A：乙酰辅酶A酰基转移酶、3-羟酰基辅酶A脱氢酶和己二酰辅酶A合成酶；或3-羟酰基辅酶A脱氢酶、3-羟基己二酰辅酶A脱水酶和己二酰辅酶A：乙酰辅酶A转移酶等，只要所需生物合成途径的酶的组合导致产生相应的所需产品。在6-氨基己酸生产的情况下，至少三种外源核酸可以编码以下酶的组合：例如，4-羟基-2-氧代庚烷-1,7-二酸(HODH)醛缩酶、2-氧代庚-4-烯-1,7-二酸(OHED)水合酶和2-氧代庚烷-1,7-二酸(2-OHD)脱羧酶；或2-氧代庚-4-烯-1,7-二酸(OHED)水合酶、2-氨基庚-4-烯-1,7-二酸(2-AHE)还原酶和2-氨基庚烷-1,7-二酸(2-AHD)脱羧酶；或3-羟基己二酰辅酶A脱水酶、2,3-脱氢己二酰辅酶A还原酶和己二酰辅酶A脱氢酶；或6-氨基-7-羧基庚-2-烯酰辅酶A还原酶、6-氨基庚二酰辅酶A水解酶和2-氨基庚二酸脱羧酶；或戊二酰辅酶Aβ-酮硫解酶、3-氨化氧化还原酶和2-氨基庚二酸脱羧酶；或3-氧代己二酰辅酶A硫解酶、5-羧基-2-戊烯酸还原酶和己二酸还原酶。在己二胺生产的情况下，至少三种外源核酸可以编码以下酶的组合，例如：6-氨基己酸激酶、[(6-氨基己酰基)氧基]膦酸(6-AHOP)氧化还原酶和6-氨基己酸半缩醛氨基转移酶；或6-氨基己酸N-乙酰转移酶、6-乙酰氨基己酸激酶和[(6-乙酰氨基己酰基)氧基]膦酸(6-AAHOP)氧化还原酶；或6-氨基己酸N-乙酰转移酶、[(6-乙酰氨基己酰基)氧基]膦酸(6-AAHOP)酰基转移酶和6-乙酰氨基己酰基辅酶A氧化还原酶；或3-氧代-6-氨基庚二酰辅酶A氧化还原酶、3-羟基-6-氨基庚二酰辅酶A脱水酶和高赖氨酸脱羧酶；或2-氧代-4-羟基-7-氨基庚酸醛缩酶、2-氧代-7-氨基庚-3-烯酸还原酶和高赖氨酸脱羧酶；或6-乙酰氨基己酸还原酶、6-乙酰氨基己醛氨基转移酶和6-乙酰氨基己胺N-乙酰转移酶。类似地，如需要，在非天然存在的微生物有机体中可以包括本文公开的生物合成途径的四种或更多种酶的任何组合，只要所需生物合成途径的酶的组合导致产生相应的所需产品。
除了如本文所述的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成之外，非天然存在的微生物有机体和方法也可以以各种相互组合以及与本领域熟知的通过其他途径实现产物生物合成的其他微生物有机体和方法一起使用。例如，除了使用6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产者之外的一种产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的替代方案是通过添加另一种能够将己二酸、6-氨基己酸或己内酰胺途径中间体转化为6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的微生物有机体。一种这样的过程包括，例如，对产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体的微生物有机体进行发酵。然后，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体可用作第二微生物有机体的底物，该第二微生物有机体将6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体转化为6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体可以直接添加到第二有机体的另一培养物中，或者6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中间体生产者的原始培养物中可以使这些微生物有机体耗尽(例如，通过细胞分离)，然后将第二有机体添加到发酵液中可用于生产最终产品，而无需中间纯化步骤。
在其他实施方案中，非天然存在的微生物有机体和方法可以在多种亚途径中组装以实现例如6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成。在这些实施方案中，所需产物的生物合成途径可以分离到不同的微生物有机体中，并且可以对不同的微生物有机体进行共培养以产生最终产物。在这样的生物合成方案中，一种微生物有机体的产物是第二微生物有机体的底物，直到合成最终产物。例如，可以通过构建包含用于将一种途径中间体转化为另一种途径中间体或产物的生物合成途径的微生物有机体来完成6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成。可替换地，也可以通过在同一容器中使用两种有机体进行共培养或共发酵而从微生物有机体生物合成产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸，其中第一微生物有机体产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸中间体，第二微生物有机体将中间体转化为6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。
鉴于本文提供的教导和指导，本领域技术人员将理解非天然存在的微生物有机体和方法以及其他微生物有机体，与具有亚途径的其他非天然存在的微生物有机体的共培养以及与本领域熟知的其他化学和/或生化过程一起存在多种组合和排列以产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。
类似地，本领域技术人员理解，宿主有机体可以基于引入一种或多种基因破坏以增加6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生产的所需特征来选择。因此，应理解，如果要讲遗传修饰引入宿主有机体中以破坏基因，可以相似地破坏催化相似但不完全相同的代谢反应的任何同源物、直系同源物或旁系同源物以确保充分破坏所需的代谢反应。由于不同有机体之间的代谢网络存在某些差异，因此本领域技术人员将理解在给定有机体中被破坏的实际基因在有机体之间可能不同。然而，鉴于本文提供的教导和指导，本领域技术人员将理解，该方法可应用于任何合适的宿主微生物体以鉴定在目标物种中构建将增加6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成的有机体所需的同源代谢改变。在具体实施方案中，如果需要并如本文所公开，增加的生产将使6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成与有机体的生长耦合，并且可以将6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生产与有机体的生长强制性地耦合。
6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶的编码核酸的来源可以包括，例如，其中编码的基因产物能够催化所提及的反应的任何物种。此类物种包括原核有机体和真核有机体，包括但不限于细菌(包括古细菌和真细菌)，以及真核生物，包括酵母、植物、昆虫、动物和哺乳动物(包括人)。在一些实施方案中，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶的编码核酸的来源显示在表1中。在一些实施方案中，醛脱氢酶的编码核酸的来源显示在表1中.在其他实施方案中，醛脱氢酶的编码核酸的来源是：Acidaminococcus、Collinsella、Peptostreptococcaceae或Romboustsia。在一些实施方案中，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶的编码核酸的来源为例如以下物质：Escherichia coli、Escherichia coli str.K12、Escherichia coli C、Escherichia coli W、Pseudomonassp、Pseudomonas knackmussii、Pseudomonas sp.Strain B13、Pseudomonas putida、Pseudomonas fluorescens、Pseudomonas stutzeri、Pseudomonas mendocina、Rhodopseudomonas palustris、Mycobacterium tuberculosis、Vibrio cholera、Heliobacter pylori、Klebsiella pneumoniae、Serratia proteamaculans、Streptomycessp.2065、Pseudomonas aeruginosa、Pseudomonas aeruginosa PAO1、Ralstoniaeutropha、Ralstonia eutropha H16、Clostridium acetobutylicum、Euglena gracilis、Treponema denticola、Clostridium kluyveri、Homo sapiens、Rattus norvegicus、Acinetobacter sp.ADP1、Acinetobacter sp.Strain M-1、Streptomyces coelicolor、Eubacterium barkeri、Peptostreptococcus asaccharolyticus、Clostridiumbotulinum、Clostridium botulinum A3 str、Clostridium tyrobutyricum、Clostridiumpasteurianum、Clostridium thermoaceticum(Moorella thermoaceticum)、Moorellathermoacetica Acinetobacter calcoaceticus、Mus musculus、Sus scrofa、Flavobacterium sp、Arthrobacter aurescens、Penicillium chrysogenum、Aspergillusniger、Aspergillus nidulans、Bacillus subtilis、Saccharomyces cerevisiae、Zymomonas mobilis、Mannheimia succiniciproducens、Clostridium ljungdahlii、Clostridium carboxydivorans、Geobacillus stearothermophilus、Agrobacteriumtumefaciens、Achromobacter denitrificans、Arabidopsis thaliana、Haemophilusinfluenzae、Acidaminococcus fermentans、Clostridium sp.M62/1、Fusobacteriumnucleatum、Bos taurus、Zoogloea ramigera、Rhodobacter sphaeroides、Clostridiumbeijerinckii、Metallosphaera sedula、Thermoanaerobacter species、Thermoanaerobacter brockii、Acinetobacter baylyi、Porphyromonas gingivalis、Leuconostoc mesenteroides、Sulfolobus tokodaii、Sulfolobus tokodaii 7、Sulfolobus solfataricus、Sulfolobus solfataricus、Sulfolobus acidocaldarius、Salmonella typhimurium、Salmonella enterica、Thermotoga maritima、Halobacteriumsalinarum、Bacillus cereus、Clostridium difficile、Alkaliphilusmetalliredigenes、Thermoanaerobacter tengcongensis、Saccharomyces kluyveri、Helicobacter pylori、Corynebacterium glutamicum、Clostridiumsaccharoperbutylacetonicum、Pseudomonas chlororaphis、Streptomycesclavuligerus、Campylobacter jejuni、Thermus thermophilus、Pelotomaculumthermopropionicum、Bacteroides capillosus、Anaerotruncuscolihominis、Natranaerobius thermophilius、Archaeoglobus fulgidus、Archaeoglobusfulgidus DSM 4304、Haloarcula marismortui、Pyrobaculum aerophilum、Pyrobaculumaerophilum str.IM2、Nicotiana tabacum、Menthe piperita、Pinus taeda、Hordeumvulgare、Zea mays、Rhodococcus opacus、Cupriavidus necator、Bradyrhizobiumjaponicum、Bradyrhizobium japonicum USDA110,Ascarius suum、butyrate-producingbacterium L2-50、Bacillus megaterium、Methanococcus maripaludis、Methanosarcinamazei、Methanosarcina mazei、Methanocarcina barkeri、Methanocaldococcusjannaschii、Caenorhabditis elegans、Leishmania major、Methylomicrobiumalcaliphilum20Z、Chromohalobacter salexigens、Archaeglubus fulgidus、Chlamydomonas reinhardtii、trichomonas vaginalis G3、Trypanosoma brucei、Mycoplana ramose、Micrococcus luteas、Acetobacter pasteurians、Kluyveromyceslactis、Mesorhizobium loti、Lactococcus lactis、Lysinibacillus sphaericus、Candida boidinii、Candida albicans SC5314、Burkholderia ambifaria AMMD、Ascarissuun、Acinetobacter baumanii、Acinetobacter calcoaceticus、Burkholderiaphymatum、Candida albicans、Clostridium subterminale、Cupriavidus taiwanensis、Flavobacterium lutescens、Lachancea kluyveri、Lactobacillus sp.30a、Leptospirainterrogans、Moorella thermoacetica、Myxococcus xanthus、Nicotiana glutinosa、Nocardia iowensis(sp.NRRL5646)、Pseudomonas reinekei MT1、Ralstonia eutrophaJMP134、Ralstonia metallidurans、Rhodococcus jostii、Schizosaccharomyces pombe、Selenomonas ruminantium、Streptomyces clavuligenus、Syntrophus aciditrophicus、Vibrio parahaemolyticus、Vibrio vulnificus，以及本文公开的或可用作相应基因的源有机体的其他示例性物种(参见实施例)。然而，由于现在可获得超过550个物种的完整基因组序列(其中超过一半可在公共数据库如NCBI中获得)，包括395个微生物基因组和各种酵母、真菌、植物和哺乳动物基因组，针对相关或远缘物种中一种或多种基因，鉴定编码所需的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成活性的基因，包括例如已知基因的同源物、直系同源物、旁系同源物和非直系同源基因置换，以及有机体之间遗传改变的互换，是本领域常规的和熟知的。因此，提及特别的有机体比如大肠杆菌的本文所述的能够进行6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成的代谢改变可以容易地应用于其他微生物体，包括原核有机体和真核有机体等。鉴于本文提供的教导和指导，本领域技术人员将知道在一种有机体中举例说明的代谢改变可以同样地应用于其他有机体。
在一些情况下，例如当6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径存在于不相关物种中时，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成可通过以下方式赋予在宿主物种上例如，通过来自不相关物种的催化相似但不完全相同的代谢反应以取代所提及的反应的一种旁系同源物或多种旁系同源物的外源表达。由于不同有机体之间的代谢网络存在一定的差异，因此本领域技术人员会理解，不同有机体之间的实际基因用途可能会有所不同。然而，鉴于本文提供的教导和指导，本领域技术人员还将理解，该教导和方法可应用于使用与本文示例的那些同源的代谢改变的所有微生物有机体以在目的物种中构建会合成6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的微生物有机体。
宿主微生物有机体可以选自例如细菌、酵母、真菌或适用于发酵过程的多种其他微生物中的任一种，并且非天然存在的微生物体在例如细菌、酵母、真菌或适用于发酵过程的多种其他微生物中的任一种中产生。示例性细菌包括选自以下的物种：Escherichiacoli、Klebsiella oxytoca、Anaerobiospirillum succiniciproducens、Actinobacillussuccinogenes、Mannheimia succiniciproducens、Rhizobium etli、Bacillus subtilis、Corynebacterium glutamicum、Gluconobacter oxydans、Zymomonas mobilis、Lactococcus lactis、Lactobacillus plantarum、Streptomyces coelicolor、Clostridium acetobutylicum、Pseudomonas fluorescens和Pseudomonas putida。示例性酵母或真菌包括选自以下的物种：Saccharomyces cerevisiae、Schizosaccharomycespombe、Kluyveromyces lactis、Kluyveromyces marxianus、Aspergillus terreus、Aspergillus niger、Pichia pastoris、Rhizopus arrhizus、Rhizobus oryzae等。例如，大肠杆菌是一种特别有用的宿主有机体，因为它是一种已充分表征的适合进行基因改造的微生物有机体。其他特别有用的宿主有机体包括酵母，例如Saccharomyces cerevisiae。应理解，任何合适的微生物宿主有机体均可用于引入代谢和/或遗传修饰以产生所需产物。
构建和测试非天然存在的产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的宿主的表达水平的方法可以例如通过本领域熟知的重组和检测方法进行。可以在例如Sambrooket al.,Molecular Cloning:A Laboratory Manual,Third Ed.,Cold Spring HarborLaboratory,New York(2001)和Ausubel et al.,Current Protocols in MolecularBiology,John Wiley and Sons,Baltimore,MD(1999)中找到此类方法。
可以使用本领域熟知的技术，包括但不限于接合、电穿孔、化学转化、转导、转染和超声转化，将参与产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的途径的外源核酸序列稳定或瞬时引入宿主细胞中。对于在大肠杆菌或其他原核细胞中的外源表达，真核核酸的基因或cDNA中的一些核酸序列可以编码靶向信号，例如N末端线粒体或其他靶向信号，如果需要，该靶向信号可以在转化到原核宿主细胞之前去除。例如，去除线粒体前导序列导致在E.coli中的表达增加(Hoffmeister et al.,J.Biol.Chem.280:4329-4338(2005))。对于在酵母或其他真核细胞中的外源表达，基因可以在不添加前导序列的情况下在胞质溶胶中表达，或者可以通过添加合适的靶向序列(例如适合宿主细胞的线粒体靶向或分泌信号)，靶向线粒体或其他细胞器，或者靶向分泌。因此，应当理解，对核酸序列进行的用以去除或包括靶向序列的适当修饰可以掺入到外源核酸序列中以赋予所需特性。此外，可以使用本领域熟知的技术对基因进行密码子优化以实现蛋白质的优化表达。
可以构建一种或多种表达载体以包括可操作地连接至在宿主有机体中起作用的表达控制序列的、如本文所例举的一种或多种6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径编码核酸。适用于微生物宿主有机体的表达载体包括，例如，质粒、噬菌体载体、病毒载体、附加体和人工染色体，包括可操作用于稳定整合到宿主染色体中的载体和选择序列或标记。此外，表达载体可以包括一种或多种选择性标记基因和合适的表达控制序列。还可以包括这样的选择性标记基因，该选择性标记基因例如提供对抗生素或毒素的抗性、补充营养缺陷型的缺陷或提供培养基中未含的关键营养物。表达控制序列可以包括本领域熟知的组成型和诱导型启动子、转录增强子、转录终止子等。当共表达两种或更多种外源编码核酸时，可以将两种核酸插入例如单个表达载体或单独的表达载体中。对于单个载体表达，编码核酸可以可操作地连接至一种共同的表达控制序列或连接至不同的表达控制序列，例如一种诱导型启动子和一种组成型启动子。可以使用本领域公知的方法来确认参与代谢或合成途径的外源核酸序列的转化。此类方法包括，例如核酸分析，例如，RNA的Northern印迹或聚合酶链反应(PCR)扩增、或用于表达基因产物的免疫印迹、或用于测试引入的核酸序列或其相应的基因产物的表达的其他合适的分析方法。本领域技术人员理解，外源核酸以足以产生所需产物的量表达，并且进一步理解，可以使用本领域熟知且如在此公开的方法优化表达水平以获得足够的表达。
在一些实施方案中，是生产所需中间体或产物例如己二酸、6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的方法。例如，用于生产己二酸的方法可以包括在产生己二酸的条件下培养具有己二酸途径的非天然存在的微生物足够的时间段，该途径包括编码己二酸途径酶的至少一种外源核酸，该己二酸途径酶按足以产生己二酸的量来表达，该己二酸途径包括琥珀酰辅酶A：乙酰辅酶A酰基转移酶、3-羟酰基辅酶A脱氢酶、3-羟己二酰辅酶A脱水酶、5-羧基-2-戊烯酰辅酶A还原酶和己二酰辅酶A合成酶或磷酸反式己二酰酶/己二酸激酶或己二酰辅酶A：乙酰辅酶A转移酶或己二酰辅酶A水解酶。此外，用于产生己二酸的方法可以包括在产生己二酸的条件下培养具有己二酸途径的非天然存在的微生物有机体足够的时间段，该途径包括编码己二酸途径酶的至少一种外源核酸，该己二酸途径酶按足以产生己二酸的量来表达，该己二酸途径包括琥珀酰辅酶A：乙酰辅酶A酰基转移酶、3-氧代己二酰辅酶A转移酶、3-氧代己二酸还原酶、3-羟基己二酸脱水酶和2-烯酸还原酶。
此外，用于生产6-氨基己酸的方法可包括在产生6-氨基己酸的条件下培养具有6-氨基己酸途径的非天然存在的微生物有机体足够的时间段，该途径包括编码6-氨基己酸途径酶的至少一种外源核酸，该6-氨基己酸途径酶按足以产生6-氨基己酸的量来表达，该6-氨基己酸途径包括辅酶A依赖性醛脱氢酶和转氨酶或6-氨基己酸脱氢酶。此外，用于产生己内酰胺的方法可包括在产生己内酰胺的条件下培养具有己内酰胺途径的非天然存在的微生物有机体足够的时间段，该途径包括编码己内酰胺途径酶的至少一种外源核酸，该己内酰胺途径酶按足以产生己内酰胺的量来表达，该己内酰胺途径包括辅酶A依赖性醛脱氢酶、转氨酶或6-氨基己酸脱氢酶和酰胺水解酶。
可以使用熟知的方法进行合适的纯化和/或测定以测试6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的产生。可以针对每种要测试的工程菌株培养合适的复制品，例如一式三份培养物。例如，可以监控工程生产宿主中的产物和副产物形成。可以通过如HPLC(高效液相色谱)、GC-MS(气相色谱-质谱)和LC-MS(液相色谱-质谱)的方法或使用本领域熟知的常规过程的其他合适的分析方法来分析最终产物和中间体以及其他有机化合物。也可以用培养物上清液来测试产物在发酵液中的释放。可以通过HPLC，例如使用用于葡萄糖和醇的折光率检测器和用于有机酸的UV检测器(Lin et al.,Biotechnol.Bioeng.90:775-779(2005))，或本领域熟知的其他合适的测定和检测方法，对副产物和残余葡萄糖进行定量。也可以使用本领域公知的方法来测定来自外源DNA序列的单个酶活性。
可以使用本领域熟知的多种方法将]6-氨基己酸、己内酰胺、己二胺或乙酰丙酸与培养物中的其他组分分离。此类分离方法包括例如萃取法以及包括连续液-液萃取、渗透蒸发、膜过滤、膜分离、反渗透、电渗析、蒸馏、结晶、离心、萃取过滤、离子交换色谱、尺寸排阻色谱、吸附色谱和超滤的方法。所有上述方法是本领域公知的。
可以培养本文所述的任何非天然存在的微生物有机体以产生和/或分泌生物合成产物。例如，可培养6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产者以进行6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成生产。
为了生产6-氨基己酸、己内酰胺、己二胺或乙酰丙酸，在具有碳源和其他必需营养物的培养基中培养重组菌株。有时需要并且可能非常需要在发酵罐中保持厌氧条件以降低整个过程的成本。例如，这样的条件可以通过首先用氮气使培养基鼓泡然后用带隔膜压合盖(septum and crimp-cap)对烧瓶进行密封来获得。对于在厌氧下未观察到生长的菌株，可以通过在隔膜上开小孔以进行有限的曝气来应用微需氧或基本厌氧条件。示例性厌氧条件先前已经进行了描述并且是本领域公知的。例如，2011年5月24日发行的专利号为7,947,483的美国专利中描述了示例性需氧和厌氧条件。如本文所公开的，发酵可以以分批、补料分批或连续方式进行。
如果需要，可以根据将培养基保持在理想的pH值的需要，通过添加碱(例如NaOH或其他碱)或酸，将培养基的pH值维持在所需的pH值，特别是中性pH值，例如约7的pH值。可以通过使用分光光度计(600nm)测量光密度来确定生长速率，并通过监测碳源随时间的消耗来确定葡萄糖摄取率。
生长培养基可以包括，例如，可以为非天然存在的微生物体供应碳源的任何碳水化合物来源。此类来源包括例如糖，例如葡萄糖、木糖、阿拉伯糖、半乳糖、甘露糖、果糖、蔗糖和淀粉。碳水化合物的其他来源包括例如可再生原料和生物质。可用作方法中的原料的示例性的生物质类型包括纤维素生物质、半纤维素生物质和木质素原料或部分原料。此类生物质原料包含例如可用作碳源的碳水化合物底物，例如葡萄糖、木糖、阿拉伯糖、半乳糖、甘露糖、果糖和淀粉。鉴于本文提供的教导和指导，本领域技术人员将理解，除了以上示例的那些之外的可再生原料和生物质也可用于培养微生物以产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。
除了可再生原料例如上面示例的那些之外，也可以对6-氨基己酸、己内酰胺、己二胺或乙酰丙酸微生物有机体进行修饰从而在作为其碳源的合成气上生长。在该特定实施方案中，在产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的有机体中表达一种或多种蛋白质或酶，以提供利用合成气或其他气态碳源的代谢途径。
合成气体，也称为合成气或发生炉气(producer gas)，是煤和含碳材料如生物质材料(包括农作物和残余物)的主要气化产物。合成气主要是H2和CO的混合物，可以从包括但不限于煤、煤油、天然气、生物质和废弃有机物的任何有机原料的气化中获得。气化通常在高燃料氧气比下进行。虽然主要是H2和CO，但合成气也可以包括少量的CO2和其他气体。因此，合成气提供了具有成本效益的气态碳源，例如CO和另外的CO2。
Wood-Ljungdahl途径催化CO和H2转化为乙酰辅酶A和其他产物例如乙酸盐。能够利用CO和合成气的有机体通常也具有通过Wood-Ljungdahl途径所包含的相同的基础酶组和转化来利用CO2和CO2/H2混合物的能力。早在发现CO也可以被相同的有机体使用并且涉及相同的途径之前，就已认识到微生物体依赖H2将CO2转化为乙酸盐。许多产乙酸菌已表现出只要存在氢气以提供必要的还原当量，就会在CO2存在下生长并产生化合物如乙酸盐(例如，参见Drake,Acetogenesis,pp.3-60 Chapman and Hall,New York,(1994))。这可以用以下等式来概括：
2CO2+4H2+n ADP+n Pi→CH3COOH+2H2O+n ATP
因此，具有Wood-Ljungdahl途径的非天然存在的微生物体也可以利用CO2和H2混合物来生产乙酰辅酶A和其他所需产物。
Wood-Ljungdahl途径是本领域熟知的并且由12个反应组成，这些反应可以分成两个分支：(1)甲基分支和(2)羰基分支。甲基分支将合成气转化为甲基四氢叶酸(甲基-THF)，而羰基分支将甲基-THF转化为乙酰辅酶A。甲基分支中的反应依次被下列酶催化：铁氧还蛋白氧化还原酶、甲酸脱氢酶、甲酰四氢叶酸合成酶、甲川四氢叶酸环化脱水酶、亚甲基四氢叶酸脱氢酶和亚甲基四氢叶酸还原酶。羰基分支中的反应依次被下列酶或蛋白质催化：钴胺素类咕啉/铁硫蛋白(cobalamide corrinoid/iron-sulfur protein)、甲基转移酶、一氧化碳脱氢酶、乙酰辅酶A合酶、乙酰辅酶A合酶二硫化物还原酶和氢化酶，这些酶也可以被称为甲基四氢叶酸：类咕啉蛋白甲基转移酶(例如AcsE)、类咕啉铁硫蛋白、镍蛋白组装蛋白(例如AcsF)、铁氧还蛋白、乙酰辅酶A合酶、一氧化碳脱氢酶和镍蛋白组装蛋白(例如，CooC)。按照本文提供的用于引入足量的编码核酸以产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径的教导和指导，本领域技术人员将理解，也可以就至少引入编码宿主有机体中不存在的Wood-Ljungdahl酶或蛋白质的核酸方面进行相同的工程设计。因此，将一种或多种编码核酸引入微生物有机体来使得经修饰的有机体包含完整的Wood-Ljungdahl途径，将赋予合成气利用能力。
此外，与一氧化碳脱氢酶和/或氢化酶活性耦合的还原性(反向)三羧酸循环还可用于将CO、CO2和/或H2转化为乙酰辅酶A和其他产物例如乙酸盐。能够通过还原性TCA途径固定碳的有机体可以利用以下酶中的一种或多种：ATP柠檬酸裂解酶、柠檬酸裂解酶、顺乌头酸酶、异柠檬酸脱氢酶、α-酮戊二酸：铁氧还蛋白氧化还原酶、琥珀酰辅酶A合成酶、琥珀酰辅酶A转移酶、延胡索酸还原酶、延胡索酸酶、苹果酸脱氢酶、NAD(P)铁氧还蛋白氧化还原酶、一氧化碳脱氢酶和氢化酶。具体地，通过一氧化碳脱氢酶和氢化酶从CO和/或H2中提取的还原当量用于通过还原性TCA循环将CO2固定为乙酰辅酶A或乙酸盐。乙酸可通过酶，比如乙酰辅酶A转移酶、乙酸激酶/磷酸转乙酰酶和乙酰辅酶A合成酶，转化为乙酰辅酶A。乙酰辅酶A可通过丙酮酸：铁氧还蛋白氧化还原酶和糖异生的酶转化为对甲苯甲酸、对苯二甲酸或(2-羟基-3-甲基-4-氧代丁氧基)膦酸前体、3-磷酸甘油醛、磷酸烯醇丙酮酸和丙酮酸。按照本文提供的关于引入足量的编码核酸以产生对甲苯甲酸、对苯二甲酸或(2-羟基-3-甲基-4-氧代丁氧基)膦酸途径的教导和指导，本领域技术人员将理解还可以针对至少引入编码宿主有机体中不存在的还原性TCA途径酶或蛋白质的核酸进行相同的工程设计。因此，将一种或多种编码核酸引入微生物有机体使得经修饰的有机体包含完整的还原性TCA途径将赋予合成气利用能力。
鉴于本文提供的教导和指导，本领域技术人员将理解可以产生在碳源例如碳水化合物上生长时分泌生物合成的化合物的非天然存在的微生物有机体。此类化合物包括例如6-氨基己酸、己内酰胺、己二胺或乙酰丙酸以及6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中的任何中间代谢物。所需要的只是改造一种或多种所需的酶活性以实现所需化合物或中间体的生物合成，这包括例如纳入6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生物合成途径中的一些或全部。因此，一些实施方案提供非天然存在的微生物有机体，该非天然存在的微生物有机体当在碳水化合物上生长时产生和/或分泌6-氨基己酸、己内酰胺、己二胺或乙酰丙酸并且产生和/或分泌6-氨基己酸、己内酰胺、己二胺或乙酰丙酸且当在碳水化合物上生长时产生和/或分泌6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径中的示出的任何中间代谢物。例如，如预期，产生己二酸的微生物有机体可以从中间体开始合成，该中间体例如，3-氧代己二酰辅酶A、3-羟基己二酰辅酶A、5-羧基-2-戊烯酰辅酶A或己二酰辅酶A(见图1)。此外，产生己二酸的微生物有机体可以从中间体开始合成，该中间体例如，3-氧代己二酰辅酶A、3-氧代己二酸、3-羟基己二酸或六-2-烯二酸。产生6-氨基己酸的微生物有机体可以从中间体例如己二酸半缩醛开始合成。根据需要，产生己内酰胺的微生物有机体可以从中间体(例如，己二酸半缩醛或6-氨基己酸)开始合成(见图1)。
在一些实施方案中，非天然存在的微生物有机体还包括编码反式烯酰辅酶A还原酶(TER)的外源表达的核酸。TER与5-羧基-2-戊烯酰辅酶A反应生成己二酰辅酶A。在一些实施方案中，TER可以是已知的TER，而在其他实施方案中，TER酶是经改造的。在一些实施方案中，改造的反式烯酰辅酶A还原酶具有与SEQ ID NO:189的氨基酸序列具有至少50％一致性的氨基酸序列，其中改造的反式烯酰辅酶A还原酶包含表2中所示的变体的任何氨基酸序列改变。
表2
Figure BDA0003433304930000451
Figure BDA0003433304930000461
Figure BDA0003433304930000471
Figure BDA0003433304930000481
在一些实施方案中，非天然存在的微生物有机体具有己二胺途径，该己二胺途径包括(i)6-氨基己酰辅酶A转移酶，(ii)6-氨基己酰辅酶A合酶，(iii)6-氨基己酰辅酶A还原酶，(iv)己二胺转氨酶，(v)己二胺脱氢酶，(v)或一种或多种酶(i)-(v)的组合。在其他实施方案中，非天然存在的微生物有机体具有己二胺途径，该己二胺途径包括3-氧代己二酰辅酶A硫解酶(Thl)、3-氧代己二酰辅酶A脱氢酶(Hbd)和3-氧代己二酰辅酶A脱水酶(“巴豆酸酶”或Crt)、5-羧基-2-戊烯酰-辅酶A还原酶(Ter)、转氨酶(HMD TA)和羧酸还原酶(CAR)。
使用本领域熟知的方法如本文所示例构建非天然存在的微生物有机体，以按照足以产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的量进行编码6-氨基己酸、己内酰胺、己二胺或乙酰丙酸途径酶的至少一种核酸的外源表达。应当理解，微生物有机体在足以产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的条件下培养。按照本文提供的教导和指导，非天然存在的微生物有机体可以实现6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成，从而导致细胞内浓度在约0.1mM-200mM之间或更高。通常，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的细胞内浓度在约3mM-150mM之间，特别地在约5mM-125mM之间，更特别地在约8mM-100mM之间，包括约10mM、20mM、50mM、80mM或更多。也可以从非天然存在的微生物有机体中实现在这些示例性范围中的每一个之间和这些示例性范围中的每一个之上的细胞内浓度。
在一些实施例中，培养条件包括厌氧或基本厌氧的生长或维持条件。示例性厌氧条件先前已经描述并且是本领域熟知的。发酵过程的示例性厌氧条件在本文中进行了描述，并在例如2011年5月24日发布的专利号为7,947,483的美国专利中进行了描述。这些条件中的任何一个都可以与非天然存在的微生物有机体以及本领域熟知的其他厌氧条件一起使用。在这种厌氧条件下，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产者可以合成细胞内浓度为5mM-10mM或更高以及本文示例的所有其他浓度的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。应理解，即使以上描述是指细胞内浓度，但产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的微生物有机体可以在细胞内产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸和/或将产物分泌到培养基。
培养条件可以包括，例如，液体培养过程以及发酵和其他大规模培养过程。如本文所述，可在厌氧或基本厌氧培养条件下获得生物合成产物的特别有用的产量。
如本文所述，用于实现6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成的一种示例性生长条件包括厌氧培养或发酵条件。在某些实施方案中，非天然存在的微生物可以在厌氧或基本厌氧条件下维持、培养或发酵。简而言之，厌氧条件是指没有氧气的环境。基本厌氧条件包括例如使得培养基中的溶解氧浓度保持在0至10％的饱和度之间的培养、分批发酵或连续发酵。基本厌氧条件还包括使细胞在液体培养基或固体琼脂上保持低于1％氧气气氛的密封室内生长或静息。氧气的百分比可以通过例如用N2/CO2混合物或其他合适的一种或多种非氧气气体对培养物进行鼓泡来保持。
本文所述的培养条件可以按比例放大并连续生长以生产6-氨基己酸、己内酰胺、己二胺或乙酰丙酸。示例性的生长过程包括，例如：补料分批发酵和分批分离；补料分批发酵和连续分离；或连续发酵和连续分离。所有这些工艺都是本领域熟知的。发酵过程对于商用量的6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成生产特别有用。通常，与非连续培养过程一样，6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的连续和/或接近连续生产将包括在足够的营养和培养基中培养产生6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的非天然存在的有机体以维持和/或几乎维持指数阶段的生长。在此类条件下的连续培养可包括例如1天、2天、3天、4天、5天、6天或7天或更长时间。可替换地，连续培养可包括1周、2周、3周、4周或5周或更多周以及直到数月。可替换地，如果适合特定应用，则可以培养有机体数小时。应当理解，连续和/或接近连续培养条件还可以包括这些示例性时段之间的所有时间间隔。还应理解，培养微生物有机体的时间是足够长以产生足够量用于所需目的的产品的时间段。
本领域熟知发酵过程。简而言之，用于6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成生产的发酵可用于例如补料分批发酵和分批分离、补料分批发酵和连续分离、或连续发酵和连续分离。本领域熟知分批和连续发酵步骤的实例。
除了使用6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产者连续生产大量6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的上述发酵过程之外，如果需要，也可以例如同时对6-氨基己酸、己内酰胺、己二胺或乙酰丙酸生产者进行化学合成过程以将产物转化为其他化合物，或者可以将产物从发酵培养物中分离并继而进行化学转化以将产物转化为其他化合物。如本文所述，利用3-氧代己二酸、六-2-烯二酸的己二酸途径中的中间体可以例如通过在铂催化剂上的化学氢化而转化为己二酸。
如本文所述，用于实现6-氨基己酸、己内酰胺、己二胺或乙酰丙酸的生物合成的示例性生长条件包括向培养条件添加渗透保护剂。在某些实施方案中，非天然存在的微生物有机体可以在渗透保护剂的存在下如上所述进行维持、培养或发酵。简而言之，渗透保护剂是指充当渗透剂并帮助本文所述的微生物有机体在渗透胁迫中存活的化合物。渗透保护剂包括但不限于：甜菜碱、氨基酸和糖海藻糖。这类的非限制性实例是甘氨酸甜菜碱、脯氨酸甜菜碱、二甲基噻亭、二甲基磺酰丙酸、3-二甲基磺酰-2-甲基丙酸、哌可酸、二甲基磺酰乙酸、胆碱、L-肉碱和四氢嘧啶。一方面，渗透保护剂是甘氨酸甜菜碱。本领域普通技术人员应理解，适用于保护本文描述的微生物有机体免受渗透胁迫的渗透保护剂的量和类型将取决于所使用的微生物有机体。例如，如实施例XXII中所述，在存在不同量的6-氨基己酸的情况下，大肠杆菌在2mM甘氨酸甜菜碱存在下合适地生长。培养条件中渗透保护剂的量可以是例如不超过约0.1mM、不超过约0.5mM、不超过约1.0mM、不超过约1.5mM、不超过约2.0mM、不超过约2.5mM、不超过约3.0mM、不超过约5.0mM、不超过约7.0mM、不超过约10mM、不超过约50mM、不超过约100mM或不超过约500mM。
成功地改造途径涉及鉴定出合适的具有足够活性和特异性的酶组。这需要鉴定出合适的酶组，将其相应的基因克隆到生产宿主中，优化发酵条件，并测定发酵后的产物形成。为了改造用于生产6-氨基己酸或己内酰胺的生产宿主，可以在宿主微生物体中表达一种或多种外源DNA序列。此外，微生物体可以使内源基因功能性缺失。这些修改将允许使用可再生原料来生产6-氨基己酸或己内酰胺。
在一些实施方案中，在将6-氨基己酸转化为HMDA期间使环状亚胺或己内酰胺的形成最小化或甚至消除需要向6-氨基己酸的胺基团添加官能团(例如，乙酰基、琥珀酰基)以防止其环化。这类似于在大肠杆菌中由L-谷氨酸形成鸟氨酸。具体地，谷氨酸首先通过N-乙酰谷氨酸合酶转化为N-乙酰基-L-谷氨酸。然后，N-乙酰基-L-谷氨酸被活化为N-乙酰谷氨酰-磷酸，N-乙酰基谷氨酰基-磷酸被还原并转氨以形成N-乙酰基-L-鸟氨酸。然后通过N-乙酰基-L-鸟氨酸脱乙酰酶从N-乙酰基-L-鸟氨酸中除去乙酰基形成L-鸟氨酸。这种途径是必要的，因为由谷氨酸形成谷氨酸5-磷酸，然后还原为谷氨酸-5-半缩醛导致形成(S)-1-吡咯啉-5-羧酸，(S)-1-吡咯啉-5-羧酸为由谷氨酸-5-半缩醛自发形成的环状亚胺。在由6-氨基己酸形成HMDA的情况下，这些步骤可包括将6-氨基己酸乙酰化为乙酰基-6-氨基己酸、用辅酶A或磷酸基团活化羧酸基团、还原、胺化和脱乙酰化。
实施例
实施例1.针对在己二酰辅酶A上的活性筛选候选醛脱氢酶
在多个物种的基因组中对编码候选醛脱氢酶(Ald)的基因进行生物信息学鉴定(表1)。合成编码每种醛脱氢酶的基因，在大肠杆菌中表达，并评估Ald活性。
将编码表1的Ald酶候选物的基因克隆到低拷贝载体中在组成型启动子下，并使用标准技术将构建体转化到大肠杆菌中。转化体在存在抗生素的LB培养基中于35℃培养过夜，然后在室温下以15000rpm的速度收集细胞。为了制备裂解物，将细胞重新悬浮在含有溶菌酶、核酸酶和10mM DTT的化学裂解溶液中，并在室温下孵育至少30分钟。所得裂解物用于测试醛脱氢酶活性。
将裂解物(5μl)添加到测定混合物中，以产生20μL的总体积，最终浓度为0.1MTris-HCl(pH7.5)、2.5mM己二酰辅酶A(AdCoA)、和0.5mM NADH或0.5mM NADPH。该测定用于筛选所有Ald酶候选物。还使用琥珀酰辅酶A(SuCoA)或乙酰辅酶A(AcCoA)作为底物测试了一些Ald候选物。AdCoA、SuCoA和AcCoA从商业供应商获得。在辅酶A底物存在下，通过NADH或NADPH的荧光线性降低来监测活性。在表3中，将使用NADH或NADPH对己二酰辅酶A具有显著活性的Ald标示为正号(+)，而将几乎没有活性或没有活性的那些标示为负号(-)。
表3.醛脱氢酶在己二酰辅酶A上的活性
Figure BDA0003433304930000531
Figure BDA0003433304930000541
Figure BDA0003433304930000551
Figure BDA0003433304930000561
Figure BDA0003433304930000571
Figure BDA0003433304930000581
实施例2.进行醛脱氢酶测定以确定底物特异性
为了确定若干种醛脱氢酶的底物偏好，使用琥珀酰辅酶A和己二酰辅酶A底物来应用底物辅酶A消耗测定。在该测定中，底物溶液含有0.1M Tris-HCl(pH7.5)、1mM己二酰辅酶A、0.2mM琥珀酰辅酶A和0.2mM乙酰辅酶A，以及过量的1.5mM NADH或NADPH辅因子。通过将裂解物加入测定缓冲液中来引发反应并在室温下孵育2小时。用1％甲酸淬灭反应，然后通过LC/MS分析方法进行评估以对每种残留底物辅酶A进行定量。Ald活性测量为每种辅酶A底物的消耗百分比。相对于测定中存在的另一种辅酶A底物，特定辅酶A底物的消耗百分比更高表明对特定底物辅酶A的偏好。图2显示了Peptostreptococcaceae bacterium oral醛脱氢酶(SEQ ID NO:7)、Acidaminococcus massiliensis醛脱氢酶(SEQ ID NO:28)、Collinsella sp.GD7醛脱氢酶(SEQ ID NO:60)和Romboutsia lituseburensis DSM醛脱氢酶(SEQ ID NO:107)从测定混合物中消耗的己二酰辅酶A比琥珀酰辅酶A多得多，因此被描述为偏好己二酰辅酶A。来自Porphyromonas gingivalis W83(SEQ ID NO:2)的醛脱氢酶被发现偏好琥珀酰辅酶A。
实施例3.醛脱氢酶的体内测定
还在体内测定中测试了证明具有己二酰辅酶A底物偏好的醛脱氢酶，其中用包括醛脱氢酶基因的构建体转化表达编码3-氧代己二酰辅酶A硫解酶(Thl)、3-氧代己二酰辅酶A脱氢酶(Hbd)和3-氧代己二酰辅酶A脱水酶(“巴豆酶酶”或Crt)、5-羧基-2-戊烯酰辅酶A还原酶(Ter)和转氨酶(TA)的基因的大肠杆菌菌株。Thl、Hbd、Crt、Ter、TA大肠杆菌菌株包括生产6-氨基己酸(6ACA)所需的所有途径酶，Ald酶除外。编码Porphyromonas gingivalisW83Ald(SEQ ID NO:2)、Peptostreptococcaceae bacterium oralAld(SEQ ID NO:7)、Acidaminococcus massiliensis Ald(SEQ ID NO:28)、Collinsella sp.GD7Ald(SEQ IDNO:60)和Romboutsia lituseburensis DSM Ald(SEQ ID NO:107)的基因分别克隆到低拷贝数质粒载体中在组成型启动子下。使用标准技术将表达Ald基因的质粒转化到Thl/Hbd/Crt/Ter/TA菌株中。然后测试包含任何一种Ald基因的转化体的6-氨基己酸(6ACA)产量。在基本培养基中向工程E.coli细胞供给2％葡萄糖，于35℃孵育18小时后，收集细胞，并通过分析型HPLC或标准LS/MS分析方法评估上清液的6ACA。如表4所示，E.coli中编码Ald酶的基因(包括Thl、Hbd、Crt、Ter和TA基因)的表达导致这些菌株产生6ACA。
表4.ACA途径中醛脱氢酶的体内活性。
Figure BDA0003433304930000601
实施例4.醛脱氢酶的动力学表征
在与实施例1中描述的裂解物筛选相似的条件下进行动力学表征；然而，在这种情况下，使用纯化的蛋白质代替细胞裂解物。使用亲和色谱来纯化Acidaminococcusmassiliensis Ald(SEQ ID NO:28)、Collinsella sp.GD7Ald(SEQ ID NO:60)和Romboutsia lituseburensis DSM Ald(SEQ ID NO:107)。在这些测定中，改变每种底物辅酶A的浓度以确定转换数(kcat)、酶对酶的底物的亲和力(KM)、针对每种底物的每种选定的Ald酶的催化效率(kcat/KM)并示于下表5中。
表5：醛脱氢酶与不同底物的动力学参数
Figure BDA0003433304930000602
使用各种底物的各种醛脱氢酶的催化效率(kcat/KM)绘制在条形图中以进行比较(图3A)。按照己二酰辅酶A的kcat/KM相对于琥珀酰辅酶A的kcat/KM的比率来计算Ald同系物对己二酰辅酶A与琥珀酰辅酶A的催化效率(kcat/KM)。图3B显示被测定的所有三种Ald酶对己二酰辅酶A比对琥珀酰辅酶A具有更高的催化效率。图3C显示被测定的所有三种Ald酶对己二酰辅酶A比对乙酰辅酶A也具有更高的催化效率。
实施例5醛脱氢酶的体内测定
在体内测定中测试了实施例3描述的大肠杆菌菌株中被证明具有己二酰辅酶A底物偏好的醛脱氢酶，该大肠杆菌菌株表达编码3-氧代己二酰辅酶A硫解酶(Thl)、3-氧代己二酰辅酶A脱氢酶(Hbd)和3-氧代己二酰辅酶A脱水酶(“巴豆酸酶”或Crt)、5-羧基-2-戊烯酰辅酶A还原酶(Ter)和转氨酶(TA)的基因，又用包含两个两外的基因(羧酸还原酶(CAR-WP_003872682.1)和另外的TA基因(HMD-TA WP_001301395.1))的构建体转化，连同Ald基因一起整合到大肠杆菌染色体中。编码Porphyromonas gingivalis W83 Ald(SEQ ID NO:2)、Peptostreptococcaceae bacterium oralAld(SEQ ID NO:7)、Acidaminococcusmassiliensis Ald(SEQ ID NO:28)、Collinsella sp.GD7 Ald(SEQ ID NO:60)和Romboutsia lituseburensis DSM Ald(SEQ ID NO:107)的基因分别克隆到低拷贝数质粒载体中在组成型启动子下。使用标准技术将用于表达Ald基因的质粒转化到Thl/Hbd/Crt/Ter/TA/CAR菌株中。使这些构建体经历与实施例3中针对6ACA生产所述相同的条件和测试。根据通过实施例3中所述的LC/MS分析方法进行的检测，该构建体显示出产生HMD。
序列表
<110> 基因组股份公司
阿米特·M·沙阿
哈里什·纳加拉让
<120> 用于改进醛脱氢酶活性的工程微生物体和方法
<130> GNO0099/WO
<150> 62/837,888
<151> 2019-04-24
<160> 190
<170> PatentIn version 3.5
<210> 1
<211> 453
<212> PRT
<213> Clostridium kluyveri DSM555
<400> 1
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420 425 430
Thr Ile Ala Thr Pro Thr Gly Glu Gly Thr Thr Ser Ala Lys Thr Phe
435 440 445
Ala Arg Ser Arg Arg Cys Val Leu Thr Asn Gly Phe Ser Ile Arg
450 455 460
<210> 5
<400> 5
000
<210> 6
<400> 6
000
<210> 7
<211> 455
<212> PRT
<213> Peptostreptococcaceae bacterium oral
<400> 7
Met Leu Asp Pro Asn Ser Met Val Asn Glu Leu Ile Arg Arg Ala Arg
1 5 10 15
Thr Ala Gln Thr Glu Phe Glu Thr Tyr Ser Gln Glu Arg Val Asp Lys
20 25 30
Ala Val Arg Ala Ile Gly Lys Ser Ile Tyr Asp His Gly Asp Glu Leu
35 40 45
Ala Lys Met Gly Ala Glu Glu Ser Gly Met Gly Arg Tyr Glu Asp Lys
50 55 60
Ile Val Lys Asn Gln Gly Lys Ser Lys Met Thr Trp Trp Arg Leu Lys
65 70 75 80
Gly Val Lys Ser Arg Gly Ile Ile Asn Ile Asp Arg Glu Lys Gln Ile
85 90 95
Tyr Glu Ile Ala Lys Pro Ile Gly Val Leu Gly Val Val Thr Pro Ala
100 105 110
Thr Asn Pro Thr Met Thr Pro Val His Asn Ala Met Ile Ala Leu Lys
115 120 125
Gly Ala Asn Ala Val Ile Ile Cys Pro His Pro Lys Thr Arg Lys Thr
130 135 140
Thr Ser Lys Thr Val Glu Tyr Met Arg Leu Ala Leu Lys Asp Ile Ser
145 150 155 160
Val Pro Glu Asp Leu Ile Gln Ile Val Asp Asp Pro Ser Ile Glu Val
165 170 175
Ser Gln Ala Leu Met Ala Phe Cys Asp Thr Thr Ile Ser Thr Gly Gly
180 185 190
Pro Gly Met Val Lys Ser Ala Tyr Ser Ser Gly Lys Pro Ala Ile Gly
195 200 205
Val Gly Pro Gly Asn Val Gln Cys Leu Val Gly Asp Asp Ala Asp Ile
210 215 220
Asp Ala Ile Val Pro Lys Ile Met Lys Gly Arg Thr Tyr Asp Asn Gly
225 230 235 240
Val Leu Cys Thr Cys Glu Gln Ser Ile Ile Cys Ala Glu Asn Leu Tyr
245 250 255
Asp Arg Leu Val Lys Gly Leu Val Asp Asn Gly Ala Tyr Phe Val Lys
260 265 270
Glu Asp Glu Val Glu Lys Leu Arg Asn Gly Phe Phe Pro Gly Gly Val
275 280 285
Met Asn Lys Asn Leu Val Gly Ser Ser Pro Phe Glu Ile Ala Lys Ala
290 295 300
Ser Gly Phe Glu Val Gln Glu Glu Ser Lys Ile Leu Leu Val Pro Val
