CN107690482B - 用于2,4-二羟基丁酸的优化生产的经修饰的微生物 - Google Patents

用于2,4-二羟基丁酸的优化生产的经修饰的微生物 Download PDF

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CN107690482B
CN107690482B CN201580080660.8A CN201580080660A CN107690482B CN 107690482 B CN107690482 B CN 107690482B CN 201580080660 A CN201580080660 A CN 201580080660A CN 107690482 B CN107690482 B CN 107690482B
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P·索卡耶
G·贝斯泰尔-卡尔
L·迪蒙-赛格诺维特
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Abstract

本发明涉及用于经由1,2,4‑丁三醇中间体通过代谢转化木糖来生产2,4‑二羟基丁酸的经遗传修饰的微生物。本发明还涉及用于生产2,4‑二羟基丁酸的方法，其通过在发酵培养基中培养所述经遗传修饰的微生物并从所述培养基中回收2,4‑DHB进行。

Description

用于2,4-二羟基丁酸的优化生产的经修饰的微生物
导言
本发明涉及经遗传修饰的微生物，用于通过经由1,2,4-丁三醇中间体代谢转化木糖来生产2,4-二羟基丁酸。本发明还涉及生产2,4-二羟基丁酸的方法，其通过在发酵培养基中培养所述经遗传修饰的微生物并从所述培养基中回收2,4-DHB进行。
2,4-二羟基丁酸(即2,4-DHB或DHB)，也称为2,4-二羟基丁(烷)酸或2,4-二羟基丁酸酯/盐，是一种具有高经济利益的化合物，因为它可充当用于化学合成2-羟基-4-(甲基硫代)-丁酸酯/盐(HMTB)(一种甲硫氨酸类似物，在动物营养中，尤其是家禽中广泛使用的食品添加剂)的前体。实际上，HMTB可作为甲硫氨酸的低价氨基酸替代物包括在动物饲料中，以显著增加肉产率(US2009/318715)。
常规地，通过在甲硫醇和碱性或酸性催化剂存在下采用α-羟基-γ-丁内酯(2HBL)的化学合成方法生产HMTB。在工业上在三个步骤中从苹果酸(DE19735575A1,AU2004200948A)，或者在两个步骤中从γ-丁内酯(WO2008/022953A1)以已知的方式制备2HBL。这两条路线引起的问题在于分离2HBL的困难以及非常显著的盐的产生，所述盐然后必须被除去。此外，HMTB的石油化学合成在经济上不可行，因为它依赖于使用有害的材料和条件以及昂贵的材料和试剂。
最近，已显示可通过调节适当的pH在水性介质中将2,4-DHB转化成α-羟基-γ-丁内酯(2HBL)(US2013/0204016)。
因此，即使没有用于2,4-二羟基丁酸的生化生产的天然代谢途径，微生物中用于DHB产生的合成代谢途径代表一种有吸引力的替代方案，因为它减轻了许多上文提及的问题。事实上，最近开发了依赖于微生物中特定酶的重组表达的代谢工程方法：WO2012/056318、WO2013/160762、WO2014/009435和EP14306564.7特别描述了通过经由不同代谢途径在经遗传修饰的微生物中发酵葡萄糖来生产2,4-DHB。通过基于作用于空间上相似的同族底物的候选酶的结构和机械知识的合理工程化，或通过筛选天然酶并通过合理设计进一步改善来获得这些专利申请中鉴定的大多数酶。更具体地，WO2012/056318已鉴定了三种非天然存在的酶(苹果酸激酶、苹果酸半醛脱氢酶和DHB脱氢酶，均被突变)，它们可在微生物中过表达以将代谢中间体(L)-苹果酸转化成2,4-DHB；WO2013/160762要求多种酶的异源表达，其中一些经突变而改善酶活性和/或底物亲和性(苹果酰CoA合成酶或琥珀酰CoA:(L)-苹果酸-CoA转移酶和/或苹果酰-CoA裂合酶；苹果酰CoA还原酶；和DHB脱氢酶)以将代谢中间体苹果酸或琥珀酰-CoA或乙醇酰-CoA转化成2,4-DHB；而WO2014/009435的方法依赖于将代谢中间体(L)-高丝氨酸转化成2-氧代-4-羟基丁酸(OHB)以及通过重组表达两种突变酶(高丝氨酸转氨酶和DHB还原酶)而在2,4-DHB中还原OHB。
本发明提出了用于在两个单一步骤中从1,2,4-丁三醇代谢中间体以微生物生产2,4-二羟基丁酸的备选的合成途径，所述步骤需要连续地将1,2,4-丁三醇氧化成2,4-二羟基丁醛，然后将2,4-二羟基丁醛氧化成2,4-DHB。
本发明人确实令人惊讶地发现，可通过在能够将碳源(更特别是木糖)转化成1,2,4-丁三醇的微生物中对上述两个反应进行遗传工程改造而大大改善2,4-DHB的总产量。
因此，本发明在本文中提供了备选的微生物，其经遗传修饰用于从木糖优化生产2,4-二羟基丁酸。经遗传工程改造而将木糖转化成1,2,4-丁三醇的此微生物包含进一步的遗传修饰以：
-将1,2,4-丁三醇氧化成2,4-二羟基丁醛，并且
-将2,4-二羟基丁醛氧化成2,4-DHB。
本发明还涉及通过发酵生产2,4-二羟基丁酸的方法，包括在含有木糖的培养基中培养本发明的微生物，并从培养基中回收产生的2,4-DHB。
发明详述
应该理解的是，下面的详细描述不是限制性的，并且可在不背离本发明的范围的情况下进行各种修改、替换、省略和改变。还应该理解，本文使用的术语是为了描述本发明的具体实施方案的目的，而并不意图为限制性的。
本文引用的所有出版物、专利和专利申请(无论是上文还是下文)均通过提述以其整体由此并入。
此外，除非另有说明，本文使用的所有技术和科学术语具有与本领域普通技术人员通常理解的相同的含义。常规的微生物学和分子生物学技术也是本领域中公知和常用的那些。此类技术对于本领域技术人员来说是公知的，并且在文献中充分地解释(参见例如Sambrook等，2012)。
尽管如此，就贯穿本说明书的不同术语的使用而言，下述定义更特别适用。
除非上下文另有明确说明，单数形式“一种”、“一个”和“该/所述”在本文中包括复数指称。因此，例如，提及“微生物”包括多种此类微生物，并且提及“内源基因”是提及一个或多个内源基因，等等。
术语“包含”、“含有”、“涉及”或“包括”或者变型在本文中以包含的意义使用，即以详明所述特征的存在，但是不排除本发明的各个实施方案中其它特征的存在或添加。
如本文所用，术语“微生物”指活的微观生物体，其可为单细胞或多细胞生物，并通常可在自然界中找到。在本发明的上下文中，微生物优选是细菌、酵母或真菌。更优选地，本发明的微生物选自肠杆菌科(Enterobacteriaceae)、芽孢杆菌科(Bacillaceae)、梭菌科(Clostridiaceae)、链霉菌科(Streptomycetaceae)、棒杆菌科(Corynebacteriaceae)和酵母。甚至更优选地，本发明的微生物是埃希氏菌属(Escherichia)、克雷伯氏菌属(Klebsiella)、热厌氧杆菌属(Thermoanaerobacterium)、梭菌属(Clostridium)、棒杆菌属(Corynebacterium)或酵母属的菌种。还甚至更优选地，本发明的微生物选自大肠杆菌(Escherichia coli)、肺炎克雷伯氏菌(Klebsiella pneumoniae)、热解糖热厌氧杆菌(Thermoanaerobacterium thermosaccharolyticum)、丙酮丁醇梭菌(Clostridiumacetobutylicum)、谷氨酸棒杆菌(Corynebacterium glutamicum)和酿酒酵母(Saccharomyces cerevisiae)。最优选地，本发明的微生物是大肠杆菌。
如本文所用，术语“重组微生物”、“经遗传修饰的微生物”或“遗传工程化微生物”是指如上文定义的微生物，所述微生物在自然界中找不到，因此在遗传上与其天然对应物不同。换言之，它是指通过导入和/或通过缺失和/或通过修饰其遗传元件而经修饰的微生物。此类修饰可通过基因工程、通过在特定选择压力下培养微生物而迫使新代谢途径的形成和进化、或通过组合这两种方法进行(参见例如WO2005/073364或WO2008/116852)。
