CN116249778A - 编码氨基酸序列、编码氧化还原酶的多核苷酸 - Google Patents

编码氨基酸序列、编码氧化还原酶的多核苷酸 Download PDF

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CN116249778A
CN116249778A CN202180060850.9A CN202180060850A CN116249778A CN 116249778 A CN116249778 A CN 116249778A CN 202180060850 A CN202180060850 A CN 202180060850A CN 116249778 A CN116249778 A CN 116249778A
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S·沙费尔
T·布拉德特
S·奥斯瓦尔特
R·玛萨
B·R·斯文森
D·T·洛根
R·潘查伊坦
M·恩格福冲
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Abstract

本发明属于用于生产精细化学品的发酵方法领域，并且本发明涵盖了编码氨基酸序列的多核苷酸和使用氧化还原酶的发酵方法。

Description

编码氨基酸序列、编码氧化还原酶的多核苷酸
本发明属于用于生产精细化学品的发酵方法领域，并且本发明涵盖了多核苷酸，其编码氨基酸序列、编码氧化还原酶，以及使用氧化还原酶的发酵方法。
患有帕金森病的患者会出现震颤、僵硬或运动迟缓等症状，这是由于大脑基底神经节中的细胞死亡引起的低水平多巴胺导致的。由于多巴胺不能通过血脑屏障，其前体分子L-DOPA(L-二羟基苯丙氨酸)被用作减轻患者症状的药物。自其1977年首次推出以来，L-DOPA(左旋多巴)已发展成为排在第一个的帕金森治疗药物，全球市场量为600MT。
现有的生产方法基于化学合成，特别是不对称氢化或氢化和手性拆分。这是一个由来已久的制造方法，但转化率和整体效率都很差。或者，L-DOPA通过酶促转化产生，即丙酮酸和儿茶酚的酶促偶联。然而，这种酶促转化方法具有原料成本高的缺点。
文献中记载了使用来自大肠杆菌(Escherichia coli)(E.coli)的4-羟基苯乙酸3-单加氧酶HpaB和同源的4-羟基苯乙酸3-单加氧酶还原酶HpaC的发酵方法，并且由于更高的转化率、对映选择性和更经济的方法而可能是有利的。
为了实现具有经济上有吸引力的方法，至关重要的是，一方面优化酪氨酸生产力，另一方面通过增加HpaB活性将L-酪氨酸转化为L-DOPA。
Prieto等人,1993中记载了4-羟基苯乙酸单加氧酶的广泛的底物谱，并指出，与非天然底物L-酪氨酸的酶活性仅为与天然底物4-HPA(4-羟基苯乙酸)的酶活性的5％。因此，酶建模被认为是增加酶活性的有希望的选择。
Shen等人,Scientific Reports(2019)9:7087/https://doi.org/10.1038/s41598-019-43577-w,p.1-11记载了对来自大肠杆菌的黄素依赖性羟化酶(HpaB)的催化多功能性的结构见解。Shen等人提到来自大肠杆菌的4-羟基苯乙酸3-羟化酶(EcHpaB)能够有效地对多种酚类化合物进行邻羟基化，并显示出广泛的化学酶学应用的巨大潜力。为了了解其催化多功能性的结构和机制基础，Shen等人通过X射线晶体学阐明了EcHpaB的晶体结构，其揭示了覆盖活性位点的独特环状结构。Shen等人进一步对该环进行了诱变研究，以探究其在底物特异性和催化活性中的作用。他们的结果不仅表明该环具有很强的可塑性和对充分诱变的强抗性，而且还提出了一种柔性环，所述柔性环使底物能够进入并稳定结合活性位点，其可能是酶催化多功能性的关键因素。报告了不同EcHpaB突变体的环序列和结构。然而，没有记载对于L-DOPA的选择性得到增强。
Fordjour等人,Microbial Cell Factories(2019)18:74/https//doi.org/10.1186/s12934-019-1122-0,p.1-10记载了用于从D-葡萄糖从头生产L-DOPA的E.coliBL21(DE3)的代谢工程改造。通过缺失tyrR、ptsG、crr、pheA和pykF同时通过过表达galP和glk来引导碳流来工程改造E.coli BL21(DE3)。还过表达TktA和ppsA以增强E4P和PEP的积累。对HpaB应用位点定向诱变以优化其活性。鉴定了三个突变体G883R、G883A和L1231M，它们相对于野生型hpaB的L-DOPA产量分别显示出3.03倍、2.9倍和2.56倍增加的活性改善。使用菌株LP-8导致摇瓶和5L生物反应器中L-DOPA的产量分别为691.24mg/L和25.53g/L。
EP3150712A1(Symrise)记载了用于提供3,4-二羟基苯基化合物及其甲基化变体的生物技术方法。EP3150712A1因此涉及通过合理设计原型酶4-羟基苯乙酸3-羟化酶(4HPA3H)的活性位点结合袋获得的基因修饰的酶，用于羟基化4-羟基苯基化合物以产生3,4-二羟基苯基化合物，并涉及包括使用所述酶或其催化活性片段的体内和体外方法的生物技术方法。
CN107541483A(Tianjin)记载了用于重组生产左旋多巴的大肠杆菌菌株、其构建方法及应用。CN107541483A因此提供了一种采用大肠杆菌重组菌株T002生产L-DOPA的方法，其中实施aroE基因的上调以增强3-脱氢酶莽草酸脱氢酶的表达，其能够进而生产L-DOPA。
本发明的一个目的是提供一种经济上有吸引力的用于生产L-DOPA的发酵方法，具有优化的酪氨酸产量和L-酪氨酸到L-DOPA的高转化率。需要具有改进的产量或更高的细胞内和/或培养基中产物的最终浓度的L-DOPA的生产方法。该方法需要可扩展，以确保L-DOPA的高效生产。
本发明的又一目的是提供一种细胞，其以使得它能够产生大量的L-DOPA的方式被修饰。
本发明的发明人已经出乎意料地确定，氧化还原酶HpaB的突变导致L-DOPA的产量增加和L-酪氨酸到L-DOPA的转化率更高。
本发明涉及一种多核苷酸，其编码氨基酸序列、编码氧化还原酶，，所编码的氨基酸序列或氧化还原酶与SEQ ID NO:1(地芽孢杆菌属的一个种(Geobacillus sp.)PA9)、SEQID NO:3(嗜热栖热菌(Thermus thermophilus))、SEQ ID NO:4(球孢链霉菌(Streptomycesglobisporus))、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌(Clostridium aminobutyricum))、SEQ ID NO:6(洋葱伯克霍尔德菌(Burkholderai cepacia))、SEQ ID NO:8(颤藻属的一个种(Oscillatoria sp.)PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌(Paraburkholderiaphymatum))的氨基酸序列至少≥50％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQID NO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换，其中氨基酸交换不是A210S或S212A。
根据本发明，在SEQ ID NO:1的位置210处或在SEQ ID NO:3、SEQ ID NO:4、SEQ IDNO:5、SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处从丙氨酸到丝氨酸的氨基酸交换和在SEQ ID NO:1的位置212处或在SEQ ID NO:3、SEQ ID NO:4、SEQ IDNO:5、SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处从丝氨酸到丙氨酸的氨基酸交换被排除并且不应成为本发明的一部分。
在备选配置中，多核苷酸具有在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、211、213、214中的一个或多个位置处或者在SEQ ID NO:3、SEQ ID NO:4、SEQ IDNO:5、SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换。
在优选实施方案中，氧化还原酶不是SEQ ID NO:2的4-羟基苯乙酸3-单加氧酶。
在优选实施方案中，氨基酸交换导致L-DOPA的产量增加和/或L-酪氨酸到L-DOPA的更高的转化率。
在优选实施方案中，由核苷酸序列编码的氧化还原酶是4-羟基苯乙酸3-单加氧酶。
根据本发明由SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ IDNO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9和SEQ ID NO:10编码的酶是4-羟基苯乙酸3-单加氧酶。
选择来自地芽孢杆菌属的一个种PA-9的同源物作为合理酶设计的起点，并在标准化的全细胞筛选系统中测试了一组突变。将筛选系统获得的有希望的结果转移到1L发酵规模上证实了这些结果，并且显示与表达大肠杆菌和地芽孢杆菌属的一个种PA-9野生型基因的参考菌株相比，突变酶的性能显著提高。
表述“SEQ ID NO:1的对应位置”或“与氨基酸序列的位置202、203、204、205、206、207、208、209、210、211、213、214可比较的位置”是指这样的事实，通过在编码多肽的N端区域(基于SEQ ID NO:1的位置202-214)插入或缺失一个编码氨基酸的密码子，位置说明和长度说明在插入的情况下正式增加一个单位或者在缺失的情况下减少一个单位。以相同的方式，通过在编码多肽的C端区域(基于位置202-214)插入或缺失一个编码氨基酸的密码子，长度说明在插入的情况下正式增加一个单位或者在缺失的情况下减少一个单位。这种可比较的位置可以很容易地通过以“比对”的形式通过比较氨基酸序列来进行鉴定，例如使用Clustal W Programme(Thompson等人,Nucleic Acids Research 22,4637-4680(1994))或MAFFT Programme(Katoh等人,Genome Information 2005；16(1),22-33)。当提到来自不同物种的不同SEQ ID NO:(例如SEQ ID NO:3–SEQ ID NO:10)，表述“SEQ ID NO:1的对应位置”是指待比较的晶体结构内的同源位置。这样的同源位置也可以通过如上所述的以“比对”的形式比较氨基酸序列或基于结构预测来进行鉴定。
在本发明的情况下，SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214对应于各种序列的以下位置：
SEQ ID NO:3的位置194、195、196、197、198、199、200、201、202、203、204、205；
SEQ ID NO:4的位置212、213、214、215、216、217、218、219、220、221、222；
SEQ ID NO:5的位置202、203、204、205、206、207、208、209、210；
SEQ ID NO:6的位置202、203、204、205、206、207、208、209、210、211、212；
SEQ ID NO:7的位置203、204、205、206、207、208、209、210、211、212、213；
SEQ ID NO:8的位置209、210、211、212、213、214、215、216、217；
SEQ ID NO:9的位置192、193、194、195、196、197、198、199、200、201；
SEQ ID NO:10的位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217。
关于本发明的氧化还原酶，用Clustal Omega比对了同源物HpaB酶的序列并且在第二步，预测了HpaB序列的二级结构并且使用二级结构预测重新比对了序列。
这样的插入和缺失基本上没有影响酶促活性。“基本上没有影响”意味着所述变体的酶促活性与具有SEQ ID NO:1的氨基酸序列的多肽的活性相差最多10％、最多7.5％、最多5％、最多2.5％或最多1％。
优选地，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ ID NO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:7(钩虫贪铜菌(Cupriavidus necator))、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)的氨基酸序列至少≥65％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换。
更优选地，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ ID NO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ IDNO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:7(钩虫贪铜菌)、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)、SEQID NO:10(皮氏罗尔斯顿菌(Ralstonia pickettii))的氨基酸序列至少≥90％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10的氨基酸序列的对应位置处的氨基酸交换。
优选地，编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶选自：
·SEQ ID NO:1(地芽孢杆菌属的一个种PA9)，其中在位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置处具有氨基酸交换；
·SEQ ID NO:3(嗜热栖热菌)，其中在位置194、195、196、197、198、199、200、201、202、203、204、205中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:21；
·SEQ ID NO:4(球孢链霉菌)，其中在位置212、213、214、215、216、217、218、219、220、221、222中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:22；
·SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)，其中在位置202、203、204、205、206、207、208、209、210中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:23；
·SEQ ID NO:6(洋葱伯克霍尔德菌)，其中在位置202、203、204、205、206、207、208、209、210、211、212中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:24；
·SEQ ID NO:7(钩虫贪铜菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:25；
·SEQ ID NO:8(颤藻属的一个种PCC 6506)，其中在位置209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:26；
·SEQ ID NO:9(瘤状副伯克霍尔德菌)，其中在位置192、193、194、195、196、197、198、199、200、201中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:27；
·SEQ ID NO:10(皮氏罗尔斯顿菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ IDNO:28。
这些不同序列中的那些位置应当是意味着根据本发明的“氨基酸序列的对应位置”。
编码氨基酸序列的多核苷酸编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13或SEQ ID NO:14的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同，其中SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13和SEQID NO:14在位置207或氨基酸序列的对应位置处具有除L-缬氨酸以外的蛋白质氨基酸。
多核苷酸优选是这样的序列，其中该多核苷酸是编码来自地芽孢杆菌属微生物的4-羟基苯乙酸3-单加氧酶的可复制核苷酸序列，其中由此编码的蛋白质序列在对应于SEQID NO:1的位置207的位置处含有除L-缬氨酸以外的蛋白质氨基酸。
在本发明的有利配置中，由该多核苷酸编码的氨基酸序列在位置207或对应位置处具有选自苏氨酸、亮氨酸、谷氨酰胺和甘氨酸的氨基酸。
特别优选的是，当由该多核苷酸编码的蛋白质序列
-在位置206或氨基酸序列的对应位置处含有除L-苏氨酸以外的蛋白质氨基酸，优选L-甲硫氨酸或L-丙氨酸；或者
-在位置208或氨基酸序列的对应位置处含有除L-赖氨酸以外的蛋白质氨基酸，优选L-精氨酸。
在优选配置中，编码氨基酸序列的多核苷酸编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:15、SEQ ID NO:16.、SEQ ID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ IDNO:20、SEQ ID NO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ ID NO:32的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同。
本发明相应地还涉及这样的多核苷酸和核酸分子，其包含这样的序列并编码SEQID NO:1至20和SEQ ID NO:29至32的多肽变体，其含有一个或多个插入或缺失。优选地，多肽含有最多5个、最多4个、最多3个或最多2个氨基酸的插入或缺失。
本发明还涉及包含由根据本发明的核苷酸序列编码的氨基酸序列的多肽。
本发明还涉及这样的埃希菌(Escherichia)、假单胞菌(Pseudomonas)或棒状杆菌(Corynebacterium)属的微生物，其包含根据本发明的多核苷酸、载体和/或多肽，并且在所述微生物中，编码4-羟基苯乙酸单加氧酶的酶的核苷酸序列优选以过表达的形式存在。
优选地，由多核苷酸编码的多肽可以在位置207处或对应位置处显示这样的氨基酸，其选自丝氨酸、亮氨酸、谷氨酰胺和甘氨酸。
由此编码的蛋白质序列可以在位置206或氨基酸序列的相应位置处含有除L-苏氨酸以外的蛋白质氨基酸，优选L-甲硫氨酸。
由此编码的蛋白质序列还可以在位置208或氨基酸序列的相应位置处含有除L-赖氨酸以外的蛋白质氨基酸，优选L-精氨酸。
本发明还涉及这样的质粒和载体，其包含根据本发明的多核苷酸并且任选地在棒状杆菌、假单胞菌或埃希菌属的微生物中复制或适用于其中。
本发明还涉及这样的棒状杆菌、假单胞菌或埃希菌属的微生物，其包含根据本发明的多核苷酸、载体和多肽。
本发明还涉及根据本发明的微生物，其特征在于根据本发明的多核苷酸整合在染色体中。同源重组允许使用根据本发明的载体将由载体转运进入细胞的根据本发明的多核苷酸交换染色体上的DNA部分(section)。为了在载体的环型DNA分子和染色体上的靶DNA之间进行有效的重组，提供了在末端具有与靶位点同源的核苷酸序列的含有根据本发明的多核苷酸的待交换DNA区域；这些决定了载体的整合和DNA交换的位点。
例如，根据本发明的多核苷酸可以在染色体中的天然基因位点处交换天然hpaB基因或在进一步的基因位点处整合。
本发明提供了大肠杆菌、恶臭假单胞菌(P.putida)或谷氨酸棒状杆菌(C.glutamicum)种的微生物，其包含要求保护的任意多核苷酸或要求保护的任意多肽或要求保护的任意载体。
微生物可以是这样的微生物，其中多核苷酸以过表达的形式存在。
微生物的特征可以在于微生物具有生产精细化学品的能力。精细化学品优选为L-二羟基苯丙氨酸(L-DOPA)。
过表达通常意味着，与起始菌株(亲本菌株)或野生型菌株(如果这是起始菌株)相比，核糖核酸、蛋白质(多肽)或酶的细胞内浓度或活性增加。起始菌株(亲本菌株)是指这样的菌株，对所述菌株进行导致过表达的措施。
在过表达中，重组过表达的方法是优选的。这些包括使用体外提供的DNA分子生产微生物的所有方法。这样的DNA分子包括，例如，启动子、表达盒、基因、等位基因、编码区等。这些通过转化、接合、转导或类似方法转化为所需的微生物。
可以通过测量基因转录的mRNA的量、测定多肽的量以及测定酶活性来确定表达或过表达的程度。
本发明公开了生产精细化学品的发酵方法，其包括以下步骤：
a)在培养基中发酵包含编码氨基酸序列的多核苷酸的微生物，所述多核苷酸编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:1或SEQ ID NO:3至SEQ ID NO:32的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同，
b)在培养基中积累精细化学品，其中获得发酵液。
如实施例3所示，根据本发明的这种方法的使用导致与相应的起始菌株相比，产品浓度和L-DOPA:L-酪氨酸的比率显著增加。
待使用的培养基或发酵培养基必须适当地满足相应菌株的需求。在AmericanSociety for Bacteriology的手册"Manual of Methods for General Bacteriology"(Washington D.C.,USA,1981)中含有各种微生物的培养基的说明。术语培养基和发酵培养基或培养基是可以互换的。
可以使用以下作为碳源：糖和碳水化合物，例如葡萄糖、蔗糖、乳糖、果糖、麦芽糖、糖蜜、来自甜菜糖或甘蔗加工的含蔗糖的溶液、淀粉、淀粉水解物和纤维素，油和脂肪，例如大豆油、葵花籽油、花生油和椰子脂，脂肪酸，例如棕榈酸、硬脂酸和亚油酸，醇类例如甘油、甲醇和乙醇，以及有机酸，例如乙酸或乳酸。
可以使用以下作为氮源：有机氮化合物，例如蛋白胨、酵母提取物、肉提取物、麦芽提取物、玉米浆、豆粕和尿素或无机化合物，例如硫酸铵、氯化铵、磷酸铵、碳酸铵和硝酸铵。氮源可以单独使用，也可以作为混合物使用。
可以使用磷酸、磷酸二氢钾或磷酸氢二钾或相应的含钠盐作为磷源。
此外，培养基必须含有盐，例如金属的氯化物或硫酸盐形式，所述金属例如钠、钾、镁、钙和铁，例如硫酸镁或硫酸铁，这是生长所必需的。最后，除上述物质外，还可以使用，必需的生长物质如氨基酸，例如高丝氨酸和维生素，例如硫胺素、生物素或泛酸。
所述起始原料可以以单批的形式加入到培养物中或在培养期间以适当的方式提供。