305 310 315 320
Ser Lys Thr Gly Lys Asp Glu Phe Leu Ala Lys Glu Lys Leu Ala Pro
325 330 335
Ile Leu Ala Leu Tyr Lys Tyr Ser Glu Trp Lys Glu Ala Val Asp Ile
340 345 350
Ala Leu Lys Asn Leu Leu Asn Glu Gly Arg Gly His Ser Val Val Ile
355 360 365
His Ser Ala Asn Lys Thr Asn Ile Glu Tyr Ala Ala Asn Ile Leu Pro
370 375 380
Val Ser Arg Val Gly Val Gly Met Val Gly Ser Ser Gly Leu Gly Gly
385 390 395 400
Gly Phe Asp Asn Gly Phe Met Pro Thr Ala Thr Leu Gly Cys Gly Ser
405 410 415
Trp Gly Asn Asn Ser Ile Ala Gly Asn Val Trp Trp Asn His Leu Val
420 425 430
Asn Ile Thr Lys Leu Ala Tyr Val Leu Asn Asp Val Ser Ile Pro Thr
435 440 445
Asp Glu Glu Ile Trp Ala Glu
450 455
<210> 8
<400> 8
000
<210> 9
<400> 9
000
<210> 10
<400> 10
000
<210> 11
<211> 452
<212> PRT
<213> Anaerocolumna jejuensis
<400> 11
Met Asn Gln Ile Ile Gln Ser Leu Val Glu Arg Ser Arg Lys Ala Gln
1 5 10 15
Gln Ile Leu Tyr Thr Tyr Asn Gln Glu Lys Thr Asp Glu Ile Val Glu
20 25 30
Met Phe Ala Ser Val Val Phe Asn His Ala Glu Pro Leu Ala Arg Met
35 40 45
Ala Val Glu Glu Ser Arg Met Gly Val Tyr Glu Asp Lys Ile Thr Lys
50 55 60
Asn Lys Glu Lys Ala Lys Thr Ile Trp Asn Ser Leu Lys Gly Lys Lys
65 70 75 80
Ser Ile Gly Ile Ile Gly Arg Glu Glu Glu Ala Gly Leu Ile Glu Ile
85 90 95
Ala Lys Pro Met Gly Val Ile Ala Ala Ala Met Pro Cys Thr Asn Pro
100 105 110
Ile Ile Thr Pro Met Cys Asn Ala Met Phe Ala Val Lys Cys Gln Asn
115 120 125
Thr Ile Ile Val Ala Pro His Pro Arg Gly Lys Lys Cys Ala Met Ala
130 135 140
Leu Ala Glu Leu Tyr Tyr Lys Glu Leu Asp Gly Met Gly Val Pro Arg
145 150 155 160
Asp Ile Phe Leu Val Val Glu Glu Pro Thr Ile Asp Leu Thr Thr Glu
165 170 175
Leu Met Ser Ala Cys Asp Thr Val Ile Ala Thr Gly Gly Met Gly Val
180 185 190
Val Lys Ser Ala Tyr Ser Ser Gly Lys Pro Ser Tyr Gly Val Gly Pro
195 200 205
Gly Asn Val Gln Gly Leu Ile Asp Glu Gly Ile Asp Tyr Arg Ala Ala
210 215 220
Ala Gly Arg Met Ile Ala Ser Arg Ile Phe Asp Asn Gly Ile Leu Cys
225 230 235 240
Thr Ser Thr Gln Ser Ile Ile Ala Pro Glu Lys Asp Tyr Glu Ser Val
245 250 255
Ile Lys Glu Phe Val Ala Gln Gly Ala Tyr Tyr Ile Asp Asp Pro Ala
260 265 270
Val Ile Ala Ser Leu Ser Glu Val Val Phe Pro Gly Gly Val Ile Asn
275 280 285
Lys Asn Val Val Gly Gln Ser Val Lys Thr Ile Ala Gly Leu Ala Gly
290 295 300
Ile Ser Ile Pro Glu Gly Thr Lys Val Ile Ile Val Lys Pro Glu Arg
305 310 315 320
His Gly Ala Gly Val Val Trp Ser Arg Glu Lys Met Cys Pro Met Met
325 330 335
Thr Ala Tyr Ser Tyr Lys Thr Trp Glu Glu Ala Val Gln Ile Ala Tyr
340 345 350
Asp Asn Leu Leu Val Glu Gly Glu Gly His Thr Ala Asp Ile Gln Ser
355 360 365
Asp Asn Gln Ala His Ile Glu Tyr Ala Gly Val Lys Leu Pro Val Ser
370 375 380
Arg Val Val Val Asn Gln Ser Cys Ser Val Met Ala Gly Gly Ala Phe
385 390 395 400
Gly Asn Ala Leu Asn Pro Ser Ala Thr Leu Gly Cys Gly Ser Trp Gly
405 410 415
Asn Asn Ala Ile Ser Glu Asn Leu Phe Tyr Thr His Leu Met Asn Lys
420 425 430
Ser Arg Ile Ala Phe Val Arg Lys Asn Trp Lys Gln Pro Ser Asp Glu
435 440 445
Glu Ile Phe Ala
450
<210> 12
<400> 12
000
<210> 13
<400> 13
000
<210> 14
<400> 14
000
<210> 15
<211> 475
<212> PRT
<213> Bacillus soli
<400> 15
Met Gln Ile Asn Glu Thr Asp Ile Lys Lys Met Val Glu Gln Val Leu
1 5 10 15
Lys Gln Leu Gly Glu Ser Gln Pro Ala Ser Ala Pro Ala Ala Ser Leu
20 25 30
Lys Asp Val Ser Tyr Gly Asp Gly Val Phe Ala Thr Val Asp Glu Ala
35 40 45
Ala Glu Ala Ala Arg Leu Ala Trp Glu Lys Leu Arg Lys Leu Pro Leu
50 55 60
Ala Ala Arg Arg Gln Met Ile Glu Asn Met Arg Glu Val Ser Arg Gln
65 70 75 80
His Val Asn Glu Leu Ala Thr Leu Ala Val Glu Glu Thr Lys Leu Gly
85 90 95
Arg Val Glu Asp Lys Val Ala Lys Ile Leu Leu Ala Val Asn Lys Thr
100 105 110
Pro Gly Val Glu Asp Leu Val Ser Thr Ala Phe Ser Gly Asp Asp Gly
115 120 125
Leu Thr Leu Val Glu Tyr Ala Pro Ile Gly Val Phe Gly Ser Ile Thr
130 135 140
Pro Ser Thr Asn Pro Ala Ala Thr Ile Ile Asn Asn Ser Ile Ser Leu
145 150 155 160
Val Ala Ala Gly Asn Thr Val Val Tyr Asn Pro His Pro Ser Ala Lys
165 170 175
Arg Val Ser Ile Lys Thr Leu Gln Leu Leu Asn Gln Ala Ile Val Ala
180 185 190
Ala Gly Gly Pro Glu Asn Thr Leu Thr Ser Val Ala Ala Pro Asn Leu
195 200 205
Glu Thr Ser Ala Gln Val Met Asn His Pro Lys Val His Ala Leu Val
210 215 220
Val Thr Gly Gly Gly Pro Val Val Lys Ala Ala Met Ala Val Gly Lys
225 230 235 240
Lys Val Ile Ala Ala Gly Pro Gly Asn Pro Pro Val Val Val Asp Glu
245 250 255
Thr Ala Ile Ile Ser Lys Ala Ala Ala Asp Ile Val Gln Gly Ala Ser
260 265 270
Phe Asp Asn Asn Val Leu Cys Thr Ala Glu Lys Glu Val Phe Val Val
275 280 285
Asp Lys Val Ala Asn Ala Leu Lys Ala Glu Met Val Lys Ser Gly Ala
290 295 300
Met Glu Leu Lys Gly Phe Gln Leu Glu Lys Leu Leu Glu Lys Val Leu
305 310 315 320
Val Lys Lys Asn Asp Lys Phe Tyr Pro Asn Arg Asp Leu Ile Gly Lys
325 330 335
Asp Ala Ala Val Ile Leu Gln Ala Ala Gly Ile Gln Ala Ser Pro Ser
340 345 350
Val Lys Leu Ile Ile Ala Glu Thr Thr Lys Asp His Pro Leu Val Met
355 360 365
Thr Glu Met Leu Met Pro Ile Leu Pro Ile Val Arg Val Ser Asn Val
370 375 380
Asp Gln Ala Ile Glu Leu Ala Val Ile Ala Glu Lys Gly Asn Arg His
385 390 395 400
Thr Ala Val Met His Ser Gln Asn Ile Thr Asn Leu Thr Lys Met Ala
405 410 415
Gln Glu Ile Gln Ala Thr Ile Phe Val Lys Asn Gly Pro Ser Val Ala
420 425 430
Gly Leu Gly Phe Glu Ser Glu Gly Phe Thr Thr Leu Thr Ile Ala Gly
435 440 445
Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys Thr Phe Thr Arg Gln Arg
450 455 460
Arg Cys Val Leu Val Asp Gly Leu Arg Ile Ile
465 470 475
<210> 16
<400> 16
000
<210> 17
<211> 496
<212> PRT
<213> Desnuesiella massiliensis
<400> 17
Met Asn Ile Thr Glu Asn Asp Ile Glu Lys Ile Ile Gln Gln Val Leu
1 5 10 15
Val Asn Ile Thr Ser Lys Pro Ser Glu Asp Val Lys Lys Asp Ala Thr
20 25 30
Pro Glu Val Lys Ala Glu Ala Thr Pro Leu Arg Lys Lys Tyr Leu Gly
35 40 45
Val Phe Glu Lys Ala Glu Asp Ala Ile Glu Ala Ala Ser Lys Ala Gln
50 55 60
Lys Lys Leu Leu Lys Glu Phe Lys Ile Glu Asp Arg Glu Arg Phe Ile
65 70 75 80
Ile Ser Ile Lys Lys Ala Thr Val Ala Asn Ala Glu Ile Leu Ala Arg
85 90 95
Met Ile Ile Asp Glu Thr Gly Met Gly Lys Tyr Glu Asp Lys Val Leu
100 105 110
Lys His Lys Leu Val Ser Glu Lys Thr Pro Gly Thr Asp Ile Leu Thr
115 120 125
Thr Glu Ala Trp Ser Gly Asp Asn Gly Leu Thr Ile Val Glu Met Ala
130 135 140
Pro Tyr Gly Val Ile Gly Ala Val Thr Pro Ser Thr Asn Pro Ser Glu
145 150 155 160
Thr Ala Ile Cys Asn Ser Ile Gly Met Ile Gly Ala Gly Asn Ser Val
165 170 175
Val Phe Asn Ala His Pro Gly Ala Lys Glu Cys Val Ala Tyr Ala Val
180 185 190
Asp Met Met Asn Lys Ala Ile Val Glu Ala Gly Gly Pro Glu Asn Leu
195 200 205
Ile Thr Met Val Ala Glu Pro Thr Met Glu Ser Leu Glu Ala Ile Met
210 215 220
Lys His Pro Glu Ile Arg Leu Leu Cys Gly Thr Gly Gly Pro Gly Leu
225 230 235 240
Val Lys Thr Leu Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
245 250 255
Gly Asn Pro Pro Val Ile Val Asp Asp Thr Ala Asn Val Lys Lys Ala
260 265 270
Gly Lys Asp Ile Ile Glu Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys
275 280 285
Ile Ala Glu Lys Glu Val Phe Val Phe Glu Asn Val Ala Asp Asp Leu
290 295 300
Ile Tyr His Met Leu Gln Asn Lys Ala Tyr Met Leu Thr Lys Asn Gln
305 310 315 320
Val Glu Glu Leu Val Lys Ile Val Leu His Glu Asn Ile Glu Glu Lys
325 330 335
Ala Val Gly Cys Ser Leu Asp Arg Lys Arg His Tyr Val Ile Asn Lys
340 345 350
Lys Trp Val Gly Lys Asp Ala Ala Leu Tyr Leu Lys Ala Leu Gly Ile
355 360 365
Glu Gly Lys Asp Asp Ile Gln Cys Leu Ile Cys Glu Val Asp Leu Asp
370 375 380
His Pro Phe Val Met Thr Glu Leu Met Met Pro Ile Leu Pro Ile Val
385 390 395 400
Arg Val Lys Gly Ile Asp Gln Ala Ile Ala Tyr Ala Lys Lys Ala Glu
405 410 415
His Gly Asn Arg His Ser Ala His Met His Ser Lys Asn Val Asp Asn
420 425 430
Leu Thr Arg Phe Ala Arg Glu Ile Glu Thr Thr Ile Phe Val Lys Asn
435 440 445
Ala Lys Ser Phe Ala Gly Val Gly Phe Gly Gly Glu Gly Phe Thr Thr
450 455 460
Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Ile Thr Ser Ala Arg Thr
465 470 475 480
Phe Thr Arg Gln Arg Arg Cys Val Leu Ala Glu Gly Phe Ser Ile Ile
485 490 495
<210> 18
<400> 18
000
<210> 19
<211> 469
<212> PRT
<213> Caldanaerobius polysaccharolyticus
<400> 19
Met Ala Gly Ile Arg Glu Glu Asp Ile Glu Leu Ile Val Arg Arg Val
1 5 10 15
Leu Ser Asn Leu Asp Leu Lys Asn Leu Lys Ala Ala Val Lys Lys Asp
20 25 30
Ile Gly Val Phe Glu Asp Met Lys Gln Ala Ile Ser Ala Ala Lys Lys
35 40 45
Ala Gln Lys Glu Leu Lys Ser Met Ser Ile Glu Phe Arg Glu Lys Ile
50 55 60
Ile Gln Asn Ile Arg Lys Lys Thr Leu Glu Asn Ala Arg Ile Met Ala
65 70 75 80
Glu Met Gly Val Gln Glu Thr Gly Met Gly Lys Val Glu His Lys Val
85 90 95
Leu Lys His Glu Leu Val Ala Arg Lys Thr Pro Gly Thr Glu Asp Ile
100 105 110
Ile Thr Thr Ala Trp Ser Gly Asp Lys Gly Leu Thr Leu Val Glu Met
115 120 125
Gly Pro Trp Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Ser
130 135 140
Glu Thr Val Ile Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ser
145 150 155 160
Val Val Phe Asn Pro His Pro Gly Ala Val Gly Val Ser Asn Tyr Ala
165 170 175
Val Arg Leu Ile Asn Glu Ala Val Val Glu Ala Gly Gly Pro Pro Asn
180 185 190
Leu Ala Val Ser Val Ala Lys Pro Thr Leu Glu Thr Ala Glu Ile Met
195 200 205
Phe Lys His Pro Asp Ile Asn Leu Leu Val Ala Thr Gly Gly Pro Gly
210 215 220
Val Val Thr Ala Val Leu Ser Thr Gly Lys Arg Ala Ile Gly Ala Gly
225 230 235 240
Ala Gly Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Arg Lys
245 250 255
Ala Ala Lys Asp Ile Val Asp Gly Ala Thr Phe Asp Asn Asn Leu Pro
260 265 270
Cys Ile Ala Glu Lys Glu Val Ile Ala Val Asn Lys Val Ala Asp Glu
275 280 285
Leu Ile Tyr Tyr Met Lys Gln Asn Gly Cys Tyr Met Ala Ser Lys Glu
290 295 300
Glu Ile Glu Glu Leu Lys Ala Met Val Leu Gln Thr Arg Asp Gly Lys
305 310 315 320
Tyr Tyr Leu Asn Arg Lys Trp Val Gly Lys Asp Ala Ser Thr Leu Leu
325 330 335
Lys Gly Ile Gly Val Asp Val Asp Asp Lys Val Arg Cys Ile Ile Phe
340 345 350
Glu Ala Thr Lys Asp His Pro Phe Val Val Glu Glu Leu Met Met Pro
355 360 365
Ile Leu Gly Ile Ile Arg Ala Glu Asn Val Asp Glu Ala Ile Ala Ile
370 375 380
Ala Val Glu Leu Glu His Gly Phe Arg His Ser Ala His Met His Ser
385 390 395 400
Lys Asn Val Asp Asn Leu Thr Lys Phe Ala Arg Ala Ile Asp Thr Ala
405 410 415
Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Ala Ile Gly Phe Gly Gly
420 425 430
Glu Gly Tyr Cys Thr Phe Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu
435 440 445
Thr Ser Ala Arg Thr Phe Thr Lys Ser Arg Arg Cys Val Leu Ala Asp
450 455 460
Gly Leu Ser Ile Arg
465
<210> 20
<400> 20
000
<210> 21
<400> 21
000
<210> 22
<400> 22
000
<210> 23
<400> 23
000
<210> 24
<211> 472
<212> PRT
<213> Cellulosilyticum sp. I15G10I2
<400> 24
Met Asn Glu Ile Glu Leu Lys Gln Val Val Glu Glu Val Val Arg Lys
1 5 10 15
Leu Gly Val Pro Ser Ala Thr Ala Pro Lys Thr Ala Pro Thr Ile Gly
20 25 30
Leu Gly Gln Gly Val Phe Glu Ser Met Asp Glu Ala Ile Thr Ala Ala
35 40 45
Lys Ala Ala Gln Glu Asp Leu His Met Met Pro Leu Glu Phe Arg Glu
50 55 60
Lys Ile Ile Ala Arg Ile Arg Glu Lys Ile Met Ala Asn Lys Glu Thr
65 70 75 80
Leu Ala Lys Met Ala Val His Glu Thr Gly Met Gly Lys Ile Gly His
85 90 95
Lys Ile Leu Lys His Glu Leu Thr Ala Lys Lys Thr Pro Gly Thr Glu
100 105 110
Cys Ile Lys Thr Arg Ala Trp Ser Gly Asp Gln Gly Leu Thr Val Ile
115 120 125
Glu Ser Gly Pro Phe Gly Val Val Gly Ala Ile Thr Pro Ser Thr Asn
130 135 140
Pro Ser Glu Thr Val Phe Cys Asn Ala Ile Gly Met Ile Ala Ala Gly
145 150 155 160
Asn Thr Val Val Phe Asn Ser His Pro Asn Ala Ala Arg Thr Ser Asn
165 170 175
Phe Ala Val Gln Leu Val Asn Glu Ala Ala Val Glu Val Gly Gly Phe
180 185 190
Glu Asn Leu Ala Thr Ser Val Leu Lys Pro Thr Val Glu Ser Gly Asn
195 200 205
Thr Leu Phe Lys His Pro Asp Ile Gln Leu Leu Val Ala Thr Gly Gly
210 215 220
Pro Gly Val Val Lys Ala Ile Leu Gln Ser Gly Lys Arg Gly Ile Ala
225 230 235 240
Ala Gly Ala Gly Asn Pro Pro Val Leu Val Asp Glu Thr Ala Asn Ile
245 250 255
Lys Lys Ala Ala Ala Asp Ile Ile Asn Gly Ala Thr Phe Asp Asn Asn
260 265 270
Leu Pro Cys Ile Ala Glu Lys Glu Val Ile Val Val Asn Glu Val Ala
275 280 285
Asp Glu Leu Ile His Tyr Met Thr Ser Glu Asn Asp Cys Tyr Met Leu
290 295 300
Lys Gly Glu Gln Ile Glu Lys Leu Ala Gln Thr Ile Leu Val Glu Lys
305 310 315 320
Asn Gly His Tyr Ile Val Asn Arg Asp Tyr Val Gly Arg Asp Ala His
325 330 335
Val Ile Leu Lys Gly Ile Gly Ile Glu Ala Pro Glu Ser Ile Arg Cys
340 345 350
Ile Ile Phe Glu Ala Ser Lys Glu His Ile Leu Val Val Glu Glu Leu
355 360 365
Met Met Pro Val Leu Gly Ile Val Arg Val Ala Asn Val Asp Glu Gly
370 375 380
Ile Ala Val Ala Lys Val Leu Glu Gly Gly Asn Arg His Ser Ala His
385 390 395 400
Met His Ser Ser Asn Val Tyr Asn Leu Thr Lys Tyr Gly Arg Ala Leu
405 410 415
Asp Thr Ala Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Gly Ile Gly
420 425 430
Phe Gly Gly Glu Gly Phe Ala Thr Phe Thr Ile Ala Ser Lys Thr Gly
435 440 445
Glu Gly Leu Thr Asn Ala Ala Ser Phe Thr Lys Ser Arg Arg Cys Val
450 455 460
Met Ala Asp Ala Leu Tyr Ile Arg
465 470
<210> 25
<211> 455
<212> PRT
<213> Geosporobacter ferrireducens
<400> 25
Met Ile Val Lys Lys Ile Leu Thr Glu Ile Thr Leu Lys Asn Glu Ala
1 5 10 15
Thr Asp Ser Ala Tyr Gly Ile Phe Asp His Met Glu Glu Ala Ile Glu
20 25 30
Ala Ala Trp Ile Ala Gln Lys Glu Leu Val Lys Tyr Ser Leu Glu Cys
35 40 45
Arg Gly Lys Phe Ile Ala Ala Met Arg Ala Ala Ala Arg Lys Asn Ile
50 55 60
Glu Leu Phe Ser Lys Met Ala Val Glu Glu Thr Gly Met Gly Arg Tyr
65 70 75 80
Glu His Lys Val Met Lys Asn Thr Val Ala Ile Glu Lys Thr Pro Gly
85 90 95
Ile Glu Asp Leu Lys Pro Asp Ala Val Ser Gly Asp His Gly Leu Thr
100 105 110
Val Phe Glu Leu Ser Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Thr
115 120 125
Thr Asn Pro Thr Glu Thr Val Ile Cys Asn Ala Ile Gly Met Ile Ala
130 135 140
Ala Gly Asn Ala Val Val Phe Ala Pro His Pro Arg Ala Lys Asn Thr
145 150 155 160
Ser Arg Lys Ala Ile Glu Ile Leu Asn Gln Ala Ile Ile Glu Ala Gly
165 170 175
Gly Pro Ala Asn Leu Ile Thr Ala Ile Lys Glu Pro Thr Ile Glu Ser
180 185 190
Ala Asn Ile Met Met Gln His Lys Lys Ile Lys Met Leu Val Ala Thr
195 200 205
Gly Gly Pro Asp Val Val Arg Thr Val Leu Ser Ser Gly Lys Lys Ala
210 215 220
Ile Gly Ala Gly Ala Gly Asn Pro Pro Ala Val Val Asp Glu Thr Ala
225 230 235 240
Asp Ile Glu Lys Ala Ala Lys Asp Ile Ile Asp Gly Cys Ser Phe Asp
245 250 255
Asn Asn Leu Pro Cys Val Ala Glu Lys Glu Val Ile Val Val Asp Ser
260 265 270
Val Ala Asp Tyr Leu Ile Phe Asn Met Gln Lys His Asn Ala Tyr Leu
275 280 285
Leu Ser Asp Glu Asn Leu Ile Lys Lys Leu Glu Lys Leu Val Phe Asn
290 295 300
Asp Lys Gly His Leu Asn Arg Asp Leu Val Gly Lys Asp Ala Asp Tyr
305 310 315 320
Ile Leu Arg Lys Ile Gly Val Asp Cys Asp Pro Ser Ile Arg Ala Ile
325 330 335
Ile Val Glu Thr Asp Lys Asn His Asp Phe Val Gln Glu Glu Leu Met
340 345 350
Met Pro Ile Leu Pro Ile Val Arg Val Lys Asp Val Asn Glu Ala Ile
355 360 365
Glu Leu Ala Val Glu Val Glu His Gly Tyr Arg His Thr Ala Ile Ile
370 375 380
His Ser Lys Asn Ile Asp Asn Leu Ser Lys Met Ala Lys Glu Ile Gln
385 390 395 400
Thr Thr Ile Phe Val Lys Asn Gly Pro Ser Tyr Ala Gly Ile Gly Val
405 410 415
Gly Gly Glu Gly Tyr Ser Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu
420 425 430
Gly Leu Thr Thr Ala Lys Ser Phe Thr Arg Ser Arg Arg Cys Val Leu
435 440 445
Val Asp Gly Phe Ser Ile Arg
450 455
<210> 26
<400> 26
000
<210> 27
<211> 465
<212> PRT
<213> Bacillus korlensis
<400> 27
Met Ile Glu Val Lys Gln Ile Glu Asp Ile Val Met Gln Val Leu Ala
1 5 10 15
Gly Leu Asn Asn His Glu Asp Pro Pro Leu Asp Gly Glu Asn Gly Leu
20 25 30
Tyr Ser Glu Met Asn Asp Ala Ile Asp Ala Ala Phe Val Ala Gln Lys
35 40 45
Glu Leu Val Lys Leu Ser Leu Ala Glu Arg Gly Arg Ile Ile Glu Ser
50 55 60
Ile Arg Thr Glu Phe Arg Lys His Ile Glu Leu Leu Ser Glu Met Ala
65 70 75 80
Val Glu Glu Thr Gly Met Gly Arg Val Lys Asp Lys Ile Asn Lys Asn
85 90 95
Leu Val Ala Val Asn Asn Thr Pro Gly Ile Glu Asp Leu Thr Thr Ala
100 105 110
Ala Cys Ser Gly Asp Asn Gly Leu Thr Val Glu Glu Leu Ser Pro Tyr
115 120 125
Gly Val Ile Gly Ser Ile Thr Pro Thr Thr Asn Pro Ser Glu Thr Ile
130 135 140
Ile Cys Asn Thr Ile Gly Met Leu Ala Ala Gly Asn Ala Ile Val Phe
145 150 155 160
Ser Pro His Pro Thr Ala Lys Arg Thr Ser Ile Glu Thr Ile Lys Ile
165 170 175
Ile Ser Lys Ala Ile Ser Lys Ala Gly Gly Pro Lys Asn Leu Val Val
180 185 190
Ser Thr Leu Gln Pro Ser Ile Glu Gln Ala Asn Ile Met Met Asn His
195 200 205
Lys Lys Val Arg Met Leu Val Ala Thr Gly Gly Pro Ala Val Val Lys
210 215 220
Ala Val Leu Ser Thr Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Glu Lys Ala Ala Lys
245 250 255
Asp Ile Ile Asp Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys Val Ala
260 265 270
Glu Lys Glu Val Ile Ala Val Asp Cys Ile Ala Asp Cys Leu Ile Glu
275 280 285
Asn Met Lys Asn Asn Gly Ala Tyr Gln Leu Thr Asp Pro Val Gln Ile
290 295 300
Gln Arg Leu Val Asp Leu Val Val Arg Asn Gly His Ala Asn Lys Asp
305 310 315 320
Phe Val Gly Lys Asn Ala Asp Phe Ile Leu Arg Gln Leu Gly Ile Glu
325 330 335
Val Gly Pro Glu Val Arg Val Val Ile Val Asp Val Lys Tyr Glu Gly
340 345 350
Arg His Pro Leu Val Leu Ala Glu Leu Met Met Pro Val Leu Pro Ile
355 360 365
Val Arg Val Asn Asn Val Asp Glu Gly Ile Asp Leu Ala Val Glu Val
370 375 380
Glu His Gly Phe Arg His Thr Ala Ile Met His Ser Lys Asn Ile Asp
385 390 395 400
Asn Leu Thr Lys Phe Ala Lys Glu Ile Gln Thr Thr Ile Phe Val Lys
405 410 415
Asn Gly Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Val Gly Tyr Thr
420 425 430
Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys
435 440 445
His Phe Ala Arg Lys Arg Arg Cys Val Leu Val Asp Gly Leu Ser Ile
450 455 460
Arg
465
<210> 28
<211> 459
<212> PRT
<213> Acidaminococcus massiliensis
<400> 28
Met Glu Gln Ala Val Lys Asp Tyr Leu Asp Lys Met Val Ala Ala Ser
1 5 10 15
Arg Ile Ala Gln Gln Glu Phe Ala Thr Tyr Pro Gln Glu Thr Val Asp
20 25 30
Lys Ala Val Arg Thr Val Gly Lys Ala Ile Tyr Asp Asn Ala Glu Leu
35 40 45
Leu Ala His Met Ala Val Asp Glu Thr Lys Met Gly Asn Tyr Ala Asp
50 55 60
Lys Ile Ala Lys Cys Val Asn Lys Ser Lys Ser Val Trp Trp Arg Met
65 70 75 80
Lys Asp Lys Lys Ser Arg Gly Ile Ile Lys Arg Ile Pro Glu Leu Gly
85 90 95
Leu Val Glu Val Ala Lys Pro Ile Gly Val Ile Gly Cys Val Ala Pro
100 105 110
Thr Thr Asn Pro Val Ile Asn Val Met Gln Asn Ala Met Cys Ala Leu
115 120 125
Lys Cys Gly Asn Ser Met Ile Val Ser Pro His Pro Arg Ala Lys His
130 135 140
Ser Ser Val Lys Thr Val Glu Val Ile Asn Glu Ala Leu Ala Ala Leu
145 150 155 160
Gly Met Pro Lys Asn Leu Ile Gln Val Ile Thr Glu Pro Ser Met Glu
165 170 175
Leu Ser Ala Gly Leu Met Ser Ala Val Asp Leu Cys Ile Cys Thr Gly
180 185 190
Gly Pro Gly Leu Val Lys Ala Ala Tyr Ser Ser Gly Lys Pro Ala Ile
195 200 205
Gly Val Gly Gln Gly Asn Val Gln Val Leu Val Asp Arg Asp Ala Asp
210 215 220
Leu Asp Gln Val Ala Ala Met Val Ile Lys Gly Arg Thr Phe Asp Asn
225 230 235 240
Gly Val Leu Cys Thr Cys Glu Gln Asn Val Ile Cys Pro Glu Asp Lys
245 250 255
Lys Glu Glu Met Ile Ala Ala Leu Lys Lys His Gly Ala Tyr Tyr Ile
260 265 270
Gly Asn Ser Glu Asp Ala Ala Lys Leu Arg Asp Thr Ala Phe Pro Asn
275 280 285
Gly Gly Pro Val Ser Lys Glu Tyr Pro Gly Ala Ser Val Lys Lys Ile
290 295 300
Ala Gln Leu Ser Gly Ile Gln Gly Ile Pro Glu Asp Ala Lys Val Ile
305 310 315 320
Val Ser Cys Thr Arg Gly Tyr Gly Lys Asp Glu Pro Leu Ala Lys Glu
325 330 335
Lys Leu Phe Pro Val Leu Ala Phe Phe Thr Tyr Asp Lys Trp Glu Asp
340 345 350
Ala Ile His Ile Ala Lys Thr Asn Leu Glu Met Glu Gly Ile Gly His
355 360 365
Ser Val Val Ile His Ser Asn Thr Pro Glu His Ile Glu Ala Val Ala
370 375 380
Glu Ala Ile Pro Val Ser Arg Phe Ala Val Asn Gln Val Gly Gly Thr
385 390 395 400
Asn Leu Gly Gly Ala Met Asp Asn Gly Leu Asn Pro Thr Thr Thr Leu
405 410 415
Gly Cys Gly Thr Trp Gly Asn Asn Ser Ile Ser Glu Asn Phe Thr Tyr
420 425 430
Tyr His Leu Met Asn Leu Thr Arg Val Ser Tyr Arg Val Pro Asp Met
435 440 445
Tyr Ile Pro Thr Asp Glu Glu Ile Trp Ala Glu
450 455
<210> 29
<400> 29
000
<210> 30
<400> 30
000
<210> 31
<211> 462
<212> PRT
<213> Lachnospiraceae bacterium 32
<400> 31
Val Ser Val Asn Glu Lys Met Val Gln Asp Val Val Lys Glu Val Met
1 5 10 15
Ala Lys Leu Gln Leu Ala Ala Gly Ala Ser Glu Gly Lys Gly Ile Phe
20 25 30
Ala Asp Met Asn Asp Ala Ile Ala Ala Ala Lys Lys Ala Gln Arg Tyr
35 40 45
Ile His Arg Met Ser Met Asp Gln Arg Glu Gln Ile Ile Ser Asn Ile
50 55 60
Arg Arg Lys Thr Lys Glu Asn Ala Glu Ile Leu Ala Arg Met Gly Val
65 70 75 80
Glu Glu Thr Gly Met Gly Asn Val Pro His Lys Ile Leu Lys His Gln
85 90 95
Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Lys Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Ile Ile
130 135 140
Cys Asn Thr Ile Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Val Lys Thr Ser Gln Phe Ala Val Asn Met Leu
165 170 175
Asn Glu Ala Ser Ile Glu Ala Gly Gly Pro Glu Asn Ile Ala Cys Thr
180 185 190
Val Gly Lys Pro Thr Met Glu Ser Ser Asn Ile Met Met Lys His Lys
195 200 205
Asp Ile Gln Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Arg Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Gly Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Asp Ser Val Val Ser Glu Leu Met His Tyr
275 280 285
Met Val Asn Glu Gln Asp Cys Tyr Leu Ala Ser Lys Glu Glu Gln Asp
290 295 300
Lys Leu Thr Ala Thr Val Leu Thr Pro Lys Gly Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Lys Thr Leu Leu Gly Met Ile Gly Val Thr Val
325 330 335
Pro Asp Asn Ile Arg Cys Ile Val Phe Glu Gly Glu Lys Glu His Pro
340 345 350
Leu Ile Ala Thr Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Lys Asp Phe Glu Asp Ala Val Glu Lys Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Ile Asp Asn Ile Thr
385 390 395 400
Arg Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Ala Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Phe Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Ser Glu Ser Leu Cys Ile Arg
450 455 460
<210> 32
<211> 462
<212> PRT
<213> Eubacterium plexicaudatum
<400> 32
Val Ser Val Asn Asp Gln Met Val Gln Asp Ile Val Arg Gln Val Leu
1 5 10 15
Ala Asn Met Arg Ile Ser Ser Asp Ala Ser Gly Ser Arg Gly Val Phe
20 25 30
Ser Asp Met Asn Glu Ala Val Glu Ala Ala Lys Lys Ala Gln Ala Val
35 40 45
Ile Gly Lys Met Pro Met Asp His Arg Glu Lys Ile Ile Ser Ser Ile
50 55 60
Arg Ala Lys Ile Met Glu Asn Ala Glu Ile Leu Ala Arg Met Gly Val
65 70 75 80
Lys Glu Thr Gly Met Gly Asn Val Gly His Lys Ile Leu Lys His Gln
85 90 95
Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Lys Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Ile Leu
130 135 140
Cys Asn Thr Ile Gly Met Val Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Phe Ala Val Asn Leu Val
165 170 175
Asn Glu Ala Ser Val Glu Ala Gly Gly Pro Asp Asn Ile Ala Cys Thr
180 185 190
Val Glu His Pro Thr Leu Asp Thr Ser Ala Ile Met Met Lys His Lys
195 200 205
Asp Ile His Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Glu Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Asp Ser Ile Ala Asp Glu Leu Met His Tyr
275 280 285
Met Ile Ser Glu Gln Gly Cys Tyr Leu Ala Ser Ala Lys Glu Gln Glu
290 295 300
Ala Leu Ile Ser Val Val Leu Lys Gly Gly Gln Leu Asn Arg Asp Cys
305 310 315 320
Val Gly Arg Asp Ala Lys Thr Leu Leu Gly Met Ile Gly Val Gln Ala
325 330 335
Pro Asp Asn Ile Arg Cys Ile Thr Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Thr Glu Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Asp Ser Phe Glu Asp Ala Val Glu Lys Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Val Asp His Ile Thr
385 390 395 400
Thr Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Gly Pro
405 410 415
Ser Tyr Ala Ala Ile Gly Phe Gly Gly Glu Gly Tyr Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Ala Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Cys Asp Ser Leu Cys Ile Arg
450 455 460
<210> 33
<211> 462
<212> PRT
<213> Clostridium sp. KNHs205
<400> 33
Val Asn Leu Lys Glu Ala Gln Val Lys Asp Ile Val Arg Lys Val Leu
1 5 10 15
Leu Gln Met Glu Ala Ser Asn Lys Glu Glu Gln Lys Leu Ser Gly Ile
20 25 30
Phe Thr Glu Met Asn Asp Ala Ile Gly Ala Ser Ile Lys Ala Gln Lys
35 40 45
Val Met Gln Gln Leu Ser Met Asp Ser Arg Glu Lys Ile Ile Ser Asn
50 55 60
Ile Arg Lys Lys Thr Leu Glu Asn Ala Glu Leu Phe Ala Arg Met Gly
65 70 75 80
Val Glu Glu Thr Gly Met Gly Asn Val Gly His Lys Ile Leu Lys His
85 90 95
Gln Leu Leu Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Ser Thr Val
100 105 110
Ala Trp Ser Gly Asp Arg Gly Leu Thr Leu Val Glu Met Gly Pro Phe
115 120 125
Gly Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Ile
130 135 140
Leu Cys Asn Ser Ile Gly Met Ile Ala Gly Gly Asn Thr Val Val Phe
145 150 155 160
Asn Pro His Pro Ala Ala Ile Gly Val Ser Asn Leu Ala Val His Met
165 170 175
Val Asn Glu Ala Ser Arg Glu Ala Gly Gly Pro Asp Asn Ile Ala Val
180 185 190
Ser Val Val Lys Pro Thr Leu Ala Ser Gly Asp Ile Met Met Lys His
195 200 205
Gln Asn Ile Pro Leu Ile Val Ala Thr Gly Gly Pro Gly Val Val Thr
210 215 220
Thr Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Val Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Met
245 250 255
Asp Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala
260 265 270
Glu Lys Glu Val Val Ala Val Gly Lys Ile Met Asp Glu Leu Leu His
275 280 285
Tyr Leu Ile Glu Asn Gly Cys Tyr Val Ile Ser Lys Glu Glu Gln Glu
290 295 300
Lys Leu Thr Ala Val Val Leu Lys Asp Asn Arg Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Lys Asp Ala Arg Thr Ile Leu Ser Met Ile Gly Ile Glu Thr
325 330 335
Pro Glu Asn Ile Arg Cys Ile Ile Phe Glu Gly Glu Lys Glu His Pro
340 345 350
Leu Ile Ala Glu Glu Leu Met Met Pro Ile Leu Gly Ile Val Arg Ala
355 360 365
Lys Asp Ile Asp Asp Ala Ile Glu Lys Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Met His Ser Lys Asn Val Asp Asn Leu Thr
385 390 395 400
Arg Phe Gly Lys Ala Val Asp Thr Ala Ile Phe Val Lys Asn Ala Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Phe Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Arg Thr Phe Thr
435 440 445
Lys Gln Arg Arg Cys Val Met Ala Asp Ser Leu Cys Ile Arg
450 455 460
<210> 34
<400> 34
000
<210> 35
<400> 35
000
<210> 36
<211> 470
<212> PRT
<213> Robinsoniella peoriensis
<400> 36
Met Ala Ile Asn Glu Gln Glu Ile Gln Asp Ile Val Arg Ser Val Leu
1 5 10 15
Lys Gly Met Gly Thr Thr Ala Asp Lys Pro Ala Gly Ser Ser Lys Lys
20 25 30
Leu Leu Gly Val Phe Asp Asp Ile Asn Asp Ala Ile Ala Ala Ala Lys
35 40 45
Glu Ala Gln Lys Glu Ile Gln Pro Met Pro Leu Glu Phe Arg Glu Lys
50 55 60
Ile Ile Ser Asn Ile Arg Lys Lys Thr Leu Glu Asn Ala Lys Met Phe
65 70 75 80
Ala Glu Leu Gly Val Glu Glu Thr Gly Met Gly Asn Val Gly His Lys
85 90 95
Ile Leu Lys His Gln Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp
100 105 110
Leu Ser Thr Val Ala Trp Ser Gly Asp Arg Gly Leu Thr Leu Val Glu
115 120 125
Met Gly Pro Phe Gly Val Ile Gly Ala Val Cys Pro Ser Thr Asn Pro
130 135 140
Thr Glu Thr Val Val Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn
145 150 155 160
Thr Val Val Phe Ala Pro His Pro Ser Ala Lys Asn Val Ser Asn Leu
165 170 175
Ala Ile Asp Met Ile Asn Arg Ala Ser Val Glu Val Gly Gly Pro Glu
180 185 190
Asn Ile Ala Val Ala Val Lys Glu Pro Thr Met Glu Val Ser Lys Val
195 200 205
Ile Phe Ser His Lys Asp Ile Ser Leu Leu Val Ala Thr Gly Gly Pro
210 215 220
Gly Val Val Thr Thr Val Leu Ser Ser Gly Lys Arg Ala Met Gly Ala
225 230 235 240
Gly Ala Gly Asn Pro Pro Val Leu Val Asp Glu Thr Ala Asn Ile Pro
245 250 255
Lys Ala Ala Glu Asp Ile Ile Asn Gly Cys Thr Phe Asp Asn Asn Leu
260 265 270
Pro Cys Ile Ala Glu Lys Glu Val Val Ala Val Asp Met Ile Ala Asp
275 280 285
Glu Leu Ile Tyr His Met Glu Gln Val Gly Cys Tyr His Ala Asn Ala
290 295 300
Glu Glu Val Gln Lys Leu Ile Gln Thr Val Phe Ile Glu Asn Asn Gly
305 310 315 320
Lys Arg Thr Leu Asn Arg Gln Cys Val Gly Arg Ser Ala Lys Val Leu
325 330 335
Leu Gly Lys Ile Gly Val Thr Val Gly Asp Glu Ile Arg Cys Ile Ile
340 345 350
Phe Glu Gly Glu Lys Thr Asn Pro Met Ile Trp Glu Glu Leu Met Met
355 360 365
Pro Ile Leu Gly Ile Val Arg Val Lys Asn Val Glu Glu Gly Met Gly
370 375 380
Ile Ala Leu Glu Leu Glu His Gly Asn Arg His Ser Ala His Met His
385 390 395 400
Ser Thr Asn Val Asn Asn Leu Thr Lys Phe Gly Lys Met Ile Asp Thr
405 410 415
Ala Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Ala Leu Gly Phe Gly
420 425 430
Gly Glu Gly Tyr Pro Thr Phe Thr Ile Cys Ser Arg Thr Gly Glu Gly
435 440 445
Leu Thr Ser Ala Lys Asn Phe Thr Lys Ser Arg Arg Cys Val Met Gly
450 455 460
Asp Ala Leu Cys Ile Arg
465 470
<210> 37
<400> 37
000
<210> 38
<211> 469
<212> PRT
<213> Caldithrix abyssi
<400> 38
Met His Leu Asp Asp Lys Gln Ile Ala Gln Ile Val Glu Thr Val Leu
1 5 10 15
Ser Arg Leu Glu Arg Asn Glu Ser Arg Thr Gly Arg Ser Arg His Pro
20 25 30
Gln Gly Val Phe Glu Thr Leu Asp Glu Ala Val Glu Ala Ala Arg Gln
35 40 45
Ala Gln Lys Lys Ile Arg Lys Leu Glu Leu Arg Ala Lys Ile Ile Gln
50 55 60
Ala Ile Arg Gln Ala Gly Val Lys His Ala Arg Glu Leu Ala Glu Met
65 70 75 80
Ala Val Gln Glu Thr Gly Met Gly Arg Val Glu Asp Lys Ile Ala Lys
85 90 95
Asn Ile Ser Gln Ala Glu Lys Thr Pro Gly Ile Glu Asp Leu Gln Pro
100 105 110
Leu Ala Leu Ser Gly Asp His Gly Leu Thr Leu Ile Glu Asn Ala Ala
115 120 125
Trp Gly Val Ile Ala Ser Val Thr Pro Ser Thr Asn Pro Gly Ala Thr
130 135 140
Val Ile Asn Asn Ser Ile Ser Met Ile Ala Ala Gly Asn Ala Val Val
145 150 155 160
Tyr Ala Pro His Pro Ala Ala Lys Lys Val Ser Gln Arg Ala Ile Glu
165 170 175
Ile Leu Asn Lys Ala Ile Glu Ala Ala Gly Gly Pro Ala Thr Leu Leu
180 185 190
Thr Thr Val Ala Glu Pro Ser Ile Glu Thr Ala Gln Lys Leu Phe Val
195 200 205
Tyr Pro Gly Ile Asp Leu Leu Val Val Thr Gly Gly Glu Ala Val Val
210 215 220
Lys Ala Ala Arg Lys Val Thr Asp Lys Arg Leu Met Ala Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Ala Lys Ala
245 250 255
Ala Arg Asp Ile Val Trp Gly Ala Ser Phe Asp Asn Asn Ile Val Cys
260 265 270
Ala Asp Glu Lys Glu Ile Ile Ala Val Asp Ala Ile Ala Asp Arg Leu
275 280 285
Lys Glu Glu Met Lys Lys His Gln Ala Val Glu Leu Thr Pro Gln Gln
290 295 300
Gly Glu Glu Leu Ala Gln Ile Ile Leu Glu Asp Tyr Pro Gly Pro Asn
305 310 315 320
Ala Arg Ile Asn Arg Lys Trp Val Gly Lys Asp Ala Tyr Lys Phe Ala
325 330 335
Arg Glu Ile Gly Leu Asn Val Ser Lys Glu Thr Arg Leu Leu Phe Val
340 345 350
Glu Ala Asp Lys Asp His Pro Phe Ala Gln Leu Glu Leu Met Met Pro
355 360 365
Val Ile Pro Leu Ile Arg Ala Ala Asp Ala Asp Lys Ala Ile Asp Leu
370 375 380
Ala Ile Glu Leu Glu His Gly Tyr Arg His Thr Ala Ala Met His Ser
385 390 395 400
Arg His Ile Asp His Met Asp Arg Met Ala Asn Glu Ile Asn Thr Ser
405 410 415
Ile Phe Val Lys Asn Gly Pro Cys Leu Ala Gly Leu Gly Phe Gly Gly
420 425 430
Glu Gly Trp Thr Ser Met Thr Ile Thr Thr Pro Thr Gly Glu Gly Val
435 440 445
Thr Ser Ala Arg Ser Phe Val Arg Leu Arg Arg Cys Val Val Val Asp
450 455 460
His Phe Arg Ile Val
465
<210> 39
<400> 39
000
<210> 40
<211> 479
<212> PRT
<213> Sporomusa sphaeroides
<400> 40
Met Thr Ile Asp Pro Asn Leu Ile Ala Lys Ile Ala Ala Glu Val Met
1 5 10 15
Ala Arg Val Gln Glu Arg Gln Pro Glu Thr Val Ser Ala Gly Glu Gly
20 25 30
Ile Phe Pro Thr Val Asp Glu Ala Val Ala Ala Ala Arg Ala Ala Gln
35 40 45
Lys Gln Leu Lys Lys Leu Ser Ile Glu Lys Arg Glu Glu Leu Ile Gln
50 55 60
Ala Met Arg Gln Ala Ala Cys Asp Asn Ala Glu Leu Leu Ala Glu Met
65 70 75 80
Gly Val Ser Glu Ser Gly Met Gly Arg Val Ser Asp Lys Val Ile Lys
85 90 95
Asn Arg Leu Ala Ala Thr Lys Thr Pro Gly Thr Glu Asp Leu Lys Ser
100 105 110
Glu Ala Trp Ser Gly Asp Arg Gly Leu Thr Leu Ile Glu Met Gly Pro
115 120 125
Tyr Gly Val Ile Gly Ser Ile Thr Pro Thr Thr Asn Pro Ser Glu Thr
130 135 140
Val Ile Cys Asn Gly Ile Gly Met Ile Ala Ala Gly Asn Ala Val Val
145 150 155 160
Phe Ser Pro His Pro Thr Ala Lys Asn Thr Ser Leu Val Thr Ile Lys
165 170 175
Leu Leu Asn Lys Ala Ile Ile Gln Ala Gly Gly Pro Pro Asn Leu Leu
180 185 190
Thr Ala Val Ala Glu Pro Ser Leu Ala Ala Thr Asn Ala Met Met Gln
195 200 205
His Pro Asp Ile Asn Met Leu Val Ala Thr Gly Gly Pro Ala Val Val
210 215 220
Lys Ala Val Met Ser Cys Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly
225 230 235 240
Asn Pro Pro Ala Val Val Asp Glu Thr Ala Asp Ile Glu Lys Ala Ala
245 250 255
Lys Asp Ile Ile Asp Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys Ile
260 265 270
Ala Glu Lys Glu Val Ile Val Val Gly Ser Val Ala Asp Lys Leu Met
275 280 285
Ala Tyr Met Gln Arg Tyr Gly Ala Tyr Leu Ile Ser Gly Pro Asp Val
290 295 300
Asp Arg Leu Ala Lys Val Ile Leu Thr Glu Lys Ala Glu Leu Ala Ala
305 310 315 320
Ala Gly Cys Thr Glu Lys Pro Lys Lys Ser Tyr Ala Val Asn Lys Asn
325 330 335
Tyr Val Gly Lys Asp Ala Arg Tyr Ile Leu Ser Gln Ile Gly Ile Gln
340 345 350
Val Pro Asp Ser Ile Arg Ala Val Ile Cys Glu Thr Pro Ala Asp His
355 360 365
Pro Phe Val Val Glu Glu Leu Met Met Pro Val Leu Pro Val Val Gln
370 375 380
Val Lys Asp Ile Asp Ala Ala Ile Glu Leu Ala Val Lys Val Glu His
385 390 395 400
Gly Asn Arg His Thr Ala Ile Met His Ser Lys Asn Val Asp Asn Leu
405 410 415
Thr Lys Leu Ala Lys Ala Ile Glu Thr Thr Ile Phe Val Lys Asn Ala
420 425 430
Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Phe Thr Thr Phe
435 440 445
Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Pro Arg Ser Phe
450 455 460
Thr Arg Gln Arg Arg Cys Val Leu Val Asp Ala Leu Ser Ile Val
465 470 475
<210> 41
<211> 524
<212> PRT
<213> Bacillus sp. FJAT-25547
<400> 41
Met Gly Val Asn Met Ser Glu Gln Asp Ile Gln Lys Ile Ile Gln Ser
1 5 10 15
Val Leu Gln Asn Ile Glu Ala Val Ser Glu Gln Asn Ser Gly His Gln
20 25 30
Val Leu His Ser Asn Asp Asn Thr Asn Pro Pro Lys Pro Leu Lys Met
35 40 45
Lys Arg Val Leu Pro Leu Ser Gln Gln Ile Asn Thr Ala Glu Leu Ser
50 55 60
His Gln Val Asn Glu Pro Gly Ala Asn Gly Val Phe Val Arg Ile Glu
65 70 75 80
Asp Ala Ile Glu Ala Gly Tyr Ile Ala Gln Leu Asn Tyr Val Lys His
85 90 95
Phe Gln Leu Lys Asp Arg Glu Lys Ile Ile Ala Ala Ile Arg Glu Ala
100 105 110
Val Ile Glu Asn Lys Glu Lys Leu Ala Gln Met Val Phe Glu Glu Thr
115 120 125
Lys Leu Gly Arg Tyr Glu Asp Lys Ile Ala Lys His Glu Leu Val Ala
130 135 140
Ser Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Ala Ala Phe Ser Gly
145 150 155 160
Asp Glu Gly Leu Thr Ile Val Glu Gln Ala Pro Phe Gly Leu Val Gly
165 170 175
Ala Val Thr Pro Val Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn Ser
180 185 190
Ile Ser Leu Leu Ala Ala Gly Asn Ala Val Val Leu Asn Val His Pro
195 200 205
Ser Ser Lys Ala Ser Cys Ala Phe Val Val Asn Leu Ile Asn Gln Ala
210 215 220
Ile Gln Asp Ala Gly Gly Pro Lys Asn Leu Val Ser Met Val Lys Asp
225 230 235 240
Pro Thr Leu Glu Thr Leu Asn Arg Ile Ile Glu Ser Pro Lys Val Lys
245 250 255
Leu Leu Val Gly Thr Gly Gly Pro Gly Met Val Lys Thr Leu Leu Lys
260 265 270
Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile
275 280 285
Val Asp Glu Thr Ala Asp Leu Lys Gln Ala Ala Lys Ser Ile Ile Glu
290 295 300
Gly Ala Ser Phe Asp Asn Asn Leu Leu Cys Ile Ala Glu Lys Glu Leu
305 310 315 320
Phe Val Ile Asp Ser Val Ala Asp Asp Leu Ile Phe Gln Met Leu Asn
325 330 335
Glu Gly Ala Tyr Met Leu Asp Gln Gln Gln Leu Ser Lys Leu Met Ser
340 345 350
Phe Ala Leu Glu Glu Asn Val His Gln Glu Ala Gly Gly Cys Ser Leu
355 360 365
Asp Asn Lys Arg Glu Tyr His Val Ser Lys Asp Trp Val Gly Lys Asp
370 375 380
Ala Ala Ser Phe Leu Arg Gln Ile Gly Val Ala Cys Glu Glu Asn Ile
385 390 395 400
Lys Leu Leu Ile Cys Glu Val Asp Phe Asp His Pro Phe Val Gln Leu
405 410 415
Glu Gln Met Met Pro Val Phe Pro Ile Val Arg Val Gly Asp Leu Asp
420 425 430
Glu Ala Ile Glu Met Ala Leu Leu Ala Glu His Gly Asn Arg His Thr
435 440 445
Ala Ile Met His Ser Lys Asn Val Asp His Leu Thr Lys Phe Ala Arg
450 455 460
Ala Ile Glu Thr Thr Ile Phe Val Lys Asn Ala Ser Ser Leu Ala Gly
465 470 475 480
Val Gly Phe Gly Gly Glu Gly His Thr Thr Met Thr Ile Ala Gly Pro
485 490 495
Thr Gly Glu Gly Ile Thr Ser Ala Lys Thr Phe Thr Arg Gln Arg Arg
500 505 510
Cys Val Leu Ala Glu Gly Gly Phe Arg Ile Ile Gly
515 520
<210> 42
<211> 471
<212> PRT
<213> Dorea sp. D27
<400> 42
Met Glu Ile Ser Thr Ser Gln Ile Ser Arg Tyr Ile Leu Asp Leu Gln
1 5 10 15
Asn Glu Leu Lys Gly Asp Ser Pro Ser Pro Ala His Met Ser Ala Gly
20 25 30
Glu His Gly Ile Phe Gln Asp Ala Glu Cys Ala Ile Met Ala Ala Ser
35 40 45
Gln Ala Gln Lys Arg Leu Met Glu Tyr Ser Leu Lys Glu Arg Glu Thr
50 55 60
Phe Ile Glu Ala Met Arg Ala Ala Ala Arg Glu Asn Ala Arg Lys Leu
65 70 75 80
Ala Glu Thr Ala His Asp Glu Thr Gly Tyr Gly His Val Glu Asp Lys
85 90 95
Val Ala Lys Asn Val Leu Ala Ala Asp Lys Thr Pro Gly Ile Glu Asp
100 105 110
Leu Asn Thr Met Ala Val Ser Gly Asp Ala Gly Leu Met Leu Thr Glu
115 120 125
Met Ala Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro
130 135 140
Thr Ala Thr Val Ile Asn Asn Gly Ile Gly Met Ile Ala Gly Gly Asn
145 150 155 160
Ala Val Val Phe Asn Pro His Pro Gly Ala Lys Lys Ala Ser Leu Leu
165 170 175
Thr Ile Lys Leu Met Asn Glu Ala Ile Val Gly Ala Gly Gly Pro Asp
180 185 190
Asn Leu Leu Cys Ala Pro Glu Glu Pro Thr Leu Asp Thr Ser Ser Val
195 200 205
Ile Met Ser His Pro Leu Val Lys Leu Leu Val Val Thr Gly Gly Glu
210 215 220
Ala Val Val Arg Thr Ala Met Lys Thr Gly Lys Lys Cys Ile Ala Ala
225 230 235 240
Gly Pro Gly Asn Pro Pro Val Val Val Asp Gly Thr Ala Asp Ile Lys
245 250 255
Arg Ala Ala Ala Asp Ile Val Lys Gly Ala His Tyr Glu Asn Cys Ile
260 265 270
Leu Cys Ile Ala Glu Lys Glu Ile Leu Val Glu Ser Cys Val Ala Asp
275 280 285
Glu Leu Ile Arg Glu Met Val Lys Glu Gly Ala Tyr Leu Ala Asp Glu
290 295 300
Lys Glu Leu Ser Ala Ile Val Gly Lys Val Met Ile Thr Ala Lys Asp
305 310 315 320
Gly Ser Tyr Ala Pro Asn Lys Lys Tyr Val Gly Arg Asp Ala Thr Tyr
325 330 335
Ile Leu Lys Glu Ala Gly Ile Cys Val Asp Arg Glu Ala Lys Ile Ile
340 345 350
Ile Ala Glu Val Pro Phe Gly His Pro Leu Val Met Thr Glu Met Leu
355 360 365
Met Pro Val Ile Pro Val Thr Arg Val Ala Thr Val Glu Glu Ala Ile
370 375 380
Glu Lys Ala Val Ile Ala Glu Asn Gly Cys His His Thr Ala Met Met
385 390 395 400
His Ser Glu Asn Val Ser Asn Leu Thr Lys Met Ala Arg Ala Ala Asp
405 410 415
Thr Thr Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Gly Leu Gly Ile
420 425 430
Asp Gly Glu Gly Tyr Thr Thr Leu Thr Ile Ala Thr Pro Thr Gly Glu
435 440 445
Gly Leu Thr Ser Ala Arg Asn Phe Thr Arg Ser Arg Arg Cys Thr Leu
450 455 460
His Gly Ser Phe Arg Ile Val
465 470
<210> 43
<400> 43
000
<210> 44
<211> 448
<212> PRT
<213> Enterococcus phoeniculicola
<400> 44
Ile Met Asn Thr Leu Ser Asp Lys Ile Leu Arg Gly Arg Gln Ala Met
1 5 10 15
Gln Ser Ile Ser Asn Tyr Thr Gln Glu Gln Val Asp Glu Met Leu Ser
20 25 30
Val Ile Ser Lys Thr Ile Phe Asp His Ala Glu Glu Leu Ala Lys Glu
35 40 45
Ala Val Glu Glu Thr Gly Leu Gly Asn Tyr Glu His Lys Ile Gly Lys
50 55 60
Asn Gln Asn Met Ala Ile Asn Ile Phe Ser His Leu Lys Gly Lys Lys
65 70 75 80
Ser Val Gly Ile Ile Gln Thr Leu Lys Glu Glu Gly Val Val Glu Ile
85 90 95
Ala His Pro Val Gly Val Ile Gly Ser Val Thr Pro Thr Thr Asn Pro
100 105 110
Thr Ile Thr Pro Leu Gly Asn Gly Leu Met Ala Leu Lys Gly Lys Asn
115 120 125
Ala Met Ile Val Ser Pro His Pro Arg Ala Lys Lys Thr Thr Lys His
130 135 140
Thr Ile Asp Leu Met Arg Ser Ala Leu Glu Ser Ile His Ala Pro Lys
145 150 155 160
Asp Leu Leu Gln Val Ile Glu Glu Pro Ser Leu Glu Leu Ser Gln Gln
165 170 175
Leu Met Arg Glu Ser Asp Val Ile Val Ala Thr Gly Gly Pro Gly Leu
180 185 190
Val Arg Ala Ala Tyr Ser Ser Gly Lys Pro Ala Phe Gly Val Gly Pro
195 200 205
Gly Asn Val Gln Ala Ile Leu Asp Asp Asp Phe Asp Ile Asn Leu Ala
210 215 220
Ala Glu Leu Thr Val Ile Gly Arg Ser Phe Asp Asn Gly Ile Val Cys
225 230 235 240
Ala Cys Gln Gln Ser Leu Leu Tyr Pro Glu Lys Lys Glu Glu Glu Leu
245 250 255
Phe Gln Ala Leu Glu Asn Asn Lys Ala Tyr Ile Ile Lys Glu Glu Ile
260 265 270
Asp Val Gln Lys Met Arg Glu Leu Leu Phe Pro Gly Gly Lys Ser Asn
275 280 285
Pro Asp Leu Val Gly Gln Thr Ala Thr Phe Ile Ala Glu Lys Ala Gly
290 295 300
Ile Lys Val Pro Glu Asp Thr Ile Ile Leu Ala Val Lys Val Thr Thr
305 310 315 320
Ser Gly Gln Glu Glu Leu Leu Val Lys Glu Lys Met Asn Pro Val Leu
325 330 335
Val Val Lys Gly Cys Glu Ser Phe Glu Glu Ala Leu Leu Asp Ala Lys
340 345 350
Asn Asn Leu Trp Val Glu Gly Ala Gly His Ser Thr Gly Ile Phe Ser
355 360 365
Asn Asn Glu Gln His Ile Leu Ser Ala Gly Glu Thr Leu Pro Val Ser
370 375 380
Arg Val Val Val Asn Gln Pro Thr Ile Asp Ala Gly Gly Ser Pro Thr
385 390 395 400
Asn Gly Leu Asn Pro Thr Val Ser Leu Gly Cys Gly Ser Trp Gly Asn
405 410 415
Asn Ser Ile Ser Glu Asn Leu Ser Tyr His His Leu Ile Asn Ile Ser
420 425 430
Arg Ile Ala Tyr Pro Ile Ser Pro Lys His Thr Glu Thr Pro Trp Asn
435 440 445
<210> 45
<211> 462
<212> PRT
<213> Blautia schinkii
<400> 45
Met Pro Ile Ser Asp Ser Met Val Gln Glu Ile Val Gln Glu Val Met
1 5 10 15
Ala Lys Met Gln Ile Ala Asp Ala Pro Ala Gly Lys His Gly Val Phe
20 25 30
Lys Asp Met Asn Glu Ala Ile Glu Ala Ala Lys Lys Thr Glu Asn Ile
35 40 45
Val Lys Arg Met Ser Met Asp Gln Arg Glu Lys Ile Ile Thr Cys Ile
50 55 60
Arg Lys Ser Ile Lys Lys Asn Ala Glu Ile Met Ala Arg Met Gly Val
65 70 75 80
Asp Glu Thr Gly Met Gly Asn Val Gly Asp Lys Ile Leu Lys His His
85 90 95
Leu Val Ala Asp Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Val Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Ile Leu
130 135 140
Cys Asn Thr Met Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Tyr Ala Val Asn Leu Leu
165 170 175
Asn Glu Ala Ser Leu Glu Ala Gly Gly Pro Asp Asn Ile Ala Val Thr
180 185 190
Val Glu Gln Pro Thr Leu Glu Thr Ser Asn Ile Met Met Lys His Lys
195 200 205
Asp Ile Pro Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Val Arg Lys Ala Ala Gln Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Ser Pro Ile Val Asp Glu Leu Met His Tyr
275 280 285
Leu Val Ser Glu Asn Asp Cys Tyr Leu Ala Ser Lys Glu Glu Gln Asp
290 295 300
Lys Leu Thr Glu Val Val Leu Ala Gly Gly Arg Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Arg Thr Leu Leu Ser Met Ile Gly Val Asn Val
325 330 335
Pro Ala Asn Ile Arg Cys Ile Val Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Ala Thr Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Lys Asp Phe Asp Asp Ala Val Glu Gln Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Ile Asp Asn Ile Thr
385 390 395 400
Lys Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Ala Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Tyr Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Ser Asp Ser Leu Cys Ile Arg
450 455 460
<210> 46
<400> 46
000
<210> 47
<211> 472
<212> PRT
<213> Clostridium intestinale
<400> 47
Met Ser Ile Asp Ala Thr Leu Val Glu Lys Leu Val Arg Gln Ala Ile
1 5 10 15
Glu Glu Ala Lys Ser Lys Asn Leu Ile Ser Phe Asn Lys Val Glu Thr
20 25 30
Leu Asn Asn Tyr Gly Ile Phe Asn Thr Met Asp Glu Ala Ile Glu Ala
35 40 45
Ser Asp Val Ala Gln Lys Glu Leu Leu Asn Thr Ser Met Ala Asn Arg
50 55 60
Gln Lys Tyr Ile Asn Ile Ile Lys Ser Thr Val Leu Lys Arg Glu Asn
65 70 75 80
Leu Glu Leu Ile Ser Arg Met Ala Val Glu Glu Thr Glu Ile Gly Arg
85 90 95
Tyr Glu His Lys Leu Ile Lys Asn Arg Val Ala Ala Glu Lys Thr Pro
100 105 110
Gly Thr Glu Asp Leu Val Thr Glu Ala Ile Thr Gly Asp Asn Gly Ile
115 120 125
Thr Leu Ile Glu Tyr Cys Pro Phe Gly Val Ile Gly Ser Ile Thr Pro
130 135 140
Thr Thr Asn Pro Thr Glu Thr Ile Ile Cys Asn Ser Met Ser Met Ile
145 150 155 160
Ala Gly Gly Asn Thr Val Val Phe Ser Pro His Pro Arg Ala Lys Asn
165 170 175
Val Ser Ile Lys Leu Ile Thr Met Leu Asn Lys Ala Leu Glu Glu Ala
180 185 190
Gly Ala Pro Lys Asn Leu Ile Val Thr Val Lys Glu Pro Ser Ile Glu
195 200 205
Asn Thr Asn Ala Met Met Asp His Pro Lys Val Arg Val Leu Val Ala
210 215 220
Thr Gly Gly Pro Ala Ile Val Lys Lys Val Met Ser Thr Gly Lys Lys
225 230 235 240
Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Val Val Asp Glu Thr
245 250 255
Ala Asn Val Glu Lys Ala Ala Ile Asp Ile Val Asn Gly Cys Ser Phe
260 265 270
Asp Asn Asn Val Pro Cys Val Ala Glu Lys Glu Val Phe Ala Val Asp
275 280 285
Gln Ile Cys Asp Tyr Leu Ile His Tyr Met Lys Leu Asn Gly Ala Tyr
290 295 300
Glu Ile Lys Asp Arg Asn Thr Ile Gln Lys Leu Leu Glu Leu Val Thr
305 310 315 320
Asn Glu Asn Gly Gly Pro Lys Val Ser Phe Val Gly Lys Asn Ala Ser
325 330 335
Tyr Ile Leu Ser Lys Leu Gly Ile Asn Val Asp Asp Asn Ile Lys Ile
340 345 350
Ile Ile Met Glu Val Asp Lys Asp His His Phe Val Lys Glu Glu Met
355 360 365
Met Met Pro Ile Leu Pro Ile Val Arg Thr Arg Asp Val Asp Glu Ala
370 375 380
Ile Glu Tyr Ala Tyr Val Ala Glu Asn Gly Asn Arg His Thr Ala Ile
385 390 395 400
Met His Ser Lys Asn Val Asp Lys Leu Thr Lys Met Ala Arg Leu Leu
405 410 415
Glu Thr Thr Ile Phe Val Lys Asn Ala Pro Ser Phe Ala Gly Leu Gly
420 425 430
Val Gly Gly Glu Gly Asn Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly
435 440 445
Glu Gly Leu Thr Thr Ala Lys Ser Phe Cys Arg Lys Arg Arg Cys Ile
450 455 460
Met Val Asp Ala Phe Asn Ile Arg
465 470
<210> 48
<211> 483
<212> PRT
<213> Massilioclostridium coli
<400> 48
Met Val Phe Ser Gln Asn Gln Ile Asp Ser Ile Val Gln Ser Val Val
1 5 10 15
Ala Gln Met Gln Gly Thr Thr Pro Thr Ser Ala Pro Ala Tyr Asp Ser
20 25 30
Thr Gln Tyr Asn Gly Arg Gln Tyr Leu Gly Val Tyr Ala Thr Met Glu
35 40 45
Glu Gly Ile Asp Ala Ala Ala Asp Ser Tyr Lys Val Ile Arg Asn Met
50 55 60
Ser Val Glu Gln Arg Glu Lys Ile Ile Thr Glu Ile Arg Lys Leu Thr
65 70 75 80
Arg Ala Glu Ala Glu Ile Met Ala Lys Leu Gly Val Glu Glu Thr Lys
85 90 95
Met Gly Arg Val Glu His Lys Thr Leu Lys His Ile Leu Val Ala Asp
100 105 110
Lys Thr Pro Gly Thr Glu Asp Ile Gln Thr Glu Ala Gln Ser Gly Asp
115 120 125
Gly Gly Leu Thr Leu Val Glu Met Ala Pro Phe Gly Ile Ile Gly Ala
130 135 140
Ile Thr Pro Ser Thr Asn Pro Ser Glu Thr Val Ile Cys Asn Ser Ile
145 150 155 160
Ala Met Ile Ala Ala Gly Asn Ala Val Val Phe Asn Pro His Pro Gly
165 170 175
Ala Ile Lys Val Ser Asn Tyr Ala Val Asp Leu Val Asn Arg Ala Ser
180 185 190
Leu Ala Ala Gly Gly Pro Ala Ser Leu Val Cys Ser Met Val Lys Pro
195 200 205
Thr Met Gln Thr Ala Asp Val Met Tyr Lys Asp Pro Arg Val Arg Met
210 215 220
Leu Val Cys Thr Gly Gly Pro Gly Val Val Lys Ser Val Leu Ser Ser
225 230 235 240
Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val
245 250 255
Asp Asp Thr Ala Asp Ile Lys Lys Ala Ala Lys Asp Ile Ile Asp Gly
260 265 270
Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu Val Phe
275 280 285
Ala Phe Ser Asn Ile Ala Asp Glu Leu Met Tyr Asn Met Gln Gln Asn
290 295 300
Gly Ala Tyr Phe Ile Thr Ala Ala Gln Ala Asp Glu Leu Ala Lys Ile
305 310 315 320
Val Leu Val Glu Lys Lys Asn Glu Lys Thr Gly Lys Ile Thr Tyr Ser
325 330 335
Val Ser Arg Asp Trp Val Gly Arg Asp Ala Lys Lys Phe Ala Ala Ala
340 345 350
Leu Gly Ile Glu Val Asp Asp Ser Val Arg Cys Leu Ile Cys Glu Val
355 360 365
Glu Glu Asp His Leu Phe Val Gln Thr Glu Leu Met Met Pro Ile Leu
370 375 380
Ala Val Val Arg Val Lys Asp Ile Asp Glu Ala Ile Glu Lys Ala Val
385 390 395 400
Arg Ala Glu His Gly Asn Arg His Ser Ala His Met His Ser Lys Asn
405 410 415
Ile Glu Asn Leu Ser Lys Phe Ala Lys Ala Ile Glu Thr Thr Ile Phe
420 425 430
Val Lys Asn Ala Pro Ser Tyr Ala Gly Ile Gly Phe Gly Ala Glu Gly
435 440 445
His Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser
450 455 460
Ala Arg Ser Phe Thr Arg Lys Arg Arg Cys Val Met Lys Asp Met Phe
465 470 475 480
His Ile Ile
<210> 49
<211> 466
<212> PRT
<213> Cloacibacillus porcorum
<400> 49
Met Asn Ile Asp Ala Ala Leu Ile Glu Gly Ile Val Lys Gly Val Met
1 5 10 15
Arg Lys Ile Asp Glu Ser Glu Asn Asn Ser Ala Gly Ser Cys Gly Ile
20 25 30
Phe Ala Asp Met Asn Asp Ala Ile Glu Ala Ala Ala Ala Ala Gln Arg
35 40 45
Arg Tyr Leu Asp Cys Ser Met Ala Asp Arg Ala Arg Phe Val Glu Ala
50 55 60
Ile Arg Gly Thr Val Leu Asn Glu Glu Asn Leu Lys Phe Met Ser Leu
65 70 75 80
Ser Thr Ile Glu Glu Thr Gly Met Gly Asn Tyr Glu His Lys Leu Val
85 90 95
Lys Asn Arg Leu Ala Ala Thr Lys Thr Pro Gly Ile Glu Asp Leu Thr
100 105 110
Thr Asp Ala Ile Thr Gly Asp Asp Gly Leu Thr Ile Val Glu Tyr Ser
115 120 125
Pro Phe Gly Val Ile Gly Ala Ile Thr Pro Thr Thr Asn Pro Thr Glu
130 135 140
Thr Ile Ile Cys Asn Ser Ile Gly Met Leu Ala Ala Gly Asn Thr Val
145 150 155 160
Val Phe Ser Pro His Pro Arg Ala Lys Lys Val Ser Leu Trp Leu Val
165 170 175
Ser Glu Leu Asn Arg Ala Leu Ala Ala Ala Gly Ala Pro Ala Asn Leu
180 185 190
Ile Val Thr Val Ser Glu Pro Ser Ile Glu Asn Thr Asn Leu Met Met
195 200 205
Ala His Pro Lys Val Arg Met Leu Val Ala Thr Gly Gly Pro Ala Ile
210 215 220
Val Lys Thr Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Ala Val Val Asp Glu Ser Ala Asn Ile Glu Lys Ala
245 250 255
Ala Lys Asp Ile Val Asp Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys
260 265 270
Ile Ala Glu Lys Glu Val Ile Val Val Asp Ser Ala Ala Asp Tyr Leu
275 280 285
Ile Phe Asn Met Lys Lys Asn Gly Ala Phe Glu Val Lys Asp Pro Ala
290 295 300
Val Ile Glu Arg Leu Val Gly Leu Val Thr Lys Glu Gly Lys Ser Pro
305 310 315 320
Lys Thr Glu Phe Val Gly Lys Ser Ala Lys Tyr Ile Leu Glu Lys Ala
325 330 335
Gly Val Glu Ala Pro Glu Asp Thr Arg Val Ile Ile Met Glu Ala Arg
340 345 350
Glu Glu His Pro Phe Val Gln Val Glu Leu Met Met Pro Ile Leu Pro
355 360 365
Ile Val Arg Ala Asp Asn Val Asn Glu Ala Ile Glu Met Ala Val Arg
370 375 380
Val Glu His Gly Asn Arg His Thr Ala Met Met His Ser Arg Asn Val
385 390 395 400
Asp Ser Leu Thr Lys Met Ala Lys Leu Ile Gln Thr Thr Ile Phe Val
405 410 415
Lys Asn Gly Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Met Gly His
420 425 430
Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala
435 440 445
Lys Thr Phe Ala Arg Arg Arg Arg Cys Val Leu Val Gly Gly Met Asp
450 455 460
Ile Arg
465
<210> 50
<400> 50
000
<210> 51
<400> 51
000
<210> 52
<400> 52
000
<210> 53
<211> 500
<212> PRT
<213> Sporosarcina globispora
<400> 53
Met Gln Glu Met Arg Asp Ala Val Lys Arg Ala Lys Glu Ala Gln Leu
1 5 10 15
Glu Tyr Met Ala Phe Thr Gln Glu Gln Val Asp Glu Ile Val Lys Asn
20 25 30
Ala Ala Asp Ala Ala Tyr Ala Lys Ser Leu Tyr Leu Ala Gln Met Ala
35 40 45
Val Glu Glu Thr Gly Met Gly Ile Val Glu His Lys Lys Ile Lys Asn
50 55 60
Glu Val Gly Ser Lys Ala Val Tyr Glu Ser Ile Lys Asp Glu Lys Thr
65 70 75 80
Val Gly Ile Ile Arg Glu Asp Arg Val Asn Lys Val Thr Glu Ile Ala