因此，根据本发明的经遗传修饰用于产生2,4-DHB的微生物意指所述微生物是能够产生2,4-DHB的如上文定义的重组微生物。换言之，所述微生物已经经遗传修饰而允许产生2,4-DHB。
在本发明的上下文中，可通过调节一个或多个内源基因的表达水平来遗传修饰微生物。“调节”在本文中指所述基因的表达水平与其天然表达水平相比被上调、下调或甚至完全消除。因此，此类调节可导致基因产物活性的增强，或者可替换地导致内源基因产物的较低活性或无活性。
术语“内源基因”在本文中是指天然存在于微生物中的基因。
可通过引入除内源调控元件外有利于上调的异源序列或通过用此类异源序列替换那些内源调控元件，或者通过将内源基因的一个或多个补充拷贝引入微生物内的染色体或质粒来过表达内源基因。也可通过将突变引入其编码序列中以修饰基因产物来修饰内源基因活性和/或表达水平。还可进行内源基因的缺失以完全抑制其在微生物内的表达。调节内源基因表达的另一种方式是用更强或更弱的启动子交换其启动子(即野生型启动子)以上调或下调此基因的表达水平。适于此类目的的启动子可为同源的或异源的并且是本领域中公知的。选择适当的启动子来调节内源基因的表达在本领域技术人员的能力范围内。
另外/或者，微生物可经遗传修饰而表达一个或多个外源基因，条件是将所述基因与对于其在宿主微生物中表达必需的所有调节元件一起导入微生物中。用外源DNA修饰或“转化”微生物是本领域技术人员的常规任务。在本发明的上下文中，术语“过表达”在本文中也就外源基因在微生物中的表达而言使用。
“外源基因”或“异源基因”在本文中指所述基因在微生物中不是天然存在的。为了在微生物中表达外源基因，可将此类基因直接整合入微生物染色体中，或者通过微生物内的质粒或载体在染色体外表达。多种质粒(在其复制起点和其在细胞中的拷贝数方面不同)在本领域中是公知的，并可由熟练技术人员容易地选择用于此目的。根据本发明的外源基因有利地为同源基因。
在本发明的上下文中，术语“同源基因”或“同源物”不仅指通过理论上共同的遗传祖先由两个物种(即微生物物种)遗传的基因，而且包括可为遗传上无关但已进化以编码进行相似功能和/或具有相似结构的蛋白质的基因(即功能同系物)。因此，术语“功能同源物”在本文中指编码功能同源蛋白质的基因。
使用数据库，如Uniprot(用于蛋白质)、Genbank(用于基因)或NCBI(用于蛋白质或基因)中可用的信息，本领域技术人员可容易地确定微生物的特定蛋白质和/或基因的序列，并基于此序列鉴定另一种微生物中的等同基因或同源物之一。此常规工作可通过一种微生物的特定基因序列与其它微生物的基因序列或基因组(其可在上文提及的数据库中找到)的序列比对来进行。有利地，可使用Altschul等(1990)开发的BLAST算法进行此类序列比对。一旦在那些序列之间建立了序列同源性，可得出共有序列并用于设计简并探针以克隆相关微生物的相应同源物基因。分子生物学的这些常规方法是本领域技术人员公知的。
应当进一步理解的是，在本发明的上下文中，如果在特定的微生物中表达编码感兴趣的蛋白质的外源基因，则优选通过将非优选的密码子或不太优选的密码子替换为编码相同氨基酸的所述微生物的优选密码子来构建此基因的合成形式。本领域确实公知密码子选择在微生物物种之间变化，这可影响感兴趣的蛋白质的重组表达水平。为了克服此问题，已开发了密码子优化方法，并由Graf等(2000),Deml等(2001)和Davis&Olsen(2011)广泛描述。值得注意地，已为了密码子优化测定而开发出几个软件，如
Figure BDA0001493181400000051
软件(Lifetechnologies)或(GenScript)的OptimumGeneTM软件。换言之，编码感兴趣蛋白质的外源基因优选进行密码子优化用于在特定微生物中表达。
根据本发明的微生物也可经遗传修饰而增加或降低一个或多个蛋白质的活性。
可通过改善蛋白质催化效率、通过降低蛋白质周转率、通过降低信使RNA(mRNA)周转率、通过增加基因的转录、或通过增加mRNA的翻译获得增加此类活性。
改善蛋白质催化效率意指对于给定底物和/或给定辅因子，增加kcat和/或降低Km，和/或对于给定抑制剂，增加Ki。kcat、Km和Ki是本领域技术人员能够确定的Michaelis-Menten常数(Segel，1993)。减少蛋白质周转率意指稳定蛋白质。改善蛋白质催化效率和/或降低蛋白质周转率的方法是本领域技术人员公知的。那些包括合理的工程化改造及序列和/或结构分析和定向诱变，以及随机诱变和筛选。可通过常规方法如聚合酶链式反应(PCR)进行定点诱变，通过随机诱变技术例如经由诱变剂(紫外线或化学试剂如亚硝基胍(NTG)或乙基甲磺酸(EMS))或DNA改组或易错PCR引入突变。稳定蛋白质也可通过在蛋白质的N-末端或C-末端添加“标签”肽序列来实现。此类标签在本领域中是公知的，并且包括谷胱甘肽-S-转移酶(GST)等等。
可通过修饰5’-非翻译区(5’-UTR)和/或编码区和/或3’-UTR的基因序列减少mRNA周转率(Carrier和Keasling，1999)。
增加基因的转录(无论是内源的还是外源的)可通过增加微生物内其拷贝的数目和/或通过使用与野生型启动子相比导致更高的基因表达水平的启动子来实现。在本发明的上下文中，术语“过表达”也用于指微生物中基因转录的增加。
如上文指示，为了增加微生物中基因的拷贝数，可在染色体上或在染色体外编码所述基因。当要在染色体上编码感兴趣的基因时，可通过本领域公知的遗传重组方法(例如基因置换)在染色体上引入基因的几个拷贝。当要在微生物中在染色体外编码基因时，它可由不同类型的质粒携带，所述质粒就其复制起点而言取决于它们可在其中复制的微生物以及它们在细胞中的拷贝数而不同。根据质粒的性质，由所述质粒转化的微生物可含有质粒的1至5个拷贝，或其约20个拷贝，或甚至其多达500个拷贝。可在大肠杆菌中复制的低拷贝数质粒的实例包括但不限于pSC101质粒(紧密复制)、RK2质粒(紧密复制)以及pACYC和pRSF1010质粒，而可在大肠杆菌中复制的高拷贝数质粒的例子是pSK bluescript II。
可增加基因表达水平的启动子也是本领域技术人员公知的，并且可为同源的(起源于相同的物种)或异源的(起源于不同的物种)。此类启动子的实例包括但不限于启动子Ptrc、Ptac、Plac和λ启动子cI。这些启动子也可由特定的化合物或由特定的外部条件如温度或光诱导(“诱导型启动子”)。
可通过修饰核糖体结合位点(RBS)增加mRNA的翻译。RBS是启动蛋白质翻译时与核糖体结合的mRNA上的序列。它可为真核生物中mRNA的5’帽、原核生物中起始密码子AUG上游6-7个核苷酸区(称为Shine-Dalgarno序列)、或病毒中的内部核糖体进入位点(IRES)。通过修饰此序列，可改变蛋白质翻译起始速率，按比例改变其生产速率，和控制其在细胞内的活性。也可通过使用软件RBS CALCULATOR(Salis，2011)来优化RBS序列的强度以达到靶向的翻译起始速率。基于mRNA的性质选择RBS序列在本领域技术人员的能力范围内。
降低蛋白质的活性可意指通过突变编码所述蛋白质的基因来降低其特异的催化活性，从而通过突变核苷酸序列或通过缺失所述基因的编码区来改变相应的氨基酸序列和/或降低细胞中蛋白质的浓度。
如本文所用，术语“氧化还原酶”指能够根据操作条件可逆地或不可逆地还原或氧化其底物的酶。更具体地说，此类酶通过催化电子从其底物(电子供体)转移到称为氧化剂(电子受体)的另一种分子而起作用。在本发明的上下文中，选择的氧化还原酶氧化它们各自的底物(1,2,4-丁三醇氧化成2,4-二羟基丁醛，2,4-二羟基丁醛氧化成2,4-DHB)，更优选以不可逆的方式进行。适应操作条件以使本发明的氧化还原酶氧化它们的底物在本领域技术人员的能力范围内。