碱性化合物如氢氧化钠、氢氧化钾、氨或氨水，或酸化合物如磷酸或硫酸，以合适的方式用于培养物的pH控制。pH值一般调节至6.0至8.5，优选6.5至8。为了控制泡沫的形成，可以使用消泡剂，例如脂肪酸的聚乙二醇酯。为了维持质粒的稳定性，可以将合适的选择性作用物质例如抗生素添加到培养基中。优选在好氧条件下进行发酵。为了维持所述好氧条件，将氧气或含氧气体混合物例如空气引入培养物中。同样可以使用富含过氧化氢的液体。任选地，发酵在低于大气压的压力下进行，例如在0.03至0.2MPa的低于大气压的压力下进行。培养的温度通常为20℃至45℃，优选25℃至40℃，特别优选30℃至37℃。在分批或补料分批方法的情况下，优选继续培养，直到形成足以用于测量获得所需有机化合物的量。此目标通常在10小时到160小时内达到。在连续方法中，可以延长培养时间。由于微生物的活性，发酵培养基和/或微生物细胞中精细化学品的富集(积累)发生。
除其他外，合适的发酵培养基的实例可以在专利文献US 5,770,409、US 5,990,350、US 5,275,940、WO 2007/012078、US 5,827,698、WO 2009/043803、US 5,756,345或US7,138,266中找到；可以任选地根据所用菌株的要求进行适当的修改。
该方法的特征可以在于它是这样的方法，其选自分批方法、分批补料方法、重复分批补料方法和连续方法。
该方法的特征还可以在于精细化学品或含有精细化学品的液体或固体产品是从含有精细化学品的发酵液中获得的。
在优选配置中，精细化学品是L-二羟基苯丙氨酸(L-DOPA)。
基于其中存在根据本发明的启动子变体的微生物的方法或发酵方法，根据本发明的方法或发酵方法的性能就选自以下的参数中的一个或多个增加了至少0.5％、至少1％、至少1.5％或至少2％：浓度(每体积形成的化合物)、产率(每碳源消耗形成的化合物)、体积生产率(每体积和时间形成的化合物)和生物质特异性生产率(每细胞干质量(dry mass)或生物干质量和时间形成的化合物或每细胞蛋白质和时间形成的化合物)或其他方法参数及其组合。
由于发酵措施，获得含有所需精细化学品、优选氨基酸或有机酸的发酵液。
然后，提供或生产或获得含有精细化学品的液体或固体形式的产品。
在优选实施方案中，发酵液是指微生物在其中在一定时间和一定温度下培养的发酵培养基或营养培养基。发酵培养基或发酵过程中使用的培养基包含确保生产所需化合物的所有物质或成分，并且通常确保生长和/或活力。
发酵完成后，所得发酵液相应地含有
a)由微生物细胞的生长产生的微生物的生物质(细胞团块(cell mass))，
b)在发酵过程中形成的所需精细化学品，
c)在发酵过程中可能形成的有机副产品，以及
d)发酵未消耗的所使用的发酵培养基或起始原料的成分，例如，例如，维生素例如生物素，或盐，例如硫酸镁。
有机副产品包括除了发酵中使用的微生物所需的相应化合物之外产生的物质，并且可能分泌这些物质。
从培养容器或发酵容器中取出发酵液，任选地收集，并用于提供含有精细化学品的液体或固体形式的产品。因此，其还使用了表述“获得含精细化学品的产品”。在最简单的情况下，从发酵容器中取出的含有精细化学品的发酵液本身就是获得的产品。
通过选自以下措施中的一种或多种
a)部分(>0％至<80％)至完全(100％)或几乎完全(≥80％、≥90％、≥95％、≥96％、≥97％、≥98％、≥99％)去除水，
b)部分(>0％至<80％)至完全(100％)或几乎完全(≥80％、≥90％、≥95％、≥96％、≥97％、≥98％、≥99％)去除生物质，其中任选地在去除之前是失活的，
c)部分(>0％至<80％)至完全(100％)或几乎完全(≥80％、≥90％、≥95％、≥96％、≥97％、≥98％、≥99％、≥99.3％、≥99.7％)去除发酵过程中形成的有机副产品，以及
d)部分(>0％)至完全(100％)或几乎完全(≥80％、≥90％、≥95％、≥96％、≥97％、≥98％、≥99％、≥99.3％、≥99.7％)去除发酵未消耗的所使用的发酵培养基的成分或起始原料，
从发酵液中实现所需有机化合物的浓度或纯化。以这种方式，分离具有所需化合物含量的产品。
部分(>0％至<80％)至完全(100％)或几乎完全(≥80％至<100％)去除水(措施a))也称为干燥。
在该方法的变形中，通过完全或几乎完全去除水、生物质、有机副产品和未消耗的所使用的发酵培养基的成分，成功实现纯(≥80％按重量计，≥90％按重量计)或高纯度(≥95％按重量计，≥97％按重量计，≥99％按重量计)产品形式的所需有机化合物、优选氨基酸、更优选L-DOPA。对于根据a)、b)、c)或d)的措施，现有技术中提供了大量的各种各样的技术说明。
在生产L-DOPA的方法的情况下，优选获得不含发酵液任何成分的产品的工艺。这些产品尤其用于人类医学、制药行业和食品行业。
根据本发明的方法用于发酵生产L-DOPA。
本发明最后涉及根据本发明的微生物的用于发酵生产L-DOPA的用途。
在下文中概括了本发明的进一步的优选实施方案：
本发明涉及编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)的氨基酸序列至少≥50％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ IDNO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换，其中氨基酸交换不是A210S或S212A。
在优选实施方案中，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ IDNO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ IDNO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)的氨基酸序列至少≥50％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中，或在SEQ ID NO:3的位置194、195、196、197、198、199、200、201、202、203、204、205处，或在SEQ ID NO:4的位置212、213、214、215、216、217、218、219、220、221、222处，或在SEQ ID NO:5的位置202、203、204、205、206、207、208、209、210处，或在SEQ ID NO:6的位置202、203、204、205、206、207、208、209、210、211、212处，或在SEQ IDNO:8的位置209、210、211、212、213、214、215、216、217处，或在SEQ ID NO:9的位置192、193、194、195、196、197、198、199、200、201处的氨基酸交换，其中氨基酸交换不是A210S或S212A。
在进一步的优选实施方案中，排除SEQ ID NO:1的位置210和212处的氨基交换。
在优选实施方案中，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ IDNO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ IDNO:7(钩虫贪铜菌)、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)的氨基酸序列至少≥65％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中，或在SEQ ID NO:3的位置194、195、196、197、198、199、200、201、202、203、204、205处，或在SEQ ID NO:4的位置212、213、214、215、216、217、218、219、220、221、222处，或在SEQ ID NO:5的位置202、203、204、205、206、207、208、209、210处，或在SEQ ID NO:6的位置202、203、204、205、206、207、208、209、210、211、212处，或在SEQ IDNO:7的位置203、204、205、206、207、208、209、210、211、212、213处，或在SEQ ID NO:8的位置209、210、211、212、213、214、215、216、217处，或在SEQ ID NO:9的位置192、193、194、195、196、197、198、199、200、201处的氨基酸交换，其中氨基酸交换不是A210S或S212A。
在进一步的优选实施方案中，排除SEQ ID NO:1的位置210和212处的氨基交换。
在优选实施方案中，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ IDNO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ IDNO:7(钩虫贪铜菌)、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)、SEQ ID NO:10(皮氏罗尔斯顿菌)的氨基酸序列至少≥90％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中，或在SEQ ID NO:3的位置194、195、196、197、198、199、200、201、202、203、204、205处，或在SEQ ID NO:4的位置212、213、214、215、216、217、218、219、220、221、222处，或在SEQ ID NO:5的位置202、203、204、205、206、207、208、209、210处，或在SEQ ID NO:6的位置202、203、204、205、206、207、208、209、210、211、212处，或在SEQ IDNO:7的位置203、204、205、206、207、208、209、210、211、212、213处，或在SEQ ID NO:8的位置209、210、211、212、213、214、215、216、217处，或在SEQ ID NO:9的位置192、193、194、195、196、197、198、199、200、201处或在SEQ ID NO:10的位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217处的氨基酸交换，其中氨基酸交换不是A210S或S212A。
在进一步的优选实施方案中，排除SEQ ID NO:1的位置210和212处的氨基交换。
优选实施方案涉及编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与以下的氨基酸序列至少≥30％相同：
·SEQ ID NO:1(地芽孢杆菌属的一个种PA9)，其中在位置202、203、204、205、206、207、208、209、210、211、213、214中的一个或多个位置处具有氨基酸交换；
·SEQ ID NO:3(嗜热栖热菌)，其中在位置194、195、196、197、198、199、200、201、202、203、204、205中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:21；
·SEQ ID NO:4(球孢链霉菌)，其中在位置212、213、214、215、216、217、218、219、220、221、222中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:22；
·SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)，其中在位置202、203、204、205、206、207、208、209、210中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:23；
·SEQ ID NO:6(洋葱伯克霍尔德菌)，其中在位置202、203、204、205、206、207、208、209、210、211、212中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:24；
·SEQ ID NO:7(钩虫贪铜菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:25；
·SEQ ID NO:8(颤藻属的一个种PCC 6506)，其中在位置209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:26；
·SEQ ID NO:9(瘤状副伯克霍尔德菌)，其中在位置192、193、194、195、196、197、198、199、200、201中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:27；
·SEQ ID NO:10(皮氏罗尔斯顿菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ IDNO:28，其中氨基酸交换不是A210S或S212A。
优选实施方案涉及编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与以下的氨基酸序列至少≥40％或至少≥50％、或至少≥60％、或至少≥70％、或至少≥80％、或至少≥90％、或至少≥95％、或至少≥97％、或至少≥98％、或至少≥99％相同：
·SEQ ID NO:1(地芽孢杆菌属的一个种PA9)，其中在位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置处具有氨基酸交换；
·SEQ ID NO:3(嗜热栖热菌)，其中在位置194、195、196、197、198、199、200、201、202、203、204、205中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:21；
·SEQ ID NO:4(球孢链霉菌)，其中在位置212、213、214、215、216、217、218、219、220、221、222中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:22；
·SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)，其中在位置202、203、204、205、206、207、208、209、210中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:23；
·SEQ ID NO:6(洋葱伯克霍尔德菌)，其中在位置202、203、204、205、206、207、208、209、210、211、212中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:24；
·SEQ ID NO:7(钩虫贪铜菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:25；
·SEQ ID NO:8(颤藻属的一个种PCC 6506)，其中在位置209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:26；
·SEQ ID NO:9(瘤状副伯克霍尔德菌)，其中在位置192、193、194、195、196、197、198、199、200、201中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:27；
·SEQ ID NO:10(皮氏罗尔斯顿菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ IDNO:28，其中氨基酸交换不是A210S或S212A。
优选实施方案涉及编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13或SEQ ID NO:14的氨基酸序列至少≥30％、≥40％、≥50％、≥60％、≥70％、≥80％、≥90％相同，其中SEQ ID NO:11、SEQID NO:12、SEQ ID NO:13和SEQ ID NO:14在位置207或氨基酸序列的对应位置处具有除L-缬氨酸以外的蛋白质氨基酸。
优选实施方案是编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13或SEQ ID NO:14的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同，其中SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13和SEQ ID NO:14在位置207或氨基酸序列的对应位置处具有除L-缬氨酸以外的蛋白质氨基酸。
在优选实施方案中，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ IDNO:11、SEQ ID NO:12、SEQ ID NO:13or SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16.、SEQID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:29、SEQ ID NO:30、SEQID NO:31、SEQ ID NO:32的氨基酸序列至少≥30％、≥40％、≥50％、≥60％、≥70％、≥80％、≥90％相同。
在优选实施方案中，编码氨基酸序列的多核苷酸编码氧化还原酶，其与SEQ IDNO:11、SEQ ID NO:12、SEQ ID NO:13or SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16.、SEQID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:29、SEQ ID NO:30、SEQID NO:31、SEQ ID NO:32至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同。
实施例
下面将参考示例性实施方案更详细地描述本发明。
实施例1：生产具有HpaB(地芽孢杆菌)突变体的大肠杆菌菌株
先前在DE102004043748A1中描述的pOM17c-质粒被用作起点，其中公开了质粒的完整序列(含有来自斯氏假单胞菌(Pseudomonas stutzeri)AK61的cyanidase基因的序列)。保留该质粒上复制起点和氨苄青霉素抗性基因的序列，添加野生型地芽孢杆菌属的一个种PA-9hpaB基因、野生型大肠杆菌hpaC基因、食油假单胞菌(Pseudomonas oleovorans)PalkB启动子和大肠杆菌rrnB终止序列以及alkS转录调节基因(Yuste等人,JBacteriol.1998Oct；180(19):5218-26)。使用地芽孢杆菌属的一个种野生型单切限制性内切酶AgeI和PmlI[New England Biolabs GmbH,Brüningstraβe 50,Geb.B852,65926Frankfurt am Main]消化该质粒。使用
Figure BDA0004113813240000201
HiFi DNA Assembly MasterMix[New England Biolabs GmbH Brüningstrasse50,Geb.B852 65926Frankfurt amMain]根据制造商的说明书用质粒主链克隆合成的DNA片段[Eurofins Genomics GermanyGmbH Anzinger Str.7a DE-85560Ebersberg]。因此，该质粒含有大肠杆菌hpaC和不同hpaB变体。转化/>
Figure BDA0004113813240000202
5-alpha Electrocompetent E.coli[New England Biolabs GmbH Brüningstrasse 50,Geb.B852 65926Frankfurt am Main]之后，将细胞铺板于含有100μg/ml的氨苄青霉素的LB-Agar上。进行限制性分析和测序以正确选择克隆的质粒。如果提及，将质粒稳定性毒素-抗毒素序列hok/sok(Thisted,EMBO J.1994Apr 15；13(8):1950-9)和cer决定因子(Summers&Sherratt,1984,DOI:10.1016/0092-8674(84)90060-6)添加到质粒以增加质粒稳定性。
产L-酪氨酸菌株DPD6021-S用于转化所选质粒。DPD6021-S是DPD4145的S期变体，其记载于US 7700328 B2并且其中另外去除了pMT100质粒。为了进行质粒转化，将菌株接种到OD600为0.05的来自LB过夜培养物，并培养至OD600为0.7。冰上孵育30min之后，收获细胞(10min,5500g；4℃)，用50ml的H2Odemin洗涤两次。在用1ml预冷的10％(v/v)甘油进行额外的洗涤步骤后，将细胞重悬于200μl预冷的10％(v/v)甘油中，并使用40μl等分试样进行转化。因此，将100ng质粒转移到电穿孔比色皿中，与40μl细胞溶液混合，并在Gene Pulser XcellElectorporation System[Bio-Rad LaboratoriesGmbH,Kapellenstraβe 12,D-85622Feldkirchen]中在2500V,200Ω&25μF进行负载。在加入1ml SOC培养基并在37℃下再生45分钟后，将100μl悬浮液铺板至含有100μg/ml氨苄青霉素的LB-琼脂。从抗性菌落中分离出质粒，并通过限制性分析证实了质粒的真实性。
产生了以下菌株：
hpaBC_Ec:大肠杆菌hpaB(野生型；参考)；
hpaB_Gs:地芽孢杆菌属的一个种PA-9hpaB(野生型)；
hpaB_V207L_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变V207L)；
hpaB_V207T_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变V207T)；
hpaB_V207Q_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变V207Q)；
hpaB_V207G_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变V207G)；
hpaB_V207T_K208R_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变V207T_K208R)；
hpaB_T206M_V207T_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变T206M_V207T)；
hpaB_T206A_V207T_Gs:地芽孢杆菌属的一个种PA-9hpaB(突变T206A_V207T).