85 90 95
Tyr Pro Tyr Gly Val Val Ala Gly Ile Ile Pro Thr Thr Asn Pro Thr
100 105 110
Ser Thr Ala Ile Phe Lys Ala Leu Ile Ser Leu Lys Thr Arg Asn Ala
115 120 125
Ile Val Val Ser Pro His Pro Arg Ala Val Lys Cys Thr Val Glu Ala
130 135 140
Leu Lys Ile Val Asn Glu Ala Ala Ile Gln Ala Gly Ala Pro Glu Gly
145 150 155 160
Leu Ile Gly Trp Ile Ser Lys Pro Ser Met Gly Ala Thr Asn Glu Leu
165 170 175
Met Lys His Arg Asp Ile Ser Leu Ile Leu Ala Thr Gly Gly Gly Gly
180 185 190
Leu Val Arg Ala Ala Tyr Ser Ser Gly Lys Pro Ala Tyr Gly Val Gly
195 200 205
Pro Gly Asn Val Pro Cys Tyr Ile Glu Lys Thr Ala Lys Val Ala Gln
210 215 220
Ser Val Lys Met Ile Ile Asp Ser Lys Ser Phe Asp Asn Gly Thr Ile
225 230 235 240
Cys Ala Thr Glu Gln Ser Ile Val Ala Asp Arg Asn Ile Lys Glu Met
245 250 255
Ala Met Arg Glu Leu Lys Asn Asn Gly Ala Tyr Ile Leu Asn Ser Asp
260 265 270
Glu Lys Ala Ala Leu Glu Lys Ile Ile Ser Pro Ser Pro Gly Lys Leu
275 280 285
Asn Pro Asp Ile Val Gly Gln Ser Ala Val Lys Ile Ala Ala Met Ala
290 295 300
Gly Ile Gln Val Pro Asn Asp Thr Arg Val Leu Ile Ala Glu Glu Thr
305 310 315 320
Lys Val Gly Lys Asp Ile Pro Phe Ser Ile Glu Lys Leu Ser Pro Ile
325 330 335
Phe Ala Phe Tyr Thr Ala Glu Ser Tyr Gln Asp Ala Lys Glu Ile Cys
340 345 350
Leu Gln Leu Leu Asn Leu Gly Gly Arg Gly His Ser Leu Ser Leu His
355 360 365
Thr Asn Asp Asp Ala Val Ala Lys Asp Phe Ala Leu Glu Met Pro Val
370 375 380
Ser Arg Ile Leu Val Asn Thr Leu Ser Ser Ile Gly Ala Val Gly Ala
385 390 395 400
Thr Thr Gly Leu Met Pro Ser Leu Thr Leu Gly Cys Gly Ser Phe Gly
405 410 415
Gly Asn Ile Thr Ser Asp Asn Val Thr Ala Arg His Leu Ile Asn Thr
420 425 430
Lys Arg Met Ala Tyr Gly Thr Lys Glu Val Thr Val Pro Lys Pro Ala
435 440 445
Ala Ser Ser Ser Ile Ala Glu Lys Glu Gln Ala Gly Ser Gln Asp Val
450 455 460
Asp His Ile Val Ser Gln Val Leu Gln Gln Val Ser Pro Gly Gly Glu
465 470 475 480
Val Asp Ala Lys Met Ile Ala Asp Met Val Asn Gln Val Met Lys Lys
485 490 495
Tyr Gln Thr Asn
500
<210> 54
<400> 54
000
<210> 55
<400> 55
000
<210> 56
<400> 56
000
<210> 57
<400> 57
000
<210> 58
<211> 528
<212> PRT
<213> Rhodobacter aestuarii
<400> 58
Met Lys Asp Ile Asp Ile Glu Asn Ala Val Ala Arg Val Leu Ser Gly
1 5 10 15
Tyr Thr Gly Pro Ala Glu Thr Pro Ala Pro Ala Pro Thr Ser Lys Pro
20 25 30
Gly Thr Thr Gly Cys Val Trp Glu Pro Val Lys Ala Val Asp Pro Val
35 40 45
Asp Asp Ile Ile Gly Gly Met Leu Thr Arg Ala Leu Gly Glu Arg Asn
50 55 60
Cys Ser Asn Cys Lys Ala Gly Asp Cys Gln Gly Lys Ala Gly Cys Leu
65 70 75 80
Ser Ile Ser Asp Ala Glu Ala Leu Glu Leu Gly Asp Gly Val Phe Ala
85 90 95
Thr Met Asp Glu Ala Val Asn Ala Ala Ala Glu Ala Gln Arg Lys Tyr
100 105 110
Leu Phe Cys Thr Met Gly Asp Arg Lys Arg Phe Val Glu Gly Ile Arg
115 120 125
Ala Ile Phe Thr Asp Glu Ala Val Leu Glu Arg Ile Ser Arg Leu Thr
130 135 140
Val Glu Gln Thr Gly Met Gly Asn Leu Ala His Lys Ile Ile Lys Asn
145 150 155 160
Arg Leu Ala Ala Glu Lys Thr Pro Gly Val Glu Asp Leu Thr Thr Glu
165 170 175
Ala Gln Ser Gly Asp Asp Gly Leu Thr Leu Val Glu Leu Ser Pro Phe
180 185 190
Gly Val Ile Gly Ala Ile Thr Pro Thr Thr Asn Pro Thr Glu Thr Val
195 200 205
Ile Cys Asn Ser Ile Gly Met Leu Ala Ala Gly Asn Ala Ala Val Phe
210 215 220
Ser Pro His Pro Arg Ala Lys Gly Val Ser Leu Leu Ala Ile Lys Leu
225 230 235 240
Ile Asn Arg Lys Leu Ala Ala Leu Gly Ala Pro Ala Asn Leu Val Val
245 250 255
Thr Val Gln Ala Pro Ser Ile Asp Asn Thr Asn Ala Met Met Ala His
260 265 270
Pro Gln Val Arg Met Leu Val Ala Thr Gly Gly Pro Gly Ile Val Arg
275 280 285
Thr Val Met Ser Thr Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn
290 295 300
Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Pro Lys Ala Ala Gln
305 310 315 320
Asp Ile Val Asn Gly Ala Ser Phe Asp Asn Asn Met Pro Cys Ile Ala
325 330 335
Glu Lys Glu Val Ile Val Val Asp Gln Val Ala Asp Phe Leu Ile Ser
340 345 350
Glu Met Gln Arg Asn Gly Ala Trp Leu Ala Ser Asp Pro Ser Val Val
355 360 365
Glu Arg Leu Ala Gln Leu Val Leu Thr Glu Lys Gly Gly Pro Gln Thr
370 375 380
Gly Cys Val Gly Lys Ser Ala Ala Trp Leu Leu Gly Gln Ile Gly Ile
385 390 395 400
Gln Val Gly Pro Asp Val Arg Leu Ile Ile Leu Glu Thr Thr Lys Asp
405 410 415
His Pro Phe Val Gln Glu Glu Leu Met Met Pro Ile Leu Pro Val Val
420 425 430
Arg Val Pro Asp Val Asp Thr Ala Ile Asp Leu Ala Val Asp Leu Glu
435 440 445
His Gly Asn Arg His Thr Ala Met Met His Ser Thr Asn Val Arg Lys
450 455 460
Leu Thr Lys Met Ala Lys Leu Ile Gln Thr Thr Ile Phe Val Lys Asn
465 470 475 480
Gly Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Tyr Thr Thr
485 490 495
Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Pro Arg Ser
500 505 510
Phe Ala Arg Arg Arg Lys Cys Val Met Val Glu Ala Leu Asn Val Arg
515 520 525
<210> 59
<211> 468
<212> PRT
<213> Clostridium grantii
<400> 59
Met Ala Ile Asn Glu Ser Gln Ile Glu Glu Ile Val Lys Gln Val Leu
1 5 10 15
Leu Asn Val Ser Gly Thr Thr Lys Val Lys Asn Glu Asn Lys Ala Ile
20 25 30
Gly Ile Phe Glu Asp Ile Glu Glu Ala Ile Asp Ala Ala Lys Ile Ala
35 40 45
Gln Lys Lys Ile Lys Lys Met Ser Met Glu Gln Arg Glu Lys Ile Ile
50 55 60
Thr Arg Ile Arg Glu Lys Thr Arg Glu Asn Ala Lys Ile Met Ser Glu
65 70 75 80
Met Ala Val Glu Glu Thr Gly Met Gly Arg Val Asp His Lys Ile Leu
85 90 95
Lys His Leu Leu Val Ala Asp Lys Thr Pro Gly Thr Glu Asp Ile Thr
100 105 110
Thr Thr Ala Trp Ser Gly Asp Asn Gly Leu Thr Leu Ile Glu Met Gly
115 120 125
Ala Phe Gly Val Ile Gly Gly Ile Thr Pro Ser Thr Asn Pro Ser Cys
130 135 140
Thr Val Leu Cys Asn Ser Ile Gly Met Ile Ala Gly Gly Asn Thr Val
145 150 155 160
Val Phe Asn Pro His Pro Gly Ala Val Lys Val Ser Asn Tyr Ala Val
165 170 175
Thr Leu Val Asn Glu Ala Ser Val Glu Cys Gly Gly Pro Glu Asn Ile
180 185 190
Ala Cys Ser Val Thr Lys Pro Thr Leu Asp Ser Gly Lys Ile Leu Met
195 200 205
Thr His Lys Asp Ile Ala Leu Leu Ala Val Thr Gly Gly Pro Gly Val
210 215 220
Val Thr Ala Ala Leu Lys Ser Gly Lys Arg Ala Leu Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Leu Gln Ser Ala
245 250 255
Ala Lys His Ile Val Asp Gly Ala Thr Phe Asp Asn Asn Leu Pro Cys
260 265 270
Ile Ala Glu Lys Glu Val Val Ala Val Glu Ser Ile Val Glu Glu Leu
275 280 285
Lys Tyr His Met Ile Asn Asn Gly Cys Tyr Glu Leu Lys Gly Ser Asp
290 295 300
Ile Asp Lys Leu Val Asn Thr Val Leu Ile Asn Asn Asn Gly Ile Ile
305 310 315 320
Gly Leu Asn Arg Asp Cys Val Gly Lys Asp Ala Lys Val Ile Leu Lys
325 330 335
Lys Leu Gly Ile Glu Val Asp Asp Ser Ile Arg Cys Ile Ile Phe Asp
340 345 350
Ala Asp Glu Asp His Ile Leu Val Leu Glu Glu Leu Met Met Pro Ile
355 360 365
Leu Gly Ile Val Lys Val Glu Asn Val Asp Glu Ala Ile Lys Leu Ala
370 375 380
Val Arg Tyr Glu His Gly Asn Arg His Ser Ala His Met His Ser Lys
385 390 395 400
Asn Ile Asp Asn Leu Thr Lys Tyr Gly Arg Glu Ile Asp Thr Ala Ile
405 410 415
Phe Val Lys Asn Ala Pro Ser Tyr Ser Ala Leu Gly Phe Asn Gly Glu
420 425 430
Gly Tyr Cys Thr Phe Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr
435 440 445
Ser Gly Lys Thr Phe Thr Lys Ser Arg Arg Cys Val Leu Ser Asp Gly
450 455 460
Leu Ser Ile Arg
465
<210> 60
<211> 449
<212> PRT
<213> Collinsella sp. GD7
<400> 60
Val Ala Glu Phe Ile Glu Arg Ala Arg Val Ala Gln Ala Glu Phe Glu
1 5 10 15
Thr Tyr Ser Gln Glu Glu Val Asp Arg Ala Val Arg Ala Ile Gly Lys
20 25 30
Ala Val Phe Asp Ala Ala Glu Pro Leu Ala Lys Leu Ala Val Glu Glu
35 40 45
Thr Arg Met Gly Arg Tyr Glu Asp Lys Ile Ala Lys Asn Ser Gly Lys
50 55 60
Thr Lys Ile Thr Trp Asp Arg Leu Lys Gly Val Lys Ser Arg Gly Ile
65 70 75 80
Ile Ala Arg His Glu Asp Glu Gly Ile Val Glu Val Ala Lys Pro Met
85 90 95
Gly Val Ile Gly Cys Ile Pro Pro Thr Thr Asn Pro Thr Met Thr Pro
100 105 110
Ala His Asn Ala Met Cys Ala Leu Lys Gly Gly Asn Ala Leu Leu Ile
115 120 125
Ser Pro His Pro Arg Ala Lys Lys Thr Gly Val Glu Thr Val Arg Ile
130 135 140
Met Arg Glu Ala Leu Glu Ala Met Gly Ala Pro Ala Asp Leu Ile Gln
145 150 155 160
Ile Ile Pro Asp Pro Thr Leu Glu Ile Ser Ser Leu Val Met Ser Met
165 170 175
Cys Asp Cys Thr Ile Ala Thr Gly Gly Pro Gly Met Val Lys Ala Val
180 185 190
Tyr Ser Ser Gly Lys Pro Ala Phe Gly Val Gly Ala Gly Asn Val Gln
195 200 205
Thr Ile Val Asp Thr Asp Ala Asp Leu Glu Leu Ser Ala Gln Gln Ile
210 215 220
Val Arg Ser Arg Thr Tyr Asp Asn Gly Val Leu Cys Thr Cys Glu Gln
225 230 235 240
Cys Ile His Val Gln Glu Asp Ile Tyr Gly Glu Met Val Arg Leu Phe
245 250 255
Gln Gln Glu Gly Ala Phe Tyr Ile Ser Glu Gln Ala Asp Val Asp Ala
260 265 270
Leu Arg Ala Ala Leu Phe Pro Asn Gly Ala Ile Asn Lys Asp Ala Val
275 280 285
Gly Ala Ser Pro Gln Phe Ile Gly Ser Leu Ala Gly Leu Asp Val Pro
290 295 300
Glu Asp Ala Lys Leu Leu Met Val Lys Val Asp Ala Tyr Gly Ala Asp
305 310 315 320
Glu Leu Leu Cys Lys Glu Lys Leu Cys Pro Val Met Cys Val Ala Ser
325 330 335
Tyr Gly Thr Trp Glu Glu Gly Val Ala Asn Ala Lys Thr Asn Leu Leu
340 345 350
His Glu Gly Ala Gly His Ser Ala Ile Val Arg Ser His Thr Ala Glu
355 360 365
His Val Asp Tyr Ala Gly Glu Gln Leu Pro Val Ser Arg Ile Gly Val
370 375 380
Asn Met Ile Gly Ser Ser Gly Leu Gly Gly Ala Phe Asp Asn Gly Leu
385 390 395 400
Asn Pro Thr Ala Thr Leu Gly Cys Gly Ser Trp Gly Asn Asn Ser Ile
405 410 415
Ser Glu Asn Leu Trp Trp His His Leu Val Asn Ile Ala Arg Ile Ala
420 425 430
Val Ala Leu Pro Asp Val Gln Val Pro Ser Asp Glu Glu Val Trp Gly
435 440 445
Glu
<210> 61
<211> 471
<212> PRT
<213> Clostridium estertheticum
<400> 61
Met Glu Ile Lys Asn Asp Glu Ile Ser Ala Met Val Glu Lys Val Leu
1 5 10 15
Gln Glu Met Asn Arg Arg Asp Leu Asn Val Ser Glu Ser Asp Gly Val
20 25 30
Phe Asp Asp Met Asp Glu Ala Ile Glu Ala Ala Ser Ile Ala Gln Lys
35 40 45
Glu Leu Ile Cys Met Ser Ile Ser Gln Arg Glu Glu Leu Ile Ser Ala
50 55 60
Met Arg Lys Ala Ile Leu Asp Asn Ala Thr Lys Ile Ala Asp Ile Cys
65 70 75 80
Val Glu Asp Thr Gly Met Gly Arg Lys Asp His Lys Tyr Leu Lys Leu
85 90 95
Lys Leu Val Ala Asn Lys Thr Pro Gly Thr Glu Val Leu Lys Thr Met
100 105 110
Ala Ile Ser Gly Asp Lys Gly Leu Thr Leu Ile Glu Met Gly Pro Phe
115 120 125
Gly Val Ile Gly Gly Ile Thr Pro Ser Thr Asn Pro Ser Ala Thr Val
130 135 140
Met Cys Asn Ser Ile Gly Met Ile Ala Ser Gly Asn Ala Ala Val Phe
145 150 155 160
Ser Pro His Pro Gly Ala Ile Glu Ser Cys Leu Ile Ser Val Arg Val
165 170 175
Leu Asn Lys Ala Ile Thr Asp Ala Gly Gly Pro Arg Asn Leu Ile Thr
180 185 190
Thr Leu Arg Lys Pro Ser Leu Glu Ser Thr Asp Thr Met Ile Asn Asn
195 200 205
Pro Lys Ile Arg Leu Val Val Ala Thr Gly Gly Pro Phe Ile Val Lys
210 215 220
Lys Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Val Lys Ala Ala Arg
245 250 255
Asp Ile Ile Ala Gly Cys Cys Phe Asp Asn Asn Leu Pro Cys Ile Ala
260 265 270
Glu Lys Glu Ala Ile Val Val Glu Ser Val Tyr Glu Lys Leu Ile Ala
275 280 285
Glu Met Leu Lys Asn Gly Asn Val Tyr Glu Leu Asp Glu Gln Gln Lys
290 295 300
Gln Lys Val Leu Asp Val Val Met Asn Lys Thr Glu Lys Gly Gly Lys
305 310 315 320
Ile Lys Tyr Gly Val Asn Lys Asn Phe Val Gly Lys Asp Ala Ser Val
325 330 335
Ile Leu Ala Ala Ala Gly Ile Glu Ala Pro Lys Gly Val Glu Cys Leu
340 345 350
Ile Cys Arg Ala Glu Asn Leu His Pro Phe Val Gln Glu Glu Leu Met
355 360 365
Met Pro Ile Leu Ala Ile Val Lys Val Lys Asp Val Asp Glu Ala Ile
370 375 380
Asn Thr Ala Val Leu Asp Glu His Gly Asn Arg His Thr Ala Met Met
385 390 395 400
His Ser Lys Asn Ile Asp Asn Leu Thr Lys Met Ser Arg Leu Ile Asp
405 410 415
Thr Thr Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Gly Ile Gly Phe
420 425 430
Gly Gly Glu Gly Trp Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu
435 440 445
Gly Ile Thr Asn Ala Thr Ser Phe Thr Arg Gln Arg Arg Cys Thr Met
450 455 460
Val Asp Ser Phe Arg Ile Ile
465 470
<210> 62
<211> 483
<212> PRT
<213> bacterium MS4
<400> 62
Met Asp Ile Asp Ala Asn Leu Ile Glu Lys Met Val Lys Gln Val Leu
1 5 10 15
Asn Glu Ile Asp Ala Gly Lys Ala Glu Lys Thr Ala Ala Ala Glu Ile
20 25 30
Lys Lys Glu Glu Lys Gly Gly Ala Tyr Gly Ile Phe Asn Thr Met Glu
35 40 45
Glu Ala Ile Asp Ala Cys Asp Ile Ala Gln Lys Gln Tyr Leu Phe Cys
50 55 60
Ser Met Ala Glu Arg Gln Lys Tyr Val Gln Thr Leu Arg Asp Val Val
65 70 75 80
Leu Lys Gln Glu Asn Leu Glu Leu Ile Ser Arg Leu Ala Val Glu Glu
85 90 95
Thr Gly Met Gly Asn Tyr Pro His Lys Leu Ile Lys Asn Arg Leu Ala
100 105 110
Ala Glu Lys Ser Pro Gly Ile Glu Asp Leu Glu Thr Thr Ala Leu Ser
115 120 125
Gly Asp Asp Gly Leu Thr Leu Val Glu Tyr Cys Pro Phe Gly Val Ile
130 135 140
Gly Ala Ile Thr Pro Ala Thr Asn Pro Thr Glu Thr Ile Ile Cys Asn
145 150 155 160
Ser Ile Gly Met Leu Ala Ala Gly Asn Ser Ile Val Phe Ser Pro His
165 170 175
Pro Arg Ala Lys Asp Val Thr Ile Arg Leu Val Thr Met Ile Asn Arg
180 185 190
Ala Leu Glu Glu Thr Gly Ala Pro Lys Asn Leu Ile Val Thr Val Met
195 200 205
Glu Pro Ser Ile Glu Asn Thr Asn Val Met Met Lys His Pro Lys Ile
210 215 220
Arg Met Leu Val Ala Thr Gly Gly Pro Gly Ile Val Lys Leu Val Met
225 230 235 240
Ser Thr Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val
245 250 255
Val Val Asp Glu Thr Ala Asp Ile Lys Lys Ala Ala Ile Asp Ile Val
260 265 270
Asn Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu
275 280 285
Val Ile Ala Val Asp Arg Ile Thr Asp Glu Leu Ile Arg Ser Met Arg
290 295 300
Glu Asn Gly Ala Tyr Gln Val Thr Asp Pro Ala Val Ile Gln Lys Leu
305 310 315 320
Ala Asp Leu Val Arg Lys Glu Gly Gly Gly Pro Lys Thr Ser Phe Val
325 330 335
Gly Lys Ser Ala Ile Tyr Ile Leu Asp Lys Ile Gly Ile Gln Ala Gly
340 345 350
Pro Glu Val Lys Val Ile Ile Met Glu Thr Pro Lys Asp His Pro Phe
355 360 365
Val Met Glu Glu Leu Met Met Pro Ile Leu Pro Ile Val Arg Thr Arg
370 375 380
Asn Val Asp Glu Ala Ile Asp Leu Ala Leu Ile Ala Glu Arg Gly Asn
385 390 395 400
Arg His Thr Ala Met Met His Ser Lys Asn Val Asp Lys Leu Thr Lys
405 410 415
Met Ala Lys Leu Leu Gln Thr Thr Ile Phe Val Lys Asn Ala Pro Ser
420 425 430
Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly His Thr Thr Phe Thr Ile
435 440 445
Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys Ser Phe Cys Arg
450 455 460
Lys Arg Arg Cys Val Leu Ser Asp Ala Phe His Ile Arg Asp Phe Ser
465 470 475 480
Lys Gly Leu
<210> 63
<211> 462
<212> PRT
<213> Clostridium glycyrrhizinilyticum
<400> 63
Val Ser Val Asn Glu Gln Met Val Gln Asp Ile Val Gln Glu Val Met
1 5 10 15
Ala Lys Met Gln Ile Thr Ser Asp Val Ser Gly Ser His Gly Val Phe
20 25 30
Lys Asp Met Asn Glu Ala Ile Ala Ala Ala Lys Lys Thr Gln Lys Ile
35 40 45
Val Gly Lys Met Ser Met Asp Gln Arg Glu Lys Ile Ile Ser Asn Ile
50 55 60
Arg Thr Lys Ile Lys Glu Asn Ala Glu Ile Met Ala Arg Met Gly Val
65 70 75 80
Gln Glu Thr Gly Met Gly Asn Val Gly His Lys Ile Leu Lys His Val
85 90 95
Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Val Leu
130 135 140
Cys Asn Thr Ile Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Phe Ala Ile Asn Leu Leu
165 170 175
Asn Glu Ala Ser Leu Glu Ala Gly Gly Pro Asp Asn Ile Ala Cys Thr
180 185 190
Val Glu Lys Pro Thr Leu Ala Ser Ser Asp Ile Met Met Lys His Lys
195 200 205
Asp Ile Pro Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Glu Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Asp Ser Ile Ala Asp Glu Leu Met Tyr Tyr
275 280 285
Met Val Ser Glu Gln Gly Cys Tyr Lys Ile Thr Lys Glu Glu Gln Asp
290 295 300
Ala Leu Thr Ala Val Val Leu Lys Asp Gly Lys Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Lys Thr Leu Leu Gly Met Ile Gly Val Thr Val
325 330 335
Pro Asp Asn Ile Arg Cys Ile Thr Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Ala Glu Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Lys Asp Phe Asp Asp Ala Val Glu Gln Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Val Asp Asn Ile Thr
385 390 395 400
Thr Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Gly Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Tyr Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Thr Asp Ser Leu Cys Ile Arg
450 455 460
<210> 64
<400> 64
000
<210> 65
<211> 482
<212> PRT
<213> Thermincola ferriacetica
<400> 65
Met Ala Ile Glu Ala Tyr Gln Ile Glu Lys Ile Val Glu Glu Val Met
1 5 10 15
Lys Lys Met Val Ser Gly Gly Ser Gly Asp Ser Phe Ala Gly Lys Ala
20 25 30
Lys Gly Ile Phe Glu Ser Val Asp Glu Ala Val Lys Ala Ala Lys Ala
35 40 45
Ala Gln Lys Glu Leu Val Ala Met Arg Ile Glu Lys Arg Glu Met Leu
50 55 60
Leu Lys Ala Met Arg Glu Ala Ala Ile Ala His Ala Glu Glu Leu Ala
65 70 75 80
Arg Leu Ala Val Glu Glu Thr Gly Met Gly Arg Val Thr Asp Lys Ile
85 90 95
Ile Lys Asn Arg Val Ala Ala Glu Lys Thr Pro Gly Thr Glu Asn Leu
100 105 110
Gln Pro Ser Ala Val Thr Gly Asp Arg Gly Leu Thr Leu Ile Glu Arg
115 120 125
Ala Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Cys
130 135 140
Ala Thr Val Ile Asn Asn Ser Ile Ser Met Val Ala Ala Gly Asn Ser
145 150 155 160
Val Val Phe Ser Val His Pro Gly Ala Lys Lys Ala Ser Leu Leu Thr
165 170 175
Val Glu Ile Leu Asn Glu Ala Ile Glu Lys Ala Gly Gly Pro Ala Asn
180 185 190
Val Leu Thr Ala Val Ala Ser Pro Ser Leu Glu Asn Thr Asn Ala Leu
195 200 205
Met Lys His Pro Asp Ile Lys Leu Leu Val Ala Thr Gly Gly Pro Gly
210 215 220
Leu Val Lys Ala Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly
225 230 235 240
Ala Gly Asn Pro Pro Ala Leu Val Asp Glu Thr Ala Asp Leu Glu Arg
245 250 255
Ala Ala Lys Ser Ile Val Ala Gly Ala Ser Phe Asp Asn Asn Leu Pro
260 265 270
Cys Ile Ala Glu Lys Glu Val Ile Val Val Asp Tyr Val Ala Asn Gln
275 280 285
Leu Ile Ser Tyr Met Lys Gln Asn Gly Ala Tyr Leu Ala Asn Asp Arg
290 295 300
Glu Ile Lys Ala Leu Met Asp Leu Val Leu Thr Lys Asn Glu Asn Leu
305 310 315 320
Lys Ala Glu Gly Cys Thr Val Lys Pro Glu Lys Leu Tyr Gly Gly Ile
325 330 335
Asn Lys Glu Tyr Val Gly Lys Asp Ala Ala Tyr Ile Met Lys Lys Ile
340 345 350
Gly Val Asp Ile Pro Glu Asp Thr Lys Leu Ile Ile Cys Glu Val Asp
355 360 365
Glu Asp His Pro Phe Val Leu Glu Glu Leu Met Met Pro Ile Leu Pro
370 375 380
Ile Val Arg Val Pro Asn Val Gln Lys Ala Ile Glu Val Gly Val Arg
385 390 395 400
Val Glu His Gly Asn Arg His Thr Ala Val Met His Ser Gln Asn Ile
405 410 415
Asp Asn Leu Ser Ala Phe Ala Arg Ala Ile Gln Thr Thr Ile Phe Val
420 425 430
Lys Asn Gly Pro Ser Tyr Ala Gly Ile Gly Ile Gly Gly Glu Gly Tyr
435 440 445
Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ala Ala
450 455 460
Ser Ser Phe Thr Arg Gln Arg Arg Cys Val Leu Val Asp Gly Phe Ser
465 470 475 480
Ile Val
<210> 66
<211> 461
<212> PRT
<213> Lachnospiraceae bacterium AC3007
<400> 66
Met Asn Glu Lys Leu Val Gln Glu Ile Val Arg Arg Val Met Ala Asp
1 5 10 15
Ile Asn Asp Glu Gly Gly Ala Asp Gly Met His Gly Val Phe Ser Asp
20 25 30
Met Asn Asp Ala Ile Glu His Ala Leu Lys Ala Gln Glu Lys Val Arg
35 40 45
Val Met Thr Leu Asp Gln Arg Glu Lys Ile Ile Ser Ala Ile Arg Arg
50 55 60
Lys Thr Asn Glu Asn Val Glu Thr Ile Ala Arg Met Gly Val Glu Glu
65 70 75 80
Thr Gly Met Gly Asn Val Gly Asp Lys Ile Leu Lys His Lys Leu Thr
85 90 95
Ala Asp Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala Trp Ser
100 105 110
Gly Asp Arg Gly Leu Thr Leu Val Glu Met Gly Pro Phe Gly Val Ile
115 120 125
Gly Ala Ile Thr Pro Ala Thr Asn Pro Ser Glu Thr Val Ile Cys Asn
130 135 140
Ser Ile Gly Met Ile Ala Gly Gly Asn Thr Val Val Phe Asn Pro His
145 150 155 160
Pro Asn Ala Lys Lys Thr Thr Ile Tyr Thr Ile Asn Met Ile Asn Glu
165 170 175
Ala Ser Leu Glu Ala Gly Gly Pro Asp Asn Ile Ala Cys Thr Val Gln
180 185 190
Glu Pro Thr Met Glu Thr Ser Ala Ile Met Met Lys His Pro Lys Ile
195 200 205
Pro Leu Leu Val Ala Thr Gly Gly Pro Gly Val Val Thr Ala Val Leu
210 215 220
Ser Ser Gly Lys Arg Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Ala
225 230 235 240
Leu Val Asp Glu Thr Ala Asp Ile Glu Lys Ala Ala Arg Asp Ile Ile
245 250 255
Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu
260 265 270
Val Val Ala Val Asp Ala Ile Phe Asp Glu Leu Met Arg His Phe Glu
275 280 285
Glu Glu Asn Gly Cys Tyr Arg Ala Ser Arg Glu Ile Gln Asp Lys Leu
290 295 300
Ile Ala Thr Val Ile Thr Pro Lys Gly Ala Leu Asn Arg Lys Cys Val
305 310 315 320
Gly Arg Asp Ala Lys Thr Leu Leu Lys Met Val Gly Val Asp Ala Pro
325 330 335
Ala Asp Thr Arg Cys Ile Ile Phe Glu Gly Glu Lys Glu His Pro Leu
340 345 350
Ile Ala Thr Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Val Lys
355 360 365
Asp Phe Arg Glu Gly Val Glu Thr Ala Val Trp Leu Glu His Gly Asn
370 375 380
Arg His Ser Ala His Ile His Ser Lys Asn Val Asp Arg Ile Thr Glu
385 390 395 400
Tyr Ala Arg Ala Leu Asp Thr Ala Ile Leu Val Lys Asn Gly Pro Ser
405 410 415
Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Tyr Pro Thr Phe Thr Ile
420 425 430
Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr Lys
435 440 445
Arg Arg Arg Cys Val Met Thr Asp Ser Leu Cys Ile Arg
450 455 460
<210> 67
<211> 471
<212> PRT
<213> Eubacterium sp. 14-2
<400> 67
Met Asn Ile Asp Glu Arg Val Val Ala Ser Ile Val Asn Ala Val Leu
1 5 10 15
Gly Arg Leu Asp Asp Val Ser Ser Pro Ala Ala Glu Ala Gly Gly Gly
20 25 30
Asn Trp Gly Ile Phe Glu Ser Met Asn Asp Ala Val Glu Ala Ala Ala
35 40 45
Ala Ala Gln Lys Lys Tyr Ile Asn Cys Thr Met His Asp Arg Ala Ala
50 55 60
Tyr Val Gln Ala Ile Arg Asp Val Val Leu Lys Gln Glu Asn Leu Glu
65 70 75 80
Tyr Ile Ser Arg Gln Ser Ala Glu Glu Thr Gly Met Gly Asn Tyr Glu
85 90 95
His Lys Leu Ile Lys Asn Arg Leu Ala Ala Thr Lys Thr Pro Gly Thr
100 105 110
Glu Asp Leu Thr Thr Asp Ala Met Ser Gly Asp Asp Gly Leu Thr Leu
115 120 125
Val Glu Tyr Ser Pro Phe Gly Val Ile Gly Ala Ile Thr Pro Thr Thr
130 135 140
Asn Pro Thr Glu Thr Ile Ile Cys Asn Ser Ile Gly Met Leu Ala Ala
145 150 155 160
Gly Asn Ser Val Val Phe Ser Pro His Pro Arg Ala Lys Asn Val Ser
165 170 175
Leu His Leu Ile Arg Leu Ile Asn Arg Ala Leu Ala Glu Ala Gly Ala
180 185 190
Pro Ala Asn Leu Val Val Thr Val Ser Gln Pro Ser Ile Glu Asn Thr
195 200 205
Asn Ala Met Met Ser His Pro Met Val Arg Met Leu Val Ala Thr Gly
210 215 220
Gly Pro Gly Ile Val Lys Thr Val Leu Ser Ser Gly Lys Lys Ala Ile
225 230 235 240
Gly Ala Gly Ala Gly Asn Pro Pro Val Val Val Asp Glu Thr Ala Asn
245 250 255
Ile Glu Lys Ala Gly Lys Asp Ile Ile Asp Gly Cys Cys Phe Asp Asn
260 265 270
Asn Leu Pro Cys Ile Ala Glu Lys Glu Val Ile Val Val Asp Ser Ala
275 280 285
Ala Asp Tyr Leu Ile Phe Asn Met Lys Lys Asn Gly Ala Tyr Glu Val
290 295 300
Lys Asp Pro Glu Ile Ile Asp Arg Ile Val Lys Leu Val Val Gln Glu
305 310 315 320
Asn Gly Lys Ser Pro Val Thr Ser Phe Val Gly Lys Ser Ala Lys Tyr
325 330 335
Ile Leu Glu Gln Ala Gly Val His Val Asp Asp Asp Val Arg Val Ile
340 345 350
Ile Ala Gln Thr Gly Glu Asp His Pro Phe Val Gln Val Glu Leu Met
355 360 365
Met Pro Ile Leu Pro Ile Val Arg Val Pro Asp Val Asp Ala Gly Ile
370 375 380
Glu Met Ala Val Arg Val Glu His Gly Asn Arg His Thr Ala Met Met
385 390 395 400
His Ser Arg Asn Val Asp Lys Leu Thr Lys Met Ala Lys Leu Ile Gln
405 410 415
Thr Thr Ile Phe Val Lys Asn Gly Pro Ser Tyr Ala Gly Ile Gly Val
420 425 430
Gly Gly Glu Gly Tyr Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu
435 440 445
Gly Leu Thr Ser Ala Lys Ser Phe Ala Arg Arg Arg Arg Cys Val Leu
450 455 460
Val Gly Gly Met Asp Val Arg
465 470
<210> 68
<400> 68
000
<210> 69
<400> 69
000
<210> 70
<400> 70
000
<210> 71
<400> 71
000
<210> 72
<400> 72
000
<210> 73
<400> 73
000
<210> 74
<211> 469
<212> PRT
<213> Anaerosalibacter massiliensis
<400> 74
Met Glu Leu Asp Lys Met Asp Leu Glu Gln Ile Val Asn Leu Val Val
1 5 10 15
Glu Gln Leu Lys Gly Glu Asp Thr Ser Ser Tyr Cys Lys Glu Glu Ser
20 25 30
Lys Asn Gly Val Phe Asn Asn Met Asn Glu Ala Ile Glu Lys Ala Tyr
35 40 45
Ile Ala Gln Lys Asp Phe Phe Lys Asn Tyr Asn Leu Glu Asp Arg Arg
50 55 60
Arg Ile Ile Lys Thr Ile Arg Lys Glu Leu Met Glu Asp Val Glu Leu
65 70 75 80
Leu Ala Lys Leu Gly Val Glu Asp Thr Gly Met Gly Arg Tyr Glu Asp
85 90 95
Lys Leu Lys Lys Asn Lys Leu Val Ile Glu Lys Thr Pro Gly Val Glu
100 105 110
Asp Leu Asn Ser Glu Val Phe Thr Gly Asp Asn Gly Leu Thr Leu Val
115 120 125
Glu Leu Ser Pro Tyr Gly Val Ile Gly Ala Ile Ala Pro Ser Thr Asn
130 135 140
Pro Ser Glu Thr Val Ile Cys Asn Ser Ile Gly Met Ile Ala Ala Gly
145 150 155 160
Asn Ser Val Val Phe Ser Pro His Pro Gly Ala Lys Asn Ile Ser Met
165 170 175
Lys Thr Val Glu Leu Ile Asn Lys Ala Ile Glu Lys Ala Gly Gly Pro
180 185 190
Lys Asn Leu Val Val Thr Thr Ser Asn Pro Ser Ile Glu Asn Ala Glu
195 200 205
Ile Met Met Lys His Glu Lys Ile Lys Met Ile Val Ala Thr Gly Gly
210 215 220
Pro Gly Val Val Lys Ser Ala Leu Ser Gln Gly Lys Lys Ala Ile Gly
225 230 235 240
Ala Gly Ala Gly Asn Pro Pro Ala Val Ile Asp Glu Thr Ala Asp Ile
245 250 255
Glu Lys Ala Ala Arg Asp Ile Ile Ala Gly Cys Ser Phe Asp Asn Asn
260 265 270
Leu Pro Cys Ile Ala Glu Lys Glu Val Ile Val Val Asp Ser Val Ala
275 280 285
Asp Tyr Leu Ile Phe Ser Met Asn Lys Asn Asn Val Tyr His Leu Lys
290 295 300
Asp Glu Glu Lys Ile Asp Lys Leu Ala Ser Met Val Ile Asp Lys Asn
305 310 315 320
Gly Arg Ile Asn Arg Lys Phe Val Gly Lys Asp Ala Lys Val Ile Leu
325 330 335
Lys Ala Val Asp Ile Glu Cys Glu His Asp Val Arg Ala Ile Ile Val
340 345 350
Glu Thr Glu Lys Asp His Pro Phe Val Val Thr Glu Leu Met Met Pro
355 360 365
Ile Leu Pro Ile Val Arg Val Lys Asp Ile Asp Glu Ala Ile Lys Leu