通常将氧化还原酶分类为酶的酶委员会(EC)编号分类中的EC1，其分组超过20种酶的亚类(如EC1.1，包括作用于供体的CH-OH基团的氧化还原酶，也称为醇氧化还原酶；EC1.2，包括作用于供体的醛基或氧代基团的氧化还原酶；EC1.3，包括作用于供体的CH-CH基团的氧化还原酶，也称为CH-CH氧化还原酶；EC1.4，包括作用于供体的CH-NH2基团的氧化还原酶供体，也称为氨基酸氧化还原酶或单胺氧化酶；EC1.5，包括作用于供体的CH-NH基团的氧化还原酶；EC1.6，包括作用于NADH或NADPH的氧化还原酶；EC1.7，包括作用于作为供体的其它含氮化合物的氧化还原酶；EC1.8，包括作用于供体的硫基团的氧化还原酶；EC1.9，包括作用于供体的血红素基团的氧化还原酶；EC1.10，包括作用于作为供体的二酚和相关物质的氧化还原酶；EC1.11，包括作用于作为受体的过氧化物的氧化还原酶，也称为过氧化物酶；EC 1.12，包括作用于作为供体的氢的氧化还原酶；EC 1.13，包括作用于单一供体及并入分子氧的氧化还原酶，也称为加氧酶；EC 1.14，包括作用于配对的供体及并入分子氧的氧化还原酶；EC 1.15，包括作用于作为受体的超氧化物自由基的氧化还原酶；EC 1.16，包括氧化金属离子的氧化还原酶；EC1.17，包括作用于CH或CH2基团的氧化还原酶；EC 1.18，包括作用于作为供体的铁硫蛋白的氧化还原酶；EC 1.19，包括作用于作为供体的还原性黄素氧还蛋白的氧化还原酶；EC 1.20，包括作用于供体中的磷或砷的氧化还原酶；EC 1.21，包括作用于X-H和Y-H以形成X-Y键的氧化还原酶；和包括其它氧化还原酶的EC 1.97)。这些氧化还原酶中的一些能够使用NADP或NAD+作为辅因子，并且因此称为NAD+/NADP+氧化还原酶。
根据本发明的术语“碳的来源”、“碳源”或“碳底物”指能够被微生物代谢且含有至少一个碳原子的任何碳源。碳源可源自可再生原料。可再生原料定义为某些工业过程所需要的原料，其可在短暂的延迟内再生并且处于足够的量以允许其转化成期望的产物。植物生物量(经处理或未处理)是一种令人感兴趣的可再生碳源。在本发明的上下文中，木糖是碳源，并且除木糖外的碳源有利地是碳水化合物。
术语“碳水化合物”在本文中是指能够被微生物代谢且含有至少一个碳原子、两个氢原子和一个氧原子的任何碳源。本发明的碳水化合物优选选自下组：单糖如葡萄糖、果糖、甘露糖、半乳糖等，二糖如蔗糖、纤维二糖、麦芽糖、乳糖等，寡糖如棉子糖、水苏糖、麦芽糖糊精等，和多糖诸如纤维素、半纤维素、淀粉等、甲醇、甲醛和甘油。根据本发明的优选实施方案，除木糖外的碳源有利地是包含3、6或12个碳原子的碳水化合物，或其任何组合。在本发明的一个更优选的实施方案中，除木糖外的碳源选自甘油、葡萄糖、半乳糖、果糖、乳糖、麦芽糖、蔗糖及其任何组合。更优选地，除木糖外的碳源选自甘油、葡萄糖、蔗糖及其任何组合，并且最优选是葡萄糖。
在本发明的第一方面，本发明涉及微生物，所述微生物经遗传修饰而通过将木糖转化成1,2,4-丁三醇来产生2,4-二羟基丁酸，其中所述微生物进一步经遗传修饰用于：
i)将1,2,4-丁三醇氧化成2,4-二羟基丁醛；和
ii)将2,4-二羟基丁醛氧化成2,4-二羟基丁酸。
更优选地，氧化步骤i)和ii)分别涉及排他性地将1,2,4-丁三醇转化成2,4-二羟基丁醛和将2,4-二羟基丁醛转化成2,4-二羟基丁酸。换言之，特别优选的是，在将1,2,4-丁三醇氧化成2,4-二羟基丁醛和将2,4-二羟基丁醛氧化成2,4-DHB的过程中，微生物不产生代谢中间体以优化2,4-DHB产生。此类排他性转化可通过使用下面进一步描述的酶来实现。
根据优选的实施方案，遗传修饰i)是编码作用于供体的CH-OH基团的氧化还原酶(EC1.1酶)的至少一个基因的过表达，和/或遗传修饰ii)是编码作用于供体的醛基或氧代基团的氧化还原酶(EC1.2酶)的至少一个基因的过表达。
作用于供体的CH-OH基团的氧化还原酶的实例包括但不限于EC 1.1.1酶(用NAD+或NADP+作为受体的氧化还原酶，也称为NAD+/NADP+氧化还原酶)、EC 1.1.2酶(用细胞色素作为受体的氧化还原酶)、EC 1.1.3酶(用氧作为受体的氧化还原酶)、EC 1.1.4酶(用二硫化物作为受体的氧化还原酶)、EC1.1.5酶(用醌或类似化合物作为受体的氧化还原酶)、EC1.1.98酶(用其它已知受体的氧化还原酶)和EC 1.1.99酶(用其它受体的氧化还原酶)。
作用于供体的醛基或氧代基团的氧化还原酶的实例包括但不限于EC1.2.1酶(用NAD+或NADP+作为受体的氧化还原酶，也称为NAD+/NADP+氧化还原酶)、EC1.2.2酶(用细胞色素作为受体的氧化还原酶)、EC 1.2.3酶(用氧作为受体的氧化还原酶)、EC 1.2.4酶(用二硫化物作为受体的氧化还原酶)、EC 1.2.5酶(用醌或类似化合物作为受体的氧化还原酶)、EC 1.2.7(用铁硫蛋白作为受体的氧化还原酶)和EC 1.2.99酶(用其它受体的氧化还原酶)。
显示上述活性的酶在本领域中是公知的，并且可由本领域技术人员容易地鉴定，例如从公众可得的数据库如BRENDA鉴定。
为了将1,2,4-丁三醇氧化成2,4-二羟基丁醛，作用于供体的CH-OH基团的氧化还原酶(EC1.1酶)优选选自下组：醇脱氢酶(即醛还原酶)、醛脱氢酶、乳醛还原酶、乙醛酸还原酶、二脱氢葡糖酸还原酶及其任何组合。更优选地，所述EC1.1氧化还原酶是醇脱氢酶(即醛还原酶)或乳醛还原酶。还更优选地，所述EC1.1氧化还原酶是作用于供体的CH-OH基团的NAD+/NADP+氧化还原酶(即EC1.1.1酶)，或以其它受体作用于供体的CH-OH基团的氧化还原酶(即EC1.1.99酶)。最优选地，所述EC 1.1酶是EC 1.1.1酶。选择适于本发明目的的EC1.1酶并且鉴定其相应的基因序列在本领域技术人员的能力范围内。
作用于供体的CH-OH基团的特别优选的氧化还原酶(EC1.1酶)根据其在数据库中的序列标识和登录号在下面的表1中描述：醇脱氢酶包括但不限于序列SEQ ID NO:1至SEQID NO:40；乳醛还原酶包括但不限于序列SEQ ID NO:41的酶；乙醛酸还原酶包括但不限于序列SEQ ID NO:42和SEQ ID NO:43的酶，并且二脱氢葡糖酸还原酶包括但不限于序列SEQID NO:44和SEQ ID NO:45的酶。
在本发明的特别优选的实施方案中，根据如图1中所述的1,2,4-丁三醇转化成2,4-二羟基丁醛的转化代谢途径，作用于供体的CH-OH基团的氧化还原酶(EC1.1酶)是序列SEQ ID NO:36或SEQ ID NO:41的酶。
编码上文提及的氧化还原酶的基因在本领域中也是公知的，并在下面的表1中描述：
-编码醇脱氢酶的基因包括但不限于大肠杆菌基因frmA(也称为adhC)、adhP(也称为yddN)、yiaY、eutG(也称为yffV)、yqhD、yeaE、ydhF、yhdN、ybdR、ybdH、ybjJ、ydjL、ahr、yahK、tas(也称为ygdS)和gldA；丙酮丁醇梭菌基因bdhB、bdhA和CA_C3392，酿酒酵母基因ADH1、ADH2、ADH3、ADH4和bdh1；枯草芽孢杆菌基因yhdN和bdhA；氧化葡糖杆菌(Gluconobacter oxydans)基因GOX1615；拟南芥基因AKR4C9；杜氏利什曼原虫(Leishmaniadonovani)基因A4UTP6；产乙醇热厌氧杆菌(Thermoanaerobacter ethanolicus)基因adhA；谷氨酸棒杆菌(Corynebacterium glutamicum)基因butA和budC；肺炎克雷伯氏菌(Klebsiella pneumoniae)基因budC；粘质沙雷氏菌(Serratia marcescens)基因slaC；红褐肉座菌(Hypocrea jecorina)基因gld2；丁酸梭菌(Clostridium butyricum)基因dhaT；弗氏柠檬酸杆菌(Citrobacter Freundii)基因dhaT；肺炎克雷伯氏菌(Klebsiellapneumoniae)基因dhaT；和编码NMDA依赖性醇脱氢酶的红串红球菌(Rhodoccuserythropolis)基因；和编码NMDA依赖性醇脱氢酶的甲醇拟无枝酸菌(Amycolatopsismethanolica)基因；
-编码乳醛还原酶的基因包括但不限于大肠杆菌基因fucO；
-编码乙醛酸还原酶的基因包括但不限于大肠杆菌基因ghrA(也称为ycdW)和ghrB(也称为yiaE)；和