hpaB_V207T_A210S_T211N_S212T:地芽孢杆菌属的一个种PA-9hpaB(突变V207T_A210S_T211N_S212T)
hpaB_V207T_A210V_T211M_S212N:地芽孢杆菌属的一个种PA-9hpaB(突变V207T_A210V_T211M_S212N)
hpaB_V207T_G213A_E214Q_D215N:地芽孢杆菌属的一个种PA-9hpaB(突变V207T_G213A_E214Q_D215N)
hpaB_I152L_V207T:地芽孢杆菌属的一个种PA-9hpaB(突变I152L_V207T)
实施例2：生产L-DOPA和L-酪氨酸(使用BioLector筛选)
在BioLector[m2p-labs；Arnold-Sommerfeld-Ring 2,52499Baesweiler]小规模测试系统中测试了具有表达地芽孢杆菌属的一个种PA-9基因的野生型和突变的变体的质粒的DPD6021-S菌株以及表达野生型基因的参考菌株。
因此，将菌株在振荡摇瓶中在10ml LB培养基(100μg/ml氨苄青霉素)中在37℃、200rpm培养18小时。将培养物接种到含有1mlLB培养基、pH 5.5(补充有7.5mM的L-酪氨酸；100μg/ml氨苄青霉素；0.25％(v/v)DCPK)的BioLector Flowerplates中，以得到0.1的起始OD600。在37℃、1200rpm和85％的相对湿度下进行培养24小时，直到该过程停止，使用高效液相色谱(HPLC)测量L-DOPA和L-酪氨酸浓度。
在Agilent 1200(Agilent Technologies,Palo Alto,Calif.)上进行HPLC。使用Inertsil ODS-3,5μm,4.6x150 mm的柱子(Agilent Technologies)。所使用的方法要求柱流速为1.00ml/min，停止时间为18分钟。流动相由如表1所示的溶剂A(2.72g/L的KH2PO4、2.5ml/L浓磷酸、40ml/L的乙腈)和溶剂B(乙腈)的比例组成。
表1：HPLC分析的流动相组成
时间[min] 溶剂A[％] 溶剂B[％]
0 100 0
6.5 100 0
10 75 25
13 75 25
13.01 100 0
18 100 0
光谱从100nm扫描到380nm，在290nm处记录了L-DOPA的信号，保留时间为2.8min，在278nm处记录了L-酪氨酸的信号，保留时间为3.9分钟。
表2：使用BioLector筛选用单个突变体生产L-DOPA和L-酪氨酸
L-DOPA[mM] L-酪氨酸[mM]
LD-EC-12[hpaBC_Ec] 2.7 5.4
hpaB_Gs 3.6 4.5
hpaB_V207Q_Gs 4.0 4.0
hpaB_V207T_Gs 4.3 3.6
hpaB_V207L_Gs 4.4 3.8
表3：使用BioLector筛选用单个和多个突变体生产L-DOPA和L-酪氨酸(质粒含有质粒稳定性元件)
L-DOPA[mM] L-酪氨酸[mM]
hpaB_V207T_Gs 5.2 3.5
hpaB_T206M_V207T_Gs 6.4 2.2
hpaB_V207T_K208R_Gs 5.4 3.6
hpaB_V207G_Gs 4.4 4.4
hpaB_T206A_V207T_Gs 4.7 3.8
hpaB_V207T_A210S_T211N_S212T 4.0 4.5
hpaB_V207T_A210V_T211M_S212N 4.0 4.6
hpaB_V207T_G213A_E214Q_D215N 3.8 4.7
hpaB_I152L_V207T 4.0 5.0
筛选结果总结在表2和表3中，并可视化于图1和图2中。图1显示了使用BioLector筛选用单个突变体生产L-DOPA和L-酪氨酸，图2显示了使用BioLector筛选用单个和多个突变体生产L-DOPA和L-酪氨酸(质粒含有质粒稳定性元件)。
实施例3：生产L-DOPA和L-酪氨酸(发酵方法)
如US 7700328 B2的实施例8所述进行菌株DPD4145的发酵，并评估菌株的L-酪氨酸和L-DOPA的产量。与US 7700328 B2的实施例8不同，没有用IPTG而是用二环丙基酮(DCPK)诱导发酵，并且在氨苄青霉素(100mg/L)存在下在pH 6.8进行发酵。定期从发酵器中取出样品，并分析L-酪氨酸、L-DOPA和生物质浓度。结果总结在表4-6中，表4显示了48h内L-DOPA的产量，表5显示了相同发酵过程48h内的L-酪氨酸的产量，表6显示了L-DOPA/L-酪氨酸的比例。结果可视化于相应的图3-4中。
图3显示了使用发酵方法用大肠杆菌HpaB(上)和地芽孢杆菌属的具体种HpaB(下)48h内生产L-DOPA和L-酪氨酸，图4显示了使用发酵方法用地芽孢杆菌属的具体种HpaBV207T突变体(上)和地芽孢杆菌属的具体种HpaB V207L突变体(下)48h内生产L-DOPA和L-酪氨酸。
表4：用发酵方法生产L-DOPA的产量
Figure BDA0004113813240000241
表5：用发酵方法生产L-酪氨酸的产量
Figure BDA0004113813240000242
表6：用发酵方法生产的L-DOPA/L-酪氨酸的比例
Figure BDA0004113813240000243
通过使用来自地芽孢杆菌属的HpaB作为野生型酶，并且用不同的突变以发酵生产L-DOPA 48h，可以清楚地表明，L-DOPA/L-酪氨酸的比例高于使用大肠杆菌野生型酶时(表6)。地芽孢杆菌属HpaB酶的突变体甚至具有更高的L-DOPA/L-酪氨酸的比例值。
还在发酵方法中测试了其他突变体Gs_V207T-T206M和Gs_V207T-K208R，并获得了L-DOPA/L-酪氨酸的比例值较高的相似结果。
实施例5：结构同源性分析
关于本发明的氧化还原酶，地芽孢杆菌属的一个种HpaB序列用于在Protein DataBank(PDB)中搜索结构同源物。使用MATT(Menke,M.,Berger,B.&Cowen,L.Matt:LocalFlexibility Aids Protein Multiple Structure Alignment.PLoS Comput.Biol.4,e10(2008))构建结构比对。
本发明的氧化还原酶的这样的比对显示在图5中，其显示了地芽孢杆菌属的一个种HpaB和2yyj(来自嗜热栖热菌的氧化还原酶)的比对。
图6：地芽孢杆菌属的一个种HpaB和4oo2(来自球孢链霉菌的氧化还原酶)的比对
图7：地芽孢杆菌属的一个种HpaB和1uv8(来自氨基丁酸梭状芽孢杆菌的氧化还原酶)的比对
图8：地芽孢杆菌属的一个种HpaB和3hwc(来自洋葱伯克霍尔德菌的氧化还原酶)的比对
图9：地芽孢杆菌属的一个种HpaB和4g5e(来自钩虫贪铜菌的氧化还原酶)的比对
图10：地芽孢杆菌属的一个种HpaB和4irn(来自颤藻属的一个种PCC 6506的氧化还原酶)的比对
图11：地芽孢杆菌属的一个种HpaB和5idu(来自瘤状副伯克霍尔德菌的氧化还原酶)的比对
图12：地芽孢杆菌属的一个种HpaB和6jhm(来自皮氏罗尔斯顿菌的氧化还原酶)的比对
建模HpaB结构的重合显示这些蛋白比对得很好。
实施例6：生产具有不同的氧化还原酶突变体的大肠杆菌菌株
用Enyzme EarI[New England Biolabs GmbH Brüningstrasse 50,Geb.B85265926Frankfurt am Main]对带有野生型地芽孢杆菌属的一个种PA-9hpaB基因和野生型大肠杆菌hpaC基因的实施例1中所述的pOM17c质粒进行限制性酶切以去除野生型地芽孢杆菌属的一个种PA-9hpaB基因以及PalkB,alkS序列和一部分大肠杆菌hpaC序列。将所得质粒主链用合成的DNA片段进行克隆，所述合成的DNA片段含有先前去除的PalkB和alkS序列和一部分大肠杆菌hpaC序列以及编码具有以下的氨基酸序列的酶的基因的野生型序列：SEQ IDNO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:7(钩虫贪铜菌)、SEQ ID NO:8(颤藻属的一个种PCC 6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)、SEQ ID NO:10(皮氏罗尔斯顿菌)。
另外，将所述质粒主链用合成的DNA片段进行克隆，所述合成的DNA片段含有先前去除的PalkB和alkS序列和一部分大肠杆菌hpaC序列以及得到活性增加的酶变体的基因的突变体序列，所述酶变体具有SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28的氨基酸序列。
所得质粒用于转化如实施例1中所述的产L-酪氨酸菌株DPD6021-S。
所得菌株在如实施例2所述的小规模筛选系统中培养或如实施例3所述发酵，以评估表达突变酶变体的菌株的转化率。如实施例1所述定量L-DOPA的形成。检测到所有变体的L-DOPA产生。
蛋白序列
SEQ ID NO:1地芽孢杆菌属的一个种PA9 HpaB
SEQ ID NO:2大肠杆菌(HpaB)4-羟基苯乙酸3-单加氧酶加氧酶成分
SEQ ID NO:3嗜热栖热菌
SEQ ID NO:4球孢链霉菌
SEQ ID NO:5氨基丁酸梭状芽孢杆菌
SEQ ID NO:6洋葱伯克霍尔德菌
SEQ ID NO:7钩虫贪铜菌
SEQ ID NO:8颤藻属的一个种PCC 6506
SEQ ID NO:9瘤状副伯克霍尔德菌
SEQ ID NO:10皮氏罗尔斯顿菌
SEQ ID NO:11地芽孢杆菌属的一个种PA9 HpaB V207L
SEQ ID NO:12地芽孢杆菌属的一个种PA9 HpaB V207T
SEQ ID NO:13地芽孢杆菌属的一个种PA9 HpaB V207Q
SEQ ID NO:14地芽孢杆菌属的一个种PA9 HpaB V207G
SEQ ID NO:15地芽孢杆菌属的一个种PA9 HpaB T206M
SEQ ID NO:16地芽孢杆菌属的一个种PA9 HpaB T206A
SEQ ID NO:17地芽孢杆菌属的一个种PA9 HpaB K208R
SEQ ID NO:18地芽孢杆菌属的一个种PA9 HpaB V207T_K208R
SEQ ID NO:19地芽孢杆菌属的一个种PA9 HpaB T206M_V207T
SEQ ID NO:20地芽孢杆菌属的一个种PA9 HpaB T206A_V207T
SEQ ID NO:21嗜热栖热菌(已突变)
SEQ ID NO:22球孢链霉菌(已突变)
SEQ ID NO:23氨基丁酸梭状芽孢杆菌(已突变)
SEQ ID NO:24洋葱伯克霍尔德菌(已突变)
SEQ ID NO:25钩虫贪铜菌(已突变)
SEQ ID NO:26颤藻属的一个种PCC 6506(已突变)
SEQ ID NO:27瘤状副伯克霍尔德菌(已突变)
SEQ ID NO:28皮氏罗尔斯顿菌(已突变)
SEQ ID NO:29地芽孢杆菌属的一个种PA9 HpaB V207T_A210S_T211N_S212T
SEQ ID NO:30地芽孢杆菌属的一个种PA9 HpaB V207T_A210V_T211M_S212N
SEQ ID NO:31地芽孢杆菌属的一个种PA9 HpaB V207T_G213A_E214Q_D215N
SEQ ID NO:32地芽孢杆菌属的一个种PA9 HpaB I152L_V207T
序列表
<110> 赢创运营有限公司
<120> 编码氨基酸序列、编码氧化还原酶的多核苷酸
<130> 2020p00038wo
<160> 45
<170> PatentIn version 3.5
<210> 1
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种（Geobacillus sp.）
<400> 1
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 2
<211> 520
<212> PRT
<213> 大肠杆菌
<400> 2
Met Lys Pro Glu Asp Phe Arg Ala Ser Thr Gln Arg Pro Phe Thr Gly
1 5 10 15
Glu Glu Tyr Leu Lys Ser Leu Gln Asp Gly Arg Glu Ile Tyr Ile Tyr
20 25 30
Gly Glu Arg Val Lys Asp Val Thr Thr His Pro Ala Phe Arg Asn Ala
35 40 45
Ala Ala Ser Val Ala Gln Leu Tyr Asp Ala Leu His Lys Pro Glu Met
50 55 60
Gln Asp Ser Leu Cys Trp Asn Thr Asp Thr Gly Ser Gly Gly Tyr Thr
65 70 75 80
His Lys Phe Phe Arg Val Ala Lys Ser Ala Asp Asp Leu Arg His Glu
85 90 95
Arg Asp Ala Ile Ala Glu Trp Ser Arg Leu Ser Tyr Gly Trp Met Gly
100 105 110
Arg Thr Pro Asp Tyr Lys Ala Ala Phe Gly Cys Ala Leu Gly Gly Thr
115 120 125
Pro Gly Phe Tyr Gly Gln Phe Glu Gln Asn Ala Arg Asn Trp Tyr Thr
130 135 140
Arg Ile Gln Glu Thr Gly Leu Tyr Phe Asn His Ala Ile Val Asn Pro
145 150 155 160
Pro Ile Asp Arg His Leu Pro Thr Asp Lys Val Lys Asp Val Tyr Ile
165 170 175
Lys Leu Glu Lys Glu Thr Asp Ala Gly Ile Ile Val Ser Gly Ala Lys
180 185 190
Val Val Ala Thr Asn Ser Ala Leu Thr His Tyr Asn Met Ile Gly Phe
195 200 205
Gly Ser Ala Gln Val Met Gly Glu Asn Pro Asp Phe Ala Leu Met Phe
210 215 220
Val Ala Pro Met Asp Ala Asp Gly Val Lys Leu Ile Ser Arg Ala Ser
225 230 235 240
Tyr Glu Met Val Ala Gly Ala Thr Gly Ser Pro Tyr Asp Tyr Pro Leu
245 250 255
Ser Ser Arg Phe Asp Glu Asn Asp Ala Ile Leu Val Met Asp Asn Val
260 265 270
Leu Ile Pro Trp Glu Asn Val Leu Leu Tyr Arg Asp Phe Asp Arg Cys
275 280 285
Arg Arg Trp Thr Met Glu Gly Gly Phe Ala Arg Met Tyr Pro Leu Gln
290 295 300
Ala Cys Val Arg Leu Ala Val Lys Leu Asp Phe Ile Thr Ala Leu Leu
305 310 315 320
Lys Lys Ser Leu Glu Cys Thr Gly Thr Leu Glu Phe Arg Gly Val Gln
325 330 335
Ala Asp Leu Gly Glu Val Val Ala Trp Arg Asn Thr Phe Trp Ala Leu
340 345 350
Ser Asp Ser Met Cys Ser Glu Ala Thr Pro Trp Val Asn Gly Ala Tyr
355 360 365
Leu Pro Asp His Ala Ala Leu Gln Thr Tyr Arg Val Leu Ala Pro Met
370 375 380
Ala Tyr Ala Lys Ile Lys Asn Ile Ile Glu Arg Asn Val Thr Ser Gly
385 390 395 400
Leu Ile Tyr Leu Pro Ser Ser Ala Arg Asp Leu Asn Asn Pro Gln Ile
405 410 415
Asp Gln Tyr Leu Ala Lys Tyr Val Arg Gly Ser Asn Gly Met Asp His
420 425 430
Val Gln Arg Ile Lys Ile Leu Lys Leu Met Trp Asp Ala Ile Gly Ser
435 440 445
Glu Phe Gly Gly Arg His Glu Leu Tyr Glu Ile Asn Tyr Ser Gly Ser
450 455 460
Gln Asp Glu Ile Arg Leu Gln Cys Leu Arg Gln Ala Gln Ser Ser Gly
465 470 475 480
Asn Met Asp Lys Met Met Ala Met Val Asp Arg Cys Leu Ser Glu Tyr
485 490 495
Asp Gln Asn Gly Trp Thr Val Pro His Leu His Asn Asn Asp Asp Ile
500 505 510
Asn Met Leu Asp Lys Leu Leu Lys
515 520
<210> 3
<211> 481
<212> PRT
<213> 嗜热栖热菌（Thermus thermophilus）
<400> 3
Met Ala Arg Thr Gly Ala Glu Tyr Ile Glu Ala Leu Lys Thr Arg Pro
1 5 10 15
Pro Asn Leu Trp Tyr Lys Gly Glu Lys Val Glu Asp Pro Thr Thr His
20 25 30
Pro Val Phe Arg Gly Ile Val Arg Thr Met Ala Ala Leu Tyr Asp Leu
35 40 45
Gln His Asp Pro Arg Tyr Arg Glu Val Leu Thr Tyr Glu Glu Glu Gly
50 55 60
Lys Arg His Gly Met Ser Phe Leu Ile Pro Lys Thr Lys Glu Asp Leu
65 70 75 80
Lys Arg Arg Gly Gln Ala Tyr Lys Leu Trp Ala Asp Gln Asn Leu Gly
85 90 95
Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Val Val Met Ala Tyr
100 105 110
Ala Ala Ser Ala Asp Tyr Phe Gly Glu Phe Ala Glu Asn Val Arg Asn
115 120 125
Tyr Tyr Arg Tyr Leu Arg Asp Gln Asp Leu Ala Thr Thr His Ala Leu
130 135 140
Thr Asn Pro Gln Val Asn Arg Ala Arg Pro Pro Ser Gly Gln Pro Asp
145 150 155 160
Pro Tyr Ile Pro Val Gly Val Val Lys Gln Thr Glu Lys Gly Ile Val
165 170 175
Val Arg Gly Ala Arg Met Thr Ala Thr Phe Pro Leu Ala Asp Glu Val
180 185 190
Leu Ile Phe Pro Ser Thr Leu Leu Gln Ala Gly Ser Glu Lys Tyr Ala
195 200 205
Leu Ala Phe Ala Leu Pro Thr Ser Thr Pro Gly Leu His Phe Val Cys
210 215 220
Arg Glu Ala Leu Val Gly Gly Asp Ser Pro Phe Asp His Pro Leu Ser
225 230 235 240
Ser Arg Val Glu Glu Met Asp Cys Leu Val Ile Phe Asp Asp Val Leu
245 250 255
Val Pro Trp Glu Arg Val Phe Ile Leu Gly Asn Val Glu Leu Cys Asn
260 265 270
Asn Ala Tyr Ala Ala Thr Gly Ala Leu Asn His Met Ala His Gln Val
275 280 285
Val Ala Leu Lys Thr Ala Lys Thr Glu Ala Phe Leu Gly Val Ala Ala
290 295 300
Leu Met Ala Glu Gly Ile Gly Ala Asp Val Tyr Gly His Val Gln Glu
305 310 315 320
Lys Ile Ala Glu Ile Ile Val Tyr Leu Glu Ala Met Arg Ala Phe Trp
325 330 335
Thr Arg Ala Glu Glu Glu Ala Lys Glu Asn Ala Tyr Gly Leu Leu Val
340 345 350
Pro Asp Arg Gly Ala Leu Asp Gly Ala Arg Asn Leu Tyr Pro Arg Leu
355 360 365
Tyr Pro Arg Ile Arg Glu Ile Leu Glu Gln Ile Gly Ala Ser Gly Leu
370 375 380
Ile Thr Leu Pro Ser Glu Lys Asp Phe Lys Gly Pro Leu Gly Pro Phe
385 390 395 400
Leu Glu Lys Phe Leu Gln Gly Ala Ala Leu Glu Ala Lys Glu Arg Val
405 410 415
Ala Leu Phe Arg Leu Ala Trp Asp Met Thr Leu Ser Gly Phe Gly Ala
420 425 430
Arg Gln Glu Leu Tyr Glu Arg Phe Phe Phe Gly Asp Pro Val Arg Met
435 440 445
Tyr Gln Thr Leu Tyr Asn Val Tyr Asn Lys Glu Pro Tyr Lys Glu Arg
450 455 460
Ile Arg Ala Phe Leu Lys Glu Ser Leu Lys Val Phe Glu Glu Val Gln
465 470 475 480
Ala
<210> 4
<211> 527
<212> PRT
<213> 球孢链霉菌（Streptomyces globisporus）
<400> 4
Met Pro His Gly Ala Glu Arg Glu Ala Ser Pro Ala Glu Glu Ser Ala
1 5 10 15
Gly Thr Arg Pro Leu Thr Gly Glu Glu Tyr Leu Glu Ser Leu Arg Asp
20 25 30
Ala Arg Glu Val Tyr Leu Asp Gly Ser Arg Val Lys Asp Val Thr Ala
35 40 45
His Pro Ala Phe His Asn Pro Ala Arg Met Thr Ala Arg Leu Tyr Asp
50 55 60
Ser Leu His Asp Pro Ala Gln Lys Ala Val Leu Thr Ala Pro Thr Asp
65 70 75 80
Ala Gly Asp Gly Phe Thr His Arg Phe Phe Thr Ala Pro Arg Ser Val
85 90 95
Asp Asp Leu Val Lys Asp Gln Ala Ala Ile Ala Ser Trp Ala Arg Lys
100 105 110
Ser Tyr Gly Trp Met Gly Arg Ser Pro Asp Tyr Lys Ala Ser Phe Leu
115 120 125
Gly Thr Leu Gly Ala Asn Ala Asp Phe Tyr Glu Pro Phe Ala Asp Asn
130 135 140
Ala Arg Arg Trp Tyr Arg Glu Ser Gln Glu Lys Val Leu Tyr Trp Asn
145 150 155 160
His Ala Phe Leu His Pro Pro Val Asp Arg Ser Leu Pro Ala Asp Glu
165 170 175
Val Gly Asp Val Phe Ile His Val Glu Arg Glu Thr Asp Ala Gly Leu
180 185 190
Val Val Ser Gly Ala Lys Val Val Ala Thr Gly Ser Ala Leu Thr His
195 200 205
Ala Ala Phe Ile Ser His Trp Gly Leu Pro Ile Lys Asp Arg Lys Phe
210 215 220
Ala Leu Val Ala Thr Val Pro Met Asp Ala Asp Gly Leu Lys Val Ile
225 230 235 240
Cys Arg Pro Ser Tyr Ser Ala Asn Ala Ala Thr Thr Gly Ser Pro Phe
245 250 255
Asp Asn Pro Leu Ser Ser Arg Leu Asp Glu Asn Asp Ala Ile Leu Val
260 265 270
Leu Asp Gln Val Leu Ile Pro Trp Glu Asn Val Phe Val Tyr Gly Asn
275 280 285
Leu Gly Lys Val His Leu Leu Ala Gly Gln Ser Gly Met Ile Glu Arg
290 295 300
Ala Thr Phe His Gly Cys Thr Arg Leu Ala Val Lys Leu Glu Phe Ile
305 310 315 320
Ala Gly Leu Leu Ala Lys Ala Leu Asp Ile Thr Gly Ala Lys Asp Phe
325 330 335
Arg Gly Val Gln Thr Arg Leu Gly Glu Val Leu Ala Trp Arg Asn Leu
340 345 350
Phe Trp Ser Leu Ser Asp Ala Ala Ala Arg Asn Pro Val Pro Trp Lys
355 360 365
Asn Gly Thr Leu Leu Pro Asn Pro Gln Ala Gly Met Ala Tyr Arg Trp
370 375 380
Phe Met Gln Ile Gly Tyr Pro Arg Val Leu Glu Ile Val Gln Gln Asp
385 390 395 400
Val Ala Ser Gly Leu Met Tyr Val Asn Ser Ser Thr Glu Asp Phe Arg
405 410 415
Asn Pro Glu Thr Gly Pro Tyr Leu Glu Lys Tyr Leu Arg Gly Ser Asp
420 425 430
Gly Ala Gly Ala Val Glu Arg Val Lys Val Met Lys Leu Leu Trp Asp
435 440 445
Ala Val Gly Ser Asp Phe Gly Gly Arg His Glu Leu Tyr Glu Arg Asn
450 455 460
Tyr Ser Gly Asn His Glu Asn Thr Arg Ile Glu Leu Leu Leu Ser Gln
465 470 475 480
Thr Ala Ser Gly Lys Leu Asp Ser Tyr Met Asp Phe Ala Gln Ala Cys
485 490 495
Met Asp Glu Tyr Asp Leu Asp Gly Trp Thr Ala Pro Asp Leu Glu Ser
500 505 510
Phe His Ala Met Arg Ser Ala Ser Arg Asp Leu Leu Gly Gly Leu
515 520 525
<210> 5
<211> 490
<212> PRT
<213> 氨基丁酸梭状芽孢杆菌（Clostridium aminobutyricum）
<400> 5
Met Leu Met Thr Ala Glu Gln Tyr Ile Glu Ser Leu Arg Lys Leu Asn
1 5 10 15
Thr Arg Val Tyr Met Phe Gly Glu Lys Ile Glu Asn Trp Val Asp His
20 25 30
Pro Met Ile Arg Pro Ser Ile Asn Cys Val Arg Met Thr Tyr Glu Leu
35 40 45
Ala Gln Asp Pro Gln Tyr Ala Asp Leu Met Thr Thr Lys Ser Asn Leu
50 55 60
Ile Gly Lys Thr Ile Asn Arg Phe Ala Asn Leu His Gln Ser Thr Asp
65 70 75 80
Asp Leu Arg Lys Lys Val Lys Met Gln Arg Leu Leu Gly Gln Lys Thr
85 90 95
Ala Ser Cys Phe Gln Arg Cys Val Gly Met Asp Ala Phe Asn Ala Val
100 105 110
Phe Ser Thr Thr Tyr Glu Ile Asp Gln Lys Tyr Gly Thr Asn Tyr His
115 120 125
Lys Asn Phe Thr Glu Tyr Leu Lys Tyr Ile Gln Glu Asn Asp Leu Ile
130 135 140
Val Asp Gly Ala Met Thr Asp Pro Lys Gly Asp Arg Gly Leu Ala Pro
145 150 155 160
Ser Ala Gln Lys Asp Pro Asp Leu Phe Leu Arg Ile Val Glu Lys Arg
165 170 175
Glu Asp Gly Ile Val Val Arg Gly Ala Lys Ala His Gln Thr Gly Ser
180 185 190
Ile Asn Ser His Glu His Ile Ile Met Pro Thr Ile Ala Met Thr Glu
195 200 205
Ala Asp Lys Asp Tyr Ala Val Ser Phe Ala Cys Pro Ser Asp Ala Asp
210 215 220
Gly Leu Phe Met Ile Tyr Gly Arg Gln Ser Cys Asp Thr Arg Lys Met
225 230 235 240
Glu Glu Gly Ala Asp Ile Asp Leu Gly Asn Lys Gln Phe Gly Gly Gln
245 250 255
Glu Ala Leu Val Val Phe Asp Asn Val Phe Ile Pro Asn Asp Arg Ile
260 265 270
Phe Leu Cys Gln Glu Tyr Asp Phe Ala Gly Met Met Val Glu Arg Phe