370 375 380
Ala Val Glu Val Glu Gln Gly Asn Arg His Thr Ala Ile Met His Ser
385 390 395 400
Lys Asn Val Asp Asn Leu Ser Arg Phe Ala Arg Glu Ile Glu Thr Thr
405 410 415
Ile Phe Val Lys Asn Ala Pro Ser Phe Ala Gly Leu Gly Phe Gly Gly
420 425 430
Glu Gly Tyr Pro Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu
435 440 445
Thr Ser Ala Arg Ser Phe Ala Arg Lys Arg Arg Cys Ser Leu Val Gly
450 455 460
Ser Phe Ser Ile Lys
465
<210> 75
<211> 473
<212> PRT
<213> Clostridium indolis DSM 755
<400> 75
Met Glu Ile Gly Ala Lys Glu Ile Glu Leu Ile Val Arg Glu Val Leu
1 5 10 15
Ala Gly Ile Glu Ser Arg Gly Ile Lys Pro Ser Tyr Thr Pro Ser Arg
20 25 30
Ser Glu Asp Gly Val Phe Glu Arg Val Glu Asp Ala Ile Glu Ala Ala
35 40 45
Tyr Ala Ala Gln Arg Glu Trp Val Glu His Tyr Arg Val Glu Asp Arg
50 55 60
Arg Arg Ile Ile Glu Ala Ile Arg Val Thr Ala Lys Ser His Ala Glu
65 70 75 80
Ser Leu Ala Lys Met Val Trp Glu Glu Thr Gly Met Gly Arg Phe Glu
85 90 95
Asp Lys Ile Gln Lys His Met Ala Val Ile Glu Lys Thr Pro Gly Val
100 105 110
Glu Cys Leu Thr Thr Glu Ala Ile Ser Gly Asp Gly Gly Leu Met Ile
115 120 125
Glu Glu Tyr Ala Pro Phe Gly Val Ile Gly Ala Ile Thr Pro Ser Thr
130 135 140
Asn Pro Thr Glu Thr Ile Ile Asn Asn Thr Ile Ser Met Ile Ala Gly
145 150 155 160
Gly Asn Ser Val Val Phe Asn Val His Pro Gly Ala Lys Arg Cys Cys
165 170 175
Ala His Cys Leu Lys Ile Leu His Gln Ala Ile Val Glu Asn Gly Gly
180 185 190
Pro Ala Ser Leu Ile Thr Met Gln Lys Glu Pro Asp Met Glu Ala Val
195 200 205
Ser Lys Leu Thr Ser Asp Pro Arg Ile Arg Leu Met Val Gly Thr Gly
210 215 220
Gly Met Pro Met Val Asn Ala Leu Leu Arg Ser Gly Lys Lys Thr Ile
225 230 235 240
Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val Asp Asp Thr Ala Asp
245 250 255
Val Ser Leu Ala Ala Arg Glu Ile Tyr Arg Gly Ala Ser Phe Asp Asn
260 265 270
Asn Ile Leu Cys Leu Ala Glu Lys Glu Val Phe Val Met Glu Arg Ala
275 280 285
Ala Asp Glu Leu Val Asn Lys Leu Ile Lys Glu Gly Ala Tyr Leu Leu
290 295 300
Ser Ser Leu Glu Leu Ser Glu Ile Leu Lys Phe Ala Met Val Glu Lys
305 310 315 320
Asn Gly Ser Tyr Glu Val Asn Lys Lys Trp Val Gly Lys Asp Ala Gly
325 330 335
Gln Phe Leu Glu Ala Ile Gly Val Ser Gly His Lys Asp Val Arg Leu
340 345 350
Leu Ile Cys Glu Thr Asp Arg Ser His Pro Phe Val Met Val Glu Gln
355 360 365
Leu Met Pro Ile Leu Pro Ile Val Arg Leu Arg Thr Phe Glu Glu Cys
370 375 380
Val Glu Ser Ala Leu Ala Ala Glu Ser Gly Asn Arg His Thr Ala Ser
385 390 395 400
Met Phe Ser Arg Asn Val Glu Asn Met Thr Lys Phe Gly Lys Ile Ile
405 410 415
Glu Thr Thr Ile Phe Thr Lys Asn Gly Ser Thr Leu Lys Gly Val Gly
420 425 430
Ile Gly Gly Glu Gly His Thr Thr Met Thr Ile Ala Gly Pro Thr Gly
435 440 445
Glu Gly Leu Thr Cys Ala Arg Ser Phe Thr Arg Arg Arg Arg Cys Met
450 455 460
Leu Ala Glu Gly Gly Leu Arg Ile Ile
465 470
<210> 76
<400> 76
000
<210> 77
<211> 467
<212> PRT
<213> Catabacter hongkongensis
<400> 77
Met Gly Leu Ser Glu Gln Gln Ile Lys Gln Ile Val Glu Glu Thr Val
1 5 10 15
Arg Asn Ile Gly Thr Gly Thr Ala Gly Ala Ala Cys Ser Gly Ser Trp
20 25 30
Met Cys Asp Asp Ala Asn Asp Ala Val Glu Asn Ala Lys Arg Ala Gln
35 40 45
Lys Gln Leu Met Thr Met Thr Leu Glu Gln Arg Gly Arg Leu Val Ser
50 55 60
Ala Met Arg Glu Ala Ala Leu Ala Asn Ser Val Lys Leu Ala Glu Met
65 70 75 80
Ala His Glu Glu Thr Gly Tyr Gly Ser Val Glu His Lys Ile Met Lys
85 90 95
Asn Glu Leu Ala Ala Lys Lys Thr Pro Gly Ile Glu Asp Leu His Thr
100 105 110
Gln Ala Phe Ser Gly Asp Asp Gly Leu Thr Ile Val Glu Gln Ala Pro
115 120 125
Phe Gly Val Ile Gly Ser Ile Thr Pro Ser Thr Asn Pro Thr Ser Thr
130 135 140
Val Ile Asn Asn Ser Ile Ser Met Val Ala Ala Gly Asn Ala Val Val
145 150 155 160
Tyr Asn Pro His Pro Ala Ala Lys Arg Ala Ser Gln Glu Ala Met Arg
165 170 175
Ile Leu Asn Glu Ala Ile Val Ser Ala Gly Gly Pro Ala Thr Leu Ile
180 185 190
Thr Thr Val Lys Glu Pro Thr Leu Glu Ser Gly Gln Val Ile Met Asn
195 200 205
His Arg Asp Ile Lys Met Leu Ser Ile Thr Gly Gly Glu Ala Val Val
210 215 220
Ala Val Ala Met Lys Thr Gly Lys Lys Val Val Ala Ala Gly Pro Gly
225 230 235 240
Asn Pro Pro Val Ile Val Asp Asp Thr Ala Val Ile Pro Lys Ala Ala
245 250 255
Lys Asp Ile Val Asp Gly Ala Ser Phe Asp Asn Asn Val Leu Cys Val
260 265 270
Ala Glu Lys Glu Val Phe Ala Phe Asp Asn Ile Thr Asp Gln Leu Met
275 280 285
Ser Glu Met Glu Lys Asn Gly Ala Tyr Arg Val Ser Gly Glu Asp Ile
290 295 300
Asn Lys Ile Val Asn Thr Val Leu Val Leu Lys Asp Gly His Tyr Val
305 310 315 320
Ile Asn Arg Lys Phe Val Gly Arg Asp Ala Thr Tyr Ile Met Gln Glu
325 330 335
Ser Gly Val Ser Tyr Thr Gly Asn Pro Arg Leu Val Ile Ala Glu Val
340 345 350
Ser Ala Asn His Pro Phe Val Thr Val Glu Met Leu Met Pro Val Leu
355 360 365
Gly Val Val Arg Val Arg Asn Ile Asp Glu Ala Val Asp Glu Ala Phe
370 375 380
Arg Ala Glu Arg Gly Cys Gln His Ser Ala Leu Ile His Ser Thr Asn
385 390 395 400
Ile Arg Asn Met Ser Lys Ala Ala Ser Thr Met Asn Thr Thr Ile Phe
405 410 415
Val Lys Asn Ala Pro Ser Tyr Ser Gly Leu Gly Phe Gly Gly Glu Gly
420 425 430
Tyr Ala Thr Leu Thr Ile Ala Thr Pro Thr Gly Glu Gly Leu Thr Ser
435 440 445
Ala Lys Thr Phe Thr Arg Ala Arg Arg Cys Val Leu Lys Gly Asp Leu
450 455 460
Arg Ile Ile
465
<210> 78
<400> 78
000
<210> 79
<400> 79
000
<210> 80
<211> 463
<212> PRT
<213> Bacillus thermotolerans
<400> 80
Met Ala Val Gln Glu Arg Asp Leu Glu Ser Ile Val Lys Lys Val Leu
1 5 10 15
Glu Glu Leu Ser Arg Lys Glu Glu Thr Pro Glu Ala Gly Gln Gly Val
20 25 30
Phe Glu Asp Met Asn Asp Ala Ile Glu Ala Ala Glu Gln Ala Gln Lys
35 40 45
Glu Leu Ile Lys Leu Ser Leu Glu Glu Arg Gly Ala Ile Ile Glu Ala
50 55 60
Ile Arg Glu Ala Ser Arg Lys His Val Glu Thr Phe Ala Arg Met Ala
65 70 75 80
Val Glu Glu Thr Gly Met Gly Asn Tyr Glu Asp Lys Val Arg Lys Asn
85 90 95
Val Leu Val Ile Asp Lys Thr Pro Gly Ile Glu Asp Leu Lys Thr Glu
100 105 110
Ala Val Ser Gly Asp Asn Gly Leu Thr Val Val Glu Leu Ser Pro Tyr
115 120 125
Gly Val Ile Gly Ser Ile Thr Pro Thr Thr Asn Pro Thr Glu Thr Ile
130 135 140
Ile Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ser Val Val Phe
145 150 155 160
Ser Pro His Pro Gly Ala Lys Asp Thr Ser Leu Lys Ala Val Glu Ile
165 170 175
Ile Asn Gln Ala Ile Val Glu Ala Gly Gly Pro Lys Asn Leu Ile Thr
180 185 190
Ser Ile Ala Glu Pro Ser Ile Asp Gln Ala Asn Ile Met Met Arg His
195 200 205
Lys Lys Val Arg Met Leu Val Ala Thr Gly Gly Pro Gly Val Val Lys
210 215 220
Ala Val Leu Thr Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Val Val Val Asp Glu Thr Ala Asp Leu Glu Lys Ala Ala Lys
245 250 255
Asp Ile Val Asp Gly Cys Ser Phe Asp Asn Asn Ile Pro Cys Val Ala
260 265 270
Glu Lys Glu Leu Phe Val Val Glu Ala Val Ala Asp Tyr Leu Val Phe
275 280 285
His Met Lys Lys His Gly Ala Phe Gln Leu Asn Asp Pro Lys His Val
290 295 300
Glu Lys Leu Thr Glu Leu Val Val Asp Asn Gly His Ala Asn Lys Glu
305 310 315 320
Phe Val Gly Lys Asp Ile Gln Tyr Ile Leu Lys Gln Ile Gly Val Asp
325 330 335
Ala Pro Gln Asp Ala Arg Ile Ala Ile Met Asp Val Gly Ala Asp His
340 345 350
Pro Leu Val Ser Ala Glu Leu Met Met Pro Ile Leu Pro Val Val Arg
355 360 365
Thr Ala Asn Val Asp Glu Ala Ile Glu Leu Ala Val Glu Ala Glu His
370 375 380
Gly Phe Arg His Thr Ser Ile Met His Ser Lys Asn Ile Asp Asn Leu
385 390 395 400
Thr Lys Phe Ala Lys Ala Ile Gln Thr Thr Ile Phe Val Lys Asn Gly
405 410 415
Pro Ser Tyr Ala Gly Leu Gly Val Gly Gly Glu Gly Tyr Thr Ser Phe
420 425 430
Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys Asp Phe
435 440 445
Ala Arg Lys Arg Lys Cys Val Leu Val Asp Ser Leu Ser Val Arg
450 455 460
<210> 81
<400> 81
000
<210> 82
<211> 511
<212> PRT
<213> Gracilibacillus kekensis
<400> 82
Met Gln Leu Asn Glu Lys Asp Ile Gln Thr Ile Ile Asp Ser Val Leu
1 5 10 15
Lys Asn Val Glu Ala Ala Val Glu Asn Arg Gln Pro Thr Gln Ala Ser
20 25 30
Gly Gln Ser Ser Glu Gln Gln Pro Ile Lys Met Lys Gln Leu Ser Pro
35 40 45
Ser Ala Pro Ser Asn Thr Phe Asn Met Ser Ser Asn Lys Asp Gly Val
50 55 60
Phe Glu Arg Val Thr Asp Ala Ile Glu Ala Ala Ser Lys Ala Gln Glu
65 70 75 80
Val Trp Met Lys Gln Tyr Thr Leu Glu Glu Lys Glu Asn Leu Ile Asn
85 90 95
Ser Ile Arg Gln Ala Val Ala Gln Gln Val Asn His Phe Ala Lys Ser
100 105 110
Ala Leu Glu Glu Thr Gly Leu Gly Asn Tyr Glu Asp Lys Val Leu Lys
115 120 125
Leu Ser Leu Thr Val Glu Lys Thr Pro Gly Thr Glu Leu Leu Gln Thr
130 135 140
Glu Thr Phe Ser Gly Asp Asp Gly Leu Ser Phe Val Glu Gln Thr Pro
145 150 155 160
Phe Gly Val Ile Gly Ala Val Thr Pro Val Thr Asn Pro Ile Asp Thr
165 170 175
Ile Val Asn Asn Gly Ile Gly Met Ile Ala Ala Gly Asn Ala Val Val
180 185 190
Phe Asn Val His Pro Ser Ala Lys Lys Thr Ser Arg Glu Met Ile Gln
195 200 205
Leu Leu Asn Gln Thr Ile Val Asn Ala Gly Gly Pro Glu Asn Leu Leu
210 215 220
Thr Met Val Gln Glu Pro Thr Ile Glu Thr Val Gln Glu Ile Ala Asn
225 230 235 240
His Pro Ser Val Lys Leu Leu Val Gly Thr Gly Gly Pro Gly Met Val
245 250 255
Lys Ser Leu Leu Lys Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly
260 265 270
Asn Pro Pro Val Ile Val Asp Glu Thr Ala Asp Leu Lys Gln Ala Ala
275 280 285
Lys Asp Ile Ile Glu Gly Ala Ser Phe Asp Asn Asn Leu Leu Cys Ile
290 295 300
Ala Glu Lys Glu Val Phe Val Leu Asp Gln Val Ala Asp Asp Leu Ile
305 310 315 320
Phe Glu Leu Leu Asn Gln Gln Val His Met Leu Asp His Gln Gln Leu
325 330 335
Glu Lys Val Met Lys Leu Thr Leu Lys Glu Asn Thr Glu Gly Ile Pro
340 345 350
Gly Gly Cys Ser Tyr Leu Ser Arg Asp Tyr Leu Val Ser Lys Asp Trp
355 360 365
Val Gly Lys Asp Ala Thr Gln Ile Leu Glu Gln Ile Gly Val Ser Asn
370 375 380
Val Gln Thr Lys Leu Leu Ile Cys Glu Val Asp Ala Glu His Pro Tyr
385 390 395 400
Val Gln Leu Glu Gln Leu Met Pro Ile Leu Pro Ile Val Arg Val Lys
405 410 415
Ser Val Asp Glu Ala Ile Glu Lys Ala Val Lys Ala Glu His Gly Asn
420 425 430
Arg His Thr Ala Val Met His Ser Asn His Ile Lys Asn Val Thr Lys
435 440 445
Phe Ala Lys Ala Ile Gly Thr Thr Ile Phe Val Asn Asn Gly Ser Ser
450 455 460
Leu Ser Gly Val Gly Tyr Arg Gly Glu Gly Phe Thr Thr Met Thr Ile
465 470 475 480
Ala Gly Pro Thr Gly Glu Gly Val Thr Ser Ala Arg Thr Phe Thr Arg
485 490 495
Gln Arg Arg Thr Val Ile Ala Asn Gly Gly Phe Asn Ile Arg Gly
500 505 510
<210> 83
<400> 83
000
<210> 84
<211> 479
<212> PRT
<213> Propionispora sp. 2/2-37
<400> 84
Met Ile Gln Glu Gln Glu Leu Ile Ala Lys Ile Thr Ala Gln Val Ile
1 5 10 15
Ala Gln Met Gln Gln Gly Gln Ala Ala Ala Val Pro Glu His Tyr Gly
20 25 30
Val Phe Asp Ser Ile Asp Gly Ala Val Ala Ala Ala Arg Lys Ala Tyr
35 40 45
Gln Ser Leu Arg Ala Leu Pro Leu Glu Lys Arg Glu Gln Leu Val Gly
50 55 60
Ala Met Arg Lys Thr Ala Tyr Asp His Ala Glu Ile Met Ala Glu Met
65 70 75 80
Ala Val Thr Glu Ser Gly Met Gly Arg Tyr Ser Asp Lys Val Ile Lys
85 90 95
Asn Arg Thr Ala Ala Leu Lys Thr Pro Gly Thr Glu Asp Leu Lys Thr
100 105 110
Arg Ala Trp Ser Gly Asp Cys Gly Leu Thr Leu Val Glu Met Gly Pro
115 120 125
Tyr Gly Val Ile Gly Ala Ile Thr Pro Thr Thr Asn Pro Thr Glu Thr
130 135 140
Leu Ile Cys Asn Gly Ile Gly Met Ile Ala Ala Gly Asn Ala Val Phe
145 150 155 160
Phe Ser Pro His Pro Thr Ala Lys Asn Thr Ser Ile Trp Thr Ile Gln
165 170 175
Leu Leu Asn Lys Ala Leu Val Glu Ala Gly Gly Pro Pro Asn Leu Leu
180 185 190
Thr Thr Val Tyr Asn Pro Ser Ile Ala Val Ala Asn Ala Met Met Lys
195 200 205
His Pro Asp Val Asn Met Leu Val Ala Thr Gly Gly Pro Gly Val Val
210 215 220
Lys Ala Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly
225 230 235 240
Asn Pro Pro Ala Val Val Asp Glu Thr Ala Asp Leu Glu Lys Ala Ala
245 250 255
Lys Asp Ile Val Ala Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys Ile
260 265 270
Ala Glu Lys Glu Val Ile Ala Val Gly Ser Ile Ala Asp Arg Leu Met
275 280 285
Asp Tyr Met Val Arg Asn Gly Ala Tyr Lys Ile Thr Pro Gln Gln Thr
290 295 300
Ala Glu Leu Val Asn Leu Leu Leu Thr Val Lys Glu Glu Lys Met Ala
305 310 315 320
Glu Gly Cys Thr Ala Lys Thr Lys Arg Thr Tyr Gly Ile Asn Lys Asp
325 330 335
Tyr Val Gly Lys Ser Ala Gln Cys Ile Leu Ser Lys Ile Gly Val Thr
340 345 350
Val Lys Asp Asp Ile Arg Val Ile Leu Cys Glu Ala Glu Ala Asp His
355 360 365
Pro Phe Val Leu Glu Glu Leu Met Met Pro Val Leu Pro Val Val Gln
370 375 380
Val Lys Asp Val Asp Ala Ala Ile Glu Leu Ala Val Arg Val Glu His
385 390 395 400
Gly Asn Arg His Thr Ala Val Met His Ser Lys Asn Val Asp His Leu
405 410 415
Thr Arg Met Ala Arg Ala Ile Asp Thr Thr Ile Phe Val Lys Asn Ala
420 425 430
Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Tyr Cys Thr Phe
435 440 445
Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Pro Arg Ser Phe
450 455 460
Thr Arg Ala Arg Arg Cys Val Leu Val Asp Gly Phe Ser Ile Val
465 470 475
<210> 85
<400> 85
000
<210> 86
<211> 478
<212> PRT
<213> Clostridium chauvoei
<400> 86
Val Phe Ser Asp Glu Lys Ser Ile Glu Glu Ile Val Ile Lys Val Leu
1 5 10 15
Glu Glu Ile His Thr Asp Arg Lys Thr Lys Cys Asn Lys Asn Cys Asn
20 25 30
Ser Asn Cys Gly Cys Asn Lys Asp Lys Phe Ile Phe Ser Ser Val Asp
35 40 45
Asp Ala Val Ala Ala Ala Lys Lys Ser Phe Phe Glu Leu Lys Lys Leu
50 55 60
Thr Ile Arg Glu Arg Glu Glu Ile Ile Lys Asn Ile Arg Lys Lys Cys
65 70 75 80
Leu Asp Tyr Ala Asp Lys Leu Ser Ile Met Ala Val Glu Glu Thr Gly
85 90 95
Met Gly Lys Val Glu Asp Lys Val Thr Lys His Ile Leu Ile Ala Glu
100 105 110
Lys Thr Pro Gly Thr Glu Asp Leu Lys Thr Thr Ala Trp Ser Gly Asp
115 120 125
Gly Gly Leu Thr Leu Ile Glu Gln Gly Ala Phe Gly Val Ile Ala Ala
130 135 140
Ile Thr Pro Ser Thr Asn Pro Thr Ala Thr Val Leu Cys Asn Ala Ile
145 150 155 160
Gly Met Ile Ser Ala Gly Asn Thr Ile Val Phe Ala Pro His Pro Asn
165 170 175
Ala Val Lys Cys Ser Asn Leu Ala Val Lys Leu Ile Asn Glu Ala Ser
180 185 190
Lys Glu Ala Gly Gly Pro Glu Asn Ile Ala Val Ser Phe Arg Lys Pro
195 200 205
Ser Ile Asp Ile Thr Thr Glu Leu Met Lys His Lys Asp Ile Ala Leu
210 215 220
Ile Ser Ala Thr Gly Gly Pro Gly Val Val Asn Gln Ala Leu Ser Ser
225 230 235 240
Gly Lys Arg Ala Leu Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val
245 250 255
Asp Glu Thr Ala Asn Ile Glu Lys Ala Ala Lys Asp Ile Ile Asp Gly
260 265 270
Ala Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu Val Ile
275 280 285
Val Ile Asp Ser Val Ser Asn Lys Leu Ile Glu Tyr Met Ile Lys Phe
290 295 300
Gly Ala Tyr Leu Leu Lys Asp Lys Glu Gln Ile Lys Arg Leu Glu Asp
305 310 315 320
Lys Leu Leu Ile Lys Asn Gly Lys Lys Val Thr Leu Asn Arg Asp Phe
325 330 335
Val Gly Lys Asp Ala Lys Val Ile Leu Asp Ser Ile Asp Ile Leu Val
340 345 350
Asp Asp Ser Ile Lys Cys Ile Ile Phe Glu Gly Asp Lys Asp Ser Leu
355 360 365
Leu Ile Lys Glu Glu Leu Met Met Pro Ile Leu Gly Ile Val Lys Val
370 375 380
Asn Asn Phe Asp Glu Ala Val Glu Cys Ala Leu Glu Leu Glu His Gly
385 390 395 400
Asn Arg His Ser Ala His Met His Ser Lys Asn Ile Asp Asn Leu Thr
405 410 415
Thr Phe Ala Arg Val Ile Asp Thr Ala Ile Phe Val Lys Asn Ala Pro
420 425 430
Ser Tyr Ser Ala Leu Gly Val Asn Ala Glu Gly Phe Ala Thr Phe Thr
435 440 445
Ile Ala Ser Lys Thr Gly Glu Gly Leu Ser Ser Thr Lys Thr Phe Thr
450 455 460
Lys Asn Arg Arg Cys Val Leu Ser Asp Gly Leu Ser Ile Arg
465 470 475
<210> 87
<211> 467
<212> PRT
<213> 10500A Thermoanaerobacterium aotearoense
<400> 87
Met Lys Val Lys Glu Glu Asp Ile Glu Ala Ile Val Lys Lys Val Leu
1 5 10 15
Ser Glu Phe Asn Phe Glu Lys Asn Thr Lys Ser Phe Arg Asp Phe Gly
20 25 30
Val Phe Gln Asp Met Asn Asp Ala Ile Arg Ala Ala Lys Asp Ala Gln
35 40 45
Lys Lys Leu Arg Asn Met Ser Met Glu Ser Arg Glu Lys Ile Ile Gln
50 55 60
Asn Ile Arg Lys Lys Ile Met Glu Asn Lys Lys Ile Leu Ala Glu Met
65 70 75 80
Gly Val Ser Glu Thr Gly Met Gly Lys Val Glu His Lys Ile Ile Lys
85 90 95
His Glu Leu Val Ala Leu Lys Thr Pro Gly Thr Glu Asp Ile Val Thr
100 105 110
Thr Ala Trp Ser Gly Asp Lys Gly Leu Thr Leu Val Glu Met Gly Pro
115 120 125
Phe Gly Val Ile Gly Thr Ile Thr Pro Ser Thr Asn Pro Ser Glu Thr
130 135 140
Val Leu Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ser Val Val
145 150 155 160
Phe Asn Pro His Pro Gly Ala Val Asn Val Ser Asn Tyr Ala Val Lys
165 170 175
Leu Val Asn Glu Ala Val Met Glu Ala Gly Gly Pro Glu Asn Leu Val
180 185 190
Ala Ser Val Glu Lys Pro Thr Leu Glu Thr Gly Asn Ile Met Phe Lys
195 200 205
Ser Pro Asp Val Ser Leu Leu Val Ala Thr Gly Gly Pro Gly Val Val
210 215 220
Thr Ser Val Leu Ser Ser Gly Lys Arg Ala Ile Gly Ala Gly Ala Gly
225 230 235 240
Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Lys Lys Ala Ala
245 250 255
Lys Asp Ile Val Asp Gly Ala Thr Phe Asp Asn Asn Leu Pro Cys Ile
260 265 270
Ala Glu Lys Glu Val Val Ser Val Asp Lys Ile Thr Asp Glu Leu Ile
275 280 285
Tyr Tyr Met Gln Gln Asn Gly Cys Tyr Lys Ile Glu Gly Arg Glu Ile
290 295 300
Glu Lys Leu Ile Glu Leu Val Leu Asp His Lys Gly Gly Lys Ile Thr
305 310 315 320
Leu Asn Arg Lys Trp Val Gly Lys Asp Ala His Leu Ile Leu Lys Ala
325 330 335
Ile Gly Ile Asp Ala Asp Glu Ser Val Arg Cys Ile Ile Phe Glu Ala
340 345 350
Glu Lys Asp Asn Pro Leu Val Val Glu Glu Leu Met Met Pro Ile Leu
355 360 365
Gly Ile Val Arg Ala Lys Asn Val Asp Glu Ala Ile Met Ile Ala Thr
370 375 380
Glu Leu Glu His Gly Asn Arg His Ser Ala His Met His Ser Lys Asn
385 390 395 400
Val Asp Asn Leu Thr Lys Phe Gly Lys Ile Ile Asp Thr Ala Ile Phe
405 410 415
Val Lys Asn Ala Pro Ser Tyr Ala Ala Leu Gly Tyr Gly Gly Glu Gly
420 425 430
Tyr Cys Thr Phe Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser
435 440 445
Ala Arg Thr Phe Thr Lys Ser Arg Arg Cys Val Leu Ala Asp Gly Leu
450 455 460
Ser Ile Arg
465
<210> 88
<211> 462
<212> PRT
<213> Ruminococcus sp. AT10
<400> 88
Val Ser Val Asn Glu Gln Met Val Gln Asp Ile Val Gln Glu Val Leu
1 5 10 15
Ala Lys Met Gln Ile Ala Ser Asp Val Ser Gly Asn Arg Gly Val Phe
20 25 30
Ala Asp Met Asn Glu Ala Ile Ala Ala Ala Gln Lys Ala Gln Lys Val
35 40 45
Val Ala Arg Met Thr Leu Asp His Arg Glu Lys Val Ile Ser Asn Ile
50 55 60
Arg Lys Lys Ile Asn Glu Asn Ala Glu Ile Leu Ala Arg Met Gly Val
65 70 75 80
Glu Glu Thr Gly Met Gly Asn Val Gly His Lys Ile Leu Lys His Gln
85 90 95
Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Val Leu
130 135 140
Cys Asn Thr Ile Gly Met Phe Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Tyr Ala Val Asn Leu Leu
165 170 175
Asn Glu Ala Ser Val Glu Ala Gly Gly Pro Asp Asn Ile Ala Cys Thr
180 185 190
Val Glu His Pro Thr Leu Glu Thr Ser Asn Ile Met Met Lys His Lys
195 200 205
Ala Ile Gln Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Arg Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Glu Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Glu Ser Val Ala Asp Glu Leu Leu His Tyr
275 280 285
Met Ile Gln Glu Gln Gly Cys Tyr Leu Ala Ser Lys Glu Glu Gln Asp
290 295 300
Ala Leu Thr Ala Val Val Leu Lys Asp Gly Arg Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Lys Thr Leu Leu Gly Met Ile Gly Val Thr Val
325 330 335
Pro Asp Asn Ile Arg Cys Ile Thr Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Ala Thr Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Lys Asp Phe Asn Asp Ala Val Glu Gln Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Val Asp His Ile Thr
385 390 395 400
Lys Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Gly Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Phe Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Ser Asp Ser Leu Cys Ile Arg
450 455 460
<210> 89
<400> 89
000
<210> 90
<211> 469
<212> PRT
<213> Acetobacterium dehalogenans
<400> 90
Met Asn Ile Asp Thr Thr Gly Ile Glu Tyr Ile Val Lys Lys Val Met
1 5 10 15
Ala Glu Ile Asp Cys Ala Asp Ala Gly Gly Lys Pro Leu Lys Asp Gly
20 25 30
Glu Leu Gly Val Phe Asn Asp Met Glu Asn Ala Ile Asp Ala Ala Phe
35 40 45
Thr Ala Gln Lys Thr Phe Met Arg Glu Ser Leu Ala Tyr Arg Ser Lys
50 55 60
Leu Ile Ala Ala Met Arg Ala Glu Met Leu Lys Lys Glu Asn Met Glu
65 70 75 80
Met Ile Cys Gln Met Ala Val Glu Glu Thr Gly Met Gly Asn Tyr Glu
85 90 95
His Lys Leu Leu Lys His Glu Leu Ala Thr Val Lys Thr Pro Gly Val
100 105 110
Glu Asp Leu Val Ala Glu Ala Phe Thr Gly Asp Asp Gly Leu Thr Leu
115 120 125
Ile Glu Gln Ser Pro Phe Gly Val Ile Gly Ser Val Ser Pro Ser Thr
130 135 140
Asn Pro Ser Glu Thr Val Ile Cys Asn Ser Ile Gly Met Leu Ala Ala
145 150 155 160
Gly Asn Thr Val Val Phe Ala Pro His Pro Ser Ala Lys Asn Thr Ser
165 170 175
Ala Leu Thr Val Lys Leu Leu Asn Lys Ala Ile Leu Glu Ala Gly Gly
180 185 190
Pro Glu Asn Leu Ile Val Thr Thr Ala Glu Pro Thr Ile Asp Ser Ala
195 200 205
Asn Thr Met Phe Ala Ser Pro Lys Ile Thr Leu Leu Cys Ala Thr Gly
210 215 220
Gly Pro Gly Val Val Lys Thr Val Leu Gln Ser Gly Lys Lys Ala Ile
225 230 235 240
Gly Ala Gly Ala Gly Asn Pro Pro Ala Leu Val Asp Glu Thr Ala Asp
245 250 255
Ile Glu Lys Ala Gly Lys Asp Ile Ile Asp Gly Cys Cys Phe Asp Asn
260 265 270
Asn Leu Pro Cys Ile Ala Glu Lys Glu Val Val Val Val Glu Gln Val
275 280 285
Ala Asp Tyr Leu Ile Phe Asn Met Lys Lys Asn Gly Ala Tyr Glu Leu
290 295 300
Lys Asp Ala Lys Lys Ile Ala Glu Leu Glu Glu Leu Val Ile Pro Gly
305 310 315 320
Gly Arg Leu Ser Arg Asp Tyr Val Gly Arg Ser Ala Lys Val Ile Leu
325 330 335
Lys Gly Ile Gly Ile Asp Val Asp Asp Ser Ile Arg Val Ile Ile Met
340 345 350
Glu Thr Ser Lys Asp His Ile Phe Ala Val Glu Glu Leu Met Met Pro
355 360 365
Ile Leu Pro Ile Val Arg Val Lys Asn Ile Ala Glu Gly Ile Asp Leu
370 375 380
Ala Val Ala Leu Glu His Gly Asn Arg His Thr Ala Ile Met His Ser
385 390 395 400
Thr Asn Ile Asn Asn Leu Thr Glu Met Ala Lys Arg Val Gln Thr Thr
405 410 415
Ile Phe Val Lys Asn Gly Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly
420 425 430
Glu Gly Tyr Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu
435 440 445
Thr Ser Ala Lys Thr Phe Thr Arg Lys Arg Arg Cys Val Leu Val Gly
450 455 460
Gly Phe Thr Ile Lys
465
<210> 91
<211> 461
<212> PRT
<213> Spirochaeta alkalica
<400> 91
Ala Thr Leu Leu Glu Arg Ala Arg Ala Ala Gln Glu Lys Ile Ala Thr
1 5 10 15
Cys Thr Gln Arg Glu Ile Asp Asp Leu Cys Leu Ser Val Gly Trp Glu
20 25 30
Val Tyr Thr Asp Glu Asn Ile Ala Lys Leu Ala Glu Cys Ala Val Gln
35 40 45
Thr Thr Gly Met Gly Asn Val Pro Asp Lys Ile Thr Lys His Lys Val
50 55 60
Lys Val Leu Gly Val Leu Lys Asp Leu Arg Lys Ala Arg Thr Val Gly
65 70 75 80
Leu Ile Glu Arg Asp Glu Ala Arg Gly Leu Ser Lys Tyr Ala Lys Pro
85 90 95
Val Gly Val Val Gly Ala Leu Leu Pro Val Thr Asn Pro Thr Ala Thr
100 105 110
Pro Ala Ser Asn Gly Leu Ser Ile Leu Lys Gly Arg Asn Ala Val Ile
115 120 125
Phe Ala Pro His Pro Arg Gly Ala Ala Ala Ser Ala Leu Ala Val Glu
130 135 140
Phe Met Arg Arg Gly Leu Arg Arg Val Gly Ala Pro Glu Asp Leu Ile
145 150 155 160
Gln Ile Val Glu Asp Pro Ser Leu Gly Gln Thr Gly Glu Leu Met Lys
165 170 175
Gln Val Asp Leu Val Val Ala Thr Gly Gly Gly Ala Met Val Lys Ala
180 185 190
Ala Tyr Ser Ser Gly Thr Pro Ala Tyr Gly Val Gly Pro Gly Asn Ser
195 200 205
Val Gln Ile Ile Ala Glu Asp Ala Asp Leu Ala Asp Ala Ala Ala Lys
210 215 220
Ile Ala Leu Ser Lys Ala Phe Asp His Ala Thr Ser Cys Ser Ser Glu
225 230 235 240
Asn Ser Ile Ile Val Glu Asp Ser Val Tyr Glu Gly Met Ile Thr Glu
245 250 255
Leu Val Gln Asn Gln Gly Cys Tyr Leu Thr Thr Pro Arg Glu Arg Ser
260 265 270
Gln Leu Glu Ala Leu Leu Trp Arg Pro Gly Lys Thr Gly Gln Leu Ala
275 280 285
Leu Asn Pro Gly Ile Ile Ala Arg Ser Ala Ala Thr Ile Ala Ala Glu
290 295 300
Ala Gly Ile Thr Leu Pro Glu Gly Thr Arg Val Ile Leu Val Glu Gly
305 310 315 320
Gln His Pro Leu Glu Gln Asp Pro Phe Ser Gln Glu Lys Leu Cys Pro
325 330 335
Val Leu Thr Val Tyr Arg Tyr Thr Arg Trp Glu Glu Ala Val Asp Leu
340 345 350
Leu Val Arg Leu Thr Asp Gln Ala Gly Thr Gly His Ser Cys Gly Ile
355 360 365
His Thr Phe Arg Glu Asp Tyr Ile Arg His Leu Gly Glu Thr Met Arg
370 375 380
Thr Ser Arg Ile Met Val Arg Gln Ala Gln Ala Pro Ala Asn Gly Gly
385 390 395 400
Asn Phe Phe Asn Ala Met Pro Ser Thr Val Thr Leu Gly Cys Gly Thr
405 410 415
Trp Gly Gly Asn Ile Thr Thr Glu Asn Ile His Trp Lys His Phe Ile
420 425 430
Asn Val Thr Trp Val Ser Glu Pro Ile Pro Pro Asp Arg Pro Asp Asp
435 440 445
Glu Glu Ile Trp Gly Ser Phe Trp Ser Arg Tyr Ala Glu
450 455 460
<210> 92
<400> 92
000
<210> 93
<400> 93
000
<210> 94
<211> 494
<212> PRT
<213> Clostridium caminithermale DSM 15212
<400> 94
Met Gln Ile Asn Glu Leu Gln Ile Glu Lys Leu Val Ala Glu Val Leu
1 5 10 15
Ala Lys Thr Leu Gly Ala Glu Gly Asn Ser Ser Leu Val Asn Asn Asn
20 25 30
Ser Ile Gly Asn Ser Asn Glu Tyr Glu Tyr Asn Gln Ser Leu Glu Val
35 40 45
Gly Val Phe Glu Lys Met Glu Asp Ala Ile Asn Glu Ala His Arg Ala
50 55 60
Tyr Gln Gln Leu Lys Asn Tyr Ser Ile Lys Asp Arg Gln Arg Phe Ile
65 70 75 80
Asp Gly Ile Lys Glu Trp Thr Leu Arg Glu Lys Asn Ile Leu Ala Lys
85 90 95
Lys Val Val Glu Glu Thr Gly Leu Gly Asn Tyr Glu Asp Lys Ile Ile
100 105 110
Lys His Glu Leu Ala Ala Arg Thr Ala Gly Thr Glu Val Leu Ser Ser
115 120 125
Lys Val Gln Ser Gly Asp Thr Gly Leu Ala Leu Ile Glu Gln Ala Pro
130 135 140
Tyr Gly Val Val Gly Ala Thr Thr Pro Ser Thr Asn Pro Ser Glu Thr
145 150 155 160
Val Ile Ser Asn Thr Ile Ala Met Leu Ala Ala Gly Asn Thr Val Val
165 170 175
Phe Asn Val His Pro Ser Ser Lys His Val Cys Ala Tyr Thr Val Ala
180 185 190