-编码二脱氢葡糖酸还原酶的基因包括但不限于大肠杆菌基因dkgA(也称为yqhE)和dkgB(也称为yafB)。
在本发明的一个特别优选的实施方案中，作用于供体的CH-OH基团的氧化还原酶(EC1.1酶)由来自大肠杆菌的fucO基因和/或来自丁酸梭菌、弗氏柠檬酸杆菌或肺炎克雷伯氏菌的dhaT基因编码，即如图1所示根据1,2,4-丁三醇变为2,4-二羟基丁醛的转化代谢途径。
如上文指示，本发明的微生物也经遗传修饰用于ii)将2,4-二羟基丁醛氧化成2,4-二羟基丁酸。为此，可在微生物中过表达编码作用于供体的醛基或氧代基团的氧化还原酶(EC1.2酶)的至少一个基因。
因此，作用于供体的醛或氧代基团的氧化还原酶(EC 1.2酶)优选选自下组：醛脱氢酶、醛氧化酶及其任何组合。更优选地，所述EC1.2氧化还原酶是醛脱氢酶。还更优选地，所述EC1.2氧化还原酶是作用于供体的醛基或氧代基团的NAD+/NADP+氧化还原酶(即EC1.2.1酶)，或用氧作为受体作用于供体的醛基或氧代基团的氧化还原酶(即EC 1.2.3酶)。最优选地，所述EC 1.2酶是EC 1.2.1酶。选择适于本发明目的的EC1.2酶并鉴定其相应的基因序列在本领域技术人员的能力范围内。
作用于供体的醛基或氧代基团的特别优选的氧化还原酶(EC 1.2酶)根据其在数据库中的序列标识和登录号在下面的表1中描述：醛脱氢酶包括但不限于序列SEQ ID NO:46至SEQ ID NO:60的酶；醛氧化酶包括但不限于序列SEQ ID NO:61的酶。
在本发明的特别优选的实施方案中，根据如图1中所述将2,4-二羟基丁醛变为2,4-二羟基丁酸的转化代谢途径，作用于供体醛基或氧代基团的氧化还原酶(EC1.2酶)是序列SEQ ID NO:46、SEQ ID NO:54或SEQ ID NO:55的酶。
编码上文提及的氧化还原酶的基因在本领域中也是公知的，并在下面的表1中描述：
-编码醛脱氢酶的基因包括但不限于大肠杆菌基因puuC、sad、gabD、prr(也称为ydcW)、aldA、aldB(也称为yiaX)、feaB和astD；肺炎克雷伯氏菌KPN_01018；酿酒酵母基因ALD4和ALD5；恶臭假单胞菌(Pseudomonas putida)基因davD；假单胞菌属菌种基因amnC；和詹氏甲烷球菌(Methanocaldococcus jannaschii)基因MJ1411；
-编码二氧戊酸脱氢酶的基因包括但不限于地衣芽孢杆菌基因ycbD；和
-编码醛氧化酶的基因包括但不限于玉蜀黍基因AO1。
在本发明的一个特别优选的实施方案中，作用于供体的醛基或氧代基团的氧化还原酶(EC1.2酶)由来自大肠杆菌的aldA、aldB或puuC基因编码，即根据如图1中所述将2,4-二羟基丁醛变为2,4-二羟基丁酸的转化代谢途径。
必须进一步注意到，当编码上述活性的内源基因已经存在于感兴趣的微生物中时，它们有利地在所述微生物中过表达。相反，当微生物不天然地包含编码此类活性的基因时，有利地用一种或多种编码所述酶的外源基因转化微生物：认为也过表达所述外源基因。如上文解释，外源基因优选为进行了密码子优化用于其在感兴趣的微生物中表达的合成基因。
在本发明的上下文中，微生物能够将木糖转化成1,2,4-丁三醇。经遗传修饰用于将木糖转化成1,2,4-丁三醇的微生物，特别是用于排他性的将木糖转化成1,2,4-丁三醇的微生物是本领域公知的，并且已经特别通过专利申请WO2008/091288和US2013/0203141描述，其通过提述并入本文。
因此，在一个优选的实施方案中，用于将木糖转化成1,2,4-丁三醇的遗传修饰是至少一个以下基因的过表达：
-编码木糖脱氢酶的基因，
-编码木糖酸内酯酶的基因，
-编码木糖酸脱水酶的基因，
-编码3-脱氧-D-甘油-戊酮糖酸(DGP)脱羧酶的基因，
-编码1,2,4-丁三醇脱氢酶的基因，和
-其任何组合。
编码上文所述酶的基因在本领域中是公知的：
-编码木糖脱氢酶的基因公开于US2013/0203141和WO2008/091288中，通过提述并入本文，并且包括但不限于来自新月柄杆菌(Caulobacter crescentus)、死海盐盒菌(Haloarcula marismortui)、真菌伯克霍尔德氏菌(Burkholderia fugorum)LB400和沃氏富盐菌(Haloferax volcanii)DS2的xdh编码基因，
-编码木糖酸内酯酶的基因包括但不限于来自新月柄杆菌的xylC基因，
-编码D-木糖酸脱水酶的基因在US2013/0203141和WO2008/091288中公开，其通过提述并入本文，并包括但不限于来自大肠杆菌的yjhG和yagF基因，
-编码3-脱氧-D-甘油-戊酮糖酸脱羧酶的基因公开于US2013/0203141和WO2008/091288中，其通过提述并入本文，并且包括但不限于来自恶臭假单胞菌的mdlC基因，和
-编码NADPH依赖性1,2,4-丁三醇脱氢酶的基因公开于US2013/0203141和WO2008/091288中，通过提述并入本文，并且包括但不限于来自大肠杆菌的adhP和yqhD基因。
在一个优选的实施方案中，特别是当经遗传修饰的微生物是大肠杆菌时：
α)编码木糖脱氢酶、木糖酸内酯酶和3-脱氧-D-甘油-戊酮糖酸脱羧酶的基因是导入微生物中的异源基因，和
β)编码木糖酸脱水酶和1,2,4-丁三醇脱氢酶的基因是内源性基因，因此有利地是过表达的。
在本发明的一个特别优选的实施方案中，木糖脱氢酶由新月柄杆菌的xdh基因编码，木糖酸内酯酶由新月柄杆菌的xylC基因编码，木糖酸脱水酶由来自大肠杆菌的yjhG和/或yagF基因编码，3-脱氧-D-甘油-戊酮糖酸脱羧酶由来自恶臭假单胞菌的mdlC基因编码，并且1,2,4-丁三醇脱氢酶由来自大肠杆菌的adhP和/或yqhD基因编码，即根据如图1中所述将木糖变为1,2,4-丁三醇的转化代谢途径。
上文提及基因的核苷酸或由所述基因编码的氨基酸序列根据它们在数据库中的登录号和版本和/或根据它们的序列标识在下面的表1中描述。
当编码上述活性的内源基因已经存在于感兴趣的微生物中时，它们有利地在所述微生物中过表达。相反，当微生物不天然地包含编码此类活性的基因时，有利地用编码所述酶的一个或多个外源基因转化微生物：认为也过表达所述外源基因。如上文解释，外源基因优选为进行了密码子优化用于在感兴趣的微生物中表达的合成基因。
根据一个优选的实施方案，本发明的微生物进一步经遗传修饰用于：
iii)从除木糖外的碳源提供用于2,4-二羟基丁酸生产和微生物生长的还原能力和/或能量，和/或
iv)至少部分地，优选完全地抑制碳分解代谢物阻抑。
如上文所述的遗传修饰iii)是特别有利的，因为其优化了2,4-DHB生产，通过使用备选碳源而非木糖来提供还原能力和/或能量，从而可将木糖排他性地转化成2,4-二羟基丁酸。
为了从除木糖外的碳源提供1,2,4-丁三醇生产和微生物生长的还原能力和/或能量，优选通过缺失和/或弱化编码使用木糖或其它代谢物的酶的基因来遗传修饰根据本发明的微生物，所述代谢物产生与木糖变为2,4-二羟基丁酸的转化相竞争的流量(flux)。
因此，根据本发明的一个优选实施方案，遗传修饰iii)是以下基因中的至少一个的弱化和/或缺失：
-编码木糖异构酶的基因(例如来自大肠杆菌的xylA基因)，
-编码木酮糖激酶的基因(例如来自大肠杆菌的xylB基因)，
-编码3-脱氧-D-甘油-戊酮糖酸醛缩酶的基因(例如来自大肠杆菌的yjhH和/或yagE基因)，
-编码酮酸脱氢酶的基因(例如来自大肠杆菌的yiaE和/或ycdW基因)，和
-其任何组合。
这些基因以及用于缺失或弱化它们的方法在US2013/0203141中充分公开，通过提述并入本文。
上文提及的基因的核苷酸或由所述基因编码的氨基酸序列也根据它们在数据库中的登录号和版本和/或根据它们的序列标识在下面的表1中描述。