275 280 285
Ala Gly Tyr His Arg Gln Ser Tyr Gly Gly Cys Lys Val Gly Val Gly
290 295 300
Asp Val Val Ile Gly Ala Ala Ala Leu Ala Ala Asp Tyr Asn Gly Ala
305 310 315 320
Gln Lys Ala Ser His Val Lys Asp Lys Leu Ile Glu Met Thr His Leu
325 330 335
Asn Glu Thr Leu Tyr Cys Cys Gly Ile Ala Cys Ser Ala Glu Gly Tyr
340 345 350
Pro Thr Ala Ala Gly Asn Tyr Gln Ile Asp Leu Leu Leu Ala Asn Val
355 360 365
Cys Lys Gln Asn Ile Thr Arg Phe Pro Tyr Glu Ile Val Arg Leu Ala
370 375 380
Glu Asp Ile Ala Gly Gly Leu Met Val Thr Met Pro Ser Glu Ala Asp
385 390 395 400
Phe Lys Ser Glu Thr Val Val Gly Arg Asp Gly Glu Thr Ile Gly Asp
405 410 415
Phe Cys Asn Lys Phe Phe Ala Ala Ala Pro Thr Cys Thr Thr Glu Glu
420 425 430
Arg Met Arg Val Leu Arg Phe Leu Glu Asn Ile Cys Leu Gly Ala Ser
435 440 445
Ala Val Gly Tyr Arg Thr Glu Ser Met His Gly Ala Gly Ser Pro Gln
450 455 460
Ala Gln Arg Ile Met Ile Ala Arg Gln Gly Asn Ile Asn Ala Lys Lys
465 470 475 480
Glu Leu Ala Lys Ala Ile Ala Gly Ile Lys
485 490
<210> 6
<211> 515
<212> PRT
<213> 洋葱伯克霍尔德菌（Burkholderai cepacia）
<400> 6
Met Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg Val
1 5 10 15
Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro Leu
20 25 30
Thr Arg Asp Tyr Ala Glu Arg Val Ala Gln Phe Tyr Asp Leu His His
35 40 45
Arg Pro Asp Leu Gln Asp Val Leu Thr Phe Val Asp Ala Asp Gly Val
50 55 60
Arg Arg Ser Arg Gln Trp Gln Asp Pro Lys Asp Ala Ala Gly Leu Arg
65 70 75 80
Val Lys Arg Lys Tyr His Glu Thr Ile Leu Arg Glu Ile Ala Ala Gly
85 90 95
Ser Tyr Gly Arg Leu Pro Asp Ala His Asn Tyr Thr Phe Thr Thr Tyr
100 105 110
Ala Asp Asp Pro Glu Val Trp Glu Lys Gln Ser Ile Gly Ala Glu Gly
115 120 125
Arg Asn Leu Thr Gln Asn Ile His Asn Phe Leu Lys Leu Leu Arg Glu
130 135 140
Lys Asp Leu Asn Cys Pro Leu Asn Phe Val Asp Pro Gln Thr Asp Arg
145 150 155 160
Ser Ser Asp Ala Ala Gln Ala Arg Ser Pro Asn Leu Arg Ile Val Glu
165 170 175
Lys Thr Asp Asp Gly Ile Ile Val Asn Gly Val Lys Ala Val Gly Thr
180 185 190
Gly Ile Ala Phe Gly Asp Tyr Met His Ile Gly Cys Leu Tyr Arg Pro
195 200 205
Gly Ile Pro Gly Glu Gln Val Ile Phe Ala Ala Ile Pro Thr Asn Thr
210 215 220
Pro Gly Val Thr Val Phe Cys Arg Glu Ser Thr Val Lys Asn Asp Pro
225 230 235 240
Ala Glu His Pro Leu Ala Ser Gln Gly Asp Glu Leu Asp Ser Thr Thr
245 250 255
Val Phe Asp Asn Val Phe Ile Pro Trp Glu Gln Val Phe His Ile Gly
260 265 270
Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg Ile Phe Asp Trp Val
275 280 285
His Tyr His Ile Leu Ile Arg Gln Val Leu Arg Ala Glu Leu Ile Val
290 295 300
Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly Thr Ser Lys Leu Pro
305 310 315 320
Thr Val Ser Ala Arg Val Ala Lys Leu Val Ala Phe His Leu Ala Met
325 330 335
Gln Ala His Leu Ile Ala Ser Glu Glu Thr Gly Phe His Thr Lys Gly
340 345 350
Gly Arg Tyr Lys Pro Asn Pro Leu Ile Tyr Asp Phe Gly Arg Ala His
355 360 365
Phe Leu Gln Asn Gln Met Ser Val Met Tyr Glu Leu Leu Asp Leu Ala
370 375 380
Gly Arg Ser Ser Leu Met Ile Pro Ser Glu Gly Gln Trp Asp Asp Ser
385 390 395 400
Gln Ser Gly Gln Trp Phe Val Lys Leu Asn Asn Gly Pro Lys Gly Asn
405 410 415
Pro Arg Glu Arg Val Gln Ile Gly Arg Val Ile Arg Asp Leu Tyr Leu
420 425 430
Thr Asp Trp Gly Gly Arg Gln Phe Met Phe Glu Asn Phe Asn Gly Thr
435 440 445
Pro Leu Phe Ala Val Phe Ala Ala Thr Met Thr Arg Asp Asp Met Ser
450 455 460
Ala Ala Gly Thr Tyr Gly Lys Phe Ala Ser Gln Val Cys Gly Ile Glu
465 470 475 480
Phe Gly Gly Ala Glu Pro Thr Ala Tyr Ala Ala Thr Ala Asp Tyr Ala
485 490 495
Lys Ala Leu Asp Lys Gly Leu Ala Pro Glu Pro Ala Ala Ala Glu Ser
500 505 510
Ala Thr Ser
515
<210> 7
<211> 517
<212> PRT
<213> 钩虫贪铜菌（Cupriavidus necator）
<400> 7
Met Ile Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg
1 5 10 15
Asn Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro
20 25 30
Lys Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His
35 40 45
His Arg Pro Asp Leu Gln Asp Val Met Thr Phe Val Asp Lys Asp Gly
50 55 60
Glu Arg Arg Thr Met Gln Trp Phe Gly His Tyr Asp Lys Glu Gln Leu
65 70 75 80
Arg Arg Lys Arg Lys Tyr His Glu Thr Ile Met Arg Glu Met Ala Gly
85 90 95
Ala Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Gln Thr
100 105 110
Tyr Ile Asp Asp Pro Ser Pro Trp Glu Thr Gln Thr Ile Gly Ala Glu
115 120 125
Gly Lys Val Lys Ala Lys Asn Ile Val Asp Phe Val Asn Phe Ala Lys
130 135 140
Lys His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Met Asp
145 150 155 160
Arg Ser Asn Pro Asp Ala Gln Gln Arg Ser Pro Gly Leu Arg Val Ile
165 170 175
Glu Lys Asn Asp Lys Gly Ile Val Val Ser Gly Val Lys Ala Ile Gly
180 185 190
Thr Gly Val Ala Phe Ala Asp Trp Ile His Ile Gly Val Phe Phe Arg
195 200 205
Pro Gly Ile Pro Gly Asp Gln Ile Ile Phe Ala Ala Thr Pro Val Asn
210 215 220
Thr Pro Gly Val Thr Ile Val Cys Arg Glu Ser Val Val Lys Glu Asp
225 230 235 240
Pro Ile Glu His Pro Leu Ala Ser Gln Gly Asp Glu Leu Asp Gly Met
245 250 255
Thr Val Phe Asp Asn Val Phe Ile Pro Trp Ser His Val Phe His Leu
260 265 270
Gly Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg Val Phe Asp Trp
275 280 285
Leu His Tyr His Ala Leu Ile Arg Gln Ser Val Arg Ala Glu Leu Met
290 295 300
Ala Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly Thr Asn Lys Ile
305 310 315 320
Pro Ala Val Gln Thr Arg Val Ala Lys Leu Ile Gly Phe His Gln Ala
325 330 335
Met Leu Ala His Ile Val Ala Ser Glu Glu Leu Gly Phe His Thr Pro
340 345 350
Gly Gly Ala Tyr Lys Pro Asn Ile Leu Ile Tyr Asp Phe Gly Arg Ala
355 360 365
Leu Tyr Leu Glu Asn Phe Ser Gln Met Ile Tyr Glu Leu Val Asp Leu
370 375 380
Ser Gly Arg Ser Ala Leu Ile Phe Ala Ser Glu Asp Gln Trp Asn Asp
385 390 395 400
Glu Ala Leu Asn Gly Trp Phe Glu Arg Met Asn Asn Gly Pro Val Gly
405 410 415
Gln Pro His Asp Arg Val Lys Ile Gly Arg Val Ile Arg Asp Leu Phe
420 425 430
Leu Thr Asp Trp Gly Asn Arg Leu Phe Val Phe Glu Asn Phe Asn Gly
435 440 445
Thr Pro Leu Gln Ala Ile Arg Met Leu Thr Met Gln Arg Ala Glu Phe
450 455 460
Ser Ala Ala Gly Pro Tyr Gly Thr Leu Ala Arg Lys Val Cys Gly Ile
465 470 475 480
Glu Leu Thr Glu Gly His Asp Ser Glu Tyr Lys Ala Thr Ala Gly Tyr
485 490 495
Ala Gln Ala Leu Asp Ser Ala Arg His Gln Glu Lys Leu Ala Leu Ser
500 505 510
Gly Thr Met Thr Val
515
<210> 8
<211> 416
<212> PRT
<213> 颤藻属的一个种（Oscillatoria sp.）PCC 6506
<400> 8
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Ala Ser Met Thr Gly Gly Gln Gln Met Gly Arg
20 25 30
Gly Ser Glu Phe Asp Phe Ala Trp Asn Ser Gln Gln Ile Gln Phe Arg
35 40 45
Lys Lys Val Ile Gln Phe Ala Gln Gln Ser Leu Ile Ser Asp Leu Ile
50 55 60
Lys Asn Asp Lys Glu Glu Ile Phe Asn Arg Asp Ala Trp Gln Lys Cys
65 70 75 80
Ser Glu Phe Gly Val His Gly Trp Pro Ile Pro Ala Arg Tyr Gly Gly
85 90 95
Gln Glu Leu Asp Ile Leu Thr Thr Ala Tyr Ala Leu Gln Gly Leu Gly
100 105 110
Tyr Gly Cys Lys Asp Asn Gly Leu Ile Phe Ala Met Asn Ala His Ile
115 120 125
Trp Ala Cys Glu Met Pro Leu Leu Thr Phe Gly Thr Glu Glu Gln Lys
130 135 140
Glu Lys Tyr Leu Pro Leu Leu Cys Arg Gly Gly Trp Ile Ala Ser His
145 150 155 160
Ala Ala Thr Glu Pro Gln Ala Gly Ser Asp Ile Tyr Ser Leu Lys Thr
165 170 175
Thr Ala Gln Lys Asp Gly Asp Lys Tyr Ile Leu Asn Gly Tyr Lys His
180 185 190
Tyr Val Thr Asn Gly Thr Ile Ala Asp Leu Phe Ile Ile Phe Ala Thr
195 200 205
Ile Asp Pro Ser Leu Gly Lys Glu Gly Leu Thr Thr Phe Met Ile Glu
210 215 220
Lys Asp Thr Pro Gly Leu Ile Leu Ser Lys Pro Ile Ser Lys Met Gly
225 230 235 240
Met Arg Thr Ala Glu Val Pro Glu Leu Arg Leu Glu Asn Cys Glu Val
245 250 255
Ser Ala Ala Asn Arg Leu Gly Glu Glu Gly Thr Gly Leu Ala Ile Phe
260 265 270
Asn His Ser Met Glu Trp Glu Arg Gly Phe Ile Leu Ala Ala Ala Val
275 280 285
Gly Thr Met Glu Arg Leu Leu Glu Gln Ser Ile Arg Tyr Ala Arg Ser
290 295 300
His Lys Gln Phe Gly Gln Ala Ile Gly Lys Phe Gln Leu Val Ala Asn
305 310 315 320
Lys Leu Val Glu Met Lys Leu Arg Leu Glu Asn Ala Lys Ala Tyr Leu
325 330 335
Tyr Lys Val Ala Trp Met Lys Glu Asn Lys Gln Met Ala Leu Leu Glu
340 345 350
Ala Ser Met Ala Asn Leu Tyr Ile Ser Glu Ala Trp Val Gln Ser Cys
355 360 365
Leu Glu Ala Ile Glu Ile His Gly Ala Tyr Gly Tyr Leu Thr Asn Thr
370 375 380
Glu Leu Glu Arg Glu Leu Arg Asp Ala Ile Ala Ser Lys Phe Tyr Ser
385 390 395 400
Gly Thr Ser Glu Ile Gln Arg Val Val Ile Ala Lys Phe Leu Gly Leu
405 410 415
<210> 9
<211> 410
<212> PRT
<213> 瘤状副伯克霍尔德菌（Paraburkholderia phymatum）
<400> 9
Met Ala His His His His His His Met Leu Ala Ala Asn Leu Ile Asp
1 5 10 15
Pro His Gly Ala Leu Ala Trp Pro Phe Phe Glu Ala Arg His Arg Glu
20 25 30
Leu Ala Ala Gly Ile Glu Ala Trp Ala Thr Gln His Leu Ala Cys Val
35 40 45
Gln His Asp Asp Thr Asp Thr Thr Cys Arg Lys Leu Val Arg Ala Leu
50 55 60
Gly Glu Ala Gly Trp Leu Lys Tyr Gly Val Gly Gly Ala Gln Tyr Gly
65 70 75 80
Gly His Gly Asp Thr Ile Asp Thr Arg Ala Val Cys Leu Leu Arg Glu
85 90 95
Thr Leu Ala Asn His Asp Gly Leu Ala Asp Phe Ala Leu Ala Met Gln
100 105 110
Gly Leu Gly Ser Gly Ala Ile Thr Leu Ala Gly Thr His Glu Gln Lys
115 120 125
Ile Arg Tyr Leu Pro Arg Val Ser Lys Gly Glu Ala Ile Ala Ala Phe
130 135 140
Ala Leu Ser Glu Pro Asp Ala Gly Ser Asp Val Ala Ala Met Ser Leu
145 150 155 160
Gln Ala Arg Ala Glu Gly Asp Cys Tyr Val Ile Asp Gly Asp Lys Thr
165 170 175
Trp Ile Ser Asn Gly Gly Ile Ala Asp Phe Tyr Val Val Phe Ala Arg
180 185 190
Thr Gly Glu Ala Pro Gly Ala Arg Gly Ile Ser Ala Phe Ile Val Asp
195 200 205
Ala Asp Thr Pro Gly Leu Gln Ile Ala Glu Arg Ile Asp Val Ile Ala
210 215 220
Pro His Pro Leu Ala Arg Leu His Phe Asp Ser Ala Arg Val Pro Arg
225 230 235 240
Ser Gln Met Leu Gly Ala Pro Gly Glu Gly Phe Lys Ile Ala Met Arg
245 250 255
Thr Leu Asp Val Phe Arg Thr Ser Val Ala Ala Ala Ser Leu Gly Phe
260 265 270
Ala Arg Arg Ala Leu Gln Glu Gly Leu Ala Arg Ala Ala Ser Arg Lys
275 280 285
Met Phe Gly Gln Thr Leu Gly Asp Phe Gln Leu Thr Gln Thr Lys Leu
290 295 300
Ala Gln Met Ala Leu Thr Ile Asp Ser Ser Ala Leu Leu Val Tyr Arg
305 310 315 320
Ala Ala Trp Leu Arg Asp Gln Gly Glu Asn Val Thr Arg Glu Ala Ala
325 330 335
Met Ala Lys Trp His Ala Ser Glu Gly Ala Gln Gln Val Ile Asp Ala
340 345 350
Ala Val Gln Leu Trp Gly Gly Met Gly Val Gln Ser Gly Thr Thr Val
355 360 365
Glu Arg Leu Tyr Arg Glu Ile Arg Ala Leu Arg Ile Tyr Glu Gly Ala
370 375 380
Thr Glu Val Gln Gln Leu Ile Val Gly Arg Asp Leu Leu Lys Ala His
385 390 395 400
Ala Ala Gln Arg Gln Gln Glu Arg Ala Ser
405 410
<210> 10
<211> 517
<212> PRT
<213> 皮氏罗尔斯顿菌（Ralstonia pickettii）
<400> 10
Met Ile Arg Thr Gly Thr Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg
1 5 10 15
Asn Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro
20 25 30
Lys Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His
35 40 45
His Arg Pro Asp Leu Gln Asp Val Met Thr Tyr Ile Asp Glu Gly Gly
50 55 60
Gln Arg Arg Ala Met Gln Trp Phe Gly His Arg Asp Lys Glu Gln Leu
65 70 75 80
Arg Arg Lys Arg Lys Tyr His Glu Thr Val Met Arg Glu Met Ala Gly
85 90 95
Ala Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Thr Thr
100 105 110
Tyr Ile Asp Asp Pro Ala Pro Trp Glu Thr Gln Ser Ile Gly Asp Asp
115 120 125
Gly His Ile Lys Ala Gly Lys Ile Val Asp Phe Ile Arg Tyr Ala Arg
130 135 140
Glu His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Met Asp
145 150 155 160
Arg Ser Asn Pro Asp Ala Gln Glu Arg Ser Pro Gly Leu Arg Val Val
165 170 175
Glu Lys Asn Glu Lys Gly Ile Val Val Asn Gly Val Lys Ala Ile Gly
180 185 190
Thr Gly Val Ala Phe Ala Asp Trp Ile His Ile Gly Val Phe Phe Arg
195 200 205
Pro Gly Ile Pro Gly Asp Gln Val Ile Phe Ala Ala Thr Pro Val Asn
210 215 220
Thr Pro Gly Val Thr Ile Val Cys Arg Glu Ser Leu Val Lys Asp Asp
225 230 235 240
Lys Val Glu His Pro Leu Ala Ala Gln Gly Asp Glu Leu Asp Gly Met
245 250 255
Thr Val Phe Glu Asn Val Phe Ile Pro Trp Ser His Val Phe His Ile
260 265 270
Gly Asn Pro Asn His Ala Lys Leu Tyr Pro Gln Arg Val Phe Asp Trp
275 280 285
Leu His Tyr His Ala Leu Ile Arg Gln Met Val Arg Ala Glu Leu Val
290 295 300
Ala Gly Leu Ala Val Leu Ile Thr Glu His Ile Gly Thr Asn Lys Ile
305 310 315 320
Pro Ala Val Gln Thr Arg Val Ala Lys Leu Ile Gly Phe His Gln Ala
325 330 335
Met Leu Ala His Leu Ile Ala Ser Glu Glu Leu Gly Phe His Thr Pro
340 345 350
Gly Gly His Tyr Lys Pro Asn Ile Leu Ile Tyr Asp Phe Gly Arg Ala
355 360 365
Leu Tyr Leu Glu Asn Phe Ser Gln Met Ile Tyr Glu Leu Val Asp Leu
370 375 380
Ser Gly Arg Ser Ala Leu Ile Phe Ala Ser Glu Asp Gln Trp Asn Asp
385 390 395 400
Asp Lys Leu Asn Gly Trp Phe Glu Arg Met Asn Asn Gly Pro Val Gly
405 410 415
Arg Pro His Asp Arg Val Lys Ile Gly Arg Val Ile Arg Asp Leu Phe
420 425 430
Leu Thr Asp Trp Gly Ser Arg Leu Phe Val Phe Glu Asn Phe Asn Gly
435 440 445
Thr Pro Leu Gln Gly Ile Arg Met Leu Thr Met Gln Arg Ala Glu Phe
450 455 460
Ser Gly Ser Gly Pro Tyr Gly Lys Leu Ala Arg Gln Val Cys Gly Ile
465 470 475 480
Asp Ser Ala Val Thr Asp Asp Thr Glu Tyr Arg Lys Thr Ala Asp Tyr
485 490 495
Ala Lys Ala Leu Asp Ala Ala Arg His Gln Glu Glu Val Ala Leu Ala
500 505 510
Gly Ala Met Ala Ile
515
<210> 11
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 11
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Leu Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 12
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 12
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 13
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 13
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Gln Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 14
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 14
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Gly Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 15
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 15
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Met Val Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 16
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 16
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Ala Val Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 17
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 17
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Arg
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 18
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 18
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Arg
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 19
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 19