Lys Ile Asn Glu Cys Ile Met Asp Leu Gly Gly Pro Ala Asn Ile Ile
195 200 205
Thr Met Val Lys Asp Pro Thr Met Glu Ser Leu Gln Val Met Ala Asn
210 215 220
Cys Pro Lys Ile Asn Leu Leu Val Gly Thr Gly Gly Pro Gly Leu Val
225 230 235 240
Arg Ala Leu Leu Lys Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly
245 250 255
Asn Pro Pro Val Val Val Asp Ser Thr Ala Asn Ile Lys Lys Ala Ala
260 265 270
Ala Asp Ile Ile Lys Gly His Ser Phe Asp Asn Asn Ile Val Cys Ile
275 280 285
Leu Glu Lys Glu Val Phe Val Val Asp Glu Val Ala Asn Glu Leu Ile
290 295 300
Glu Asn Met Lys Ser Glu Gly Ala Phe Tyr Leu Asp Ser Ser Tyr Ile
305 310 315 320
Ser Ala Leu Thr Asp Leu Ile Ile Glu Ala Thr Asp Lys Lys Phe Phe
325 330 335
Leu Gly Asn Ser Ser Lys Thr Thr Asn Leu His Thr Lys Lys Glu Trp
340 345 350
Val Gly Lys Asp Ala Tyr Lys Ile Leu Asp Ala Leu Gly Ile Arg Tyr
355 360 365
Ser Thr Arg Pro Lys Cys Ile Ile Cys Glu Val Pro Phe Glu His Pro
370 375 380
Phe Val Gln Leu Glu Leu Leu Met Pro Val Leu Pro Ile Val Arg Val
385 390 395 400
Glu Asn Phe Val Lys Gly Val Glu Tyr Ala Val Glu Ala Glu His Gly
405 410 415
Asn Arg His Thr Ala Ile Val His Ser Gln Asn Ile Asp Asn Ile Thr
420 425 430
Tyr Tyr Ala Lys Ala Ile Asp Thr Thr Ile Phe Val Lys Asn Ala Pro
435 440 445
Ser Val Ala Gly Ile Gly Val Asp Ser Glu Ser Val Val Ser Phe Ser
450 455 460
Ile Ala Gly Pro Thr Gly Glu Gly Ile Thr Thr Ala Lys Asp Phe Thr
465 470 475 480
Arg Ala Arg His Cys Val Leu Val Asp Gly Phe Arg Ile Ile
485 490
<210> 95
<211> 464
<212> PRT
<213> Caldanaerobius fijiensis
<400> 95
Val Val Lys Glu Glu Gln Ile Glu Ala Ile Val Arg Glu Val Leu Arg
1 5 10 15
Arg Ile Asp Arg Glu Asp Ile Lys Leu Asn Glu Asp Lys His Gln Leu
20 25 30
Gly Val Phe Asp Lys Met Glu Asp Ala Ile Glu Ala Ala Lys Asp Ala
35 40 45
Phe Glu Lys Phe Ser Asn Met Thr Leu Glu Asp Arg Glu Arg Phe Ile
50 55 60
Ser Glu Ile Arg Lys Ala Thr Leu Glu Asn Ala Arg Val Leu Ala Glu
65 70 75 80
Met Gly Val Lys Glu Thr Gly Met Gly Lys Val Glu His Lys Val Leu
85 90 95
Lys His Gln Leu Val Ala Lys Lys Thr Pro Gly Thr Glu Asp Leu Lys
100 105 110
Thr Gln Ala Trp Ser Gly Asp Lys Gly Leu Thr Leu Val Glu Met Ala
115 120 125
Pro Phe Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Ser Glu
130 135 140
Thr Ile Ile Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ala Val
145 150 155 160
Val Phe Ser Pro His Pro Gly Ala Lys Arg Val Ser Asn Phe Ala Val
165 170 175
Asp Met Ile Asn Arg Ala Ile Ile Arg Ala Gly Gly Pro Glu Asn Leu
180 185 190
Val Val Ser Ile Lys Glu Pro Ser Ile Asn Thr Thr Asn Ala Met Ile
195 200 205
Lys His Pro Asp Val Lys Leu Leu Val Ala Thr Gly Gly Pro Glu Ile
210 215 220
Val Lys Ile Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Lys Lys Ala
245 250 255
Ala Lys Asp Ile Ile Asp Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys
260 265 270
Ile Ala Glu Lys Glu Val Ile Ala Val Glu Lys Ile Tyr Arg Asp Leu
275 280 285
Leu Asp Glu Ile Leu Lys Gln Gly Val Tyr Lys Leu Asn Ala Leu Gln
290 295 300
Ile Ser Lys Leu Glu Asn Leu Val Leu Met Asp Gly Lys Leu Asn Lys
305 310 315 320
Lys Leu Val Gly Lys Asp Ala Lys Val Ile Leu Asp Gln Ile Gly Ile
325 330 335
Asn Val Ser Asp Asp Ile Arg Cys Ile Ile Cys Glu Thr Asp Glu Asp
340 345 350
His Pro Phe Val Met Glu Glu Leu Met Met Pro Ile Leu Pro Ile Val
355 360 365
Lys Ala Lys Asn Ile Asp Asp Ala Ile Arg Ile Ala Val Lys Ala Glu
370 375 380
Lys Asn Asn Arg His Thr Ala His Ile His Ser Lys Asn Ile Asp Asn
385 390 395 400
Ile Thr Arg Tyr Ala Lys Ala Ile Asn Thr Thr Ile Leu Val Lys Asn
405 410 415
Ala Pro Ser Tyr Ala Gly Ile Gly Phe Gly Gly Glu Gly Phe Thr Thr
420 425 430
Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Gln Thr
435 440 445
Phe Thr Arg Met Arg Arg Cys Val Leu Ala Asp Gly Leu Arg Ile Ile
450 455 460
<210> 96
<400> 96
000
<210> 97
<211> 480
<212> PRT
<213> Pelosinus fermentans
<400> 97
Met Ser Ile Asp Gln Ala Leu Ile Glu Lys Ile Thr Leu Glu Ile Leu
1 5 10 15
Thr Lys Met Gln Thr Gly Ala Lys Ala Ala Pro Ala Gly Tyr Gly Asp
20 25 30
Gly Ile Phe Glu Thr Val Asp Glu Ala Val Ala Ala Ala Arg Lys Ala
35 40 45
Tyr Gln Glu Leu Lys Thr Leu Ser Leu Glu Lys Arg Glu Val Leu Ile
50 55 60
Lys Ala Met Arg Asp Val Ala Tyr Glu Asn Ala Thr Ile Leu Ala Gln
65 70 75 80
Met Ala Val Asp Glu Ser Gly Met Gly Arg Val Ser Asp Lys Ile Ile
85 90 95
Lys Asn Gln Val Ala Ala Leu Lys Thr Pro Gly Thr Glu Asp Leu Thr
100 105 110
Thr Gln Ala Trp Ser Gly Asp Asn Gly Leu Thr Leu Ile Glu Met Gly
115 120 125
Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Thr Thr Asn Pro Thr Glu
130 135 140
Thr Val Ile Cys Asn Gly Ile Gly Met Ile Ala Ala Gly Asn Thr Val
145 150 155 160
Phe Phe Ser Pro His Pro Thr Ala Lys Asn Thr Ser Met Lys Ile Ile
165 170 175
Thr Leu Leu Asn Gln Ala Ile Val Lys Ala Gly Gly Pro Asn Asn Leu
180 185 190
Leu Thr Ser Val Ala Asn Pro Ser Ile Lys Ala Ala Asn Glu Met Met
195 200 205
Lys His Pro Gly Ile Asn Met Leu Val Ala Thr Gly Gly Pro Gly Val
210 215 220
Val Lys Ala Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Ile Val Asp Glu Thr Ala Asp Ile Glu Lys Ala
245 250 255
Ala Arg Asp Ile Val Ala Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys
260 265 270
Ile Ala Glu Lys Glu Val Ile Ala Ile Gly Ser Ile Ala Asp Arg Leu
275 280 285
Ile Thr Tyr Met Gln Lys Tyr Gly Ala Tyr Leu Ile Ser Gly Ser Asn
290 295 300
Ile Asp Arg Leu Leu Asn Val Ile Met Thr Val Gln Glu Glu Lys Ile
305 310 315 320
Ala Glu Gly Cys Thr Asp Lys Pro Lys Arg Ser Tyr Gly Ile Asn Lys
325 330 335
Asp Tyr Val Gly Lys Asp Ala Lys Tyr Leu Leu Ser Lys Ile Gly Ile
340 345 350
Asp Val Pro Asp Ser Val Arg Val Val Leu Cys Glu Thr Pro Ala Asp
355 360 365
His Pro Phe Val Ile Glu Glu Leu Met Met Pro Val Leu Pro Val Val
370 375 380
Gln Val Lys Asp Ile Asp Glu Ala Ile Glu Val Ala Val Arg Val Glu
385 390 395 400
His Gly Asn Arg His Thr Ala Ala Met His Ser Lys Asn Val Asp His
405 410 415
Leu Thr Arg Phe Ala Arg Ala Val Glu Thr Thr Ile Phe Val Lys Asn
420 425 430
Ala Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Phe Thr Ser
435 440 445
Phe Thr Leu Ala Gly Pro Thr Gly Glu Gly Ile Thr Ser Pro Arg Ser
450 455 460
Phe Thr Arg Gln Arg Arg Cys Val Leu Val Asp Ala Phe Ser Ile Val
465 470 475 480
<210> 98
<400> 98
000
<210> 99
<400> 99
000
<210> 100
<211> 463
<212> PRT
<213> Blautia wexlerae
<400> 100
Met Pro Val Ser Glu Ser Met Val Gln Glu Ile Val Gln Gln Val Met
1 5 10 15
Ala Lys Met Gln Ile Ala Asp Ala Pro Ala Glu Lys Gln His Gly Val
20 25 30
Phe Lys Asp Met Asn Asp Ala Ile Glu Ala Ala Lys Lys Ser Gln Glu
35 40 45
Ile Val His Lys Met Ser Met Asp Gln Arg Glu Lys Ile Ile Ser Cys
50 55 60
Ile Arg Lys Lys Ile Lys Glu Asn Ala Glu Ile Met Ala Arg Met Gly
65 70 75 80
Val Glu Glu Thr Lys Met Gly Asn Val Gly Asp Lys Ile Leu Lys His
85 90 95
His Leu Val Ala Asp Lys Thr Pro Gly Thr Glu Ala Ile Thr Thr Thr
100 105 110
Ala Trp Ser Gly Asp Arg Gly Leu Thr Leu Val Glu Met Gly Pro Phe
115 120 125
Gly Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Val
130 135 140
Leu Cys Asn Thr Met Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe
145 150 155 160
Asn Pro His Pro Ala Ala Val Lys Thr Ser Leu Tyr Ala Val Asn Leu
165 170 175
Val Asn Glu Ala Ser Leu Glu Gln Gly Gly Pro Asp Asn Ile Ala Val
180 185 190
Ser Val Glu Asn Pro Thr Leu Asp Thr Ser Ser Val Met Met Lys His
195 200 205
Lys Asp Ile His Leu Leu Val Ala Thr Gly Gly Pro Gly Val Val Thr
210 215 220
Ala Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Arg
245 250 255
Asp Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala
260 265 270
Glu Lys Glu Val Val Ala Val Ser Ser Ile Met Asp Glu Leu Met His
275 280 285
Tyr Met Leu Thr Glu Asn Asp Cys Tyr Leu Ala Ser Lys Glu Glu Gln
290 295 300
Asp Lys Leu Val Glu Val Val Leu Ala Gly Gly Lys Leu Asn Arg Lys
305 310 315 320
Cys Val Gly Arg Asp Ala Arg Thr Leu Leu Ser Met Ile Gly Val Asp
325 330 335
Ala Pro Ala Asn Ile Arg Cys Ile Ile Phe Glu Gly Pro Lys Glu His
340 345 350
Pro Leu Ile Thr Thr Glu Leu Met Met Pro Ile Leu Gly Ile Val Arg
355 360 365
Ala Arg Asp Phe Glu Asp Ala Val Glu Gln Ala Val Trp Leu Glu His
370 375 380
Gly Asn Arg His Ser Ala His Ile His Ser Lys Asn Val Asp Arg Ile
385 390 395 400
Thr Thr Tyr Ala Lys Ala Ile Asp Thr Ala Ile Val Val Lys Asn Gly
405 410 415
Pro Ser Tyr Ala Ser Leu Gly Phe Gly Ser Glu Gly Tyr Thr Thr Phe
420 425 430
Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Cys Ala Ser Thr Phe
435 440 445
Thr Lys Arg Arg Arg Cys Ile Met Glu Asp Ser Leu Cys Ile Arg
450 455 460
<210> 101
<211> 500
<212> PRT
<213> Paenibacillus sp. OSY-SE
<400> 101
Ile Lys Leu Thr Glu Thr Asp Ile Gln Asn Ile Ile Gln Gly Val Leu
1 5 10 15
Lys Asn Ile Glu Gln Asn Leu Pro Gly Ala Gln Ala Ala Asp Asp Ala
20 25 30
Ala Thr Gly Gln Ala Lys Pro Glu Ser Ala Pro Val Ala Ala Ala Pro
35 40 45
Val Arg Ser Asn Gly Asp Tyr Gly Val Phe Asp Glu Ala Glu Ala Ala
50 55 60
Ile Ala Ala Ala Tyr Gln Ala Gln Arg Ala Tyr Ala His His Phe Ser
65 70 75 80
Met Gln Asp Arg Glu Arg Phe Ile Ala Ala Ile Arg Lys Ala Thr Leu
85 90 95
Glu His Lys Glu Thr Leu Ala Ser Met Val Leu Lys Glu Thr Lys Leu
100 105 110
Gly Arg Tyr Glu Asp Lys Ile Ala Lys Leu Glu Leu Thr Ala Leu Lys
115 120 125
Thr Pro Gly Thr Glu Asp Leu Glu Thr Lys Ala Phe Ser Gly Asp Asn
130 135 140
Gly Leu Thr Leu Val Lys Asp Gly Pro Phe Gly Val Ile Gly Ala Val
145 150 155 160
Thr Pro Val Thr Asn Ser Val Glu Thr Val Ile Asn Asn Ala Ile Gly
165 170 175
Met Leu Ala Ala Gly Asn Ala Val Val Tyr Asn Val His Pro Ser Ser
180 185 190
Lys Ala Cys Cys Ala Tyr Ala Val Lys Met Ile Asn Arg Ala Val Gln
195 200 205
Glu Ala Gly Gly Pro Glu His Leu Val Thr Met Val Lys Glu Pro Thr
210 215 220
Lys Glu Thr Leu Asp Ala Ile Thr Gln Ser Pro Lys Val Gln Leu Leu
225 230 235 240
Val Gly Thr Gly Gly Pro Gly Leu Val Arg Ala Leu Leu Arg Ser Gly
245 250 255
Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val Asp
260 265 270
Glu Thr Ala Asn Ile Glu Arg Ala Ala Lys Glu Ile Ile Ala Gly Ala
275 280 285
Ser Phe Glu Asn Asn Ile Leu Cys Ile Ala Glu Lys Glu Val Phe Val
290 295 300
Val Asp Lys Val Ala Asp Asp Leu Leu Phe His Met Leu Asn His Gly
305 310 315 320
Ala Tyr Arg Leu Asp Asp Arg Glu Leu Glu Gln Val Met Ser Phe Ala
325 330 335
Leu Glu Ala Asn Val Asn Glu Thr Ala Gly Gly Cys Ser Leu Asp Met
340 345 350
Lys Arg Glu Tyr His Thr Val Lys Glu Trp Ile Gly Lys Asp Ala Ala
355 360 365
Leu Phe Leu Glu Lys Ile Gly Val Thr Pro Glu Lys Glu Val Lys Leu
370 375 380
Leu Ile Cys Glu Val Asp Phe Asp His Pro Phe Val Gln Leu Glu Gln
385 390 395 400
Met Met Pro Val Leu Pro Ile Val Arg Val Ser Asp Leu Asp Glu Ala
405 410 415
Ile Arg Leu Ala Val Glu Ala Glu His Gly Asn Arg His Thr Ala Leu
420 425 430
Met His Ser Thr Asn Val Ala Asn Phe Ala Ala Phe Glu Arg Ala Ile
435 440 445
Gly Thr Thr Ile Phe Val Lys Asn Ala Ser Ser Leu Ala Gly Val Gly
450 455 460
Ala Gly Gly Glu Gly Cys Thr Thr Met Thr Ile Ala Gly Pro Thr Gly
465 470 475 480
Glu Gly Leu Thr Ser Ala Arg Thr Phe Thr Arg Lys Lys Arg Cys Val
485 490 495
Leu Ala Glu Arg
500
<210> 102
<400> 102
000
<210> 103
<211> 476
<212> PRT
<213> Spirochaetes bacterium GWC2_52_13
<400> 103
Val Ser Gln Ser Ile Glu Asp Thr Val Arg Thr Leu Val Glu Lys Leu
1 5 10 15
Val Leu Glu Tyr Ser Ala Ser Ser Val Gly Val Asp His Ile Ala Pro
20 25 30
Ser Gln Tyr Ala Ser Gly Ile Phe Pro Thr Met Asp Leu Ala Val Lys
35 40 45
Ala Ala Tyr Glu Ala Gln Arg His Leu Val Gly Leu Pro Leu Glu Lys
50 55 60
Arg Lys Glu Ile Val Gln Ala Met Arg Glu Thr Ala Met Asp His Ala
65 70 75 80
Gln Glu Phe Ala Glu Met Ala Val Gln Glu Ser Gly Arg Gly Asn Val
85 90 95
Ala Asp Lys Ile Ala Lys Asn Ile Leu Ala Ala Lys Lys Thr Pro Gly
100 105 110
Val Glu Asp Val Glu Thr Ser Ala Tyr Ser Asp Glu His Gly Leu Ser
115 120 125
Leu Val Glu Arg Ala Pro Tyr Gly Val Ile Gly Ser Ile Thr Pro Val
130 135 140
Thr Asn Pro Thr Ala Thr Ile Ile Asn Asn Gly Ile Ser Met Ile Ser
145 150 155 160
Gly Gly Asn Ser Val Val Phe Asn Pro His Pro Gly Ala Lys Asn Val
165 170 175
Ser Cys Phe Ala Ile Glu Val Leu Asn Ala Ala Ile Glu Arg Val Gly
180 185 190
Gly Pro Arg Asn Leu Leu Val Ser Leu Ala Gln Pro Thr Ile Glu Ser
195 200 205
Ala Asn Glu Met Met Gly His Gln Lys Ile Ser Leu Leu Val Val Thr
210 215 220
Gly Gly Pro Gly Val Val Lys Ala Ala Met Asn Ser Gly Lys Lys Val
225 230 235 240
Ile Ala Ala Gly Pro Gly Asn Pro Pro Cys Val Val Asp Glu Thr Ala
245 250 255
Lys Ile Gln Lys Ala Ala Lys Asp Ile Val Asp Gly Ala Ser Phe Asp
260 265 270
Asn Asn Leu Val Cys Ile Cys Glu Lys Glu Val Leu Val Val Lys Ser
275 280 285
Val Ala Asn Glu Leu Ile Gly Glu Met Gln Lys Val Gly Ala Tyr Leu
290 295 300
Leu Ser Asp Gln Gln Ala Lys Ser Leu Leu Asp Gln Ile Ile Glu Val
305 310 315 320
Pro Gly Met Met Asn Ser Glu Gly Val Val Lys Arg Glu Tyr Val Gly
325 330 335
Lys Ser Pro Ser Phe Leu Ala Ser Leu Ile Gly Val Thr Val Pro Glu
340 345 350
Ser Thr Arg Leu Leu Ile Cys Asp Val Asp Ala Gly Asn Pro Leu Val
355 360 365
Trp Thr Glu Gln Leu Met Pro Phe Leu Pro Ile Val Arg Met Glu Asn
370 375 380
Val Asp Gln Cys Ile Asp Leu Ala Val Gln Cys Glu His Gly Phe Arg
385 390 395 400
His Thr Ala Ile Met His Ser Leu Asn Val Glu Lys Leu Ser Lys Met
405 410 415
Ala Arg Gln Met Asn Cys Ser Leu Phe Val Lys Asn Gly Pro Cys Tyr
420 425 430
Ala Gly Leu Gly Asn Gly Gly Ala Gly Tyr Thr Ser Phe Thr Ile Ala
435 440 445
Ser Pro Thr Gly Glu Gly Leu Thr Arg Ala Arg Thr Phe Thr Arg Glu
450 455 460
Arg Arg Cys Thr Leu Val Asp Tyr Phe Arg Ile Ile
465 470 475
<210> 104
<400> 104
000
<210> 105
<400> 105
000
<210> 106
<400> 106
000
<210> 107
<211> 454
<212> PRT
<213> Romboutsia lituseburensis DSM
<400> 107
Met Glu Ala Arg Asp Tyr Val Leu Gln Leu Ile Asn Lys Ala Arg Ile
1 5 10 15
Ala Gln Lys Glu Phe Glu Lys Tyr Ser Gln Glu Gln Val Asp Glu Ala
20 25 30
Val Arg Ala Ile Gly Lys Ser Ile Tyr Asp Asn Gly Glu Met Leu Ala
35 40 45
Arg Met Ala Val Asp Glu Thr Lys Met Gly Val Tyr Glu Asp Lys Ile
50 55 60
Val Lys Asn Lys Gly Lys Ser Lys Ala Val Trp Asn Lys Leu Lys Gly
65 70 75 80
Val Lys Ser Arg Gly Ile Ile Lys Tyr Ile Ala Glu Glu Gly Leu Val
85 90 95
Glu Val Ala Lys Pro Ile Gly Val Val Gly Ala Val Thr Pro Thr Thr
100 105 110
Asn Pro Thr Met Thr Pro Met His Asn Ala Met Ile Ala Leu Lys Gly
115 120 125
Gly Asn Ala Ile Ile Ile Cys Pro His Pro Arg Ala Lys Asn Thr Gly
130 135 140
Val Lys Thr Val Asp Leu Met Arg Glu Ala Leu Asp Lys Val Gly Ala
145 150 155 160
Pro Lys Asp Leu Ile Gln Ile Val Asn Glu Pro Thr Val Glu Ile Ser
165 170 175
Asn Leu Val Met Gln Leu Ser Asp Val Cys Val Ser Thr Gly Gly Pro
180 185 190
Gly Met Val Lys Val Ala Tyr Ser Ser Gly Lys Pro Ala Phe Gly Val
195 200 205
Gly Ala Gly Asn Val Gln Cys Leu Ile Asp Lys Asp Ala Asn Leu Glu
210 215 220
Glu Val Val Pro Lys Val Ile Lys Gly Arg Ile Tyr Asp Asn Gly Ile
225 230 235 240
Leu Cys Thr Cys Glu Gln Ser Ala Ile Cys Pro Asp Glu Met Tyr Asn
245 250 255
Glu Phe Ile Asp Arg Leu Val Gln Ser Gly Ala Tyr Tyr Ile Glu Lys
260 265 270
Glu Glu Glu Val Lys Ser Leu Arg Lys Ala Leu Phe Pro Asp Gly Asn
275 280 285
Ile Ser Lys Asp Cys Val Gly Ala Ser Pro Tyr Glu Ile Ala Lys Met
290 295 300
Ala Ser Ile Ala Ile Pro Lys Asp Thr Lys Leu Leu Val Val Lys Val
305 310 315 320
Glu Lys Tyr Gly Thr Glu Glu Tyr Phe Ala Lys Glu Lys Met Cys Pro
325 330 335
Val Leu Ser Ala Tyr Lys Tyr Glu Lys Trp Glu Asp Ala Val Asn Ile
340 345 350
Ala Asn Gln Asn Leu Glu Tyr Glu Gly Lys Gly His Ser Ala Ile Ile
355 360 365
His Ser Tyr Thr Lys Glu Asn Ile Glu Tyr Ala Ala Asn Ile Leu Pro
370 375 380
Val Ser Arg Phe Gly Val Asn Gln Ile Gly Ser Ser Gly Leu Gly Gly
385 390 395 400
Ser Phe Leu Asn Gly Leu Asn Pro Thr Ala Thr Leu Gly Cys Gly Ser
405 410 415
Trp Gly Asn Asn Ser Ile Ser Glu Asn Leu Trp Phe Asn His Leu Ile
420 425 430
Asn Val Ser Lys Ile Ala Tyr Glu Val Pro Ser Lys Lys Ile Pro Thr
435 440 445
Asp Asp Glu Ile Trp Asn
450
<210> 108
<400> 108
000
<210> 109
<211> 485
<212> PRT
<213> Clostridium sp. CAG:448
<400> 109
Met Ala Ile Asn Trp Thr Glu Ala Gln Ile Ala Asp Ile Val Ser Lys
1 5 10 15
Val Ile Ala Gly Met Gly Glu Gln Thr Leu Val Asn Asp Lys Glu Trp
20 25 30
Asp Ala Thr Gln Tyr His Gly Arg Lys Leu Ile Gly Ile Phe Glu Thr
35 40 45
Met Glu Glu Ala Ile Asp Ala Ala Ser Ala Gly Tyr Ala Ala Ile Arg
50 55 60
Ala Met Ser Val Ala Gln Arg Glu Thr Leu Ile Ser Ser Ile Arg Thr
65 70 75 80
Tyr Cys Arg Asn Glu Ala Arg Ile Met Ala Glu Leu Gly Val Ala Glu
85 90 95
Thr His Met Gly Arg Val Asp His Lys Thr Ala Lys His Ile Leu Val
100 105 110
Ala Asp Lys Thr Pro Gly Thr Glu Asp Ile Val Ala Glu Ala Lys Thr
115 120 125
Gly Asp Cys Gly Leu Thr Leu Thr Glu Arg Ala Pro Phe Gly Val Val
130 135 140
Gly Ala Ile Thr Pro Ser Thr Asn Pro Ser Glu Thr Val Ile Cys Asn
145 150 155 160
Ser Met Gly Met Ile Ala Ala Gly Asn Gly Val Val Phe Asn Pro His
165 170 175
Pro Gly Ala Ile Ala Thr Ser Asn Tyr Ala Val Asp Leu Val Asn Arg
180 185 190
Ala Val Phe Ala Ala Gly Gly Pro Lys Val Leu Val Ala Ser Val Arg
195 200 205
Lys Pro Thr Met Asp Thr Ala Gln Val Met Tyr Lys His Pro Ala Ile
210 215 220
Arg Leu Leu Val Cys Thr Gly Gly Pro Gly Val Val Lys Ala Val Leu
225 230 235 240
Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val
245 250 255
Ile Val Asp Asp Thr Ala Asp Ile Glu Lys Ala Ala Lys Asp Ile Ile
260 265 270
Asp Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu
275 280 285
Val Phe Val Phe Asp Asn Val Ala Asp Arg Leu Ile Ala Gly Met Leu
290 295 300
Arg Asn Gly Cys Ile Lys Leu Thr Arg Glu Gln Ala Asp Glu Leu Ala
305 310 315 320
Lys Val Val Val Val Glu Lys Thr Asp Ser Lys Thr Gly Lys Val Thr
325 330 335
Arg Ser Val Asn Arg Asp Cys Val Gly Arg Asp Cys Arg Val Ile Leu
340 345 350
Lys Lys Ile Gly Ile Glu Val Gly Pro Glu Ile Arg Cys Ala Ile Ala
355 360 365
Glu Val Pro Phe Glu His Thr Phe Val Gln Thr Glu Leu Met Met Pro
370 375 380
Ile Leu Gly Ile Val Arg Val Lys Asp Ile Asp Glu Ala Ile Asp Leu
385 390 395 400
Ala Val Lys Ala Glu His Gly Asn Arg His Thr Ala His Met His Ser
405 410 415
Lys Asn Ile Asp Asn Leu Ser Arg Phe Ala Lys Ala Ile Glu Thr Thr
420 425 430
Ile Phe Val Lys Asn Ala Pro Ser Tyr Ala Gly Ile Gly Phe Gly Gly
435 440 445
Glu Gly His Thr Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Ile
450 455 460
Thr Ser Ala Lys Ser Tyr Thr Arg Leu Arg Arg Cys Val Met Ala Asp
465 470 475 480
His Phe Arg Ile Ile
485
<210> 110
<400> 110
000
<210> 111
<211> 462
<212> PRT
<213> Yersinia bercovieri ATCC 43970
<400> 111
Met Asn Thr Asn Asp Leu Glu Ser Leu Ile Arg Thr Ile Leu Thr Glu
1 5 10 15
Gln Leu Thr Pro Ala Thr Ala Ser Ala Ser Asn Ala Ile Phe Ala Ser
20 25 30
Val Asp Glu Ala Val Asn Ala Ala His Ser Ala Phe Leu Arg Tyr Gln
35 40 45
Gln Ser Pro Met Lys Thr Arg Ser Ala Ile Ile Ser Ala Leu Arg Gln
50 55 60
Gln Leu Lys Pro Gln Leu Ala Ser Leu Ser Glu Arg Gly Ala Ser Glu
65 70 75 80
Thr Gly Met Gly Asn Lys Glu Asp Lys Phe Leu Lys Asn Lys Ala Ala
85 90 95
Leu Glu Asn Thr Pro Gly Ile Glu Asp Leu Ser Thr Thr Ala Leu Thr
100 105 110
Gly Asp Gly Gly Met Val Leu Phe Glu Tyr Ser Pro Phe Gly Val Ile
115 120 125
Gly Ser Val Ala Pro Ser Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn
130 135 140
Ser Ile Ser Met Leu Ala Ala Gly Asn Ala Val Tyr Phe Ser Pro His
145 150 155 160
Pro Gly Ala Lys Ala Val Ser Leu Asp Leu Ile Ala Gln Ile Glu Ala
165 170 175
Ile Ile Phe Asn Ser Cys Gly Ile Arg Asn Leu Val Val Thr Val Gln
180 185 190
Glu Pro Ser Phe Glu Ala Thr Gln Gln Met Met Ala His Asp Lys Ile
195 200 205
Ala Leu Leu Ala Ile Thr Gly Gly Pro Ala Ile Val Ala Met Gly Met
210 215 220
Lys Ser Gly Lys Lys Val Ile Gly Ala Gly Ala Gly Asn Pro Pro Cys
225 230 235 240
Leu Val Asp Glu Thr Ala Glu Leu Ala Lys Ala Ala Gln Asp Ile Val
245 250 255
Ser Gly Ala Ser Phe Asp Tyr Asn Leu Pro Cys Ile Ala Glu Lys Ser
260 265 270
Leu Ile Val Val Glu Ser Val Ala Asp Arg Leu Leu Gln Gln Met Gln
275 280 285
Ala Phe Asp Ala Leu Leu Ile Ser Asn Pro Gln Asp Val Asp Ser Leu
290 295 300
Arg Lys Ala Cys Leu Thr Pro Gln Gly His Ala Asn Lys Asn Leu Val
305 310 315 320
Gly Lys Ser Pro Leu Glu Leu Leu Lys Ala Ala Gly Leu Thr Cys Pro
325 330 335
Ala Lys Ala Pro Arg Leu Leu Leu Val Glu Val Ala Gly Asp Asp Pro
340 345 350
Leu Val Thr Thr Glu Gln Leu Met Pro Leu Leu Pro Val Val Arg Val
355 360 365
Lys Asp Phe Asp Ala Ala Leu Thr Leu Ala Leu Gln Val Glu Gly Gly
370 375 380
Leu His His Thr Ala Thr Met His Ser Gln Asn Val Ser Arg Leu Asn
385 390 395 400
Leu Ala Ala Arg Leu Leu Gln Thr Ser Ile Phe Val Lys Asn Gly Pro
405 410 415
Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Phe Thr Thr Phe Thr
420 425 430
Ile Ala Thr Pro Thr Gly Glu Gly Thr Thr Ser Ala Arg Thr Phe Ala
435 440 445
Arg Gln Arg Arg Cys Val Leu Thr Asn Gly Phe Ser Ile Arg
450 455 460
<210> 112
<211> 511
<212> PRT
<213> Proteocatella sphenisci
<400> 112
Val Asp Ile Gly Gln Lys Asp Ile Glu Leu Ile Val Gln Gln Val Leu
1 5 10 15
Lys Asn Val Val Ser Gln Ser Ala Ala Ala Gln Ser Asn Ser Gln Pro
20 25 30
Glu Val Lys Thr Tyr Arg Pro Gly Val Pro Val Gln Glu Phe Ser Met
35 40 45
Lys Ser Gln Tyr Ala Pro Ser Ser Pro Tyr Pro Ser Ser Ser Gln Ser
50 55 60
Ser Ala Gly Asp Tyr Gly Val Phe Glu Thr Met Asp Gln Ala Val Glu
65 70 75 80
Ala Ala Tyr Gln Ala Gln Lys Ile Tyr Gln Ala Lys Phe Gln Leu Lys
85 90 95
Asp Arg Glu Arg Leu Ile Lys Ser Ile Arg Glu Thr Gly Met Lys Asn
100 105 110
Val Glu Lys Leu Ala Arg Met Ser Val Asp Glu Thr Gly Leu Gly Arg
115 120 125
Tyr Glu Asp Lys Ile Leu Lys Asn Thr Leu Val Leu Glu Arg Thr Pro
130 135 140
Gly Thr Glu Cys Leu Lys Thr Glu Ala Ile Ser Gly Asp Asp Gly Leu
145 150 155 160
Thr Ile Ile Glu His Ala Pro Tyr Gly Val Ile Gly Ser Ile Thr Pro
165 170 175
Val Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn Val Ile Ser Met Ile
180 185 190
Ala Gly Gly Asn Ser Val Val Phe Asn Val His Pro Ser Ala Lys Glu
195 200 205
Ser Cys Arg Phe Ala Val Gln Met Ile Asn Lys Ala Ile Glu Glu Val
210 215 220
Gly Gly Pro Lys Asn Leu Val Ser Met Val Lys Gln Pro Thr Leu Asp
225 230 235 240
Thr Val Ser Gln Leu Ser Lys Asn Asp Lys Val Arg Leu Met Ala Gly
245 250 255
Thr Gly Gly Met Pro Met Val Arg Ser Leu Leu Gln Ser Gly Lys Lys
260 265 270
Val Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val Asp Glu Thr
275 280 285
Ala Asp Ile Lys Arg Ala Ala Ala Glu Ile Phe Lys Gly Ala Ser Phe
290 295 300
Asp Asn Asn Val Leu Cys Leu Ala Glu Lys Glu Val Phe Ile Val Glu
305 310 315 320
Ser Val Ala Thr Asp Phe Val Tyr Asn Met Ile Gln Glu Gly Ala Phe
325 330 335
Leu Leu Asn Glu Ser Gln Leu Glu Lys Ile Met Asn Leu Val Leu Thr
340 345 350
Tyr Glu Glu Thr Pro Asn Gly Arg Glu Tyr His Thr Ser Lys Asn Trp
355 360 365
Val Gly Lys Asp Ala Gly Lys Met Leu Asp Ala Ile Gly Ile Asn Gly
370 375 380
Lys Ser Asp Cys Arg Leu Leu Ile Cys Glu Val Gly Pro Asn His Pro
385 390 395 400
Phe Val Leu Leu Glu Gln Leu Met Pro Val Leu Pro Ile Val Lys Cys
405 410 415
Lys Asn Leu Asp Glu Ala Ile Lys Phe Ala Met Ile Ala Glu His Gly
420 425 430
Asn Arg His Thr Ala Ser Met Phe Ser Gln Ser Ile Asn Asn Leu Thr
435 440 445
Arg Phe Ala Arg Glu Val Glu Thr Thr Ile Phe Val Lys Asn Ala Ala
450 455 460
Thr Leu Ala Gly Val Gly Phe Gly Gly Glu Gly His Thr Thr Met Thr
465 470 475 480
Ile Ala Gly Pro Thr Gly Glu Gly Ile Thr Asn Ala Val Ser Phe Thr
485 490 495
Arg Gln Arg Arg Cys Ala Leu Ser Glu Gly Gly Phe Arg Ile Ile
500 505 510
<210> 113
<400> 113
000
<210> 114
<400> 114
000
<210> 115
<400> 115
000
<210> 116
<400> 116
000
<210> 117
<211> 480
<212> PRT
<213> Pelosinus propionicus DSM
<400> 117
Met Ser Ile Asp Gln Ala Leu Ile Glu Lys Ile Thr Leu Glu Ile Leu
1 5 10 15
Ser Lys Met Gln Thr Gly Ala Lys Ala Ala Pro Thr Gly Tyr Gly Ser
20 25 30
Gly Ile Phe Glu Thr Val Asp Glu Ala Val Ala Ala Ala Arg Lys Ala
35 40 45
Tyr Gln Glu Leu Lys Thr Leu Ser Leu Glu Lys Arg Glu Val Leu Ile
50 55 60
Lys Ala Met Arg Asp Val Ala Tyr Glu Asn Ala Thr Ile Leu Ala Gln
65 70 75 80
Met Ala Val Asp Glu Ser Gly Met Gly Arg Val Ser Asp Lys Ile Ile
85 90 95
Lys Asn Gln Val Ala Ala Leu Lys Thr Pro Gly Thr Glu Asp Leu Thr
100 105 110
Thr Gln Ala Trp Ser Gly Asp Asn Gly Leu Thr Leu Ile Glu Met Gly
115 120 125
Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Thr Thr Asn Pro Thr Glu
130 135 140
Thr Val Ile Cys Asn Gly Ile Gly Met Ile Ala Ala Gly Asn Thr Val
145 150 155 160
Phe Phe Ser Pro His Pro Thr Ala Lys Asn Thr Ser Ile Lys Ile Ile
165 170 175
Thr Leu Leu Asn Asp Ala Ile Val Lys Ala Gly Gly Pro Asn Asn Leu
180 185 190
Leu Thr Ser Val Ala Asn Pro Ser Ile Lys Ala Ala Asn Glu Met Met
195 200 205
Lys His Pro Gly Ile Asn Met Leu Val Ala Thr Gly Gly Pro Gly Val
210 215 220
Val Lys Ala Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Ile Val Asp Glu Thr Ala Asp Ile Glu Lys Ala
245 250 255
Ala Arg Asp Ile Val Ala Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys
260 265 270
Ile Ala Glu Lys Glu Val Ile Ala Val Gly Ser Ile Ala Asp Arg Leu
275 280 285
Ile Thr Tyr Met Gln Lys Tyr Gly Ala Tyr Leu Ile Ser Gly Ser Asn
290 295 300
Ile Asp Arg Leu Leu Asp Val Ile Met Thr Val Gln Glu Glu Lys Ile
305 310 315 320
Ala Glu Gly Cys Thr Asp Lys Pro Lys Arg Ser Tyr Gly Ile Asn Lys
325 330 335
Asp Tyr Val Gly Lys Asp Ala Lys Tyr Leu Leu Ser Lys Ile Gly Ile