如果本发明的微生物如上文所述进行遗传修饰，则相应优选的是微生物使用除木糖外的碳源来提供2,4-DHB生产和微生物生长必需的还原能力和/或能量。在这方面，在本领域中公知的是，一些微生物相对于其它碳源优选某些特定的碳源。值得注意的是，大多数天然存在的微生物，特别是大肠杆菌相对于其它糖优选使用葡萄糖，即使它们能够代谢一系列单糖(Kim等，2010)。然而，一些微生物不能以有效的方式共利用葡萄糖和木糖。实际上，当两种糖存在于培养基中时，微生物中可发生二期(dizuxic)发酵模式：此调节机制在本领域中公知为分解代谢物阻抑，并且在20世纪40年代已由Monod对葡萄糖和乳糖报道。已经在肠杆菌科中广泛研究此类机制。在大肠杆菌中，已经明显认识到分解代谢物阻抑如下发生：葡萄糖借助葡萄糖特异性通透酶EIICBglc(由ptsG编码)进入磷酸烯醇丙酮酸(PEP):碳水化合物磷酸转移酶系统(PTS)；在葡萄糖转运期间，通过去磷酸化的EIIAglc(由crr编码)降低了环AMP(cAMP)的水平，这又限制了分解代谢物阻遏蛋白(CRP)和cAMP复合物(cAMP-CRP)的可用性；随后，涉及除葡萄糖外的糖的分解代谢并通常由cAMP-CRP调节的所有基因都受到阻抑。不被PTS系统转运的碳水化合物的分解代谢物阻抑也在文献中报道，但尚未得到充分理解(Hogema等，1997)。
因此，在一个优选的实施方案中，允许抑制碳分解代谢物阻抑的遗传修饰iv)选自以下的至少一种：
-缺失编码磷酸转移酶系统的葡萄糖通透酶的基因，
-缺失编码磷酸载体Hpr蛋白的基因，
-优选从不受cAMP-CRP调节的组成性或诱导型启动子表达糖输入蛋白系统中所涉及的基因和/或操纵子，其中所述糖是除木糖外的碳源，
-表达编码木糖转运蛋白，如同向转运蛋白或ABC转运蛋白的基因，优选从不受cAMP-CRP调节的组成性或诱导型启动子表达，
-过表达编码葡萄糖同向转运蛋白的基因，
-过表达编码葡萄糖易化蛋白的基因，
-过表达编码葡糖激酶的基因，
-调节牵涉cAMP水平的基因，优选编码腺苷酸环化酶的基因的表达，
-调节编码CRP和/或CRP样蛋白质的基因的表达，
-表达编码不依赖cAMP的CRP蛋白质的基因，优选从不受cAMP-CRP调节的组成性或诱导型启动子表达，以及
-其任何组合。
更优选地，为了共利用木糖和葡萄糖，缺失编码磷酸转移酶和/或磷酸载体Hpr蛋白的基因有利地与过表达编码葡萄糖通透酶或葡萄糖易化蛋白的基因连同过表达编码葡糖激酶的基因组合。
编码上文描述的蛋白质的基因在本领域是公知的：
-编码磷酸转移酶系统的葡萄糖通透酶的基因包括但不限于来自大肠杆菌的pstG基因，
-编码磷酸载体Hpr蛋白质(其为磷酸烯醇丙酮酸：糖磷酸转移酶系统(PTS糖)的两种糖非特异性蛋白质组分之一)的基因包括但不限于来自大肠杆菌的ptsH基因，
-涉及糖输入蛋白系统(PTS或其它系统)的基因/操纵子包括但不限于来自大肠杆菌的用于同化乳糖的lacY基因、来自大肠杆菌的用于同化麦芽糖的malFG基因、以及分别来自鼠伤寒沙门氏菌和大肠杆菌的用于同化蔗糖的scrKYABR或cscBKAR操纵子，
-编码木糖转运蛋白的基因包括但不限于来自大肠杆菌的xylFGH操纵子，来自大肠杆菌的xylE和araE基因，
-编码葡萄糖同向转运蛋白的基因包括但不限于来自大肠杆菌的galP基因，
-编码葡萄糖易化蛋白的基因包括但不限于运动发酵单胞菌(Zymomonasmobilis)的glf基因，
-编码葡糖激酶的基因包括但不限于来自大肠杆菌的glk基因，
-编码腺苷酸环化酶的基因包括但不限于来自大肠杆菌的cyaA基因，
-编码CRP和/或CRP样蛋白质的基因包括但不限于来自大肠杆菌的CRP基因和来自枯草芽孢杆菌和其它硬壁菌门(firmicutes)的ccpA基因，
-Hogema等(1997)已经描述了编码不依赖cAMP的CRP蛋白质的基因，包括但不限于编码在两个区中突变的CRP蛋白的基因：对应于CRP蛋白的cAMP结合域的残基53至62和对应于DNA结合域(螺旋-转角-螺旋)的近端边界的残基141至148。此类蛋白质突变体的实例包括CRP突变体Asp至His残基53、Ser至Phe残基62、Gly至Asp残基141、Arg至Asp残基142、Leu至Arg残基148及其任何组合。
更优选地，为了在大肠杆菌中共利用木糖和葡萄糖，ptsG或ptsH基因缺失可与galP或glf基因的过表达连同glk基因的过表达组合。
上文提及基因的核苷酸序列或由所述基因编码的氨基酸序列根据它们在数据库中的登录号和版本和/或根据它们的序列标识在下面的表1中描述。
如上文解释，当编码上述活性的内源基因已经存在于感兴趣的微生物中时，它们有利地在所述微生物中过表达。然而，如果微生物不天然地包含编码此类活性的基因，则有利地用一个或多个编码所述酶的外源基因转化微生物：认为也过表达所述外源基因。如上文解释，外源基因优选为进行了密码子优化用于在感兴趣的微生物中表达的合成基因。相反，可根据上文描述的任一种方法缺失内源基因或弱化其表达。
根据本发明的除木糖外的碳源如上文定义，并且包括但不限于包含3、6或12个碳原子的碳水化合物，或其任何组合。优选地，除木糖外的碳源选自下组：甘油、葡萄糖、半乳糖、果糖、乳糖、麦芽糖、蔗糖及其任何组合。
最优选地，本发明中使用的除木糖外的碳源是葡萄糖。
或者，除木糖外的碳源可为蔗糖。因此，在本发明的优选实施方案中，微生物包含进一步的遗传修饰以使用蔗糖作为除木糖外的碳源，如WO2012/004247中所述，其通过提述并入本文。为此，微生物优选包含编码PTS蔗糖利用系统和/或非PTS蔗糖利用系统的功能基因。
PTS蔗糖利用系统是基于通过磷酸烯醇丙酮酸(PEP)-依赖性蔗糖磷酸转移酶系统(蔗糖-PTS)转运蔗糖的蔗糖利用系统。磷酸转移酶系统将糖(例如蔗糖或葡萄糖)的转运与使用PEP作为磷酸供体的糖的磷酸化偶联。在转运入细胞后，蔗糖磷酸被转化酶切割成葡萄糖-6-磷酸和果糖。果糖然后被果糖激酶磷酸化成果糖-6-磷酸。编码此PTS蔗糖利用系统的基因可由调节蛋白控制。
非PTS蔗糖利用系统是基于通过独立于磷酸烯醇丙酮酸的系统转运蔗糖的蔗糖利用系统。在转运入细胞后，蔗糖被转化酶切割成葡萄糖和果糖。果糖然后被果糖激酶磷酸化成果糖-6-磷酸，且葡萄糖被葡糖激酶磷酸化成葡萄糖-6-磷酸。编码此非PTS蔗糖利用系统的基因可由调节蛋白控制。
在本发明的一个特别优选的实施方案中，微生物天然表达或已经经遗传修饰以表达来自沙门氏菌属的操纵子scrKYABR的基因，即编码果糖激酶的scrK基因、编码孔蛋白的scrY基因、编码IIBC蛋白的scrA基因、编码蔗糖-6-P转化酶的scrB基因和编码阻遏物的scrR基因，以及它们的任何组合。可使用携带scrKYABR的接合质粒pUR400转化微生物。这些基因可单独、以包含这些基因的至少两个的任何组合、或全部组合在一起在微生物中表达。可优选省略基因scrR。
在本发明的特别优选的实施方案中，微生物天然表达或已经经遗传修饰以表达来自大肠杆菌菌株EC3132的基因，即编码蔗糖:质子同向运输转运系统(cscB基因)、果糖激酶(cscK)、转化酶(cscA基因)和蔗糖特异性阻遏物(cscR基因)的操纵子cscBKAR基因。这些基因可单独、以包含这些基因的至少两个的任何组合、或全部组合在一起在微生物中表达。可优选省略基因cscR。也可使用来自其它生物的同源基因。
然而，在一个优选的实施方案中，本发明的微生物包含牵涉产生NADPH作为还原能力来源的至少一个基因的进一步遗传修饰。事实上，还原酶如脱氢酶需要微生物中可用的还原能力，特别是以NADPH的形式。增加细胞中NADPH可用性的策略在本领域中是公知的，并且已经特别由Lee等(2013)综述，并且还由US8088620、WO2012/055798和EP14305691.9描述，上述文件通过提述并入本文。
根据本发明，用于改善NADPH产生并因此改善其在微生物中的可用性的遗传修饰优选选自：
-过表达编码膜结合转氢酶的基因或操纵子，
-缺失或弱化编码可溶性转氢酶的基因，
-过表达编码产生NADPH的甘油醛-3-磷酸脱氢酶的基因，
-缺失或弱化编码磷酸葡糖异构酶的基因，
-缺失或弱化编码磷酸果糖激酶的基因，
-过表达编码葡萄糖-6-磷酸脱氢酶的基因，
-过表达编码能够产生NADPH的硫辛酰胺脱氢酶的突变体基因，
-过表达编码双功能NAD(P)H水合物修复酶的基因，和
-其任何组合。
缺失或弱化编码磷酸果糖激酶的基因更优选与过表达编码葡萄糖-6-磷酸脱氢酶的基因组合，以增加通过戊糖磷酸途径的NADPH流量。
更优选地，用于改善NADPH产生的遗传修饰选自：
-过表达编码膜结合转氢酶的基因，
-缺失或弱化编码磷酸葡糖异构酶和/或可溶性转氢酶的基因，和