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Met Thr Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 20
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 20
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Ala Thr Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 21
<211> 481
<212> PRT
<213> 嗜热栖热菌
<400> 21
Met Ala Arg Thr Gly Ala Glu Tyr Ile Glu Ala Leu Lys Thr Arg Pro
1 5 10 15
Pro Asn Leu Trp Tyr Lys Gly Glu Lys Val Glu Asp Pro Thr Thr His
20 25 30
Pro Val Phe Arg Gly Ile Val Arg Thr Met Ala Ala Leu Tyr Asp Leu
35 40 45
Gln His Asp Pro Arg Tyr Arg Glu Val Leu Thr Tyr Glu Glu Glu Gly
50 55 60
Lys Arg His Gly Met Ser Phe Leu Ile Pro Lys Thr Lys Glu Asp Leu
65 70 75 80
Lys Arg Arg Gly Gln Ala Tyr Lys Leu Trp Ala Asp Gln Asn Leu Gly
85 90 95
Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Val Val Met Ala Tyr
100 105 110
Ala Ala Ser Ala Asp Tyr Phe Gly Glu Phe Ala Glu Asn Val Arg Asn
115 120 125
Tyr Tyr Arg Tyr Leu Arg Asp Gln Asp Leu Ala Thr Thr His Ala Leu
130 135 140
Thr Asn Pro Gln Val Asn Arg Ala Arg Pro Pro Ser Gly Gln Pro Asp
145 150 155 160
Pro Tyr Ile Pro Val Gly Val Val Lys Gln Thr Glu Lys Gly Ile Val
165 170 175
Val Arg Gly Ala Arg Met Thr Ala Thr Phe Pro Leu Ala Asp Glu Val
180 185 190
Leu Ile Phe Pro Ser Thr Thr Leu Gln Ala Gly Ser Glu Lys Tyr Ala
195 200 205
Leu Ala Phe Ala Leu Pro Thr Ser Thr Pro Gly Leu His Phe Val Cys
210 215 220
Arg Glu Ala Leu Val Gly Gly Asp Ser Pro Phe Asp His Pro Leu Ser
225 230 235 240
Ser Arg Val Glu Glu Met Asp Cys Leu Val Ile Phe Asp Asp Val Leu
245 250 255
Val Pro Trp Glu Arg Val Phe Ile Leu Gly Asn Val Glu Leu Cys Asn
260 265 270
Asn Ala Tyr Ala Ala Thr Gly Ala Leu Asn His Met Ala His Gln Val
275 280 285
Val Ala Leu Lys Thr Ala Lys Thr Glu Ala Phe Leu Gly Val Ala Ala
290 295 300
Leu Met Ala Glu Gly Ile Gly Ala Asp Val Tyr Gly His Val Gln Glu
305 310 315 320
Lys Ile Ala Glu Ile Ile Val Tyr Leu Glu Ala Met Arg Ala Phe Trp
325 330 335
Thr Arg Ala Glu Glu Glu Ala Lys Glu Asn Ala Tyr Gly Leu Leu Val
340 345 350
Pro Asp Arg Gly Ala Leu Asp Gly Ala Arg Asn Leu Tyr Pro Arg Leu
355 360 365
Tyr Pro Arg Ile Arg Glu Ile Leu Glu Gln Ile Gly Ala Ser Gly Leu
370 375 380
Ile Thr Leu Pro Ser Glu Lys Asp Phe Lys Gly Pro Leu Gly Pro Phe
385 390 395 400
Leu Glu Lys Phe Leu Gln Gly Ala Ala Leu Glu Ala Lys Glu Arg Val
405 410 415
Ala Leu Phe Arg Leu Ala Trp Asp Met Thr Leu Ser Gly Phe Gly Ala
420 425 430
Arg Gln Glu Leu Tyr Glu Arg Phe Phe Phe Gly Asp Pro Val Arg Met
435 440 445
Tyr Gln Thr Leu Tyr Asn Val Tyr Asn Lys Glu Pro Tyr Lys Glu Arg
450 455 460
Ile Arg Ala Phe Leu Lys Glu Ser Leu Lys Val Phe Glu Glu Val Gln
465 470 475 480
Ala
<210> 22
<211> 527
<212> PRT
<213> 球孢链霉菌
<400> 22
Met Pro His Gly Ala Glu Arg Glu Ala Ser Pro Ala Glu Glu Ser Ala
1 5 10 15
Gly Thr Arg Pro Leu Thr Gly Glu Glu Tyr Leu Glu Ser Leu Arg Asp
20 25 30
Ala Arg Glu Val Tyr Leu Asp Gly Ser Arg Val Lys Asp Val Thr Ala
35 40 45
His Pro Ala Phe His Asn Pro Ala Arg Met Thr Ala Arg Leu Tyr Asp
50 55 60
Ser Leu His Asp Pro Ala Gln Lys Ala Val Leu Thr Ala Pro Thr Asp
65 70 75 80
Ala Gly Asp Gly Phe Thr His Arg Phe Phe Thr Ala Pro Arg Ser Val
85 90 95
Asp Asp Leu Val Lys Asp Gln Ala Ala Ile Ala Ser Trp Ala Arg Lys
100 105 110
Ser Tyr Gly Trp Met Gly Arg Ser Pro Asp Tyr Lys Ala Ser Phe Leu
115 120 125
Gly Thr Leu Gly Ala Asn Ala Asp Phe Tyr Glu Pro Phe Ala Asp Asn
130 135 140
Ala Arg Arg Trp Tyr Arg Glu Ser Gln Glu Lys Val Leu Tyr Trp Asn
145 150 155 160
His Ala Phe Leu His Pro Pro Val Asp Arg Ser Leu Pro Ala Asp Glu
165 170 175
Val Gly Asp Val Phe Ile His Val Glu Arg Glu Thr Asp Ala Gly Leu
180 185 190
Val Val Ser Gly Ala Lys Val Val Ala Thr Gly Ser Ala Leu Thr His
195 200 205
Ala Ala Phe Ile Ser His Thr Gly Leu Pro Ile Lys Asp Arg Lys Phe
210 215 220
Ala Leu Val Ala Thr Val Pro Met Asp Ala Asp Gly Leu Lys Val Ile
225 230 235 240
Cys Arg Pro Ser Tyr Ser Ala Asn Ala Ala Thr Thr Gly Ser Pro Phe
245 250 255
Asp Asn Pro Leu Ser Ser Arg Leu Asp Glu Asn Asp Ala Ile Leu Val
260 265 270
Leu Asp Gln Val Leu Ile Pro Trp Glu Asn Val Phe Val Tyr Gly Asn
275 280 285
Leu Gly Lys Val His Leu Leu Ala Gly Gln Ser Gly Met Ile Glu Arg
290 295 300
Ala Thr Phe His Gly Cys Thr Arg Leu Ala Val Lys Leu Glu Phe Ile
305 310 315 320
Ala Gly Leu Leu Ala Lys Ala Leu Asp Ile Thr Gly Ala Lys Asp Phe
325 330 335
Arg Gly Val Gln Thr Arg Leu Gly Glu Val Leu Ala Trp Arg Asn Leu
340 345 350
Phe Trp Ser Leu Ser Asp Ala Ala Ala Arg Asn Pro Val Pro Trp Lys
355 360 365
Asn Gly Thr Leu Leu Pro Asn Pro Gln Ala Gly Met Ala Tyr Arg Trp
370 375 380
Phe Met Gln Ile Gly Tyr Pro Arg Val Leu Glu Ile Val Gln Gln Asp
385 390 395 400
Val Ala Ser Gly Leu Met Tyr Val Asn Ser Ser Thr Glu Asp Phe Arg
405 410 415
Asn Pro Glu Thr Gly Pro Tyr Leu Glu Lys Tyr Leu Arg Gly Ser Asp
420 425 430
Gly Ala Gly Ala Val Glu Arg Val Lys Val Met Lys Leu Leu Trp Asp
435 440 445
Ala Val Gly Ser Asp Phe Gly Gly Arg His Glu Leu Tyr Glu Arg Asn
450 455 460
Tyr Ser Gly Asn His Glu Asn Thr Arg Ile Glu Leu Leu Leu Ser Gln
465 470 475 480
Thr Ala Ser Gly Lys Leu Asp Ser Tyr Met Asp Phe Ala Gln Ala Cys
485 490 495
Met Asp Glu Tyr Asp Leu Asp Gly Trp Thr Ala Pro Asp Leu Glu Ser
500 505 510
Phe His Ala Met Arg Ser Ala Ser Arg Asp Leu Leu Gly Gly Leu
515 520 525
<210> 23
<211> 490
<212> PRT
<213> 氨基丁酸梭状芽孢杆菌
<400> 23
Met Leu Met Thr Ala Glu Gln Tyr Ile Glu Ser Leu Arg Lys Leu Asn
1 5 10 15
Thr Arg Val Tyr Met Phe Gly Glu Lys Ile Glu Asn Trp Val Asp His
20 25 30
Pro Met Ile Arg Pro Ser Ile Asn Cys Val Arg Met Thr Tyr Glu Leu
35 40 45
Ala Gln Asp Pro Gln Tyr Ala Asp Leu Met Thr Thr Lys Ser Asn Leu
50 55 60
Ile Gly Lys Thr Ile Asn Arg Phe Ala Asn Leu His Gln Ser Thr Asp
65 70 75 80
Asp Leu Arg Lys Lys Val Lys Met Gln Arg Leu Leu Gly Gln Lys Thr
85 90 95
Ala Ser Cys Phe Gln Arg Cys Val Gly Met Asp Ala Phe Asn Ala Val
100 105 110
Phe Ser Thr Thr Tyr Glu Ile Asp Gln Lys Tyr Gly Thr Asn Tyr His
115 120 125
Lys Asn Phe Thr Glu Tyr Leu Lys Tyr Ile Gln Glu Asn Asp Leu Ile
130 135 140
Val Asp Gly Ala Met Thr Asp Pro Lys Gly Asp Arg Gly Leu Ala Pro
145 150 155 160
Ser Ala Gln Lys Asp Pro Asp Leu Phe Leu Arg Ile Val Glu Lys Arg
165 170 175
Glu Asp Gly Ile Val Val Arg Gly Ala Lys Ala His Gln Thr Gly Ser
180 185 190
Ile Asn Ser His Glu His Ile Ile Met Pro Thr Thr Ala Met Thr Glu
195 200 205
Ala Asp Lys Asp Tyr Ala Val Ser Phe Ala Cys Pro Ser Asp Ala Asp
210 215 220
Gly Leu Phe Met Ile Tyr Gly Arg Gln Ser Cys Asp Thr Arg Lys Met
225 230 235 240
Glu Glu Gly Ala Asp Ile Asp Leu Gly Asn Lys Gln Phe Gly Gly Gln
245 250 255
Glu Ala Leu Val Val Phe Asp Asn Val Phe Ile Pro Asn Asp Arg Ile
260 265 270
Phe Leu Cys Gln Glu Tyr Asp Phe Ala Gly Met Met Val Glu Arg Phe
275 280 285
Ala Gly Tyr His Arg Gln Ser Tyr Gly Gly Cys Lys Val Gly Val Gly
290 295 300
Asp Val Val Ile Gly Ala Ala Ala Leu Ala Ala Asp Tyr Asn Gly Ala
305 310 315 320
Gln Lys Ala Ser His Val Lys Asp Lys Leu Ile Glu Met Thr His Leu
325 330 335
Asn Glu Thr Leu Tyr Cys Cys Gly Ile Ala Cys Ser Ala Glu Gly Tyr
340 345 350
Pro Thr Ala Ala Gly Asn Tyr Gln Ile Asp Leu Leu Leu Ala Asn Val
355 360 365
Cys Lys Gln Asn Ile Thr Arg Phe Pro Tyr Glu Ile Val Arg Leu Ala
370 375 380
Glu Asp Ile Ala Gly Gly Leu Met Val Thr Met Pro Ser Glu Ala Asp
385 390 395 400
Phe Lys Ser Glu Thr Val Val Gly Arg Asp Gly Glu Thr Ile Gly Asp
405 410 415
Phe Cys Asn Lys Phe Phe Ala Ala Ala Pro Thr Cys Thr Thr Glu Glu
420 425 430
Arg Met Arg Val Leu Arg Phe Leu Glu Asn Ile Cys Leu Gly Ala Ser
435 440 445
Ala Val Gly Tyr Arg Thr Glu Ser Met His Gly Ala Gly Ser Pro Gln
450 455 460
Ala Gln Arg Ile Met Ile Ala Arg Gln Gly Asn Ile Asn Ala Lys Lys
465 470 475 480
Glu Leu Ala Lys Ala Ile Ala Gly Ile Lys
485 490
<210> 24
<211> 515
<212> PRT
<213> 洋葱伯克霍尔德菌
<400> 24
Met Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg Val
1 5 10 15
Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro Leu
20 25 30
Thr Arg Asp Tyr Ala Glu Arg Val Ala Gln Phe Tyr Asp Leu His His
35 40 45
Arg Pro Asp Leu Gln Asp Val Leu Thr Phe Val Asp Ala Asp Gly Val
50 55 60
Arg Arg Ser Arg Gln Trp Gln Asp Pro Lys Asp Ala Ala Gly Leu Arg
65 70 75 80
Val Lys Arg Lys Tyr His Glu Thr Ile Leu Arg Glu Ile Ala Ala Gly
85 90 95
Ser Tyr Gly Arg Leu Pro Asp Ala His Asn Tyr Thr Phe Thr Thr Tyr
100 105 110
Ala Asp Asp Pro Glu Val Trp Glu Lys Gln Ser Ile Gly Ala Glu Gly
115 120 125
Arg Asn Leu Thr Gln Asn Ile His Asn Phe Leu Lys Leu Leu Arg Glu
130 135 140
Lys Asp Leu Asn Cys Pro Leu Asn Phe Val Asp Pro Gln Thr Asp Arg
145 150 155 160
Ser Ser Asp Ala Ala Gln Ala Arg Ser Pro Asn Leu Arg Ile Val Glu
165 170 175
Lys Thr Asp Asp Gly Ile Ile Val Asn Gly Val Lys Ala Val Gly Thr
180 185 190
Gly Ile Ala Phe Gly Asp Tyr Met His Ile Gly Cys Thr Tyr Arg Pro
195 200 205
Gly Ile Pro Gly Glu Gln Val Ile Phe Ala Ala Ile Pro Thr Asn Thr
210 215 220
Pro Gly Val Thr Val Phe Cys Arg Glu Ser Thr Val Lys Asn Asp Pro
225 230 235 240
Ala Glu His Pro Leu Ala Ser Gln Gly Asp Glu Leu Asp Ser Thr Thr
245 250 255
Val Phe Asp Asn Val Phe Ile Pro Trp Glu Gln Val Phe His Ile Gly
260 265 270
Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg Ile Phe Asp Trp Val
275 280 285
His Tyr His Ile Leu Ile Arg Gln Val Leu Arg Ala Glu Leu Ile Val
290 295 300
Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly Thr Ser Lys Leu Pro
305 310 315 320
Thr Val Ser Ala Arg Val Ala Lys Leu Val Ala Phe His Leu Ala Met
325 330 335
Gln Ala His Leu Ile Ala Ser Glu Glu Thr Gly Phe His Thr Lys Gly
340 345 350
Gly Arg Tyr Lys Pro Asn Pro Leu Ile Tyr Asp Phe Gly Arg Ala His
355 360 365
Phe Leu Gln Asn Gln Met Ser Val Met Tyr Glu Leu Leu Asp Leu Ala
370 375 380
Gly Arg Ser Ser Leu Met Ile Pro Ser Glu Gly Gln Trp Asp Asp Ser
385 390 395 400
Gln Ser Gly Gln Trp Phe Val Lys Leu Asn Asn Gly Pro Lys Gly Asn
405 410 415
Pro Arg Glu Arg Val Gln Ile Gly Arg Val Ile Arg Asp Leu Tyr Leu
420 425 430
Thr Asp Trp Gly Gly Arg Gln Phe Met Phe Glu Asn Phe Asn Gly Thr
435 440 445
Pro Leu Phe Ala Val Phe Ala Ala Thr Met Thr Arg Asp Asp Met Ser
450 455 460
Ala Ala Gly Thr Tyr Gly Lys Phe Ala Ser Gln Val Cys Gly Ile Glu
465 470 475 480
Phe Gly Gly Ala Glu Pro Thr Ala Tyr Ala Ala Thr Ala Asp Tyr Ala
485 490 495
Lys Ala Leu Asp Lys Gly Leu Ala Pro Glu Pro Ala Ala Ala Glu Ser
500 505 510
Ala Thr Ser
515
<210> 25
<211> 517
<212> PRT
<213> 钩虫贪铜菌
<400> 25
Met Ile Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg
1 5 10 15
Asn Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro
20 25 30
Lys Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His
35 40 45
His Arg Pro Asp Leu Gln Asp Val Met Thr Phe Val Asp Lys Asp Gly
50 55 60
Glu Arg Arg Thr Met Gln Trp Phe Gly His Tyr Asp Lys Glu Gln Leu
65 70 75 80
Arg Arg Lys Arg Lys Tyr His Glu Thr Ile Met Arg Glu Met Ala Gly
85 90 95
Ala Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Gln Thr
100 105 110
Tyr Ile Asp Asp Pro Ser Pro Trp Glu Thr Gln Thr Ile Gly Ala Glu
115 120 125
Gly Lys Val Lys Ala Lys Asn Ile Val Asp Phe Val Asn Phe Ala Lys
130 135 140
Lys His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Met Asp
145 150 155 160
Arg Ser Asn Pro Asp Ala Gln Gln Arg Ser Pro Gly Leu Arg Val Ile
165 170 175
Glu Lys Asn Asp Lys Gly Ile Val Val Ser Gly Val Lys Ala Ile Gly
180 185 190
Thr Gly Val Ala Phe Ala Asp Trp Ile His Ile Gly Thr Phe Phe Arg
195 200 205
Pro Gly Ile Pro Gly Asp Gln Ile Ile Phe Ala Ala Thr Pro Val Asn
210 215 220
Thr Pro Gly Val Thr Ile Val Cys Arg Glu Ser Val Val Lys Glu Asp
225 230 235 240
Pro Ile Glu His Pro Leu Ala Ser Gln Gly Asp Glu Leu Asp Gly Met
245 250 255
Thr Val Phe Asp Asn Val Phe Ile Pro Trp Ser His Val Phe His Leu
260 265 270
Gly Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg Val Phe Asp Trp
275 280 285
Leu His Tyr His Ala Leu Ile Arg Gln Ser Val Arg Ala Glu Leu Met
290 295 300
Ala Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly Thr Asn Lys Ile
305 310 315 320
Pro Ala Val Gln Thr Arg Val Ala Lys Leu Ile Gly Phe His Gln Ala
325 330 335
Met Leu Ala His Ile Val Ala Ser Glu Glu Leu Gly Phe His Thr Pro
340 345 350
Gly Gly Ala Tyr Lys Pro Asn Ile Leu Ile Tyr Asp Phe Gly Arg Ala
355 360 365
Leu Tyr Leu Glu Asn Phe Ser Gln Met Ile Tyr Glu Leu Val Asp Leu
370 375 380
Ser Gly Arg Ser Ala Leu Ile Phe Ala Ser Glu Asp Gln Trp Asn Asp
385 390 395 400
Glu Ala Leu Asn Gly Trp Phe Glu Arg Met Asn Asn Gly Pro Val Gly
405 410 415
Gln Pro His Asp Arg Val Lys Ile Gly Arg Val Ile Arg Asp Leu Phe
420 425 430
Leu Thr Asp Trp Gly Asn Arg Leu Phe Val Phe Glu Asn Phe Asn Gly
435 440 445
Thr Pro Leu Gln Ala Ile Arg Met Leu Thr Met Gln Arg Ala Glu Phe
450 455 460
Ser Ala Ala Gly Pro Tyr Gly Thr Leu Ala Arg Lys Val Cys Gly Ile
465 470 475 480
Glu Leu Thr Glu Gly His Asp Ser Glu Tyr Lys Ala Thr Ala Gly Tyr
485 490 495
Ala Gln Ala Leu Asp Ser Ala Arg His Gln Glu Lys Leu Ala Leu Ser
500 505 510
Gly Thr Met Thr Val
515
<210> 26
<211> 416
<212> PRT
<213> 颤藻属的一个种PCC 6506
<400> 26
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Ala Ser Met Thr Gly Gly Gln Gln Met Gly Arg
20 25 30
Gly Ser Glu Phe Asp Phe Ala Trp Asn Ser Gln Gln Ile Gln Phe Arg
35 40 45
Lys Lys Val Ile Gln Phe Ala Gln Gln Ser Leu Ile Ser Asp Leu Ile
50 55 60
Lys Asn Asp Lys Glu Glu Ile Phe Asn Arg Asp Ala Trp Gln Lys Cys
65 70 75 80
Ser Glu Phe Gly Val His Gly Trp Pro Ile Pro Ala Arg Tyr Gly Gly
85 90 95
Gln Glu Leu Asp Ile Leu Thr Thr Ala Tyr Ala Leu Gln Gly Leu Gly
100 105 110
Tyr Gly Cys Lys Asp Asn Gly Leu Ile Phe Ala Met Asn Ala His Ile
115 120 125
Trp Ala Cys Glu Met Pro Leu Leu Thr Phe Gly Thr Glu Glu Gln Lys
130 135 140
Glu Lys Tyr Leu Pro Leu Leu Cys Arg Gly Gly Trp Ile Ala Ser His
145 150 155 160
Ala Ala Thr Glu Pro Gln Ala Gly Ser Asp Ile Tyr Ser Leu Lys Thr
165 170 175
Thr Ala Gln Lys Asp Gly Asp Lys Tyr Ile Leu Asn Gly Tyr Lys His
180 185 190
Tyr Val Thr Asn Gly Thr Ile Ala Asp Leu Phe Ile Ile Phe Ala Thr
195 200 205
Thr Asp Pro Ser Leu Gly Lys Glu Gly Leu Thr Thr Phe Met Ile Glu
210 215 220
Lys Asp Thr Pro Gly Leu Ile Leu Ser Lys Pro Ile Ser Lys Met Gly
225 230 235 240
Met Arg Thr Ala Glu Val Pro Glu Leu Arg Leu Glu Asn Cys Glu Val
245 250 255
Ser Ala Ala Asn Arg Leu Gly Glu Glu Gly Thr Gly Leu Ala Ile Phe
260 265 270
Asn His Ser Met Glu Trp Glu Arg Gly Phe Ile Leu Ala Ala Ala Val
275 280 285
Gly Thr Met Glu Arg Leu Leu Glu Gln Ser Ile Arg Tyr Ala Arg Ser