340 345 350
Asp Val Pro Asp Ser Val Lys Val Val Leu Cys Glu Thr Pro Ala Asp
355 360 365
His Pro Phe Val Ile Glu Glu Leu Met Met Pro Val Leu Pro Val Val
370 375 380
Gln Val Lys Asp Ile Asp Glu Ala Ile Glu Val Ala Val Arg Val Glu
385 390 395 400
His Gly Asn Arg His Thr Ala Ala Met His Ser Lys Asn Val Asp His
405 410 415
Leu Thr Arg Phe Ala Arg Ala Val Glu Thr Thr Ile Phe Val Lys Asn
420 425 430
Ala Pro Ser Tyr Ala Gly Ile Gly Val Gly Gly Glu Gly Phe Thr Ser
435 440 445
Phe Thr Leu Ala Gly Pro Thr Gly Glu Gly Ile Thr Ser Pro Arg Ser
450 455 460
Phe Thr Arg Gln Arg Arg Cys Val Leu Val Asp Ala Phe Ser Ile Val
465 470 475 480
<210> 118
<400> 118
000
<210> 119
<400> 119
000
<210> 120
<400> 120
000
<210> 121
<400> 121
000
<210> 122
<400> 122
000
<210> 123
<400> 123
000
<210> 124
<400> 124
000
<210> 125
<400> 125
000
<210> 126
<400> 126
000
<210> 127
<400> 127
000
<210> 128
<400> 128
000
<210> 129
<400> 129
000
<210> 130
<400> 130
000
<210> 131
<400> 131
000
<210> 132
<400> 132
000
<210> 133
<400> 133
000
<210> 134
<211> 457
<212> PRT
<213> Clostridium sp. KLE
<400> 134
Met Val Gln Asp Ile Val Lys Glu Val Val Ala Arg Met Gln Leu Ser
1 5 10 15
Gly Thr Ala Gln Ser Ala Gln His Gly Val Phe Asn Asp Met Asn Gln
20 25 30
Ala Ile Glu Ala Ala Lys Glu Ala Glu Lys Thr Val Arg Arg Met Thr
35 40 45
Met Asp Gln Arg Glu Gln Ile Val Ser Asn Ile Arg Lys Lys Thr His
50 55 60
Glu Ala Ala Glu Ile Leu Ala Arg Met Gly Val Glu Glu Thr Gly Met
65 70 75 80
Gly Asn Val Gly Asp Lys Ile Leu Lys His His Leu Leu Ala Asp Lys
85 90 95
Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala Trp Ser Gly Asp Arg
100 105 110
Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly Val Ile Gly Ala Ile
115 120 125
Thr Pro Cys Thr Asn Pro Ser Glu Thr Val Leu Cys Asn Ser Ile Gly
130 135 140
Met Ile Ala Ala Gly Asn Thr Val Val Phe Asn Pro His Pro Gln Ala
145 150 155 160
Ile Arg Thr Ser Ile Phe Ala Ile Asn Leu Val Asn Glu Ala Ser Leu
165 170 175
Glu Ala Gly Gly Pro Asp Asn Val Ala Cys Thr Val Phe Lys Pro Thr
180 185 190
Leu Glu Thr Ser Asn Ile Met Met Lys His Lys Asp Ile Pro Leu Ile
195 200 205
Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala Val Leu Ser Ser Gly
210 215 220
Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro Pro Ala Leu Val Asp
225 230 235 240
Glu Thr Ala Asp Ile Arg Lys Ala Ala Ala Asp Ile Val Asn Gly Cys
245 250 255
Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu Lys Glu Ile Val Ala
260 265 270
Val Asp Ser Ile Ala Asp Glu Leu Met Asn Tyr Met Ile Ser Glu Gln
275 280 285
Gly Cys Tyr Leu Ile Ser Lys Glu Glu Gln Asp Lys Leu Thr Ala Thr
290 295 300
Val Leu Thr Pro Lys Gly Leu Asn Arg Lys Cys Val Gly Arg Asp Ala
305 310 315 320
Arg Thr Leu Leu Ser Met Ile Gly Ile Gln Ala Pro Glu Asn Ile Arg
325 330 335
Cys Ile Val Phe Glu Gly Glu Lys Glu His Pro Leu Ile Ser Glu Glu
340 345 350
Leu Met Met Pro Ile Leu Gly Leu Val Arg Ala Lys Asp Phe Asp Asp
355 360 365
Ala Val Glu Lys Ala Val Trp Leu Glu His Gly Asn Arg His Ser Ala
370 375 380
His Ile His Ser Lys Asn Ile Asp Asn Ile Thr Lys Tyr Ala Arg Ala
385 390 395 400
Ile Asp Thr Ala Ile Leu Val Lys Asn Ala Pro Ser Tyr Ala Ala Leu
405 410 415
Gly Phe Gly Gly Glu Gly Phe Cys Thr Phe Thr Ile Ala Ser Arg Thr
420 425 430
Gly Glu Gly Leu Thr Ser Thr Ser Thr Phe Thr Lys Arg Arg Arg Cys
435 440 445
Val Met Ser Asp Ser Leu Cys Ile Arg
450 455
<210> 135
<211> 527
<212> PRT
<213> Caldalkalibacillus thermarum TA2.A1
<400> 135
Met Asn Met Thr Glu Lys Asp Ile Glu Lys Ile Val Gln Ser Val Leu
1 5 10 15
His Asn Val Glu Ser Ala Leu Gly Lys Ser Ala Ser Ala Ser Pro Ser
20 25 30
Val Ser Ala Val Ser Val Ala Ser Gly Glu Gly Ile Lys Pro Val Gln
35 40 45
Phe Lys Gln Val Pro Val Phe Gln Gln Glu Thr Val Lys Ser Pro Asn
50 55 60
Arg Asn Arg Asn Leu Gly Gly Ala Glu Glu Lys Trp Gly Val Phe Asn
65 70 75 80
His Met Glu Asp Ala Ile Glu Ala Ser Tyr Arg Ala Gln Met Glu Phe
85 90 95
Val Lys His Phe Gln Leu Lys Asp Arg Glu Lys Ile Ile Thr Ala Ile
100 105 110
Arg Glu Ala Val Leu Arg Glu Lys Glu Val Leu Ala Arg Lys Val Tyr
115 120 125
Glu Glu Thr Lys Ile Gly Arg Tyr Glu Asp Lys Val Ala Lys His Glu
130 135 140
Leu Ala Ala Leu Lys Thr Pro Gly Thr Glu Asp Leu Lys Thr Glu Ala
145 150 155 160
Phe Ser Gly Asp Asn Gly Leu Thr Ile Val Glu Arg Ala Pro Tyr Gly
165 170 175
Leu Ile Gly Ala Val Thr Pro Val Thr Asn Pro Thr Glu Thr Ile Ile
180 185 190
Asn Asn Ala Ile Gly Met Leu Ala Ala Gly Asn Ala Val Val Phe Asn
195 200 205
Val His Pro Ser Ser Lys Arg Ser Cys Ala Tyr Ala Val Gln Leu Ile
210 215 220
Asn Lys Ala Ile Thr Glu Ala Gly Gly Pro His His Leu Val Thr Met
225 230 235 240
Val Lys Glu Pro Thr Leu Asp Thr Leu Gln Thr Leu Ile Asp Ser Pro
245 250 255
Lys Val Lys Leu Leu Val Gly Thr Gly Gly Pro Gly Leu Val Gln Thr
260 265 270
Leu Leu Lys Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro
275 280 285
Pro Val Ile Val Asp Asp Thr Ala Asp Leu Glu His Ala Ala Arg Ser
290 295 300
Ile Ile Glu Gly Ala Ala Phe Asp Asn Asn Leu Leu Cys Ile Ala Glu
305 310 315 320
Lys Glu Val Phe Val Leu Glu Ser Val Ala Asp Asp Leu Ile Phe His
325 330 335
Met Leu Asn His Gly Ala Tyr Met Leu Gly Gln His Glu Val Glu Gln
340 345 350
Val Met Ala Phe Ala Leu Glu Glu Gln Gly Asn Glu Gln Asn Arg Gly
355 360 365
Cys Gly Phe Asn Pro Gln Arg His Tyr Gln Val Ser Lys Asp Trp Ile
370 375 380
Gly Gln Asp Ala Arg Leu Phe Leu Glu His Ile Gly Val Gln Pro Pro
385 390 395 400
Thr Glu Val Lys Leu Leu Ile Cys Asp Val Glu Phe Asp His Pro Phe
405 410 415
Val Gln Leu Glu Gln Met Met Pro Val Leu Pro Ile Val Arg Val Lys
420 425 430
Thr Leu Asp Glu Ala Ile Glu Lys Ala Val Met Ala Glu His Gly Asn
435 440 445
Arg His Thr Ala Ile Met His Ser Lys Asn Val Asp His Leu Thr Lys
450 455 460
Phe Ala Arg Ala Ile Gln Thr Thr Leu Phe Val Lys Asn Ala Ser Ser
465 470 475 480
Leu Ala Gly Val Gly Tyr Gly Gly Glu Gly His Thr Thr Met Thr Ile
485 490 495
Ala Gly Pro Thr Gly Glu Gly Val Thr Ser Ala Lys Thr Phe Thr Arg
500 505 510
Glu Arg Arg Cys Val Leu Ala Glu Gly Gly Phe Arg Ile Ile Gly
515 520 525
<210> 136
<400> 136
000
<210> 137
<211> 476
<212> PRT
<213> Caldalkalibacillus thermarum TA2.A1
<400> 137
Val Pro Val Phe Gln Gln Glu Thr Val Lys Ser Pro Asn Arg Asn Arg
1 5 10 15
Asn Leu Gly Gly Ala Glu Glu Lys Trp Gly Val Phe Asn His Met Glu
20 25 30
Asp Ala Ile Glu Ala Ser Tyr Arg Ala Gln Met Glu Phe Val Lys His
35 40 45
Phe Gln Leu Lys Asp Arg Glu Lys Ile Ile Thr Ala Ile Arg Glu Ala
50 55 60
Val Leu Arg Glu Lys Glu Val Leu Ala Arg Lys Val Tyr Glu Glu Thr
65 70 75 80
Lys Ile Gly Arg Tyr Glu Asp Lys Val Ala Lys His Glu Leu Ala Ala
85 90 95
Leu Lys Thr Pro Gly Thr Glu Asp Leu Lys Thr Glu Ala Phe Ser Gly
100 105 110
Asp Asn Gly Leu Thr Ile Val Glu Arg Ala Pro Tyr Gly Leu Ile Gly
115 120 125
Ala Val Thr Pro Val Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn Ala
130 135 140
Ile Gly Met Leu Ala Ala Gly Asn Ala Val Val Phe Asn Val His Pro
145 150 155 160
Ser Ser Lys Arg Ser Cys Ala Tyr Ala Val Gln Leu Ile Asn Lys Ala
165 170 175
Ile Thr Glu Ala Gly Gly Pro His His Leu Val Thr Met Val Lys Glu
180 185 190
Pro Thr Leu Asp Thr Leu Gln Thr Leu Ile Asp Ser Pro Lys Val Lys
195 200 205
Leu Leu Val Gly Thr Gly Gly Pro Gly Leu Val Gln Thr Leu Leu Lys
210 215 220
Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile
225 230 235 240
Val Asp Asp Thr Ala Asp Leu Glu His Ala Ala Arg Ser Ile Ile Glu
245 250 255
Gly Ala Ala Phe Asp Asn Asn Leu Leu Cys Ile Ala Glu Lys Glu Val
260 265 270
Phe Val Leu Glu Ser Val Ala Asp Asp Leu Ile Phe His Met Leu Asn
275 280 285
His Gly Ala Tyr Met Leu Gly Gln His Glu Val Glu Gln Val Met Ala
290 295 300
Phe Ala Leu Glu Glu Gln Gly Asn Glu Gln Asn Arg Gly Cys Gly Phe
305 310 315 320
Asn Pro Gln Arg His Tyr Gln Val Ser Lys Asp Trp Ile Gly Gln Asp
325 330 335
Ala Arg Leu Phe Leu Glu His Ile Gly Val Gln Pro Pro Thr Glu Val
340 345 350
Lys Leu Leu Ile Cys Asp Val Glu Phe Asp His Pro Phe Val Gln Leu
355 360 365
Glu Gln Met Met Pro Val Leu Pro Ile Val Arg Val Lys Thr Leu Asp
370 375 380
Glu Ala Ile Glu Lys Ala Val Met Ala Glu His Gly Asn Arg His Thr
385 390 395 400
Ala Ile Met His Ser Lys Asn Val Asp His Leu Thr Lys Phe Ala Arg
405 410 415
Ala Ile Gln Thr Thr Leu Phe Val Lys Asn Ala Ser Ser Leu Ala Gly
420 425 430
Val Gly Tyr Gly Gly Glu Gly His Thr Thr Met Thr Ile Ala Gly Pro
435 440 445
Thr Gly Glu Gly Val Thr Ser Ala Lys Thr Phe Thr Arg Glu Arg Arg
450 455 460
Cys Val Leu Ala Glu Gly Gly Phe Arg Ile Ile Gly
465 470 475
<210> 138
<400> 138
000
<210> 139
<400> 139
000
<210> 140
<400> 140
000
<210> 141
<400> 141
000
<210> 142
<400> 142
000
<210> 143
<400> 143
000
<210> 144
<400> 144
000
<210> 145
<211> 462
<212> PRT
<213> Blautia sp. CAG:257
<400> 145
Met Pro Ile Ser Glu Asn Met Val Gln Glu Ile Val Gln Glu Val Met
1 5 10 15
Ala Lys Met Gln Ile Ala Glu Ala Pro Ala Gly Lys His Gly Ile Phe
20 25 30
Lys Asp Met Asn Asp Ala Ile Glu Ala Ala Lys Lys Ala Glu Leu Ile
35 40 45
Val Lys Arg Met Ser Met Asp Gln Arg Glu Lys Ile Ile Thr Cys Ile
50 55 60
Arg Lys Lys Ile Lys Glu Asn Ala Glu Val Leu Ala Arg Met Gly Val
65 70 75 80
Glu Glu Thr Gly Met Gly Asn Val Gly Asp Lys Ile Leu Lys His His
85 90 95
Leu Val Ala Asp Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Ile Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Val Leu
130 135 140
Cys Asn Thr Met Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Tyr Ala Ile Asn Leu Leu
165 170 175
Asn Glu Ala Ser Leu Glu Ser Gly Gly Pro Asp Asn Ile Ala Val Thr
180 185 190
Val Glu Lys Pro Thr Leu Glu Thr Ser Asp Ile Met Met Lys His Lys
195 200 205
Asp Ile His Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Gln Asp
245 250 255
Ile Val Asn Gly Cys Thr Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Ser Ser Ile Ala Asp Glu Leu Met His Tyr
275 280 285
Leu Ile Thr Glu Asn Asp Cys Tyr Leu Ala Ser Lys Glu Glu Gln Asp
290 295 300
Lys Leu Thr Glu Val Val Leu Ala Gly Gly Lys Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Arg Thr Leu Leu Ser Met Ile Gly Val Asp Ala
325 330 335
Pro Ala Asn Ile Arg Cys Ile Val Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Ala Glu Glu Leu Met Met Pro Ile Leu Gly Met Val Arg Ala
355 360 365
Arg Asp Phe Asp Asp Ala Val Glu Gln Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Val Asp Asn Ile Thr
385 390 395 400
Lys Tyr Ala Arg Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Gly Pro
405 410 415
Ser Tyr Ser Ala Leu Gly Phe Gly Gly Glu Gly Phe Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Ser Asp Ser Leu Cys Ile Arg
450 455 460
<210> 146
<211> 469
<212> PRT
<213> Listeria marthii FSL
<400> 146
Met Glu Ser Leu Glu Leu Glu Gln Leu Val Lys Lys Val Leu Leu Glu
1 5 10 15
Lys Leu Ala Glu Gln Lys Glu Ala Pro Ala Lys Pro Ile Thr Gln Gly
20 25 30
Ala Lys Ser Gly Ile Phe Asp Thr Val Asp Glu Ala Val Gln Ala Ala
35 40 45
Val Ile Ala Gln Asn Cys Tyr Lys Glu Lys Ser Leu Glu Glu Arg Arg
50 55 60
Asn Val Val Lys Ala Ile Arg Glu Thr Leu Tyr Pro Glu Ile Glu Thr
65 70 75 80
Ile Ala Thr Lys Ala Val Ala Glu Thr Gly Met Gly Asn Val Ala Asp
85 90 95
Lys Ile Leu Lys Asn Thr Leu Ala Ile Glu Lys Thr Pro Gly Val Glu
100 105 110
Asp Leu Tyr Thr Glu Val Ala Thr Gly Asp Asn Gly Met Thr Leu Tyr
115 120 125
Glu Leu Ser Pro Tyr Gly Val Ile Gly Ala Val Ala Pro Ser Thr Asn
130 135 140
Pro Thr Glu Thr Leu Ile Cys Asn Thr Ile Gly Met Leu Ala Ala Gly
145 150 155 160
Asn Ala Val Phe Tyr Ser Pro His Pro Gly Ala Lys Asn Ile Ser Leu
165 170 175
Trp Leu Ile Glu Lys Leu Asn Thr Ile Val Arg Glu Ser Cys Gly Ile
180 185 190
Asp Asn Leu Val Val Thr Val Glu Lys Pro Ser Ile Gln Ala Ala Gln
195 200 205
Glu Met Met Asn His Pro Lys Val Pro Leu Leu Val Ile Thr Gly Gly
210 215 220
Pro Gly Val Val Leu Gln Ala Met Gln Ser Gly Lys Lys Val Ile Gly
225 230 235 240
Ala Gly Ala Gly Asn Pro Pro Ser Ile Val Asp Glu Thr Ala Asn Ile
245 250 255
Glu Lys Ala Ala Ala Asp Ile Val Asp Gly Ala Ser Phe Asp His Asn
260 265 270
Ile Leu Cys Ile Ala Glu Lys Ser Ile Val Ala Val Glu Ser Ile Ala
275 280 285
Asp Phe Leu Leu Phe Gln Met Glu Lys Asn Gly Ala Leu His Val Thr
290 295 300
Asn Pro Ser Asp Ile Gln Lys Leu Glu Lys Val Ala Val Thr Asp Lys
305 310 315 320
Gly Val Thr Asn Lys Lys Leu Val Gly Lys Ser Ala Ala Glu Ile Leu
325 330 335
Lys Glu Ala Gly Ile Thr Cys Asp Phe Thr Pro Arg Leu Ile Ile Val
340 345 350
Glu Thr Thr Lys Thr His Pro Phe Ala Thr Val Glu Leu Leu Met Pro
355 360 365
Ile Val Pro Leu Val Arg Val Pro Asp Phe Asp Glu Ala Leu Glu Val
370 375 380
Ala Ile Glu Leu Glu Gln Gly Leu His His Thr Ala Thr Met His Ser
385 390 395 400
Gln Asn Ile Ser Arg Leu Asn Lys Ala Ala Arg Asp Met Gln Thr Ser
405 410 415
Ile Phe Val Lys Asn Gly Pro Ser Phe Ala Gly Leu Gly Phe Arg Gly
420 425 430
Glu Gly Ser Thr Thr Phe Thr Ile Ala Thr Pro Thr Gly Glu Gly Thr
435 440 445
Thr Thr Ala Arg His Phe Ala Arg Arg Arg Arg Cys Val Leu Thr Asp
450 455 460
Gly Phe Ser Ile Arg
465
<210> 147
<400> 147
000
<210> 148
<211> 473
<212> PRT
<213> Clostridium methoxybenzovorans
<400> 148
Met Glu Ile Gly Ala Lys Glu Ile Glu Leu Ile Val Arg Glu Val Leu
1 5 10 15
Ala Gly Ile Glu Ser Arg Gly Ile Lys Pro Ser Tyr Thr Pro Ser Arg
20 25 30
Ser Glu Asp Gly Val Phe Glu Arg Val Glu Asp Ala Ile Glu Ala Ala
35 40 45
Tyr Ala Ala Gln Arg Glu Trp Val Glu His Tyr Arg Val Glu Asp Arg
50 55 60
Arg Arg Ile Ile Glu Ala Ile Arg Val Thr Ala Lys Ser His Ala Glu
65 70 75 80
Ser Leu Ala Lys Met Val Trp Glu Glu Thr Gly Met Gly Arg Phe Glu
85 90 95
Asp Lys Ile Gln Lys His Met Ala Val Ile Glu Lys Thr Pro Gly Val
100 105 110
Glu Cys Leu Thr Thr Glu Ala Ile Ser Gly Asp Gly Gly Leu Met Ile
115 120 125
Glu Glu Tyr Ala Pro Phe Gly Val Ile Gly Ala Ile Thr Pro Ser Thr
130 135 140
Asn Pro Thr Glu Thr Ile Ile Asn Asn Thr Ile Ser Met Ile Ala Gly
145 150 155 160
Gly Asn Ser Val Val Phe Asn Val His Pro Gly Ala Lys Arg Cys Cys
165 170 175
Ala His Cys Leu Lys Ile Leu His Gln Ala Ile Val Glu Asn Gly Gly
180 185 190
Pro Ala Ser Leu Ile Thr Met Gln Lys Glu Pro Asp Met Glu Ala Val
195 200 205
Ser Lys Leu Thr Ser Asp Pro Arg Ile Arg Leu Met Val Gly Thr Gly
210 215 220
Gly Met Pro Met Val Asn Ala Leu Leu Arg Ser Gly Lys Lys Thr Ile
225 230 235 240
Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val Asp Asp Thr Ala Asp
245 250 255
Val Ser Leu Ala Ala Arg Glu Ile Tyr Arg Gly Ala Ser Phe Asp Asn
260 265 270
Asn Ile Leu Cys Leu Ala Glu Lys Glu Val Phe Val Met Glu Arg Ala
275 280 285
Ala Asp Glu Leu Val Asn Lys Leu Ile Lys Glu Gly Ala Tyr Leu Leu
290 295 300
Ser Ser Met Glu Leu Ser Glu Ile Leu Lys Phe Ala Met Val Glu Lys
305 310 315 320
Asn Gly Ser Tyr Glu Val Asn Lys Lys Trp Val Gly Lys Asp Ala Gly
325 330 335
Gln Phe Leu Glu Ala Ile Gly Val Ser Gly His Lys Asp Val Arg Leu
340 345 350
Leu Ile Cys Glu Thr Asp Arg Ser His Pro Phe Val Met Val Glu Gln
355 360 365
Leu Met Pro Ile Leu Pro Ile Val Arg Leu Arg Thr Phe Glu Glu Cys
370 375 380
Val Glu Ser Ala Leu Ala Ala Glu Ser Gly Asn Arg His Thr Ala Ser
385 390 395 400
Met Phe Ser Arg Asn Val Glu Asn Met Thr Lys Phe Gly Lys Ile Ile
405 410 415
Glu Thr Thr Ile Phe Thr Lys Asn Gly Ser Thr Leu Lys Gly Val Gly
420 425 430
Ile Gly Gly Glu Gly His Thr Thr Met Thr Ile Ala Gly Pro Thr Gly
435 440 445
Glu Gly Leu Thr Cys Ala Arg Ser Phe Thr Arg Arg Arg Arg Cys Met
450 455 460
Leu Ala Glu Gly Gly Leu Arg Ile Ile
465 470
<210> 149
<211> 477
<212> PRT
<213> Bacillus sp. m3-13
<400> 149
Val Gln Ile Lys Glu Ser Asp Ile Lys Glu Met Val Ala Gln Val Leu
1 5 10 15
Ala Gln Leu Gly Asp Glu Ser Lys Gln Pro Ser Pro Ala Ser Glu Gln
20 25 30
Gly Ser Asn Glu Val Pro Leu Gly Asn Gly Val Phe Thr Thr Val Asp
35 40 45
Gln Ala Thr Glu Ala Ala Thr Glu Ala Trp Asp Lys Leu Arg Ala Thr
50 55 60
Ser Leu Glu Thr Arg Lys Asn Met Ile Glu Lys Met Arg Glu Val Ser
65 70 75 80
Arg Glu His Ala Lys Ala Leu Ala Glu Leu Ala Val Lys Glu Thr Gly
85 90 95
Leu Gly Arg Val Glu Asp Lys Val Ala Lys Asn Leu Leu Ala Ala Asp
100 105 110
Lys Thr Pro Gly Val Glu Asp Ile Val Ala Thr Thr Tyr Ser Gly Asp
115 120 125
Gly Gly Leu Thr Leu Val Glu Tyr Ser Pro Val Gly Val Tyr Gly Ala
130 135 140
Ile Thr Pro Ser Thr Asn Pro Ala Ala Thr Ile Ile Asn Asn Ser Ile
145 150 155 160
Ser Leu Val Ala Ala Gly Asn Ala Val Val Phe Asn Pro His Pro Ser
165 170 175
Ala Lys Gln Val Ser Ile Lys Thr Met Gln Leu Leu Asn Glu Ala Ile
180 185 190
Val Ala Ala Gly Gly Pro Ala Asn Thr Leu Thr Ser Val Ala Ser Pro
195 200 205
Asn Ile Glu Thr Ser Asn Glu Val Met Lys His Pro Lys Val Arg Ala
210 215 220
Leu Val Val Thr Gly Gly Gly Ile Val Val Gln Ala Ala Met Ser Ala
225 230 235 240
Gly Lys Lys Val Ile Ala Ala Gly Pro Gly Asn Pro Pro Val Val Val
245 250 255
Asp Glu Thr Ala Ile Ile Ser Lys Ala Ala Lys Asp Ile Val Thr Gly
260 265 270
Ala Ser Phe Asp Asn Asn Val Leu Cys Thr Ala Glu Lys Glu Val Phe
275 280 285
Val Val Glu Lys Val Ala Asn Thr Leu Lys Ser Glu Met Thr Lys Asn
290 295 300
Gly Ala Val Glu Leu Lys Gly Tyr Gln Leu Glu Lys Leu Leu Gly Lys
305 310 315 320
Ile Leu Val Lys Lys Gly Glu Lys Tyr Tyr Pro Asn Arg Asp Phe Ile
325 330 335
Gly Lys Asp Ala Ser Val Leu Leu Glu Ala Ala Gly Ile Arg Ser Asp
340 345 350
Ser Asn Val Lys Leu Ile Ile Ala Glu Thr Lys Glu Asp His Pro Leu
355 360 365
Val His Thr Glu Met Leu Met Pro Ile Leu Pro Ile Val Arg Val Ser
370 375 380
Asp Val Asp Lys Ala Ile Ser Leu Ala Val Lys Ala Glu Lys Gly Asn
385 390 395 400
Arg His Thr Ala Ile Met His Ser Gln Asn Val Thr Asn Leu Thr Lys
405 410 415
Met Ala Lys Glu Ile Gln Ala Thr Ile Phe Val Lys Asn Gly Pro Ser
420 425 430
Val Ala Gly Leu Gly Tyr Gln Ser Glu Gly Phe Thr Thr Leu Thr Ile
435 440 445
Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys Thr Phe Thr Arg
450 455 460
Gln Arg Arg Cys Val Leu Val Asp Gly Phe Arg Ile Ile
465 470 475
<210> 150
<211> 472
<212> PRT
<213> bacterium CG2_30_54_10
<400> 150
Met Ser Val Ser Lys Asp Glu Ile Asn Val Ile Val Gln Glu Val Leu
1 5 10 15
Lys Ala Ile Glu Thr Ser Gly Gly Leu Pro Ser Ala Ala Ser Ser Val
20 25 30
Gly Arg Ile Ser Gln Lys Gly Val Phe Glu Asn Leu Asp Asp Ala Ile
35 40 45
Lys Ala Ala Gly Gln Ala Gln Lys Lys Leu Val Glu Leu Pro Leu Lys
50 55 60
Thr Arg Gly Glu Ile Ile Ala Asn Met Arg Arg Arg Ala Ala Glu Asn
65 70 75 80
Val Glu Glu Ile Ser Arg Leu Gly His Glu Glu Thr Gly Tyr Gly Arg
85 90 95
Ile Ala Asp Lys Ile Gln Lys Asn Met Leu Ala Ile Thr Lys Thr Pro
100 105 110
Gly Ile Glu Asp Leu Gln Pro Val Ala Tyr Ser Gly Asp His Gly Leu
115 120 125
Thr Ile Val Glu Gln Ala Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro
130 135 140
Ser Thr Asn Pro Ser Glu Thr Val Ile Cys Asn Ser Ile Gly Met Ile
145 150 155 160
Ala Ala Gly Asn Ala Val Val Phe Gly Pro His Pro Ser Ala Ala Gln
165 170 175
Val Cys Leu Leu Ala Ile Ser Val Leu Asn Asp Ala Val Val Glu Ala
180 185 190
Gly Gly Pro Glu Asn Leu Met Val Ser Val Ser Lys Pro Ser Ile Gln
195 200 205
Thr Ala Gln Ala Leu Met Ala His Pro Asp Ile Arg Leu Leu Val Val
210 215 220
Thr Gly Gly Pro Ala Val Val Ala Ala Ala Ala Lys Ser Gly Lys Lys
225 230 235 240
Phe Ile Ala Ala Gly Pro Gly Asn Pro Pro Ala Val Val Asp Glu Thr
245 250 255
Ala Asp Leu Lys Lys Ala Ala Arg Asp Ile Ile Ser Gly Ala Thr Leu
260 265 270
Asp Asn Asn Ile Leu Cys Ile Ala Glu Lys Glu Ile Ile Val Val Glu
275 280 285
Ser Val Ala Asp Glu Leu Lys Arg His Leu Cys Asn Ser Gly Ala Tyr
290 295 300
Glu Ala Ser Ala Arg Glu Ile Leu Gln Leu Glu Lys Leu Val Ile Asp
305 310 315 320
Pro Arg Thr His Gly Pro Asn Arg Ser Phe Ile Gly Lys Asn Ala Ser
325 330 335
Val Ile Leu Asp Ala Ile Gly Val Lys Val Ser Asp Glu Val Arg Met
340 345 350
Val Leu Cys Glu Val Gly Pro Asp His Pro Phe Val Val Glu Glu Met
355 360 365
Met Met Pro Val Val Pro Leu Val Arg Val Arg Asp Val His Thr Ala
370 375 380
Val Asp Phe Ala Val Lys Ile Glu His Gly Cys Arg His Thr Ala Ile
385 390 395 400
Met His Ser Lys Asn Leu Asp Asn Leu His Leu Met Ala Thr Arg Cys
405 410 415
Asn Cys Ser Ile Phe Val Lys Asn Gly Pro Ser Tyr Ala Gly Leu Gly
420 425 430
Leu Gly Gly Glu Gly Phe Thr Thr Phe Thr Ile Ala Ser Pro Thr Gly
435 440 445
Glu Gly Leu Thr Ser Ala Arg Thr Phe Thr Arg Gln Arg Arg Cys Val
450 455 460
Leu Val Asp Tyr Phe Arg Ile Val
465 470
<210> 151
<400> 151
000
<210> 152
<211> 475
<212> PRT
<213> Candidatus Izimaplasma sp
<400> 152
Met Ser Thr Asn Asp Leu Ile Lys Gln Leu Thr Glu Glu Met Glu Arg
1 5 10 15
Lys Tyr Gly Asn Asp Val Val Thr Lys Pro Asn Thr Pro Thr Asn Ser
20 25 30
Tyr Asn Thr Gly Tyr Val Gly Ile Phe Glu Asn Val Glu Asp Ala Ile
35 40 45
Leu Ala Ala Lys Glu Ser Gln Lys Gln Leu Met Glu Leu Ser Met Lys
50 55 60
Lys Arg Lys Glu Ile Ile Glu Ala Met Arg Lys Ala Ser Leu Glu Asn
65 70 75 80
Ala Glu Lys Leu Ala Ile Met Ala His Glu Glu Thr Gly Phe Gly Arg
85 90 95
Val Ala Asp Lys Ile Ile Lys Asn Val Leu Ala Ala Glu Lys Thr Pro
100 105 110
Gly Thr Glu Asp Leu Ser Ser Ser Thr Phe Thr Gly Asp Asp Gly Met
115 120 125
Thr Leu Val Glu Leu Ala Pro Tyr Gly Val Ile Gly Ser Ile Thr Pro
130 135 140
Ser Thr Asn Pro Ser Ser Thr Ile Ile Asn Asn Ser Ile Ser Met Val
145 150 155 160
Ala Ala Gly Asn Gly Val Val Tyr Asn Pro His Pro Ser Ala Lys Lys
165 170 175
Val Thr Ser Glu Thr Ile Ser Ile Leu Asn Lys Ala Ile Ser Ser Val
180 185 190
Gly Gly Pro Arg Glu Leu Leu Thr Ala Pro Leu Thr Pro Thr Met Asp
195 200 205
Thr Ser Lys Val Ile Met Thr His Lys Asp Val Arg Ile Leu Val Val
210 215 220
Thr Gly Gly Glu Ala Val Val Gly Val Ala Met Lys Ser Gly Lys Lys
225 230 235 240
Val Ile Ala Ala Gly Pro Gly Asn Pro Pro Val Ile Val Asp Glu Thr
245 250 255
Ala Asn Ile Lys Lys Ala Ala Asn Asp Val Phe Arg Gly Ala Ser Phe
260 265 270
Asp Asn Asn Ile Leu Cys Ile Ala Glu Lys Glu Ala Phe Val Ile Asn
275 280 285
Ser Val Ile Asn Glu Phe Lys Gln Glu Met Val Ser Asn Gly Ala Tyr
290 295 300
Glu Leu Lys Arg His Glu Ile Asp Leu Val Thr Glu Glu Val Phe Thr
305 310 315 320
Lys Asn Lys Asn Gly Asp Thr Val Val Asn Arg Lys His Val Gly Lys
325 330 335
Ser Ala Val Glu Ile Leu Lys Ala Cys Asn Ile Met Val His Gln Asp
340 345 350
Ile Arg Leu Ile Thr Ala Glu Val Ser Glu Asn His Pro Phe Ile Thr
355 360 365
Val Glu Met Leu Met Pro Val Leu Gly Ile Val Arg Val Tyr Ser Ile
370 375 380
Asp Glu Ala Ile Glu Lys Ala Val Ile Ala Glu Asp Gly Cys Leu His
385 390 395 400
Thr Ala Ile Met His Ser Glu Ser Val Ser Asn Leu Thr Lys Ala Ala
405 410 415
Arg Ala Leu Asn Thr Ser Ile Phe Val Lys Asn Ala Pro Ser Phe Ala
420 425 430
Gly Leu Gly Ile Glu Gly Glu Gly Phe Thr Thr Leu Thr Ile Ala Thr
435 440 445
Pro Thr Gly Glu Gly Leu Thr Ser Ala Arg Ser Phe Thr Arg Ile Arg
450 455 460
Arg Cys Thr Leu Ser Gly Gly Phe Arg Ile Val
465 470 475
<210> 153
<400> 153
000
<210> 154
<400> 154
000
<210> 155
<400> 155
000
<210> 156
<400> 156
000
<210> 157
<211> 462
<212> PRT
<213> Firmicutes bacterium CAG:41
<400> 157
Val Pro Ile Asn Glu Asn Met Val Gln Asp Ile Val Gln Glu Val Leu
1 5 10 15
Ala Lys Met Gln Ile Gln Glu Ala Pro Thr Gly Lys His Gly Val Phe
20 25 30
Lys Asp Met Asn Glu Ala Ile Glu Ala Ala Lys Lys Ala Gln Gln Thr
35 40 45
Val Lys Lys Met Ser Met Asp Gln Arg Glu Lys Ile Leu Ser Ile Ile
50 55 60
Arg Lys Lys Ile Cys Glu Asn Ala Glu Thr Met Ala Arg Met Gly Val
65 70 75 80
Glu Glu Thr Gly Met Gly Asn Val Gly Asp Lys Ile Leu Lys His Arg
85 90 95
Leu Val Ala Glu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr Thr Ala
100 105 110
Trp Ser Gly Asp Arg Gly Leu Thr Leu Val Glu Met Gly Pro Phe Gly
115 120 125
Val Ile Gly Ala Ile Thr Pro Cys Thr Asn Pro Ser Glu Thr Val Leu
130 135 140
Cys Asn Thr Met Gly Met Leu Ala Gly Gly Asn Thr Val Val Phe Asn
145 150 155 160
Pro His Pro Ala Ala Ile Lys Thr Ser Ile Phe Ala Ile Asn Leu Leu
165 170 175
Asn Glu Ala Ser Leu Glu Gly Gly Gly Pro Asp Asn Ile Ala Cys Thr
180 185 190
Val Glu Asn Pro Thr Leu Glu Thr Ser Asn Ile Met Met Lys His Lys
195 200 205
Asp Ile Pro Leu Ile Ala Ala Thr Gly Gly Pro Gly Val Val Thr Ala
210 215 220
Val Leu Ser Ser Gly Lys Arg Gly Ile Gly Ala Gly Ala Gly Asn Pro
225 230 235 240
Pro Ala Leu Val Asp Glu Thr Ala Asp Ile Arg Lys Ala Ala Gln Asp
245 250 255
Ile Val Asn Gly Cys Val Phe Asp Asn Asn Leu Pro Cys Ile Ala Glu
260 265 270
Lys Glu Ile Val Ala Val Ser Ser Val Val Asp Glu Leu Met His Tyr
275 280 285
Met Val Thr Glu Gln Gly Cys Tyr Leu Ala Ser Lys Glu Glu Gln Asp
290 295 300
Ala Leu Thr Ala Val Val Leu Ala Gly Gly Arg Leu Asn Arg Lys Cys
305 310 315 320
Val Gly Arg Asp Ala Arg Thr Leu Leu Ser Met Ile Gly Val Asp Ala
325 330 335
Pro Ala Asn Ile Arg Cys Ile Thr Phe Glu Gly Pro Lys Glu His Pro
340 345 350
Leu Ile Ala Glu Glu Leu Met Met Pro Ile Leu Gly Val Val Arg Ala
355 360 365
Lys Asp Phe Glu Asp Ala Val Glu Gln Ala Val Trp Leu Glu His Gly
370 375 380
Asn Arg His Ser Ala His Ile His Ser Lys Asn Ile Asp Asn Ile Thr
385 390 395 400
Thr Tyr Ala Lys Ala Ile Asp Thr Ala Ile Leu Val Lys Asn Ala Pro
405 410 415
Ser Tyr Ala Ala Leu Gly Phe Gly Gly Glu Gly Tyr Cys Thr Phe Thr
420 425 430
Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser Ala Ser Thr Phe Thr
435 440 445
Lys Arg Arg Arg Cys Val Met Ser Asp Ser Leu Cys Ile Arg
450 455 460
<210> 158
<211> 465
<212> PRT
<213> Fusobacterium nucleatum subsp.