-过表达编码产生NADPH的甘油醛-3-磷酸脱氢酶的基因。
编码上文描述的蛋白质的基因在本领域中是公知的：
-编码膜结合转氢酶的基因或操纵子包括但不限于来自大肠杆菌的pntAB操纵子，特别由WO2012/055798A1所述，
-编码可溶性转氢酶的基因包括但不限于来自大肠杆菌的udhA基因，
-编码产生NADPH的甘油醛-3-磷酸脱氢酶的基因包括但不限于来自变异链球菌的gapN(如Centeno-Leija等，2013所述)，其可用于例如取代来自大肠杆菌的内源gapA基因，
-编码磷酸葡糖异构酶的基因包括但不限于来自大肠杆菌的pgi基因，
-编码磷酸果糖激酶的基因包括但不限于来自大肠杆菌的pfkA基因，如特别由WO2005/047498描述，
-编码葡萄糖-6-磷酸脱氢酶的基因包括但不限于来自大肠杆菌的zwf基因，如特别由Lim等(2002)描述，
-编码能够产生NADPH的硫辛酰胺脱氢酶的突变体基因包括但不限于来自大肠杆菌的突变lpd基因(lpd*)，如特别由Bocanegra等(1993)描述，和
-编码双功能NAD(P)H水合物修复酶的基因包括但不限于来自大肠杆菌的yjeF基因，如特别由Marbaix等(2011)描述。
在大肠杆菌中，如果缺失或弱化pfkA基因，则所述遗传修饰优选与过表达zwf基因组合。
更优选地，用于改善NADPH产生的遗传修饰选自：
-过表达来自大肠杆菌的pntAB操纵子，
-缺失来自大肠杆菌的pfkA基因和/或udhA基因，
-用来自变异链球菌的gapN基因取代来自大肠杆菌的gapA。
上文提及的基因的核苷酸序列或由所述基因编码的氨基酸序列根据它们在数据库中的登录号和版本和/或根据它们的序列标识在下面的表1中描述。
如上文解释，当编码上述活性的内源基因已经存在于感兴趣的微生物中时，它们有利地在所述微生物中过表达。然而，如果微生物不天然地包含编码此类活性的基因，则有利地用一个或多个编码所述酶的外源基因转化微生物：认为也过表达所述外源基因。如上文解释，外源基因优选为进行了密码子优化用于在感兴趣的微生物中表达的合成基因。相反，可根据上文描述的任一种方法缺失内源基因或弱化其表达。
表1：本发明的基因和蛋白质(ND＝未公开)
Figure BDA0001493181400000181
Figure BDA0001493181400000191
Figure BDA0001493181400000201
Figure BDA0001493181400000211
Figure BDA0001493181400000221
Figure BDA0001493181400000231
Figure BDA0001493181400000241
另一方面，本发明涉及生产2,4-二羟基丁酸的方法，其包括：
a)在包含木糖的培养基中，在允许将木糖转化成2,4-二羟基丁酸的发酵条件下培养如上文所述的本发明的经遗传修饰的微生物，以及
b)从所述培养基中回收2,4-二羟基丁酸。
发酵培养基和碳源是本领域中公知的。根据本发明，术语“发酵过程”、“发酵”或“培养”可互换使用，指允许给定微生物生长的实验条件。微生物的生长通常在具有适于所用微生物的适当生长培养基的发酵罐中进行。
“适当的培养基”在本文中指培养基(例如无菌液体培养基)，其包含对于微生物的维持和/或生长必需的或有益的营养物如碳源或碳底物；氮源，例如蛋白胨、酵母提取物、肉提取物、麦芽提取物、尿素、硫酸铵、氯化铵、硝酸铵和磷酸铵；磷源，例如磷酸二氢钾或磷酸氢二钾；微量元素(例如金属盐)，例如镁盐、钴盐和/或锰盐；以及生长因子如氨基酸和维生素。
根据一个优选的实施方案，步骤a)的培养基进一步包含除木糖外的碳源，其优选为碳水化合物。
所述碳水化合物可选自下组：单糖如葡萄糖、果糖、甘露糖、阿拉伯糖、半乳糖等，二糖如蔗糖、纤维二糖、麦芽糖、乳糖等，寡糖如棉子糖、水苏糖(stacchyose)、麦芽糖糊精等，多糖如纤维素、半纤维素、淀粉等，甲醇、甲醛和甘油。根据本发明的特别优选的碳水化合物包含3、6或12个碳原子，更优选甘油、葡萄糖、果糖、半乳糖、乳糖、麦芽糖、蔗糖及其任何组合。最优选地，碳水化合物是葡萄糖。或者，所述碳水化合物可为蔗糖。
在本发明的一个优选实施方案中，碳源(优选碳水化合物)源自可再生原料，如植物生物质。
本领域技术人员可容易地确定用于培养根据本发明的微生物必需的培养条件。特别地，公知的是可在20℃-55℃，优选在25℃-40℃的温度使细菌发酵。更特别地，可在约30℃至约37℃的温度培养大肠杆菌。
本发明的方法可以分批过程、补料分批过程或连续过程并在需氧、微需氧或厌氧条件下进行。
“在需氧条件下”的发酵意指通过将气体溶解到培养物的液相中来将氧气提供给培养物。这可通过如下实现：(1)将含氧气体(例如空气)喷入液相中，或者(2)摇动含有培养基的容器以将顶部空间中所含的氧气转移到液相中。在需氧条件下发酵的主要优点是氧作为电子受体的存在改善菌株产生更多ATP形式的能量用于细胞过程的能力，由此改善菌株的一般代谢。
微需氧条件可在本文中使用并且定义为将低百分比的氧气(例如，使用含有0.1-10％的氧气，用氮气填充至100％的气体混合物)溶解到液相中的培养条件。
相反，“厌氧条件”定义为不向培养基提供氧气的培养条件。严格厌氧条件可通过向培养基中喷入惰性气体(如氮气)来除去痕量的其它气体来获得。硝酸盐可用作电子受体以改善菌株的ATP产生并改善其代谢。
根据优选的实施方案，本发明的方法还包括纯化步骤b)的2,4-DHB的步骤c)。用于纯化羧酸和具体地羟基酸的方法在本领域中是公知的，并且特别在WO2002/090312、WO2002/022544和WO2012/153042中描述，其通过提述并入本文。优选地，通过本领域技术人员公知的手段，且特别是离子交换层析技术如离子交换树脂或固定或模拟移动床离子交换树脂，将2,4-DHB在其于培养基中回收后纯化。
附图简述
图1表示将D-木糖转化成2,4-DHB的代谢途径(*：任选缺失或弱化的基因)。
实施例
在以下实施例中进一步限定了本发明。应该理解的是，这些实施例虽然指示了本发明的优选实施方案，但仅以说明的方式给出。从上面的公开和这些实例中，本领域的技术人员可对本发明进行各种变化以使其适于各种用途和条件，而不需要修改本发明的基本手段。
描述了构建具有这些能力的微生物所需要的示例性基因和酶以及用于克隆和转化、监测产物形成和使用工程化微生物进行生产的方法。
具体地，实施例显示了经修饰的大肠杆菌菌株，但是这些修饰可容易地在相同科的其它微生物中进行。
大肠杆菌属于肠杆菌科，其包含革兰氏阴性、杆状、非孢子形成，并且长度通常为1-5μm的成员。大多数成员具有用于活动的鞭毛，但一些属是不能运动的。此科的许多成员是在人类和其它动物的肠中发现的肠道菌群的正常部分，而其它在水中或土壤中发现，或者是多种不同动物和植物上的寄生物。大肠杆菌是最重要的模式生物体之一，但肠杆菌科的其它重要成员包括克雷伯氏菌属(特别是土生克雷伯氏菌、植物克雷伯氏菌或产酸克雷伯氏菌)，泛菌属和沙门氏菌属。
方案
以下实施例中描述了用于构建2,4-二羟基丁酸生产菌株的几种方案。
在本发明中使用的方案1(通过同源重组的染色体修饰、重组体的选择和抗生素盒切除)和方案2(噬菌体P1的转导)已经在专利申请WO2013/001055中充分描述。
方案3：构建重组质粒
本领域技术人员已知充分描述和知道重组DNA技术。
简言之，使用寡核苷酸(本领域技术人员能够设计)和MG1655大肠杆菌K-12或其它微生物基因组DNA作为基质(根据待扩增的靶向基因)对DNA片段进行PCR扩增。用相容的限制酶消化DNA片段和选择的质粒，连接，然后转化到感受态细胞中。分析转化体并通过DNA测序验证感兴趣的重组质粒。
实施例1：通过在MG1655大肠杆菌中过量产生来自新途径的酶从木糖生产2,4-DHB-构建菌株1至11。
修饰大肠杆菌菌株MG1655以使用图1所示的途径从D-木糖生产2,4-二羟基丁酸(2,4-DHB)。该工作寻求使朝向2,4-DHB生产的碳流量最大化，从而除去所有涉及其它木糖消耗途径或涉及2,4-DHB-中间体化合物的转化(其代表了产物的损失)的酶。