290 295 300
His Lys Gln Phe Gly Gln Ala Ile Gly Lys Phe Gln Leu Val Ala Asn
305 310 315 320
Lys Leu Val Glu Met Lys Leu Arg Leu Glu Asn Ala Lys Ala Tyr Leu
325 330 335
Tyr Lys Val Ala Trp Met Lys Glu Asn Lys Gln Met Ala Leu Leu Glu
340 345 350
Ala Ser Met Ala Asn Leu Tyr Ile Ser Glu Ala Trp Val Gln Ser Cys
355 360 365
Leu Glu Ala Ile Glu Ile His Gly Ala Tyr Gly Tyr Leu Thr Asn Thr
370 375 380
Glu Leu Glu Arg Glu Leu Arg Asp Ala Ile Ala Ser Lys Phe Tyr Ser
385 390 395 400
Gly Thr Ser Glu Ile Gln Arg Val Val Ile Ala Lys Phe Leu Gly Leu
405 410 415
<210> 27
<211> 410
<212> PRT
<213> 瘤状副伯克霍尔德菌
<400> 27
Met Ala His His His His His His Met Leu Ala Ala Asn Leu Ile Asp
1 5 10 15
Pro His Gly Ala Leu Ala Trp Pro Phe Phe Glu Ala Arg His Arg Glu
20 25 30
Leu Ala Ala Gly Ile Glu Ala Trp Ala Thr Gln His Leu Ala Cys Val
35 40 45
Gln His Asp Asp Thr Asp Thr Thr Cys Arg Lys Leu Val Arg Ala Leu
50 55 60
Gly Glu Ala Gly Trp Leu Lys Tyr Gly Val Gly Gly Ala Gln Tyr Gly
65 70 75 80
Gly His Gly Asp Thr Ile Asp Thr Arg Ala Val Cys Leu Leu Arg Glu
85 90 95
Thr Leu Ala Asn His Asp Gly Leu Ala Asp Phe Ala Leu Ala Met Gln
100 105 110
Gly Leu Gly Ser Gly Ala Ile Thr Leu Ala Gly Thr His Glu Gln Lys
115 120 125
Ile Arg Tyr Leu Pro Arg Val Ser Lys Gly Glu Ala Ile Ala Ala Phe
130 135 140
Ala Leu Ser Glu Pro Asp Ala Gly Ser Asp Val Ala Ala Met Ser Leu
145 150 155 160
Gln Ala Arg Ala Glu Gly Asp Cys Tyr Val Ile Asp Gly Asp Lys Thr
165 170 175
Trp Ile Ser Asn Gly Gly Ile Ala Asp Phe Tyr Val Val Phe Ala Arg
180 185 190
Met Gly Glu Ala Pro Gly Ala Arg Gly Ile Ser Ala Phe Ile Val Asp
195 200 205
Ala Asp Thr Pro Gly Leu Gln Ile Ala Glu Arg Ile Asp Val Ile Ala
210 215 220
Pro His Pro Leu Ala Arg Leu His Phe Asp Ser Ala Arg Val Pro Arg
225 230 235 240
Ser Gln Met Leu Gly Ala Pro Gly Glu Gly Phe Lys Ile Ala Met Arg
245 250 255
Thr Leu Asp Val Phe Arg Thr Ser Val Ala Ala Ala Ser Leu Gly Phe
260 265 270
Ala Arg Arg Ala Leu Gln Glu Gly Leu Ala Arg Ala Ala Ser Arg Lys
275 280 285
Met Phe Gly Gln Thr Leu Gly Asp Phe Gln Leu Thr Gln Thr Lys Leu
290 295 300
Ala Gln Met Ala Leu Thr Ile Asp Ser Ser Ala Leu Leu Val Tyr Arg
305 310 315 320
Ala Ala Trp Leu Arg Asp Gln Gly Glu Asn Val Thr Arg Glu Ala Ala
325 330 335
Met Ala Lys Trp His Ala Ser Glu Gly Ala Gln Gln Val Ile Asp Ala
340 345 350
Ala Val Gln Leu Trp Gly Gly Met Gly Val Gln Ser Gly Thr Thr Val
355 360 365
Glu Arg Leu Tyr Arg Glu Ile Arg Ala Leu Arg Ile Tyr Glu Gly Ala
370 375 380
Thr Glu Val Gln Gln Leu Ile Val Gly Arg Asp Leu Leu Lys Ala His
385 390 395 400
Ala Ala Gln Arg Gln Gln Glu Arg Ala Ser
405 410
<210> 28
<211> 517
<212> PRT
<213> 皮氏罗尔斯顿菌
<400> 28
Met Ile Arg Thr Gly Thr Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg
1 5 10 15
Asn Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro
20 25 30
Lys Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His
35 40 45
His Arg Pro Asp Leu Gln Asp Val Met Thr Tyr Ile Asp Glu Gly Gly
50 55 60
Gln Arg Arg Ala Met Gln Trp Phe Gly His Arg Asp Lys Glu Gln Leu
65 70 75 80
Arg Arg Lys Arg Lys Tyr His Glu Thr Val Met Arg Glu Met Ala Gly
85 90 95
Ala Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Thr Thr
100 105 110
Tyr Ile Asp Asp Pro Ala Pro Trp Glu Thr Gln Ser Ile Gly Asp Asp
115 120 125
Gly His Ile Lys Ala Gly Lys Ile Val Asp Phe Ile Arg Tyr Ala Arg
130 135 140
Glu His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Met Asp
145 150 155 160
Arg Ser Asn Pro Asp Ala Gln Glu Arg Ser Pro Gly Leu Arg Val Val
165 170 175
Glu Lys Asn Glu Lys Gly Ile Val Val Asn Gly Val Lys Ala Ile Gly
180 185 190
Thr Gly Val Ala Phe Ala Asp Trp Ile His Ile Gly Thr Phe Phe Arg
195 200 205
Pro Gly Ile Pro Gly Asp Gln Val Ile Phe Ala Ala Thr Pro Val Asn
210 215 220
Thr Pro Gly Val Thr Ile Val Cys Arg Glu Ser Leu Val Lys Asp Asp
225 230 235 240
Lys Val Glu His Pro Leu Ala Ala Gln Gly Asp Glu Leu Asp Gly Met
245 250 255
Thr Val Phe Glu Asn Val Phe Ile Pro Trp Ser His Val Phe His Ile
260 265 270
Gly Asn Pro Asn His Ala Lys Leu Tyr Pro Gln Arg Val Phe Asp Trp
275 280 285
Leu His Tyr His Ala Leu Ile Arg Gln Met Val Arg Ala Glu Leu Val
290 295 300
Ala Gly Leu Ala Val Leu Ile Thr Glu His Ile Gly Thr Asn Lys Ile
305 310 315 320
Pro Ala Val Gln Thr Arg Val Ala Lys Leu Ile Gly Phe His Gln Ala
325 330 335
Met Leu Ala His Leu Ile Ala Ser Glu Glu Leu Gly Phe His Thr Pro
340 345 350
Gly Gly His Tyr Lys Pro Asn Ile Leu Ile Tyr Asp Phe Gly Arg Ala
355 360 365
Leu Tyr Leu Glu Asn Phe Ser Gln Met Ile Tyr Glu Leu Val Asp Leu
370 375 380
Ser Gly Arg Ser Ala Leu Ile Phe Ala Ser Glu Asp Gln Trp Asn Asp
385 390 395 400
Asp Lys Leu Asn Gly Trp Phe Glu Arg Met Asn Asn Gly Pro Val Gly
405 410 415
Arg Pro His Asp Arg Val Lys Ile Gly Arg Val Ile Arg Asp Leu Phe
420 425 430
Leu Thr Asp Trp Gly Ser Arg Leu Phe Val Phe Glu Asn Phe Asn Gly
435 440 445
Thr Pro Leu Gln Gly Ile Arg Met Leu Thr Met Gln Arg Ala Glu Phe
450 455 460
Ser Gly Ser Gly Pro Tyr Gly Lys Leu Ala Arg Gln Val Cys Gly Ile
465 470 475 480
Asp Ser Ala Val Thr Asp Asp Thr Glu Tyr Arg Lys Thr Ala Asp Tyr
485 490 495
Ala Lys Ala Leu Asp Ala Ala Arg His Gln Glu Glu Val Ala Leu Ala
500 505 510
Gly Ala Met Ala Ile
515
<210> 29
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 29
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Lys
195 200 205
Lys Ser Asn Thr Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 30
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 30
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Lys
195 200 205
Lys Val Met Asn Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 31
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 31
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Lys
195 200 205
Lys Ala Thr Ser Ala Gln Asn Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 32
<211> 494
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 32
Met Lys Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys
1 5 10 15
Gln Ala Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val
20 25 30
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
35 40 45
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
50 55 60
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
65 70 75 80
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
85 90 95
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
100 105 110
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
115 120 125
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
130 135 140
Ile Ser Leu Thr His Thr Leu Leu His Pro Gln Met Asn Arg Ala Lys
145 150 155 160
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
165 170 175
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
180 185 190
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Thr Lys
195 200 205
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
210 215 220
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
225 230 235 240
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
245 250 255
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
260 265 270
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
275 280 285
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
290 295 300
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
305 310 315 320
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
325 330 335
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
340 345 350
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
355 360 365
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
370 375 380
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
385 390 395 400
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
405 410 415
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
420 425 430
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
435 440 445
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
450 455 460
Asp Gly Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu
465 470 475 480
Arg Gly Ala Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 33
<211> 490
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 33
Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys Gln Ala Lys Ser
1 5 10 15
Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val Thr Val His Pro
20 25 30
Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu Tyr Asp Arg Gln
35 40 45
Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro Thr Thr Gly Gln
50 55 60
Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile Asp Glu Leu Ile
65 70 75 80
Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met Ser Ala Gly Met
85 90 95
Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val Met Ala Met Gly
100 105 110
Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met Phe Ala Glu Asn
115 120 125
Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp Ile Ser Leu Thr
130 135 140
His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys Ala Leu His Glu
145 150 155 160
Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu Arg Arg Lys Asp
165 170 175
Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr Gln Gly Gly Ile
180 185 190
Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys Lys Ala Thr Ser
195 200 205
Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro Asn Asn Thr Pro
210 215 220
Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr Gly Arg Ser Ala
225 230 235 240
Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly Asp Ala Ile Val
245 250 255
Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val Phe Val Cys Gly
260 265 270
Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr Asn Ala Val Val
275 280 285
His Met Ser His Gln Val Val Ala Lys Asn Ile Val Lys Thr Glu Phe
290 295 300
Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile Gly Ile Gly Glu
305 310 315 320
Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met Leu Val Leu Glu
325 330 335
Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn Ala Lys Arg Asp
340 345 350
Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu Asp Ala Ala Arg
355 360 365
Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu Ile Ile Arg Ile
370 375 380
Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu Ala Asp Phe Gln
385 390 395 400
His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met Gln Gly Ala Thr
405 410 415
Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu Ala Trp Asp Leu
420 425 430
Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr Glu Tyr Tyr Phe
435 440 445
Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe Asp Gly Tyr Glu
450 455 460
Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu Arg Gly Ala Lys
465 470 475 480
Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 34
<211> 473
<212> PRT
<213> 嗜热栖热菌
<400> 34
Ala Arg Thr Gly Ala Glu Tyr Ile Glu Ala Leu Lys Thr Arg Pro Pro
1 5 10 15
Asn Leu Trp Tyr Lys Gly Glu Lys Val Glu Asp Pro Thr Thr His Pro
20 25 30
Val Phe Arg Gly Ile Val Arg Thr Met Ala Ala Leu Tyr Asp Leu Gln
35 40 45
His Asp Pro Arg Tyr Arg Glu Val Leu Thr Tyr Glu Glu Glu Gly Lys
50 55 60
Arg His Gly Met Ser Phe Leu Ile Pro Lys Thr Lys Glu Asp Leu Lys
65 70 75 80
Arg Arg Gly Gln Ala Tyr Lys Leu Trp Ala Asp Gln Asn Leu Gly Met
85 90 95
Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Val Val Met Ala Tyr Ala
100 105 110
Ala Ser Ala Asp Tyr Phe Gly Glu Phe Ala Glu Asn Val Arg Asn Tyr
115 120 125
Tyr Arg Tyr Leu Arg Asp Gln Asp Leu Ala Thr Thr His Ala Leu Thr
130 135 140
Asn Pro Gln Val Asn Arg Ala Arg Pro Pro Ser Gly Gln Pro Asp Pro
145 150 155 160
Tyr Ile Pro Val Gly Val Val Lys Gln Thr Glu Lys Gly Ile Val Val
165 170 175
Arg Gly Ala Arg Met Thr Ala Thr Phe Pro Leu Ala Asp Glu Val Leu
180 185 190
Ile Phe Pro Ser Thr Leu Leu Gln Ala Gly Ser Glu Lys Tyr Ala Leu
195 200 205
Ala Phe Ala Leu Pro Thr Ser Thr Pro Gly Leu His Phe Val Cys Arg
210 215 220
Glu Ala Leu Val Gly Gly Asp Ser Pro Phe Asp His Pro Leu Ser Ser
225 230 235 240
Arg Val Glu Glu Met Asp Cys Leu Val Ile Phe Asp Asp Val Leu Val
245 250 255
Pro Trp Glu Arg Val Phe Ile Leu Gly Asn Val Glu Leu Cys Asn Asn
260 265 270
Ala Tyr Ala Ala Thr Gly Ala Leu Asn His Met Ala His Gln Val Val
275 280 285
Ala Leu Lys Thr Ala Lys Thr Glu Ala Phe Leu Gly Val Ala Ala Leu
290 295 300
Met Ala Glu Gly Ile Gly Ala Asp Val Tyr Gly His Val Gln Glu Lys
305 310 315 320
Ile Ala Glu Ile Ile Val Tyr Leu Glu Ala Met Arg Ala Phe Trp Thr
325 330 335
Arg Ala Glu Glu Glu Ala Lys Glu Asn Ala Tyr Gly Leu Leu Val Pro
340 345 350
Asp Arg Gly Ala Leu Asp Gly Ala Arg Asn Leu Tyr Pro Arg Leu Tyr
355 360 365
Pro Arg Ile Arg Glu Ile Leu Glu Gln Ile Gly Ala Ser Gly Leu Ile
370 375 380
Thr Leu Pro Ser Glu Lys Asp Phe Lys Gly Pro Leu Gly Pro Phe Leu
385 390 395 400
Glu Lys Phe Leu Gln Gly Ala Ala Leu Glu Ala Lys Glu Arg Val Ala
405 410 415
Leu Phe Arg Leu Ala Trp Asp Met Thr Leu Ser Gly Phe Gly Ala Arg
420 425 430
Gln Glu Leu Tyr Glu Arg Phe Phe Phe Gly Asp Pro Val Arg Met Tyr
435 440 445
Gln Thr Leu Tyr Asn Val Tyr Asn Lys Glu Pro Tyr Lys Glu Arg Ile
450 455 460
Arg Ala Phe Leu Lys Glu Ser Leu Lys
465 470
<210> 35
<211> 488
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 35
Met Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys Gln Ala
1 5 10 15
Lys Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val Thr Val
20 25 30
His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu Tyr Asp
35 40 45
Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro Thr Thr
50 55 60
Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile Asp Glu
65 70 75 80
Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met Ser Ala
85 90 95
Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val Met Ala
100 105 110
Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met Phe Ala
115 120 125
Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp Ile Ser
130 135 140
Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys Ala Leu
145 150 155 160
His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu Arg Arg
165 170 175
Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr Gln Gly
180 185 190
Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys Lys Ala
195 200 205
Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro Asn Asn
210 215 220
Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr Gly Arg
225 230 235 240
Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly Asp Ala