<400> 158
Met Glu Phe Glu Val Asn Asn Ile Glu Glu Ile Val Glu Leu Ile Met
1 5 10 15
Lys Lys Met Ala Glu Ser Asn Ile Ser Thr Ala Gly Asn Ser Lys Asn
20 25 30
Gly Val Phe Asp Asn Val Asp Gly Ala Ile Glu Glu Ala Lys Lys Ala
35 40 45
Gln Ala Ile Leu Phe Ser Ser Lys Leu Glu Leu Arg Glu Lys Ile Ile
50 55 60
Ala Ser Ile Arg Asp Thr Leu Lys Asn His Val Thr Glu Leu Ala Glu
65 70 75 80
Leu Ala Val Lys Glu Thr Gly Met Gly Arg Val Ala Asp Lys Glu Leu
85 90 95
Lys Asn Lys Ile Ala Ile Glu Lys Thr Pro Gly Leu Glu Asp Leu Lys
100 105 110
Ala Phe Ala Phe Ser Gly Asp Asp Gly Leu Thr Val Met Glu Leu Ser
115 120 125
Pro Tyr Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Ser Glu
130 135 140
Thr Val Ile Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ala Val
145 150 155 160
Ile Phe Ala Pro His Pro Gly Ala Lys Arg Thr Ser Ile Arg Thr Val
165 170 175
Glu Leu Ile Asn Glu Ala Ile Arg Lys Val Gly Gly Pro Asp Asn Leu
180 185 190
Ile Val Thr Ile Arg Glu Pro Ser Ile Glu Asn Thr Glu Lys Ile Ile
195 200 205
Ala Asn Pro Asn Ile Lys Met Leu Val Ala Thr Gly Gly Pro Gly Val
210 215 220
Val Lys Thr Val Met Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala
225 230 235 240
Gly Asn Pro Pro Val Leu Val Asp Glu Thr Ala Asp Ile Glu Lys Ala
245 250 255
Ala Lys Asp Ile Ile Ala Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys
260 265 270
Thr Ala Glu Lys Glu Val Val Ala Val Asp Ser Ile Val Asn Tyr Leu
275 280 285
Ile Phe Glu Met Gln Lys Asn Gly Ala Tyr Leu Leu Lys Asp Lys Glu
290 295 300
Leu Ile Glu Lys Leu Leu Ser Leu Val Leu Lys Asn Asn Ser Pro Asp
305 310 315 320
Arg Lys Tyr Val Gly Arg Asp Ala Lys Tyr Leu Leu Lys Gln Ile Gly
325 330 335
Ile Glu Val Gly Asp Glu Ile Lys Val Ile Ile Val Glu Thr Asp Lys
340 345 350
Asn His Pro Phe Ala Val Glu Glu Leu Leu Met Pro Ile Leu Pro Ile
355 360 365
Val Lys Val Lys Asp Ala Leu Glu Gly Ile Lys Val Ala Lys Glu Leu
370 375 380
Glu Arg Gly Leu Arg His Thr Ala Val Ile His Ser Lys Asn Ile Asp
385 390 395 400
Ile Leu Thr Lys Tyr Ala Arg Glu Met Glu Thr Thr Ile Leu Val Lys
405 410 415
Asn Gly Pro Ser Tyr Ala Gly Ile Gly Ile Gly Gly Glu Gly His Val
420 425 430
Thr Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys
435 440 445
Ser Phe Ala Arg Asn Arg Arg Cys Val Leu Val Gly Gly Phe Ser Ile
450 455 460
Lys
465
<210> 159
<211> 467
<212> PRT
<213> Thermoanaerobacterium xylanolyticum LX-11
<400> 159
Met Lys Val Lys Glu Glu Asp Ile Glu Ala Ile Val Lys Lys Val Leu
1 5 10 15
Ser Glu Phe Asn Leu Glu Lys Thr Thr Ser Lys Tyr Gly Asp Val Gly
20 25 30
Ile Phe Gln Asp Met Asn Asp Ala Ile Ser Ala Ala Lys Asp Ala Gln
35 40 45
Lys Lys Leu Arg Asn Met Pro Met Glu Ser Arg Glu Lys Ile Ile Gln
50 55 60
Asn Ile Arg Lys Lys Ile Met Glu Asn Lys Lys Ile Leu Ala Glu Met
65 70 75 80
Gly Val Arg Glu Thr Gly Met Gly Arg Val Glu His Lys Ile Val Lys
85 90 95
His Glu Leu Val Ala Leu Lys Thr Pro Gly Thr Glu Asp Ile Thr Thr
100 105 110
Thr Ala Trp Ser Gly Asp Lys Gly Leu Thr Leu Val Glu Met Gly Pro
115 120 125
Phe Gly Val Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Ser Glu Thr
130 135 140
Val Leu Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ser Val Val
145 150 155 160
Phe Asn Pro His Pro Gly Ala Val Asn Val Ser Asn Tyr Ala Val Lys
165 170 175
Leu Val Asn Glu Ala Ala Met Glu Ala Gly Gly Pro Glu Asn Leu Val
180 185 190
Val Ser Val Glu Lys Pro Thr Leu Glu Thr Gly Asn Val Met Phe Lys
195 200 205
Ser Ser Asp Val Ser Leu Leu Val Ala Thr Gly Gly Pro Gly Val Val
210 215 220
Thr Ala Val Leu Ser Ser Gly Lys Arg Ala Ile Gly Ala Gly Ala Gly
225 230 235 240
Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Lys Lys Ala Ala
245 250 255
Lys Asp Ile Ile Asp Gly Ala Thr Phe Asp Asn Asn Leu Pro Cys Ile
260 265 270
Ala Glu Lys Glu Val Val Ser Val Asp Lys Ile Thr Asp Glu Leu Ile
275 280 285
Tyr Tyr Met Gln Lys Asn Gly Cys Tyr Lys Ile Glu Gly Arg Glu Ile
290 295 300
Glu Lys Leu Ile Glu Leu Val Leu Asp His Glu Gly Gly Lys Thr Thr
305 310 315 320
Leu Asn Arg Lys Trp Val Gly Lys Asp Ala His Leu Ile Leu Lys Ala
325 330 335
Ile Gly Ile Asp Ala Asp Glu Ser Val Arg Cys Ile Ile Phe Glu Ala
340 345 350
Glu Lys Asp Asn Pro Leu Val Val Glu Glu Leu Met Met Pro Ile Leu
355 360 365
Gly Ile Val Arg Ala Lys Asn Val Asp Glu Ala Ile Met Ile Ala Thr
370 375 380
Glu Leu Glu His Gly Asn Arg His Ser Ala His Met His Ser Lys Asn
385 390 395 400
Ile Asp Asn Leu Thr Lys Phe Gly Lys Ile Ile Asp Thr Ala Ile Phe
405 410 415
Val Lys Asn Ala Pro Ser Tyr Ala Ala Leu Gly Tyr Gly Gly Glu Gly
420 425 430
Tyr Cys Thr Phe Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser
435 440 445
Ala Arg Thr Phe Thr Lys Ser Arg Arg Cys Val Leu Ala Asp Gly Leu
450 455 460
Ser Ile Arg
465
<210> 160
<400> 160
000
<210> 161
<400> 161
000
<210> 162
<400> 162
000
<210> 163
<400> 163
000
<210> 164
<211> 441
<212> PRT
<213> Listeria monocytogenes
<400> 164
Met Thr Lys Gly Ala Lys Ser Gly Val Phe Asp Thr Val Asp Glu Ala
1 5 10 15
Val Gln Ala Ala Val Ile Ala Gln Asn Ser Tyr Lys Glu Lys Ser Leu
20 25 30
Glu Glu Arg Arg Asn Val Val Lys Ala Ile Arg Glu Ala Leu Tyr Pro
35 40 45
Glu Ile Glu Ser Ile Ala Ala Arg Ala Val Ala Glu Thr Gly Met Gly
50 55 60
Asn Val Ala Asp Lys Ile Leu Lys Asn Thr Leu Ala Ile Glu Lys Thr
65 70 75 80
Pro Gly Val Glu Asp Leu Tyr Thr Glu Val Ala Thr Gly Asp Asn Gly
85 90 95
Met Thr Leu Tyr Glu Leu Ser Pro Tyr Gly Val Ile Gly Ala Val Ala
100 105 110
Pro Ser Thr Asn Pro Thr Glu Thr Leu Ile Cys Asn Thr Ile Gly Met
115 120 125
Leu Ala Ala Gly Asn Ala Val Phe Tyr Ser Pro His Pro Gly Ala Lys
130 135 140
Asn Ile Ser Leu Trp Leu Ile Glu Lys Leu Asn Thr Ile Val Arg Glu
145 150 155 160
Ser Cys Gly Val Asp Asn Leu Val Val Thr Val Glu Lys Pro Ser Ile
165 170 175
Gln Ala Ala Gln Glu Met Met Asn His Pro Lys Val Pro Leu Leu Val
180 185 190
Ile Thr Gly Gly Pro Gly Val Val Leu Gln Ala Met Gln Ser Gly Lys
195 200 205
Lys Val Ile Gly Ala Gly Ala Gly Asn Pro Pro Ser Ile Val Asp Glu
210 215 220
Thr Ala Asn Ile Glu Lys Ala Ala Ala Asp Ile Val Asp Gly Ala Ser
225 230 235 240
Phe Asp His Asn Ile Leu Cys Ile Ala Glu Lys Ser Val Val Ala Val
245 250 255
Asp Ser Ile Ala Asp Phe Leu Met Phe Gln Met Glu Lys Asn Gly Ala
260 265 270
Leu His Val Thr Asn Pro Ser Asp Ile Gln Lys Leu Glu Lys Val Ala
275 280 285
Val Thr Asp Lys Gly Val Thr Asn Lys Lys Leu Val Gly Lys Ser Ala
290 295 300
Ser Glu Ile Leu Lys Glu Ala Gly Ile Ala Cys Asp Phe Ser Pro Arg
305 310 315 320
Leu Ile Ile Val Glu Thr Glu Lys Thr His Pro Phe Ala Thr Val Glu
325 330 335
Leu Leu Met Pro Ile Val Pro Val Val Arg Val Pro Asn Phe Glu Glu
340 345 350
Ala Leu Glu Val Ala Ile Glu Leu Glu Gln Gly Leu His His Thr Ala
355 360 365
Thr Met His Ser Gln Asn Ile Ser Arg Leu Asn Lys Ala Ala Arg Asp
370 375 380
Met Gln Thr Ser Ile Phe Val Lys Asn Gly Pro Ser Phe Ala Gly Leu
385 390 395 400
Gly Phe Arg Gly Glu Gly Ser Thr Thr Phe Thr Ile Ala Thr Pro Thr
405 410 415
Gly Glu Gly Thr Thr Thr Ala Arg His Phe Ala Arg Arg Arg Arg Cys
420 425 430
Val Leu Thr Asp Gly Phe Ser Ile Arg
435 440
<210> 165
<211> 481
<212> PRT
<213> Clostridium lavalense
<400> 165
Met Glu Ile Glu Thr Arg Asp Ile Glu Arg Ile Val Arg Gln Val Met
1 5 10 15
Ala Val Met Glu Gln Gln Gly Thr Ile Ala Gly Gly Ala Tyr Pro Pro
20 25 30
Ala Pro Gly Thr Pro Ala Pro Arg Gly Asp Asn Gly Val Phe Glu Arg
35 40 45
Val Glu Asp Ala Ile Asp Ala Ala Tyr Ala Ala Gly Arg Glu Trp Ala
50 55 60
Phe His Tyr Lys Val Glu Asp Arg Arg Arg Val Ile Glu Ala Ile Arg
65 70 75 80
Val Met Ala Arg Glu Asn Ala Arg Thr Leu Ala Gln Met Val Arg Asp
85 90 95
Glu Thr Gly Met Gly Arg Met Glu Asp Lys Val Glu Lys His Leu Ala
100 105 110
Val Ala Asp Lys Thr Pro Gly Val Glu Cys Leu Thr Thr Asp Ala Ile
115 120 125
Ser Gly Asp Gly Gly Leu Met Ile Glu Glu Tyr Ala Pro Phe Gly Val
130 135 140
Ile Gly Ala Ile Thr Pro Ser Thr Asn Pro Thr Glu Thr Val Ile His
145 150 155 160
Asn Thr Ile Ser Met Ile Ala Gly Gly Asn Ser Val Val Phe Asn Val
165 170 175
His Pro Gly Ala Lys Lys Cys Cys Ala Phe Cys Leu Gln Leu Leu His
180 185 190
Lys Thr Ile Val Glu Asn Gly Gly Pro Ala Asn Leu Ile Thr Met Gln
195 200 205
Arg Glu Pro Thr Met Asp Ala Val Asn Lys Met Thr Ser Ser Pro Lys
210 215 220
Ile Arg Leu Met Val Gly Thr Gly Gly Met Gly Met Val Asn Ala Leu
225 230 235 240
Leu Arg Ser Gly Lys Lys Thr Ile Gly Ala Gly Ala Gly Asn Pro Pro
245 250 255
Val Ile Val Asp Asp Thr Ala Asp Val Lys Leu Ala Ala Arg Glu Leu
260 265 270
Tyr Trp Gly Ala Ser Phe Asp Asn Asn Leu Phe Cys Phe Ala Glu Lys
275 280 285
Glu Val Phe Val Met Glu Ala Ser Ala Asp Gly Leu Ile Arg Gly Leu
290 295 300
Val Glu Gln Gly Ala Tyr Leu Leu Thr Pro Ala Glu Thr Glu Ala Ile
305 310 315 320
Val Lys Leu Ala Leu Ile Gln Lys Asp Gly Lys Tyr Glu Val Asn Lys
325 330 335
Lys Trp Val Gly Lys Asp Ala Gly Leu Phe Leu Lys Ala Ile Gly Val
340 345 350
Ser Gly His Glu Asn Thr Arg Leu Leu Ile Cys Asp Val Pro Lys Cys
355 360 365
His Pro Tyr Val Met Val Glu Gln Leu Met Pro Val Leu Pro Ile Val
370 375 380
Arg Cys Arg Thr Phe Asp Glu Cys Ile Gln Cys Ser Val Glu Ala Glu
385 390 395 400
Gln Gly Asn Arg His Thr Ser Ser Ile Phe Ser Thr Asn Val Tyr Asn
405 410 415
Met Thr Lys Phe Gly Lys Glu Ile Glu Thr Thr Ile Tyr Val Lys Asn
420 425 430
Gly Ala Thr Leu Arg Gly Leu Gly Ile Gly Gly Glu Gly His Thr Thr
435 440 445
Met Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Cys Ala Arg Ser
450 455 460
Phe Thr Arg Arg Arg Arg Cys Met Leu Ala Glu Gly Gly Leu Arg Ile
465 470 475 480
Ile
<210> 166
<211> 474
<212> PRT
<213> Acetanaerobacterium elongatum
<400> 166
Met Glu Phe Ala Val Asn Glu Ile Ser Met Ile Val Glu Gln Val Leu
1 5 10 15
Lys Asn Leu Asp Leu Ser Lys Val Ser Ala Gly Asn Ala Pro Ala Ser
20 25 30
Pro Lys Gly Asp Tyr Gly Val Phe Glu Asn Val Glu Asp Ala Ile Glu
35 40 45
Ala Ala Tyr Gln Ala Gln Lys Ile Tyr Leu Asp Lys Phe Gln Val Lys
50 55 60
Asp Arg Gln Arg Ile Ile Ala Ala Ile Arg Lys Val Cys Arg Glu Asn
65 70 75 80
Ala Glu Thr Leu Ala Arg Met Val Arg Glu Glu Ser Lys Met Gly Arg
85 90 95
Tyr Glu Asp Lys Ile Gln Lys His Leu Ala Val Ile Asp Asn Thr Pro
100 105 110
Gly Pro Glu Cys Leu Thr Thr Asp Ala Ile Ser Gly Asp Ser Gly Leu
115 120 125
Met Leu Glu Glu Tyr Ala Pro Phe Gly Leu Ile Gly Ala Ile Thr Pro
130 135 140
Val Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn Thr Ile Ser Met Ile
145 150 155 160
Ser Gly Gly Asn Ser Val Val Phe Asn Val His Pro Ser Ala Lys Asn
165 170 175
Val Cys Ala Tyr Cys Leu Arg Leu Ile Asn Lys Thr Ile Ile Asp Asn
180 185 190
Gly Gly Pro Ala Asn Leu Ile Thr Met Ala Lys Glu Pro Thr Met Asp
195 200 205
Thr Val Lys Ala Ile Ser Ser Ser Pro Lys Val Arg Leu Met Val Gly
210 215 220
Thr Gly Gly Met Pro Met Val Asn Ala Leu Leu Arg Ser Gly Lys Lys
225 230 235 240
Val Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile Val Asp Asn Thr
245 250 255
Ala Asp Ile Lys Lys Ala Ala Lys Asp Ile Tyr Tyr Gly Ala Ser Phe
260 265 270
Asp Asn Asn Leu Leu Cys Leu Ala Glu Lys Glu Val Phe Val Leu Asp
275 280 285
Glu Val Ala Asn Gln Phe Ile Tyr Asn Met Val Glu Glu Gly Ala Tyr
290 295 300
Leu Leu Asn Gly Val Gln Leu Glu Lys Ile Leu Asn Leu Val Phe Lys
305 310 315 320
Phe Asp Gly Lys Tyr Asp Val Asn Lys Lys Trp Val Gly Gln Asp Ala
325 330 335
Gly Lys Met Leu Asp Ala Ile Gly Val Glu Gly Lys Ser Asp Thr Arg
340 345 350
Leu Leu Ile Cys Glu Val Pro His Asp His Pro Phe Val Met Val Glu
355 360 365
Gln Leu Met Pro Val Leu Pro Ile Val Arg Cys Arg Asn Leu Asp Glu
370 375 380
Ala Ile Glu Tyr Ala Tyr Ile Ala Glu Ser Gly Asn Arg His Thr Ala
385 390 395 400
Ser Met Phe Ser Lys Asn Val Asp Asn Met Thr Arg Phe Ala Arg Lys
405 410 415
Ile Glu Thr Thr Ile Phe Val Lys Asn Gly Pro Thr Leu Asn Gly Val
420 425 430
Gly Ile Gly Gly Glu Gly Tyr Ala Thr Met Thr Ile Ala Gly Pro Thr
435 440 445
Gly Glu Gly Leu Thr Cys Ala Lys Ser Phe Thr Arg Arg Arg Arg Cys
450 455 460
Met Leu Ser Asp Gly Gly Leu Arg Val Ile
465 470
<210> 167
<211> 464
<212> PRT
<213> Alkaliphilus peptidifermentans DSM
<400> 167
Met Val Glu Glu Leu Lys Ile Glu Glu Ile Ile Arg Arg Val Met Lys
1 5 10 15
Glu Ile Ser Ser Lys Asn Glu Thr Gly Glu Glu Gly Ala Tyr Gly Ile
20 25 30
Phe Gln Asp Met Asn Asp Ala Val Asp Ala Ala Tyr Ile Ala Gln Lys
35 40 45
Glu Leu Ile Gly Phe Asn Leu Glu Thr Arg Gly Lys Phe Ile Glu Ala
50 55 60
Met Arg Gln Ala Ala Arg Gln Asn Val Glu Leu Leu Ser Lys Met Ala
65 70 75 80
His Glu Glu Thr Asp Met Gly Arg Tyr Glu Asp Lys Ile Leu Lys Asn
85 90 95
Arg Leu Ala Ile Glu Lys Thr Pro Gly Ile Glu Asp Leu Gly Ser Glu
100 105 110
Val Phe Thr Gly Asp Asp Gly Leu Thr Leu Ile Glu Leu Ser Pro Tyr
115 120 125
Gly Val Ile Gly Ser Ile Ser Pro Val Thr Asn Pro Ser Glu Thr Ile
130 135 140
Ile Cys Asn Ala Ile Gly Met Ile Ala Ala Gly Asn Ala Val Ala Phe
145 150 155 160
Ser Pro His Pro Ser Ala Lys Lys Thr Ser Leu Lys Thr Ile Glu Ile
165 170 175
Leu Asn Lys Gly Ile Ile Glu Ala Gly Gly Pro Lys Asn Leu Ile Val
180 185 190
Ala Val Glu Asn Pro Ser Ile Glu Gln Ala Glu Ala Met Met Lys His
195 200 205
Lys Lys Ile Asn Met Leu Val Ala Thr Gly Gly Pro Gly Val Val Lys
210 215 220
Ser Val Leu Ser Ser Gly Lys Lys Ala Ile Gly Ala Gly Ala Gly Asn
225 230 235 240
Pro Pro Ala Val Val Asp Glu Thr Ala Asp Ile Glu Lys Ala Ala Arg
245 250 255
Asp Ile Ile Ala Gly Cys Ser Phe Asp Asn Asn Leu Pro Cys Val Ala
260 265 270
Glu Lys Glu Val Ile Val Val Asp Ser Val Ala Asp Tyr Leu Ile Phe
275 280 285
Asn Met Lys Lys Asn Gly Ala Tyr Glu Leu Lys Glu Lys Asp Leu Ile
290 295 300
Glu Gln Leu Glu Lys Leu Val Val Asn Glu Lys Gly Tyr Pro Val Lys
305 310 315 320
Glu Phe Val Gly Lys Asn Ala Asp Tyr Ile Leu Ser Lys Met Gly Ile
325 330 335
Lys Cys Asp Asp Ser Ile Arg Ala Ile Ile Val Glu Val Pro Lys Ser
340 345 350
His Pro Phe Val Val Gly Glu Leu Met Met Pro Val Leu Pro Ile Val
355 360 365
Arg Val Asn Asp Val Glu Glu Ala Ile Lys Leu Ala Val Glu Val Glu
370 375 380
His Gly Phe Lys His Thr Ala Ile Met His Ser Lys Asn Ile Asp Arg
385 390 395 400
Leu Ser Lys Phe Ala Lys Glu Ile Gln Thr Thr Ile Phe Val Lys Asn
405 410 415
Gly Pro Ser Phe Ala Gly Ile Gly Val Gly Gly Glu Gly Tyr Ala Thr
420 425 430
Phe Thr Ile Ala Gly Pro Thr Gly Glu Gly Leu Thr Ser Ala Lys Ser
435 440 445
Phe Ala Arg Arg Arg Arg Cys Thr Leu Val Gly Gly Phe Ser Ile Lys
450 455 460
<210> 168
<400> 168
000
<210> 169
<400> 169
000
<210> 170
<400> 170
000
<210> 171
<400> 171
000
<210> 172
<400> 172
000
<210> 173
<400> 173
000
<210> 174
<400> 174
000
<210> 175
<400> 175
000
<210> 176
<211> 481
<212> PRT
<213> Clostridium populeti
<400> 176
Met Asp Ile Ser Ser Gln Glu Ile Glu Ala Ile Val Arg Lys Val Ile
1 5 10 15
Ala Gly Ile Asn Pro Ala Thr Asn Val Thr Pro Asp Ile Pro Ala Ile
20 25 30
Lys Ser Pro Lys Tyr Thr Gly Asp Asn Gly Val Phe Glu Arg Val Glu
35 40 45
Glu Ala Val Glu Ala Ala Trp Lys Ala Gln Arg Asp Trp Val Thr Asn
50 55 60
Tyr Lys Val Glu Asp Arg His Arg Ile Val Glu Ala Ile Arg Arg Cys
65 70 75 80
Gly Arg Asp His Val Glu Glu Trp Ser His Leu Ile Val Glu Glu Thr
85 90 95
Gln Met Gly Arg Tyr Glu Asp Lys Val Glu Lys His Leu Ala Val Ile
100 105 110
Asn Lys Thr Pro Gly Pro Glu Cys Leu Thr Thr Glu Ala Ile Ser Gly
115 120 125
Asp Ala Gly Leu Met Ile Glu Glu Tyr Ala Pro Phe Gly Val Ile Gly
130 135 140
Ser Ile Thr Pro Thr Thr Asn Pro Thr Glu Thr Met Ile His Asn Thr
145 150 155 160
Ile Ser Met Ile Ser Gly Gly Asn Ser Ile Val Phe Asn Val His Pro
165 170 175
Arg Ala Lys Arg Val Cys Ala Glu Cys Leu Gln Ala Leu His Lys Ala
180 185 190
Ile Val Asp Ala Gly Gly Pro Ala Asn Leu Ile Thr Met Leu Arg Glu
195 200 205
Pro Thr Met Asp Thr Val Asp Met Leu Thr Ser Asn Pro Lys Val Arg
210 215 220
Leu Met Thr Gly Thr Gly Gly Met Gly Met Val Asn Ala Leu Leu Arg
225 230 235 240
Ser Gly Lys Lys Cys Ile Gly Ala Gly Ala Gly Asn Pro Pro Val Ile
245 250 255
Val Asp Glu Thr Ala Asp Val Glu Leu Ala Ala Arg Lys Ile Tyr Glu
260 265 270
Gly Ala Ser Phe Asp Asn Asn Ile Leu Cys Phe Ala Glu Lys Glu Val
275 280 285
Phe Val Val Ser Pro Asn Tyr Glu Gly Phe Ile His Asn Ile Gln Lys
290 295 300
Gln Gly Ala Tyr Leu Leu Asn Asn Ser Gln Val Glu Ala Leu Val Lys
305 310 315 320
Ile Cys Leu Glu Pro Asn Lys Asn Gln Ser Gly Tyr Glu Val Asn Lys
325 330 335
Lys Trp Val Gly Lys Asn Ala Ala Leu Ile Leu Ala Gln Ile Gly Val
340 345 350
Gln Val Glu Asp Ser Cys Arg Leu Ala Val Cys Glu Val Pro Ala Asp
355 360 365
His Pro Phe Val Leu Val Glu Gln Met Met Pro Val Leu Pro Ile Val
370 375 380
Arg Cys Ser Thr Phe Glu Glu Ala Met Glu Lys Ala Val Ile Ala Glu
385 390 395 400
Gln Gly Asn Arg His Thr Ser Ser Ile Phe Ser Lys Asp Val Asp His
405 410 415
Met Thr Arg Phe Ala Arg Leu Ile Glu Thr Thr Ile Tyr Val Lys Asn
420 425 430
Ser Cys Thr Lys Ala Gly Val Gly Ile Gly Gly Glu Gly His Cys Thr
435 440 445
Met Thr Ile Ala Gly Pro Thr Gly Glu Gly Ile Thr Asn Ala Lys Ser
450 455 460
Phe Cys Arg Arg Arg Arg Cys Met Leu Ala Glu Gly Gly Leu Arg Ile
465 470 475 480
Ile
<210> 177
<400> 177
000
<210> 178
<400> 178
000
<210> 179
<400> 179
000
<210> 180
<400> 180
000
<210> 181
<400> 181
000
<210> 182
<400> 182
000
<210> 183
<400> 183
000
<210> 184
<400> 184
000
<210> 185
<400> 185
000
<210> 186
<400> 186
000
<210> 187
<211> 450
<212> PRT
<213> Candidatus Bacteroides periocalifornicus
<400> 187
Met Thr Ile Ala Glu Met Val Ala Lys Ala Arg Val Ala Gln Ala Glu
1 5 10 15
Phe Glu Lys Asn Phe Asp Gln Ala Lys Thr Asp Ala Val Val Arg Glu
20 25 30
Ile Gly Lys Thr Val Phe Asp Asn Ala Glu Met Leu Ala Lys Met Ala
35 40 45
Val Glu Glu Thr Arg Met Gly Val Tyr Glu Asp Lys Val Ala Lys Asn
50 55 60
Lys Gly Lys Ala Arg Gly Val Trp Tyr Asp Leu Lys Gly Lys Lys Ser
65 70 75 80
Met Gly Val Leu Ser Val Asp Pro Glu Thr Asp Leu Ile Thr Met Leu
85 90 95
Lys Pro Val Gly Val Val Ala Ala Ile Thr Pro Thr Thr Asn Pro Ile
100 105 110
Val Thr Pro Met Ser Lys Ser Met Phe Ala Val Lys Gly Lys Asn Ala
115 120 125
Ile Ile Val Ala Pro His Pro Arg Ser Lys Lys Cys Thr Ala Lys Thr
130 135 140
Ile Glu Leu Ile Asn Lys Ala Ile Ala Lys Phe Gly Val Pro Lys Asp
145 150 155 160
Leu Ile Gln Val Ile Glu Glu Pro Ser Ile Pro Leu Thr Gln Glu Leu
165 170 175
Met Ala Ser Cys Asp Val Val Leu Ala Thr Gly Gly Met Gly Met Val
180 185 190
Lys Ala Ala Tyr Ser Ser Gly Lys Pro Ser Tyr Gly Val Gly Ala Gly
195 200 205
Asn Val Gln Val Ile Ile Asp Arg Gly Val Asp Tyr Asp Lys Ala Ala
210 215 220
Ala Thr Ile Ile Lys Gly Arg Ile Phe Asp Asn Gly Ile Ile Cys Ser
225 230 235 240
Gly Glu Gln Ser Phe Ile Tyr Pro Lys Asp Glu Lys Ala Lys Val Phe
245 250 255
Asp Ala Phe Lys Lys Asn Gly Ala Tyr Ile Val Ala Asp Ala Asp His
260 265 270
Asp Lys Val Val Asn Ala Leu Phe Glu Asp Gly His Ile Ala Gly Asp
275 280 285
Val Val Gly Gln Ser Val Gln Phe Val Ala Lys Lys Ala Gly Leu Asn
290 295 300
Val Pro Ala Asp Ala Arg Val Ile Val Val Glu Ala Lys Gly Val Gly
305 310 315 320
Ala Gln Asp Pro Ile Cys Lys Glu Lys Met Cys Pro Val Leu Ala Ala
325 330 335
Phe Gly Tyr Asp Lys Phe Glu Glu Ala Ile Gln Ile Ala Lys Thr Asn
340 345 350
Leu Leu Asn Glu Gly Asn Gly His Ser Ala Gly Ile His Ser Asn Asn
355 360 365
Glu Glu His Ile Arg Met Val Gly Glu Gly Leu Thr Val Ser Arg Val
370 375 380
Val Val Asn Ala Pro Val Ser Thr Thr Ala Gly Gly Ala Ile Gly Ser
385 390 395 400
Gly Leu Ala Val Thr Asn Thr Leu Gly Cys Gly Thr Trp Gly Asn Asn
405 410 415
Thr Leu Ser Glu Asn Leu Thr Tyr Lys His Leu Leu Asn Thr Thr Arg
420 425 430
Val Ala Arg Ile Ser Pro Lys Val His Gln Pro Thr Asp Glu Glu Leu
435 440 445
Trp Gly
450
<210> 188
<211> 480
<212> PRT
<213> Anaerocolumna aminovalerica
<400> 188
Met Glu Phe Gly Thr Lys Glu Ile Ser Met Ile Val Glu Gln Val Leu
1 5 10 15
Lys Asn Leu Glu Glu Asn Asn Leu Ile Ser Thr Lys Lys Thr Ser Asn
20 25 30
Ser Gly Leu Tyr Ser Asp Lys Gly Asp Tyr Gly Val Phe Glu Arg Val
35 40 45
Glu Asp Ala Ile Asp Ala Ala Tyr Glu Ala Gln Lys Ile Tyr Leu Asp
50 55 60
Asn Phe Lys Ile Lys Asp Arg Gln Arg Leu Ile Ala Ala Ile Arg Lys
65 70 75 80
Val Ser Ile Glu Asn Ala Glu Thr Leu Ala Arg Met Ile Val Glu Glu
85 90 95
Ser Lys Met Gly Arg Val Glu Asp Lys Val Lys Lys His Leu Ala Val
100 105 110
Ile Glu Asn Thr Pro Gly Pro Glu Cys Leu Thr Thr Asp Ala Ile Thr
115 120 125
Gly Asp Gly Gly Leu Met Ile Glu Glu Tyr Ala Pro Phe Gly Leu Ile
130 135 140
Gly Ala Ile Thr Pro Val Thr Asn Pro Thr Glu Thr Ile Ile Asn Asn
145 150 155 160
Thr Ile Ser Met Ile Ser Gly Gly Asn Gly Ile Val Phe Asn Val His
165 170 175
Pro Ser Ala Lys Lys Val Cys Ala Tyr Cys Leu Gln Phe Ile Asn Lys
180 185 190
Thr Ile Ile Glu Asn Gly Gly Pro Ala Asn Leu Ile Thr Met Val Lys
195 200 205
Glu Pro Thr Met Glu Thr Cys Asn Ile Ile Thr Gln Ser Pro Lys Val
210 215 220
Arg Leu Met Val Gly Thr Gly Gly Met Gly Met Val Asn Ser Leu Leu
225 230 235 240
Arg Ser Gly Lys Lys Thr Ile Gly Ala Gly Ala Gly Asn Pro Pro Val
245 250 255
Ile Val Asp Glu Thr Ala Asp Ile Lys Lys Ala Ala Lys Asp Ile Tyr
260 265 270
Tyr Gly Ala Ser Phe Asp Asn Asn Leu Leu Cys Leu Ala Glu Lys Glu
275 280 285
Val Phe Val Leu Glu Glu Val Ala Asn Asp Phe Ile Tyr Asn Met Val
290 295 300
Asp Glu Gly Ala Phe Leu Leu Asn Gly Ala Gln Leu Glu Ala Ile Thr
305 310 315 320
Asn Leu Val Leu Lys Tyr Glu Asn Gly Lys Tyr Asp Ile Asn Lys Lys
325 330 335
Trp Val Gly Gln Asp Ala Gly Lys Met Leu Glu Ala Ile Gly Ile Thr
340 345 350
Gly Lys Ser Asp Thr Arg Leu Leu Ile Cys Asp Val Pro Tyr Asp Asn
355 360 365
Pro Phe Val Leu Leu Glu Gln Leu Met Pro Val Leu Pro Ile Val Arg
370 375 380
Cys Lys Asn Leu Asn Gln Ala Ile Asp Tyr Ala Met Ile Ala Glu Ser
385 390 395 400
Gly Asn Arg His Thr Ala Ser Met Phe Ser Lys Asn Val Asp Asn Met
405 410 415
Thr Arg Phe Ala Arg Lys Ile Glu Thr Thr Ile Phe Val Lys Asn Gly
420 425 430
Cys Thr Leu Glu Gly Val Gly Ile Gly Gly Glu Gly Tyr Thr Thr Met
435 440 445
Thr Ile Ala Gly Pro Thr Gly Glu Gly Ile Thr Cys Ala Lys Ser Phe
450 455 460
Thr Arg Arg Arg Arg Cys Met Leu Ala Asp Gly Gly Leu Arg Ile Ile
465 470 475 480
<210> 189
<211> 364
<212> PRT
<213> Candida tropicalis
<400> 189
Met Ile Thr Ala Gln Ala Val Leu Tyr Thr Gln His Gly Glu Pro Lys
1 5 10 15
Asp Val Leu Phe Thr Gln Ser Phe Glu Ile Asp Asp Asp Asn Leu Ala
20 25 30
Pro Asn Glu Val Ile Val Lys Thr Leu Gly Ser Pro Val Asn Pro Ser
35 40 45
Asp Ile Asn Gln Ile Gln Gly Val Tyr Pro Ser Lys Pro Ala Lys Thr
50 55 60
Thr Gly Phe Gly Thr Thr Glu Pro Ala Ala Pro Cys Gly Asn Glu Gly
65 70 75 80
Leu Phe Glu Val Ile Lys Val Gly Ser Asn Val Leu Ser Leu Glu Ala
85 90 95
Gly Asp Trp Val Ile Pro Ser His Val Asn Phe Gly Thr Trp Arg Thr
100 105 110
His Ala Leu Gly Asn Asp Asp Asp Phe Ile Lys Leu Pro Asn Pro Ala
115 120 125
Gln Ser Lys Ala Asn Gly Lys Pro Asn Gly Leu Thr Ile Asn Gln Gly
130 135 140
Ala Thr Ile Ser Val Asn Pro Leu Thr Ala Tyr Leu Met Leu Thr His
145 150 155 160
Tyr Val Lys Leu Thr Pro Gly Lys Asp Trp Phe Ile Gln Asn Gly Gly
165 170 175
Thr Ser Ala Val Gly Lys Tyr Ala Ser Gln Ile Gly Lys Leu Leu Asn
180 185 190
Phe Asn Ser Ile Ser Val Ile Arg Asp Arg Pro Asn Leu Asp Glu Val
195 200 205
Val Ala Ser Leu Lys Glu Leu Gly Ala Thr Gln Val Ile Thr Glu Asp
210 215 220
Gln Asn Asn Ser Arg Glu Phe Gly Pro Thr Ile Lys Glu Trp Ile Lys
225 230 235 240
Gln Ser Gly Gly Glu Ala Lys Leu Ala Leu Asn Cys Val Gly Gly Lys
245 250 255
Ser Ser Thr Gly Ile Ala Arg Lys Leu Asn Asn Asn Gly Leu Met Leu
260 265 270
Thr Tyr Gly Gly Met Ser Phe Gln Pro Val Thr Ile Pro Thr Ser Leu
275 280 285
Tyr Ile Phe Lys Asn Phe Thr Ser Ala Gly Phe Trp Val Thr Glu Leu
290 295 300
Leu Lys Asn Asn Lys Glu Leu Lys Thr Leu Thr Leu Asn Gln Ile Ile
305 310 315 320
Ala Trp Tyr Glu Glu Gly Lys Leu Thr Asp Ala Lys Ser Ile Glu Thr
325 330 335
Leu Tyr Asp Gly Thr Lys Pro Leu His Glu Leu Tyr Gln Asp Gly Val
340 345 350
Ala Asn Ser Lys Asp Gly Lys Gln Leu Ile Thr Tyr
355 360
<210> 190
<211> 405
<212> PRT
<213> Euglena gracilis
<400> 190
Met Ala Met Phe Thr Thr Thr Ala Lys Val Ile Gln Pro Lys Ile Arg
1 5 10 15
Gly Phe Ile Cys Thr Thr Thr His Pro Ile Gly Cys Glu Lys Arg Val
20 25 30
Gln Glu Glu Ile Ala Tyr Ala Arg Ala His Pro Pro Thr Ser Pro Gly
35 40 45
Pro Lys Arg Val Leu Val Ile Gly Cys Ser Thr Gly Tyr Gly Leu Ser
50 55 60
Thr Arg Ile Thr Ala Ala Phe Gly Tyr Gln Ala Ala Thr Leu Gly Val
65 70 75 80
Phe Leu Ala Gly Pro Pro Thr Lys Gly Arg Pro Ala Ala Ala Gly Trp
85 90 95
Tyr Asn Thr Val Ala Phe Glu Lys Ala Ala Leu Glu Ala Gly Leu Tyr
100 105 110
Ala Arg Ser Leu Asn Gly Asp Ala Phe Asp Ser Thr Thr Lys Ala Arg
115 120 125
Thr Val Glu Ala Ile Lys Arg Asp Leu Gly Thr Val Asp Leu Val Val
130 135 140
Tyr Ser Ile Ala Ala Pro Lys Arg Thr Asp Pro Ala Thr Gly Val Leu
145 150 155 160
His Lys Ala Cys Leu Lys Pro Ile Gly Ala Thr Tyr Thr Asn Arg Thr
165 170 175
Val Asn Thr Asp Lys Ala Glu Val Thr Asp Val Ser Ile Glu Pro Ala
180 185 190
Ser Pro Glu Glu Ile Ala Asp Thr Val Lys Val Met Gly Gly Glu Asp
195 200 205
Trp Glu Leu Trp Ile Gln Ala Leu Ser Glu Ala Gly Val Leu Ala Glu
210 215 220
Gly Ala Lys Thr Val Ala Tyr Ser Tyr Ile Gly Pro Glu Met Thr Trp
225 230 235 240
Pro Val Tyr Trp Ser Gly Thr Ile Gly Glu Ala Lys Lys Asp Val Glu
245 250 255
Lys Ala Ala Lys Arg Ile Thr Gln Gln Tyr Gly Cys Pro Ala Tyr Pro
260 265 270
Val Val Ala Lys Ala Leu Val Thr Gln Ala Ser Ser Ala Ile Pro Val
275 280 285
Val Pro Leu Tyr Ile Cys Leu Leu Tyr Arg Val Met Lys Glu Lys Gly
290 295 300
Thr His Glu Gly Cys Ile Glu Gln Met Val Arg Leu Leu Thr Thr Lys
305 310 315 320
Leu Tyr Pro Glu Asn Gly Ala Pro Ile Val Asp Glu Ala Gly Arg Val
325 330 335
Arg Val Asp Asp Trp Glu Met Ala Glu Asp Val Gln Gln Ala Val Lys
340 345 350
Asp Leu Trp Ser Gln Val Ser Thr Ala Asn Leu Lys Asp Ile Ser Asp
355 360 365
Phe Ala Gly Tyr Gln Thr Glu Phe Leu Arg Leu Phe Gly Phe Gly Ile
370 375 380
Asp Gly Val Asp Tyr Asp Gln Pro Val Asp Val Glu Ala Asp Leu Pro
385 390 395 400
Ser Ala Ala Gln Gln
405

Claims (45)

1.一种非天然存在的微生物有机体，所述非天然存在的微生物有机体包含编码醛脱氢酶的至少一种外源核酸，所述醛脱氢酶与己二酰辅酶A反应形成己二酸-半缩醛，其中所述醛脱氢酶对作为底物的己二酰辅酶A比对作为底物的琥珀酰辅酶A、乙酰辅酶A或二者具有更大的催化效率，和/或所述醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物、乙酰辅酶A底物、或琥珀酰辅酶A和乙酰辅酶A具有更高的转换数。
2.根据权利要求1所述的非天然存在的微生物有机体，其中所述醛脱氢酶不包含SEQID NO：1、SEQ ID NO：2或SEQ ID NO：3的氨基酸序列。
3.根据权利要求2所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物具有更大的催化效率。
4.根据权利要求1-3中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物的催化效率是对琥珀酰辅酶A底物的特异性的至少两倍。
5.根据权利要求1-4中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物比对琥珀酰辅酶A底物具有更高的转换数。
6.根据权利要求1-5中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物比对乙酰辅酶A底物具有更大的催化效率。
7.根据权利要求1-6中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物的催化效率是对乙酰辅酶A底物的催化效率的至少五倍。
8.根据权利要求1-7中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶对己二酰辅酶A底物比对乙酰辅酶A底物具有更高的转换数。
9.根据权利要求1-8中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体比对照微生物有机体将更多的己二酰辅酶A转化为己二酸半缩醛，除了所述对照微生物有机体不包含编码醛脱氢酶的外源核酸之外，所述对照微生物有机体与所述非天然存在的微生物有机体基本相同。
10.根据权利要求1-9中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶包含与SEQ ID NO：4、7、11、15、17、19、24、25、27、28、31-33、36、38、40-42、44、45、47、53、58-60、63、65-67、74、75、77、80、82、84、86-88、90、91、94、95、97、100、101、103、107、109、111、112、117、134、135、137、145、146、148-150、152、157-159、164-167、176、187和188中的任一个的至少25个连续氨基酸具有至少约60％的氨基酸序列一致性的氨基酸序列。
11.根据权利要求1-10中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶包含与SEQ ID NO：7、28、60或107中的任一个的至少25个连续氨基酸具有至少约60％的氨基酸序列一致性的氨基酸序列。
12.根据权利要求1-11中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶包含SEQ ID NO：7、28、60或107的氨基酸序列。
13.根据权利要求1-12中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶使用NADH作为辅因子。
14.根据权利要求1-10中任一项所述的非天然存在的微生物，其中所述醛脱氢酶包含与SEQ ID NO：53、77、82、94和152中的任一个的至少25个连续氨基酸具有至少约60％的氨基酸序列一致性的氨基酸序列。
15.根据权利要求14所述的非天然存在的微生物有机体，其中所述醛脱氢酶使用NADH、NADPH或两者作为辅因子。
16.根据权利要求1-15中任一项所述的非天然存在的微生物有机体，其中编码与己二酰辅酶A反应以形成己二酸-半缩醛的醛脱氢酶的所述至少一种外源核酸对于所述微生物有机体是异源的。
17.根据权利要求1-16中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体包含6-氨基己酸途径。
18.根据权利要求17所述的非天然存在的微生物有机体，其中所述6-氨基己酸途径包括：(i)转氨酶，(ii)6-氨基己酸脱氢酶，或(iii)转氨酶和6-氨基己酸脱氢酶两者。
19.根据权利要求17-18中任一项所述的非天然存在的微生物有机体，其中所述微生物有机体还包含编码6-氨基己酸途径酶中的一种或多种的一种或多种另外的外源核酸。
20.根据权利要求19所述的非天然存在的微生物有机体，其中编码6-氨基己酸途径酶中的一种或多种的外源核酸对于所述微生物有机体是异源的。
21.根据权利要求1-20中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体包含己二胺途径。
22.根据权利要求21所述的非天然存在的微生物有机体，其中所述己二胺途径包含：(i)6-氨基己酰辅酶A转移酶、(ii)6-氨基己酰辅酶A合酶、(iii)6-氨基己酰辅酶A还原酶、(iv)己二胺转氨酶、(v)己二胺脱氢酶、(v)或一种或多种酶(i)-(v)的组合。
23.根据权利要求21-22中任一项所述的非天然存在的微生物有机体，其中所述微生物有机体还包含编码己二胺途径酶中的一种或多种的一种或多种另外的外源核酸。
24.根据权利要求23所述的非天然存在的微生物有机体，其中编码己二胺途径酶中的一种或多种的外源核酸对于所述微生物有机体是异源的。
25.根据权利要求1-24中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体包含己内酰胺途径。
26.根据权利要求25所述的非天然存在的微生物有机体，其中所述己内酰胺途径包含氨基水解酶。
27.根据权利要求25-26中任一项所述的非天然存在的微生物有机体，其中所述微生物有机体还包含编码氨基水解酶的一种或多种另外的外源核酸。
28.根据权利要求27所述的非天然存在的微生物有机体，其中编码氨基水解酶的外源核酸对于所述微生物有机体是异源的。
29.根据权利要求1-28中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶来源于原核物种。
30.根据权利要求29所述的非天然存在的微生物有机体，其中所述醛脱氢酶来源于Acidaminococcus、Collinsella、Peptostreptococcaceae或Romboustsia。
31.根据权利要求1-30中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体包括Acinetobacter、Actinobacillus、Anaerobiospirillum、Aspergillus、Bacillus、Clostridium、Corynebacterium、Escherichia、Gluconobacter、Klebsiella、Kluyveromyces、Lactococcus、Lactobacillus、Mannheimia、Pichia、Pseudomonas、Rhizobium、Rhizopus、Saccharomyces、Schizosaccharomyces、Streptomyces和Zymomonas。
32.根据权利要求1-31中任一项所述的非天然存在的微生物有机体，其中所述非天然存在的微生物有机体包含两种、三种、四种、五种、六种或七种外源核酸，所述外源核酸各自编码6-氨基己酸途径、己二胺途径、己内酰胺途径、1,6-己二醇途径、己内酯途径或两种或更多种途径的组合的酶。
33.根据权利要求1-31中任一项所述的非天然存在的微生物有机体，其中所述醛脱氢酶还与6-氨基己酰辅酶A反应以形成6-氨基己酸半缩醛。
34.产生己二酸-半缩醛的方法，所述方法包括将权利要求1-33中任一项的非天然存在的微生物有机体培养足够的时间和条件以生产己二酸-半缩醛。
35.产生6-氨基己酸(6ACA)的方法，所述方法包括在产生6ACA的条件下将权利要求1-33中任一项所述的非天然存在的微生物有机体培养足够的时间段。
36.根据权利要求35所述的方法，所述方法还包括从微生物有机体、发酵液或两者中回收6ACA。
37.产生己二胺的方法，所述方法包括在产生己二胺的条件下将权利要求1-33中任一项所述的非天然存在的微生物有机体培养足够的时间段。
38.根据权利要求37所述的方法，所述方法还包括从微生物有机体、发酵液或两者中回收己二胺。
39.根据权利要求34-38中任一项所述的方法，其中所述非天然存在的微生物有机体包含两种、三种、四种、五种、六种或七种外源核酸序列，所述外源核酸序列各自编码己二胺途径酶。
40.产生己内酰胺的方法，所述方法包括在产生己内酯、1,6-己二醇或己内酰胺的条件下将权利要求1-33中任一项所述的非天然存在的微生物有机体培养足够的时间段。
41.根据权利要求40所述的方法，所述方法还包括从非天然存在的微生物有机体、发酵液或两者中回收己内酰胺。
42.根据权利要求34-41中任一项所述的方法，其中所述非天然存在的微生物有机体包含两种、三种、四种、五种、六种或七种外源核酸，所述外源核酸各自编码己内酰胺途径酶。
43.根据权利要求34-42中任一项所述的方法，其中所述培养在包含糖的发酵液中进行。
44.根据权利要求34-43中任一项所述的方法，其中所述非天然存在的微生物有机体包括Acinetobacter、Actinobacillus、Anaerobiospirillum、Aspergillus、Bacillus、Clostridium、Corynebacterium、Escherichia、Gluconobacter、Klebsiella、Kluyveromyces、Lactococcus、Lactobacillus、Mannheimia、Pichia、Pseudomonas、Rhizobium、Rhizopus、Saccharomyces、Schizosaccharomyces、Streptomyces和Zymomonas的物种。
45.由权利要求34-44中任一项的方法合成的生物衍生的6-氨基己酸、己二胺、1,6-己二醇、己内酯或己内酰胺。
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