除由大肠杆菌天然表达的基因(序列SEQ ID NO:64的yjhG基因和序列SEQ ID NO:65的yagF基因，其编码木糖酸脱水酶；以及序列SEQ ID NO:66的adhP基因和序列SEQ IDNO:67的yqhD基因，其编码醇脱氢酶-NAD(P)H依赖性1,2,4-丁三醇脱氢酶)外，使用pCL1920质粒(Lerner&Inouye,1990)，用Ptrc人工启动子(专利WO 2007/0770441中给出的序列)及其自身的核糖体结合位点分别表达编码以下酶的基因：新月柄杆菌的木糖脱氢酶和木糖酸内酯酶(分别为xdh[CauloCyc上的CC0821，SEQ ID NO:62]和xylC[CauloCyc上的CC0820，SEQ ID NO:63])，恶臭假单胞菌的3-脱氧-D-甘油-戊酮糖酸脱羧酶(序列SEQ ID NO:68的mdlC基因)，大肠杆菌或丁酸梭菌的醇脱氢酶-NAD(P)+依赖性2,4-二羟基丁醛脱氢酶序列(分别为编码序列SEQ ID NO:41的酶的来自大肠杆菌的fucO基因或编码序列SEQ ID NO:36的酶的来自丁酸梭菌的dhaT基因)和大肠杆菌的醛脱氢酶(编码序列SEQ ID NO:46的酶的来自大肠杆菌的puuC基因或编码序列SEQ ID NO:55的酶的来自大肠杆菌的aldB基因)。实际上，首先在pCL1920质粒上克隆基因xdh、xylC、mdlC，给出质粒pDHB0001，然后在质粒pDHB0001上序贯克隆fucO或dhaT和puuC或aldB，产生4种组合，得到质粒pDHB0002至pDHB0005，如下文表中描述。
Figure BDA0001493181400000271
此外，为了阻断天然木糖分解代谢途径，使用由Datsenko&Wanner，2000所述的同源重组策略并且根据方案1，从大肠杆菌MG1655染色体中缺失编码D-木糖异构酶(序列SEQID NO:69的xylA基因)和D-木酮糖激酶(序列SEQ ID NO:70的xylB基因)的基因。更准确地，为了缺失xylAB操纵子，用SEQ ID NO:104和SEQ ID NO:105的引物产生携带抗生素抗性基因连同FRT位点的被与xylAB操纵子的上游和下游区同源的序列围绕的PCR产物，并将其导入先前已经转化有pKD46载体的大肠杆菌MG1655中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株1。
为了避免3-脱氧-D-葡糖戊酮糖酸(DGP)的降解，也使用相同的同源重组策略缺失了编码酮酸脱氢酶(序列SEQ ID NO:73的yiaE基因；和序列SEQ ID NO:74的ycdW基因)和DGP醛缩酶(序列SEQ ID NO:71的yjhH基因；序列SEQ ID NO:72的yagE基因)的基因。更准确地，为了缺失yjhH基因(SEQ ID NO:71)，用SEQ ID NO:106和SEQ ID NO:107的引物产生携带抗生素抗性基因连同FRT位点的被与yjhH基因的上游和下游区同源的序列围绕的PCR产物，并将其导入先前转化有pKD46载体的菌株1中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株2。然后，为了缺失yagE基因(SEQ ID NO:72)，产生携带抗生素抗性基因连同FRT位点的被与yagE基因(SEQ ID NO:108和SEQ ID NO:109)的上游和下游区同源的序列围绕的PCR产物，并将其导入先前转化有pKD46载体的菌株2中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株3。然后，为了缺失yiaE基因(SEQID NO:73)，用SEQ ID NO:110和SEQ ID NO:111的引物产生携带抗生素抗性基因连同FRT位点的被与yiaE基因的上游和下游区同源的序列围绕的PCR产物，并将其导入预先转化有pKD46载体的菌株3中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株4。最后，为了缺失ycdW基因(SEQ ID NO:74)，用SEQ ID NO:112和SEQ ID NO:113的引物产生携带抗生素抗性基因连同FRT位点的被与ycdW基因的上游和下游区同源的序列围绕的PCR产物，并将其导入预先转化有pKD46载体的菌株4中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株5。
由于yjhH基因与yjhG(SEQ ID NO:64)和yjhI一起属于yjhIHG操纵子，必须选择与yjhH的上游和下游区同源的序列，从而不改变周围基因的表达。对于属于yagEF操纵子的yagE基因(SED ID NO:72)是相同的。
最后，为了抑制分解代谢物阻抑，通过使用由Datsenko&Wanner，2000描述的同源重组策略(根据方案1)，如专利申请EP 14305691.9中，特别是在所述文献的实施例2中所述，使用SEQ ID NO:114和SEQ ID NO:115的引物缺失由ptsG基因(SEQ ID NO:75)编码的葡萄糖磷酸转移酶IIBC(Glc)。将适当的PCR产物导入先前转化有pKD46载体的菌株5中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株6。
每次使用卡那霉素、氯霉素、庆大霉素、四环素、杀稻瘟素或壮观霉素中不同的抗生素抗性基因。在使用菌株6之前，如由Datsenko&Wanner，2000所述(根据方案1)，使用Flp重组酶从ΔxylAB,ΔyjhH,ΔyagE,ΔyiaE,ΔycdW修饰中除去抗生素盒，产生菌株7。
然后，将上文所述的每个质粒pDHB0002、pDHB0003、pDHB0004或pDHB0005导入菌株7中，产生菌株8至11，如下表所述。
Figure BDA0001493181400000281
实施例2：通过增加生产菌株的NADPH可用性来改善2,4-二羟基-丁酸产量-构建菌株12至20。
由adhP和yqhD编码的1,2,4-丁三醇脱氢酶需要生物体中可用的还原力，特别是NADPH形式，因此过表达参与NADPH产生的基因。
通过将大肠杆菌MG1655的pntA基因的内源启动子和核糖体结合位点替换为诱导型Ptrc启动子(来自质粒pTRC99A，Amersham Pharmacia)和确定的核糖体结合位点RBS120(来自RBS Calculator软件)，如专利申请EP14305691.9中所述，特别是所述文献的实施例4中所述(本文称为SEQ ID NO:116)过量产生由pntAB操纵子(SEQ ID NO:94)编码的膜结合质子易位吡啶核苷酸转氢酶。将专利申请EP 14305691.9中描述的适当的PCR产物导入先前转化有pKD46载体的菌株7中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株12。
使用SEQ ID NO:117和SEQ ID NO:118的引物，通过使用由Datsenko&Wanner，2000描述的同源重组策略(根据方案1)并如专利申请WO2012/055798中特别是在所述申请的实施例2中所述，缺失由sthA基因(先前称为udhA，序列SEQ ID NO:95)编码的可溶性吡啶核苷酸转氢酶。将适当的PCR产物导入先前转化有pKD46载体的菌株12中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株13。
使用SEQ ID NO:119和SEQ ID NO:120的引物，通过使用由Datsenko&Wanner，2000描述的同源重组策略(根据方案1)并如专利申请EP 14305691.9中特别是在所述申请的实施例5中所述，缺失由pfkA基因(SEQ ID NO:99)编码的果糖磷酸激酶。将适当的PCR产物导入预先转化了pKD46载体的菌株13中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株14。