245 250 255
Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val Phe Val
260 265 270
Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr Asn Ala
275 280 285
Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val Lys Thr
290 295 300
Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile Gly Ile
305 310 315 320
Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met Leu Val
325 330 335
Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn Ala Lys
340 345 350
Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu Asp Ala
355 360 365
Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu Ile Ile
370 375 380
Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu Ala Asp
385 390 395 400
Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met Gln Gly
405 410 415
Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu Ala Trp
420 425 430
Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr Glu Tyr
435 440 445
Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe Asp Gly
450 455 460
Tyr Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu Arg Gly
465 470 475 480
Ala Lys Ser Val Phe Ile Pro Ala
485
<210> 36
<211> 491
<212> PRT
<213> 球孢链霉菌
<400> 36
Thr Arg Pro Leu Thr Gly Glu Glu Tyr Leu Glu Ser Leu Arg Asp Ala
1 5 10 15
Arg Glu Val Tyr Leu Asp Gly Ser Arg Val Lys Asp Val Thr Ala His
20 25 30
Pro Ala Phe His Asn Pro Ala Arg Met Thr Ala Arg Leu Tyr Asp Ser
35 40 45
Leu His Asp Pro Ala Gln Lys Ala Val Leu Thr Ala Pro Thr Asp Ala
50 55 60
Gly Asp Gly Phe Thr His Arg Phe Phe Thr Ala Pro Arg Ser Val Asp
65 70 75 80
Asp Leu Val Lys Asp Gln Ala Ala Ile Ala Ser Trp Ala Arg Lys Ser
85 90 95
Tyr Gly Trp Met Gly Arg Ser Pro Asp Tyr Lys Ala Ser Phe Leu Gly
100 105 110
Thr Leu Gly Ala Asn Ala Asp Phe Tyr Glu Pro Phe Ala Asp Asn Ala
115 120 125
Arg Arg Trp Tyr Arg Glu Ser Gln Glu Lys Val Leu Tyr Trp Asn His
130 135 140
Ala Phe Leu His Pro Pro Val Asp Arg Ser Glu Val Gly Asp Val Phe
145 150 155 160
Ile His Val Glu Arg Glu Thr Asp Ala Gly Leu Val Val Ser Gly Ala
165 170 175
Lys Val Val Ala Thr Gly Ser Ala Leu Thr His Ala Ala Phe Ile Ser
180 185 190
His Trp Gly Leu Pro Ile Lys Asp Arg Lys Phe Ala Leu Val Ala Thr
195 200 205
Val Pro Met Asp Ala Asp Gly Leu Lys Val Ile Cys Arg Pro Ser Tyr
210 215 220
Ser Ala Asn Ala Ala Thr Thr Gly Ser Pro Phe Asp Asn Pro Leu Ser
225 230 235 240
Ser Arg Leu Asp Glu Asn Asp Ala Ile Leu Val Leu Asp Gln Val Leu
245 250 255
Ile Pro Trp Glu Asn Val Phe Val Tyr Gly Asn Leu Gly Lys Val His
260 265 270
Leu Leu Ala Gly Gln Ser Gly Met Ile Glu Arg Ala Thr Phe His Gly
275 280 285
Cys Thr Arg Leu Ala Val Lys Leu Glu Phe Ile Ala Gly Leu Leu Ala
290 295 300
Lys Ala Leu Asp Ile Thr Gly Ala Lys Asp Phe Arg Gly Val Gln Thr
305 310 315 320
Arg Leu Gly Glu Val Leu Ala Trp Arg Asn Leu Phe Trp Ser Leu Ser
325 330 335
Asp Ala Ala Ala Arg Asn Pro Val Pro Trp Lys Asn Gly Thr Leu Leu
340 345 350
Pro Asn Pro Gln Ala Gly Met Ala Tyr Arg Trp Phe Met Gln Ile Gly
355 360 365
Tyr Pro Arg Val Leu Glu Ile Val Gln Gln Asp Val Ala Ser Gly Leu
370 375 380
Met Tyr Val Asn Ser Ser Thr Glu Asp Phe Arg Asn Pro Glu Thr Gly
385 390 395 400
Pro Tyr Leu Glu Lys Tyr Leu Arg Gly Ser Asp Gly Ala Gly Ala Val
405 410 415
Glu Arg Val Lys Val Met Lys Leu Leu Trp Asp Ala Val Gly Ser Asp
420 425 430
Phe Gly Gly Arg His Glu Leu Tyr Glu Arg Asn Tyr Ser Gly Asn His
435 440 445
Glu Asn Thr Arg Ile Glu Leu Leu Leu Ser Gln Thr Ala Ser Gly Lys
450 455 460
Leu Asp Ser Tyr Met Asp Phe Ala Gln Ala Cys Met Asp Glu Tyr Asp
465 470 475 480
Leu Asp Gly Trp Thr Ala Pro Asp Leu Glu Ser
485 490
<210> 37
<211> 491
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 37
Pro Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys Gln Ala Lys
1 5 10 15
Ser Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val Thr Val His
20 25 30
Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu Tyr Asp Arg
35 40 45
Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro Thr Thr Gly
50 55 60
Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile Asp Glu Leu
65 70 75 80
Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met Ser Ala Gly
85 90 95
Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val Met Ala Met
100 105 110
Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met Phe Ala Glu
115 120 125
Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp Ile Ser Leu
130 135 140
Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys Ala Leu His
145 150 155 160
Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu Arg Arg Lys
165 170 175
Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr Gln Gly Gly
180 185 190
Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys Lys Ala Thr
195 200 205
Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro Asn Asn Thr
210 215 220
Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr Gly Arg Ser
225 230 235 240
Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly Asp Ala Ile
245 250 255
Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val Phe Val Cys
260 265 270
Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr Asn Ala Val
275 280 285
Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val Lys Thr Glu
290 295 300
Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile Gly Ile Gly
305 310 315 320
Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met Leu Val Leu
325 330 335
Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn Ala Lys Arg
340 345 350
Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu Asp Ala Ala
355 360 365
Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu Ile Ile Arg
370 375 380
Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu Ala Asp Phe
385 390 395 400
Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met Gln Gly Ala
405 410 415
Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu Ala Trp Asp
420 425 430
Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr Glu Tyr Tyr
435 440 445
Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe Asp Gly Tyr
450 455 460
Glu Lys Glu Pro Tyr Lys Gln Phe Val Arg Glu Phe Leu Arg Gly Ala
465 470 475 480
Lys Ser Val Phe Ile Pro Ala Asp Asn Lys His
485 490
<210> 38
<211> 490
<212> PRT
<213> 氨基丁酸梭状芽孢杆菌
<400> 38
Met Leu Met Thr Ala Glu Gln Tyr Ile Glu Ser Leu Arg Lys Leu Asn
1 5 10 15
Thr Arg Val Tyr Met Phe Gly Glu Lys Ile Glu Asn Trp Val Asp His
20 25 30
Pro Met Ile Arg Pro Ser Ile Asn Cys Val Arg Met Thr Tyr Glu Leu
35 40 45
Ala Gln Asp Pro Gln Tyr Ala Asp Leu Met Thr Thr Lys Ser Asn Leu
50 55 60
Ile Gly Lys Thr Ile Asn Arg Phe Ala Asn Leu His Gln Ser Thr Asp
65 70 75 80
Asp Leu Arg Lys Lys Val Lys Met Gln Arg Leu Leu Gly Gln Lys Thr
85 90 95
Ala Ser Cys Phe Gln Arg Cys Val Gly Met Asp Ala Phe Asn Ala Val
100 105 110
Phe Ser Thr Thr Tyr Glu Ile Asp Gln Lys Tyr Gly Thr Asn Tyr His
115 120 125
Lys Asn Phe Thr Glu Tyr Leu Lys Tyr Ile Gln Glu Asn Asp Leu Ile
130 135 140
Val Asp Gly Ala Met Thr Asp Pro Lys Gly Asp Arg Gly Leu Ala Pro
145 150 155 160
Ser Ala Gln Lys Asp Pro Asp Leu Phe Leu Arg Ile Val Glu Lys Arg
165 170 175
Glu Asp Gly Ile Val Val Arg Gly Ala Lys Ala His Gln Thr Gly Ser
180 185 190
Ile Asn Ser His Glu His Ile Ile Met Pro Thr Ile Ala Met Thr Glu
195 200 205
Ala Asp Lys Asp Tyr Ala Val Ser Phe Ala Cys Pro Ser Asp Ala Asp
210 215 220
Gly Leu Phe Met Ile Tyr Gly Arg Gln Ser Cys Asp Thr Arg Lys Met
225 230 235 240
Glu Glu Gly Ala Asp Ile Asp Leu Gly Asn Lys Gln Phe Gly Gly Gln
245 250 255
Glu Ala Leu Val Val Phe Asp Asn Val Phe Ile Pro Asn Asp Arg Ile
260 265 270
Phe Leu Cys Gln Glu Tyr Asp Phe Ala Gly Met Met Val Glu Arg Phe
275 280 285
Ala Gly Tyr His Arg Gln Ser Tyr Gly Gly Cys Lys Val Gly Val Gly
290 295 300
Asp Val Val Ile Gly Ala Ala Ala Leu Ala Ala Asp Tyr Asn Gly Ala
305 310 315 320
Gln Lys Ala Ser His Val Lys Asp Lys Leu Ile Glu Met Thr His Leu
325 330 335
Asn Glu Thr Leu Tyr Cys Cys Gly Ile Ala Cys Ser Ala Glu Gly Tyr
340 345 350
Pro Thr Ala Ala Gly Asn Tyr Gln Ile Asp Leu Leu Leu Ala Asn Val
355 360 365
Cys Lys Gln Asn Ile Thr Arg Phe Pro Tyr Glu Ile Val Arg Leu Ala
370 375 380
Glu Asp Ile Ala Gly Gly Leu Met Val Thr Met Pro Ser Glu Ala Asp
385 390 395 400
Phe Lys Ser Glu Thr Val Val Gly Arg Asp Gly Glu Thr Ile Gly Asp
405 410 415
Phe Cys Asn Lys Phe Phe Ala Ala Ala Pro Thr Cys Thr Thr Glu Glu
420 425 430
Arg Met Arg Val Leu Arg Phe Leu Glu Asn Ile Cys Leu Gly Ala Ser
435 440 445
Ala Val Gly Tyr Arg Thr Glu Ser Met His Gly Ala Gly Ser Pro Gln
450 455 460
Ala Gln Arg Ile Met Ile Ala Arg Gln Gly Asn Ile Asn Ala Lys Lys
465 470 475 480
Glu Leu Ala Lys Ala Ile Ala Gly Ile Lys
485 490
<210> 39
<211> 480
<212> PRT
<213> 洋葱伯克霍尔德菌
<400> 39
Met Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg Val
1 5 10 15
Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro Leu
20 25 30
Thr Arg Asp Tyr Ala Glu Arg Val Ala Gln Phe Tyr Asp Leu His His
35 40 45
Arg Pro Asp Leu Gln Asp Val Leu Thr Phe Val Asp Ala Asp Gly Val
50 55 60
Arg Arg Ser Arg Gln Trp Gln Asp Pro Lys Asp Ala Ala Gly Leu Arg
65 70 75 80
Val Lys Arg Lys Tyr His Glu Thr Ile Leu Arg Glu Ile Ala Ala Gly
85 90 95
Ser Tyr Gly Arg Leu Pro Asp Ala His Asn Tyr Thr Phe Thr Thr Tyr
100 105 110
Ala Asp Asp Pro Glu Val Trp Glu Lys Gln Ser Ile Gly Ala Glu Gly
115 120 125
Arg Asn Leu Thr Gln Asn Ile His Asn Phe Leu Lys Leu Leu Arg Glu
130 135 140
Lys Asp Leu Asn Cys Pro Leu Asn Phe Val Asp Pro Gln Thr Asp Arg
145 150 155 160
Ser Ser Asp Ala Ala Gln Ala Arg Ser Pro Asn Leu Arg Ile Val Glu
165 170 175
Lys Thr Asp Asp Gly Ile Ile Val Asn Gly Val Lys Ala Val Gly Thr
180 185 190
Gly Ile Ala Phe Gly Asp Tyr Met His Ile Gly Cys Leu Tyr Arg Pro
195 200 205
Gly Ile Pro Gly Glu Gln Val Ile Phe Ala Ala Ile Pro Thr Asn Thr
210 215 220
Pro Gly Val Thr Val Phe Cys Arg Glu Ser Thr Val Lys Asn Asp Pro
225 230 235 240
Ala Glu His Pro Leu Ala Ser Gln Gly Asp Glu Leu Asp Ser Thr Thr
245 250 255
Val Phe Asp Asn Val Phe Ile Pro Trp Glu Gln Val Phe His Ile Gly
260 265 270
Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg Ile Phe Asp Trp Val
275 280 285
His Tyr His Ile Leu Ile Arg Gln Val Leu Arg Ala Glu Leu Ile Val
290 295 300
Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly Thr Ser Lys Leu Pro
305 310 315 320
Thr Val Ser Ala Arg Val Ala Lys Leu Val Ala Phe His Leu Ala Met
325 330 335
Gln Ala His Leu Ile Ala Ser Glu Glu Thr Gly Phe His Thr Lys Gly
340 345 350
Gly Arg Tyr Lys Pro Asn Pro Leu Ile Tyr Asp Phe Gly Arg Ala His
355 360 365
Phe Leu Gln Asn Gln Met Ser Val Met Tyr Glu Leu Leu Asp Leu Ala
370 375 380
Gly Arg Ser Ser Leu Met Ile Pro Ser Glu Gly Gln Trp Asp Asp Ser
385 390 395 400
Gln Ser Gly Gln Trp Phe Val Lys Leu Asn Asn Gly Pro Lys Gly Asn
405 410 415
Pro Arg Glu Arg Val Gln Ile Gly Arg Val Ile Arg Asp Leu Tyr Leu
420 425 430
Thr Asp Trp Gly Gly Arg Gln Phe Met Phe Glu Asn Phe Asn Gly Thr
435 440 445
Pro Leu Phe Ala Val Phe Ala Ala Thr Met Thr Arg Asp Asp Met Ser
450 455 460
Ala Ala Gly Thr Tyr Gly Lys Phe Ala Ser Gln Val Cys Gly Ile Glu
465 470 475 480
<210> 40
<211> 469
<212> PRT
<213> 钩虫贪铜菌
<400> 40
Ile Arg Thr Gly Lys Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg Asn
1 5 10 15
Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro Lys
20 25 30
Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His His
35 40 45
Arg Pro Asp Leu Gln Asp Val Met Thr Phe Val Asp Lys Asp Gly Glu
50 55 60
Arg Arg Thr Met Gln Trp Phe Gly His Tyr Asp Lys Glu Gln Leu Arg
65 70 75 80
Arg Lys Arg Lys Tyr His Glu Thr Ile Met Arg Glu Met Ala Gly Ala
85 90 95
Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Gln Thr Tyr
100 105 110
Ile Asp Asp Pro Ser Pro Trp Glu Thr Gln Thr Ile Gly Ala Glu Gly
115 120 125
Lys Val Lys Ala Lys Asn Ile Val Asp Phe Val Asn Phe Ala Lys Lys
130 135 140
His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Met Pro Gly
145 150 155 160
Leu Arg Val Ile Glu Lys Asn Asp Lys Gly Ile Val Val Ser Gly Val
165 170 175
Lys Ala Ile Gly Thr Gly Val Ala Phe Ala Asp Trp Ile His Ile Gly
180 185 190
Val Phe Phe Arg Pro Gly Ile Pro Gly Asp Gln Ile Ile Phe Ala Ala
195 200 205
Thr Pro Val Asn Thr Pro Gly Val Thr Ile Val Cys Arg Glu Ser Val
210 215 220
Val Lys Glu Asp Pro Ile Glu His Pro Leu Ala Ser Gln Gly Asp Glu
225 230 235 240
Leu Asp Gly Met Thr Val Phe Asp Asn Val Phe Ile Pro Trp Ser His
245 250 255
Val Phe His Leu Gly Asn Pro Glu His Ala Lys Leu Tyr Pro Gln Arg
260 265 270
Val Phe Asp Trp Leu His Tyr His Ala Leu Ile Arg Gln Ser Val Arg
275 280 285
Ala Glu Leu Met Ala Gly Leu Ala Ile Leu Ile Thr Glu His Ile Gly
290 295 300
Thr Asn Lys Ile Pro Ala Val Gln Thr Arg Val Ala Lys Leu Ile Gly
305 310 315 320
Phe His Gln Ala Met Leu Ala His Ile Val Ala Ser Glu Glu Leu Gly
325 330 335
Phe His Thr Pro Gly Gly Ala Tyr Lys Pro Asn Ile Leu Ile Tyr Asp
340 345 350
Phe Gly Arg Ala Leu Tyr Leu Glu Asn Phe Ser Gln Met Ile Tyr Glu
355 360 365
Leu Val Asp Leu Ser Gly Arg Ser Ala Leu Ile Phe Ala Ser Glu Asp
370 375 380
Gln Trp Asn Asp Glu Ala Leu Asn Gly Trp Phe Glu Arg Met Asn Asn
385 390 395 400
Gly Pro Val Gly Gln Pro His Asp Arg Val Lys Ile Gly Arg Val Ile
405 410 415
Arg Asp Leu Phe Leu Thr Asp Trp Gly Asn Arg Leu Phe Val Phe Glu
420 425 430
Asn Phe Asn Gly Thr Pro Leu Gln Ala Ile Arg Met Leu Thr Met Gln
435 440 445
Arg Ala Glu Phe Ser Ala Ala Gly Pro Tyr Gly Thr Leu Ala Arg Lys
450 455 460
Val Cys Gly Ile Glu
465
<210> 41
<211> 436
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 41
Thr Val His Pro Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu
1 5 10 15
Tyr Asp Arg Gln Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro
20 25 30
Thr Thr Gly Gln Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile
35 40 45
Asp Glu Leu Ile Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met
50 55 60
Ser Ala Gly Met Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val
65 70 75 80
Met Ala Met Gly Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met
85 90 95
Phe Ala Glu Asn Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp
100 105 110
Ile Ser Leu Thr His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys
115 120 125
Ala Leu His Glu Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu
130 135 140
Arg Arg Lys Asp Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr
145 150 155 160
Gln Gly Gly Ile Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys
165 170 175
Lys Ala Thr Ser Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro
180 185 190
Asn Asn Thr Pro Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr
195 200 205
Gly Arg Ser Ala Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly
210 215 220
Asp Ala Ile Val Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val
225 230 235 240
Phe Val Cys Gly Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr
245 250 255
Asn Ala Val Val His Met Ser His Gln Val Val Ala Lys Asn Ile Val
260 265 270
Lys Thr Glu Phe Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile
275 280 285
Gly Ile Gly Glu Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met
290 295 300
Leu Val Leu Glu Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn
305 310 315 320
Ala Lys Arg Asp Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu
325 330 335
Asp Ala Ala Arg Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu
340 345 350
Ile Ile Arg Ile Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu
355 360 365
Ala Asp Phe Gln His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met
370 375 380
Gln Gly Ala Thr Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu
385 390 395 400
Ala Trp Asp Leu Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr
405 410 415
Glu Tyr Tyr Phe Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe
420 425 430
Asp Gly Tyr Glu
435
<210> 42
<211> 377
<212> PRT
<213> 颤藻属的一个种
<400> 42
Ala Trp Asn Ser Gln Gln Ile Gln Phe Arg Lys Lys Val Ile Gln Phe
1 5 10 15
Ala Gln Gln Ser Leu Ile Ser Asp Leu Ile Lys Asn Asp Lys Glu Glu
20 25 30
Ile Phe Asn Arg Asp Ala Trp Gln Lys Cys Ser Glu Phe Gly Val His
35 40 45
Gly Trp Pro Ile Pro Ala Arg Tyr Gly Gly Gln Glu Leu Asp Ile Leu
50 55 60
Thr Thr Ala Tyr Ala Leu Gln Gly Leu Gly Tyr Gly Cys Lys Asp Asn
65 70 75 80
Gly Leu Ile Phe Ala Met Asn Ala His Ile Trp Ala Cys Glu Met Pro
85 90 95
Leu Leu Thr Phe Gly Thr Glu Glu Gln Lys Glu Lys Tyr Leu Pro Leu
100 105 110
Leu Cys Arg Gly Gly Trp Ile Ala Ser His Ala Ala Thr Glu Pro Gln
115 120 125
Ala Gly Ser Asp Ile Tyr Ser Leu Lys Thr Thr Ala Gln Lys Asp Gly
130 135 140
Asp Lys Tyr Ile Leu Asn Gly Tyr Lys His Tyr Val Thr Asn Gly Thr
145 150 155 160
Ile Ala Asp Leu Phe Ile Ile Phe Ala Thr Ile Asp Pro Ser Leu Gly
165 170 175
Lys Glu Gly Leu Thr Thr Phe Met Ile Glu Lys Asp Thr Pro Gly Leu
180 185 190
Ile Leu Ser Lys Pro Ile Ser Lys Met Gly Met Arg Thr Ala Glu Val
195 200 205
Pro Glu Leu Arg Leu Glu Asn Cys Glu Val Ser Ala Ala Asn Arg Leu
210 215 220
Gly Glu Glu Gly Thr Gly Leu Ala Ile Phe Asn His Ser Met Glu Trp
225 230 235 240
Glu Arg Gly Phe Ile Leu Ala Ala Ala Val Gly Thr Met Glu Arg Leu
245 250 255
Leu Glu Gln Ser Ile Arg Tyr Ala Arg Ser His Lys Gln Phe Gly Gln
260 265 270
Ala Ile Gly Lys Phe Gln Leu Val Ala Asn Lys Leu Val Glu Met Lys
275 280 285
Leu Arg Leu Glu Asn Ala Lys Ala Tyr Leu Tyr Lys Val Ala Trp Met
290 295 300
Lys Glu Asn Lys Gln Met Ala Leu Leu Glu Ala Ser Met Ala Asn Leu
305 310 315 320
Tyr Ile Ser Glu Ala Trp Val Gln Ser Cys Leu Glu Ala Ile Glu Ile
325 330 335
His Gly Ala Tyr Gly Tyr Leu Thr Asn Thr Glu Leu Glu Arg Glu Leu
340 345 350
Arg Asp Ala Ile Ala Ser Lys Phe Tyr Ser Gly Thr Ser Glu Ile Gln
355 360 365
Arg Val Val Ile Ala Lys Phe Leu Gly
370 375
<210> 43
<211> 470
<212> PRT
<213> 地芽孢杆菌属的一个种
<400> 43
Ala Lys Thr Gly Lys Glu Tyr Met Glu Arg Leu Lys Gln Ala Lys Ser
1 5 10 15
Ser Val Tyr Ile His Gly Glu Lys Val Glu Asp Val Thr Val His Pro
20 25 30
Ala Phe Arg Asn Val Val Arg Ser Met Ala Ala Leu Tyr Asp Arg Gln
35 40 45
Tyr Glu Lys Pro Glu Lys Met Leu Tyr Arg Ser Pro Thr Thr Gly Gln
50 55 60
Pro Val Gly Met Thr Phe Ile Gln Pro Thr Thr Ile Asp Glu Leu Ile
65 70 75 80
Ala Arg Arg Glu Ala Thr Gln Glu Trp Ala Arg Met Ser Ala Gly Met
85 90 95
Met Gly Arg Ser Pro Asp Tyr Leu Asn Ala Glu Val Met Ala Met Gly
100 105 110
Ile Ala Asn Asp Leu Phe Ala Glu Asp Asp Pro Met Phe Ala Glu Asn
115 120 125
Ala Lys Asn Tyr Tyr Glu Tyr Ala Arg Glu His Asp Ile Ser Leu Thr
130 135 140
His Thr Leu Ile His Pro Gln Met Asn Arg Ala Lys Ala Leu His Glu
145 150 155 160
Gln Asn Asp Ala Asp Val Pro Leu His Leu Val Glu Arg Arg Lys Asp
165 170 175
Gly Ile Ile Val Ser Gly Ile Arg Leu Leu Ala Thr Gln Gly Gly Ile
180 185 190
Thr Asp Glu Ile Leu Val Phe Pro Ser Thr Val Lys Lys Ala Thr Ser
195 200 205
Gly Glu Asp Pro Tyr Ala Leu Ala Phe Ala Ile Pro Asn Asn Thr Pro
210 215 220
Gly Val Lys Phe Ile Cys Arg Glu Ala Phe Asp Tyr Gly Arg Ser Ala
225 230 235 240
Trp Asp His Pro Leu Ala Ser Arg Phe Glu Glu Gly Asp Ala Ile Val
245 250 255
Ser Phe Glu Asn Val Phe Val Pro Trp Glu Arg Val Phe Val Cys Gly
260 265 270
Asn Ser Ser Ile Cys Asn Arg Thr Phe Arg Glu Thr Asn Ala Val Val
275 280 285
His Met Ser His Gln Val Val Ala Lys Asn Ile Val Lys Thr Glu Phe
290 295 300
Leu Leu Gly Val Thr Leu Cys Leu Ile Glu Ala Ile Gly Ile Gly Glu
305 310 315 320
Phe Gln His Val Lys Asp Lys Gly Ala Glu Ile Met Leu Val Leu Glu
325 330 335
Thr Met Lys Ser His Leu Tyr Arg Ala Glu His Asn Ala Lys Arg Asp
340 345 350
Arg Trp Gly Thr Met Thr Pro Asp Phe Ala Ala Leu Asp Ala Ala Arg
355 360 365
Asn Trp Tyr Pro Arg Ile Tyr Pro Arg Leu Ala Glu Ile Ile Arg Ile
370 375 380
Leu Gly Ala Ser Gly Leu Met Ala Ile Pro Thr Glu Ala Asp Phe Gln
385 390 395 400
His Glu Glu Ile Gly Asp Ile Val Arg Arg Ala Met Gln Gly Ala Thr
405 410 415
Val Asp Gly Tyr Glu Arg Val Gln Leu Phe Arg Leu Ala Trp Asp Leu
420 425 430
Thr Met Ser Ala Phe Gly Ala Arg Gln Thr His Tyr Glu Tyr Tyr Phe
435 440 445
Phe Gly Asp Pro Val Arg Met Gly Met Ala Tyr Phe Asp Gly Tyr Glu
450 455 460
Lys Glu Pro Tyr Lys Gln
465 470
<210> 44
<211> 379
<212> PRT
<213> 瘤状副伯克霍尔德菌
<400> 44
Phe Phe Glu Ala Arg His Arg Glu Leu Ala Ala Gly Ile Glu Ala Trp
1 5 10 15
Ala Thr Gln His Leu Ala Cys Val Gln His Asp Asp Thr Asp Thr Thr
20 25 30
Cys Arg Lys Leu Val Arg Ala Leu Gly Glu Ala Gly Trp Leu Lys Tyr
35 40 45
Gly Val Gly Gly Ala Gln Tyr Gly Gly His Gly Asp Thr Ile Asp Thr
50 55 60
Arg Ala Val Cys Leu Leu Arg Glu Thr Leu Ala Asn His Asp Gly Leu
65 70 75 80
Ala Asp Phe Ala Leu Ala Met Gln Gly Leu Gly Ser Gly Ala Ile Thr
85 90 95
Leu Ala Gly Thr His Glu Gln Lys Ile Arg Tyr Leu Pro Arg Val Ser
100 105 110
Lys Gly Glu Ala Ile Ala Ala Phe Ala Leu Ser Glu Pro Asp Ala Gly
115 120 125
Ser Asp Val Ala Ala Met Ser Leu Gln Ala Arg Ala Glu Gly Asp Cys
130 135 140
Tyr Val Ile Asp Gly Asp Lys Thr Trp Ile Ser Asn Gly Gly Ile Ala
145 150 155 160
Asp Phe Tyr Val Val Phe Ala Arg Thr Gly Glu Ala Pro Gly Ala Arg
165 170 175
Gly Ile Ser Ala Phe Ile Val Asp Ala Asp Thr Pro Gly Leu Gln Ile
180 185 190
Ala Glu Arg Ile Asp Val Ile Ala Pro His Pro Leu Ala Arg Leu His
195 200 205
Phe Asp Ser Ala Arg Val Pro Arg Ser Gln Met Leu Gly Ala Pro Gly
210 215 220
Glu Gly Phe Lys Ile Ala Met Arg Thr Leu Asp Val Phe Arg Thr Ser
225 230 235 240
Val Ala Ala Ala Ser Leu Gly Phe Ala Arg Arg Ala Leu Gln Glu Gly
245 250 255
Leu Ala Arg Ala Ala Ser Arg Lys Met Phe Gly Gln Thr Leu Gly Asp
260 265 270
Phe Gln Leu Thr Gln Thr Lys Leu Ala Gln Met Ala Leu Thr Ile Asp
275 280 285
Ser Ser Ala Leu Leu Val Tyr Arg Ala Ala Trp Leu Arg Asp Gln Gly
290 295 300
Glu Asn Val Thr Arg Glu Ala Ala Met Ala Lys Trp His Ala Ser Glu
305 310 315 320
Gly Ala Gln Gln Val Ile Asp Ala Ala Val Gln Leu Trp Gly Gly Met
325 330 335
Gly Val Gln Ser Gly Thr Thr Val Glu Arg Leu Tyr Arg Glu Ile Arg
340 345 350
Ala Leu Arg Ile Tyr Glu Gly Ala Thr Glu Val Gln Gln Leu Ile Val
355 360 365
Gly Arg Asp Leu Leu Lys Ala His Ala Ala Gln
370 375
<210> 45
<211> 472
<212> PRT
<213> 皮氏罗尔斯顿菌
<400> 45
Met Ile Arg Thr Gly Thr Gln Tyr Leu Glu Ser Leu Asn Asp Gly Arg
1 5 10 15
Asn Val Trp Val Gly Asn Glu Lys Ile Asp Asn Val Ala Thr His Pro
20 25 30
Lys Thr Arg Asp Tyr Ala Gln Arg His Ala Asp Phe Tyr Asp Leu His
35 40 45
His Arg Pro Asp Leu Gln Asp Val Met Thr Tyr Ile Asp Glu Gly Gly
50 55 60
Gln Arg Arg Ala Met Gln Trp Phe Gly His Arg Asp Lys Glu Gln Leu
65 70 75 80
Arg Arg Lys Arg Lys Tyr His Glu Thr Val Met Arg Glu Met Ala Gly
85 90 95
Ala Ser Phe Pro Arg Thr Pro Asp Val Asn Asn Tyr Val Leu Thr Thr
100 105 110
Tyr Ile Asp Asp Pro Ala Pro Trp Glu Thr Gln Ser Ile Gly Asp Asp
115 120 125
Gly His Ile Lys Ala Gly Lys Ile Val Asp Phe Ile Arg Tyr Ala Arg
130 135 140
Glu His Asp Leu Asn Cys Ala Pro Gln Phe Val Asp Pro Gln Glu Arg
145 150 155 160
Ser Pro Gly Leu Arg Val Val Glu Lys Asn Glu Lys Gly Ile Val Val
165 170 175
Asn Gly Val Lys Ala Ile Gly Thr Gly Val Ala Phe Ala Asp Trp Ile
180 185 190
His Ile Gly Val Phe Phe Arg Pro Gly Ile Pro Gly Asp Gln Val Ile
195 200 205
Phe Ala Ala Thr Pro Val Asn Thr Pro Gly Val Thr Ile Val Cys Arg
210 215 220
Glu Ser Leu Val Lys Asp Asp Lys Val Glu His Pro Leu Ala Ala Gln
225 230 235 240
Gly Asp Glu Leu Asp Gly Met Thr Val Phe Glu Asn Val Phe Ile Pro
245 250 255
Trp Ser His Val Phe His Ile Gly Asn Pro Asn His Ala Lys Leu Tyr
260 265 270
Pro Gln Arg Val Phe Asp Trp Leu His Tyr His Ala Leu Ile Arg Gln
275 280 285
Met Val Arg Ala Glu Leu Val Ala Gly Leu Ala Val Leu Ile Thr Glu
290 295 300
His Ile Gly Thr Asn Lys Ile Pro Ala Val Gln Thr Arg Val Ala Lys
305 310 315 320
Leu Ile Gly Phe His Gln Ala Met Leu Ala His Leu Ile Ala Ser Glu
325 330 335
Glu Leu Gly Phe His Thr Pro Gly Gly His Tyr Lys Pro Asn Ile Leu
340 345 350
Ile Tyr Asp Phe Gly Arg Ala Leu Tyr Leu Glu Asn Phe Ser Gln Met
355 360 365
Ile Tyr Glu Leu Val Asp Leu Ser Gly Arg Ser Ala Leu Ile Phe Ala
370 375 380
Ser Glu Asp Gln Trp Asn Asp Asp Lys Leu Asn Gly Trp Phe Glu Arg
385 390 395 400
Met Asn Asn Gly Pro Val Gly Arg Pro His Asp Arg Val Lys Ile Gly
405 410 415
Arg Val Ile Arg Asp Leu Phe Leu Thr Asp Trp Gly Ser Arg Leu Phe
420 425 430
Val Phe Glu Asn Phe Asn Gly Thr Pro Leu Gln Gly Ile Arg Met Leu
435 440 445
Thr Met Gln Arg Ala Glu Phe Ser Gly Ser Gly Pro Tyr Gly Lys Leu
450 455 460
Ala Arg Gln Val Cys Gly Ile Asp
465 470

Claims (18)

1.编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ IDNO:1(地芽孢杆菌属的一个种(Geobacillus sp.)PA9)、SEQ ID NO:3(嗜热栖热菌(Thermusthermophilus))、SEQ IDNO:4(球孢链霉菌(Streptomyces globisporus))、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌(Clostridium aminobutyricum))、SEQ ID NO:6(洋葱伯克霍尔德菌(Burkholderai cepacia))、SEQ ID NO:8(颤藻属的一个种(Oscillatoria sp.)PCC6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌(Paraburkholderia phymatum))的氨基酸序列至少≥50％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ IDNO:6、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换，
其中氨基酸交换不是A210S或S212A。
2.根据权利要求1所述的编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ IDNO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:7(钩虫贪铜菌(Cupriavidus necator))、SEQ IDNO:8(颤藻属的一个种PCC6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)的氨基酸序列至少≥65％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ IDNO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9的氨基酸序列的对应位置处的氨基酸交换。
3.根据前述权利要求中任一项所述的编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:1(地芽孢杆菌属的一个种PA9)、SEQ ID NO:3(嗜热栖热菌)、SEQ ID NO:4(球孢链霉菌)、SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)、SEQ ID NO:6(洋葱伯克霍尔德菌)、SEQ ID NO:7(钩虫贪铜菌)、SEQ ID NO:8(颤藻属的一个种PCC6506)、SEQ ID NO:9(瘤状副伯克霍尔德菌)、SEQ ID NO:10(皮氏罗尔斯顿菌(Ralstoniapickettii))的氨基酸序列至少≥90％相同，
其特征在于在SEQ ID NO:1的位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置中或者在SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ IDNO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10的氨基酸序列的对应位置处的氨基酸交换。
4.根据前述权利要求中任一项所述的编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶选自：
·SEQ ID NO:1(地芽孢杆菌属的一个种PA9)，其中在位置202、203、204、205、206、207、208、209、210、211、212、213、214中的一个或多个位置处具有氨基酸交换；
·SEQ ID NO:3(嗜热栖热菌)，其中在位置194、195、196、197、198、199、200、201、202、203、204、205中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:21；
·SEQ ID NO:4(球孢链霉菌)，其中在位置212、213、214、215、216、217、218、219、220、221、222中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:22；
·SEQ ID NO:5(氨基丁酸梭状芽孢杆菌)，其中在位置202、203、204、205、206、207、208、209、210中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:23；
·SEQ ID NO:6(洋葱伯克霍尔德菌)，其中在位置202、203、204、205、206、207、208、209、210、211、212中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:24；
·SEQ ID NO:7(钩虫贪铜菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:25；
·SEQ ID NO:8(颤藻属的一个种PCC 6506)，其中在位置209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:26；
·SEQ ID NO:9(瘤状副伯克霍尔德菌)，其中在位置192、193、194、195、196、197、198、199、200、201中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:27；
·SEQ ID NO:10(皮氏罗尔斯顿菌)，其中在位置203、204、205、206、207、208、209、210、211、212、213、214、215、216、217中的一个或多个位置处具有氨基酸交换；或者SEQ ID NO:28。
5.根据前述权利要求中任一项所述的编码氨基酸序列的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13或SEQ ID NO:14的氨基酸序列至少≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同，其中SEQ ID NO:11、SEQ IDNO:12、SEQ ID NO:13和SEQ ID NO:14在位置207或氨基酸序列的对应位置处具有除L-缬氨酸以外的蛋白质氨基酸。
6.根据前述权利要求中任一项所述的编码氨基酸序列的多核苷酸，其编码氧化还原酶，其中该多核苷酸是编码来自地芽孢杆菌属微生物的4-羟基苯乙酸3-单加氧酶的可复制核苷酸序列，其中由此编码的蛋白质序列在对应于SEQ ID NO:1的位置207的位置处含有除L-缬氨酸以外的蛋白质氨基酸。
7.根据前述权利要求中任一项所述的编码氨基酸的多核苷酸，其编码氧化还原酶，其中由此编码的蛋白质序列在位置207或对应位置处具有选自苏氨酸、亮氨酸、谷氨酰胺和甘氨酸的氨基酸。
8.根据前述权利要求中任一项所述的编码氨基酸的多核苷酸，其编码氧化还原酶，其中由此编码的蛋白质序列
-在位置206或氨基酸序列的对应位置处含有除L-苏氨酸以外的蛋白质氨基酸，优选L-甲硫氨酸或L-丙氨酸；或者
-在位置208或氨基酸序列的对应位置处含有除L-赖氨酸以外的蛋白质氨基酸，优选L-精氨酸。
9.根据前述权利要求中任一项所述的编码氨基酸的多核苷酸，其编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:15、SEQ IDNO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ IDNO:19、SEQ IDNO:20、SEQ ID NO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ IDNO:32的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同。
10.载体，其包含根据权利要求1至9中任一项所述的多核苷酸。
11.根据权利要求10所述的载体，其适用于在埃希菌(Escherichia)、假单胞菌(Pseudomonas)或棒状杆菌(Corynebacterium)属的微生物中复制。
12.多肽，其包含由根据权利要求1至9中任一项所述的多核苷酸编码的氨基酸序列。
13.埃希菌、假单胞菌或棒状杆菌属的微生物，其包含根据权利要求1至9中任一项所述的多核苷酸或根据权利要求10所述的多肽或根据权利要求10至11所述的载体。
14.根据权利要求13所述的微生物，其中根据权利要求1至7中任一项所述的多核苷酸以过表达的形式存在。
15.根据权利要求13至14中任一项所述的微生物，其特征在于该微生物具有生产精细化学品的能力，其中优选精细化学品为L-二羟基苯丙氨酸(L-DOPA)。
16.生产精细化学品的发酵方法，其包括以下步骤：
a)在培养基中发酵包含编码氨基酸序列的多核苷酸的微生物，所述多核苷酸编码氧化还原酶，所编码的氧化还原酶与SEQ ID NO:1或SEQ ID NO:3至SEQ ID NO:32的氨基酸序列至少≥90％、≥92％、≥94％、≥96％、≥97％、≥98％、≥99％或100％、优选≥97％、特别优选≥98％、非常特别优选≥99％并且极度优选100％相同，
b)在培养基中积累精细化学品，其中获得发酵液。
17.根据权利要求16所述的方法，其特征在于从含有精细化学品的发酵液获得精细化学品或含有精细化学品的液体或固体产品，并且所述精细化学品是L-DOPA。
18.根据权利要求13至15中任一项所述的微生物用于发酵生产L-DOPA的用途。
CN202180060850.9A 2020-07-15 2021-07-14 编码氨基酸序列、编码氧化还原酶的多核苷酸 Pending CN116249778A (zh)

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