通过使用Datsenko&Wanner，2000描述的同源重组策略(根据方案1)将由来自大肠杆菌的gapA基因(SEQ ID NO:97)编码的NAD依赖性磷酸甘油醛脱氢酶替换为编码NADP依赖性甘油醛-3-磷酸脱氢酶的来自变异链球菌的gapN基因(SEQ ID NO:96)(Centeno-Leija等，2013)。通过如下完成gapN基因对gapA基因的取代：将gapA启动子和核糖体结合位点同时取代为热诱导型PR01启动子(SEQ ID NO:121)及其关联的不耐热性阻抑物CI857(SEQ IDNO122)(如专利申请EP 2532751的实施例1中所述)和确定的核糖体结合位点RBS150(来自RBS Calculator软件，SEQ ID NO:123)。更准确地，使用SEQ ID NO:124和SEQ ID NO:125的引物产生携带CI857基因、PR01启动子、RBS150核糖体结合位点、来自变异链球菌的gapN基因和抗生素抗性基因连同FRT位点的被与gapA基因的上游和下游区同源的序列围绕的PCR产物，并将其导入先前转化为pKD46载体的菌株14中。用适当的寡核苷酸验证抗生素抗性转化体，并将保留的菌株命名为菌株15。
在使用菌株15之前，根据方案1使用Flp重组酶从pntAB、udhA、pfkA和gapA基因座除去抗生素盒，产生菌株16。最后，将上文所述的每个质粒pDHB0002、pDHB0003、pDHB0004或pDHB0005导入菌株16中，产生菌株17至20，如下表中所述。
Figure BDA0001493181400000301
实施例3：在产生2,4-二羟基-丁酸的大肠杆菌重组菌株中过量生产涉及蔗糖代谢的酶-构建菌株21至26。
为了避免当在蔗糖和木糖的混合物上培养菌株时的分解代谢物阻遏，通过使用由Datsenko&Wanner，2000描述的同源重组策略(根据方案1)将编码木糖的输入蛋白的操纵子xylFGH(XylF周质蛋白，XylG ATP结合亚基，XylH膜亚基)的天然启动子替换为人工启动子。更准确地，使用SEQ ID NO:127和SEQ ID NO:128的引物产生携带Ptrc人工启动子(SEQ IDNO:126)和卡那霉素抗性基因连同FRT位点的被与xylF基因且与染色体上xylF基因的上游区同源的序列围绕的PCR产物，并将其导入先前转化有pKD46载体的菌株16中。用适当的寡核苷酸验证卡那霉素抗性转化体，并将保留的菌株命名为菌株21。
在使用菌株21之前，根据方案1使用Flp重组酶从xylFGH操纵子除去抗生素盒，产生菌株22。
为了允许大肠杆菌在蔗糖上生长，将来自鼠伤寒沙门氏菌的质粒pUR400(Schmidt等,1982,SEQ ID NO:81)的编码果糖激酶的基因scrK、孔蛋白(porine)的基因scrY、蛋白质IIBC的基因scrA、蔗糖-6-磷酸转化酶的基因scrB和阻抑物的基因scrR克隆于pBBR1MCS质粒(Kovach等，1995)上的天然启动子下，得到质粒pBDO0003。
最后，如下表所述，将质粒pBDO0003与质粒pDHB0002、pDHB0003、pDHB0004或pDHB0005之一组合引入菌株22，产生菌株23至菌株26。
Figure BDA0001493181400000302
实施例4：2,4-二羟基丁酸的微生物生产。
使用改良的M9培养基(Anderson,1946)在500ml带挡板的锥形瓶中评价生产菌株，所述改良的M9培养基补充有30g/l MOPS、20g/L D-木糖和10g/l葡萄糖，并且调节为pH 6,8。当在预培养和培养物中有必要时添加浓度50mg.L-1的壮观霉素和5mg.L-1的四环素。在37℃在LB培养基(Sigma)中培养预培养物。在培养24小时后，使用它于30℃和以200rpm接种改良的M9培养基的50mL培养物至OD600约0.2。当培养基中的糖耗尽时，将培养物离心，并且通过LC-MS/MS对培养液分析2,4-二羟基-丁酸。
2,4-二羟基丁酸滴度/效价(titer)表示如下：
Figure BDA0001493181400000311
表2：锥形瓶中评价的每种菌株的2,4-二羟基丁酸(DHB)滴度。
符号(-)是指不及5mg/L的2,4-DHB产生，而符号(+)对应于5-15mg/L的2,4-DHB产生，并且符号(++)对应于高于15mg/L的2,4-DHB产生。
菌株 DHB滴度
MG1655 -
菌株8至11 +
菌株17至20 ++
对所有重组菌株观察2,4-二羟基丁酸产生。通过为增加细胞中NADPH汇集物而进行的遗传修饰来增强产量。
实施例5中在与上文所述相同的条件下，在木糖和蔗糖的混合物上测试菌株23至26(未在表2中呈现)，并且其2,4-DHB产生与菌株8至11相似。
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序列表
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Trp Ser Cys Leu Gly Gly Gly Arg Leu Phe Asn Asp Asp Tyr Phe Gln
210 215 220
Pro Leu Arg Asp Glu Leu Ala Val Val Ala Glu Glu Leu Asn Ala Gly
225 230 235 240
Ser Ile Glu Gln Val Val Tyr Ala Trp Val Leu Arg Leu Pro Ser Gln
245 250 255
Pro Leu Pro Ile Ile Gly Ser Gly Lys Ile Glu Arg Val Arg Ala Ala
260 265 270
Val Glu Ala Glu Thr Leu Lys Met Thr Arg Gln Gln Trp Phe Arg Ile
275 280 285
Arg Lys Ala Ala Leu Gly Tyr Asp Val Pro
290 295
<210> 8
<211> 122
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 8
Met Trp Leu Leu Asp Gln Trp Ala Glu Arg His Ile Ala Glu Ala Gln
1 5 10 15
Ala Lys Gly Glu Phe Asp Asn Leu Ala Gly Ser Gly Glu Pro Leu Ile
20 25 30
Leu Asp Asp Asp Ser His Val Pro Pro Glu Leu Arg Ala Gly Tyr Arg
35 40 45
Leu Leu Lys Asn Ala Gly Cys Leu Pro Pro Glu Leu Glu Gln Arg Arg
50 55 60
Glu Ala Ile Gln Leu Leu Asp Ile Leu Lys Gly Ile Arg His Asp Asp
65 70 75 80
Pro Gln Tyr Gln Glu Val Ser Arg Arg Leu Ser Leu Leu Glu Leu Lys
85 90 95
Leu Arg Gln Ala Gly Leu Ser Thr Asp Phe Leu Arg Gly Asp Tyr Ala
100 105 110
Asp Lys Leu Leu Asp Lys Ile Asn Asp Asn
115 120
<210> 9
<211> 412
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 9
Met Lys Ala Leu Thr Tyr His Gly Pro His His Val Gln Val Glu Asn
1 5 10 15
Val Pro Asp Pro Gly Val Glu Gln Ala Asp Asp Ile Ile Leu Arg Ile
20 25 30
Thr Ala Thr Ala Ile Cys Gly Ser Asp Leu His Leu Tyr Arg Gly Lys
35 40 45
Ile Pro Gln Val Lys His Gly Asp Ile Phe Gly His Glu Phe Met Gly
50 55 60
Glu Val Val Glu Thr Gly Lys Asp Val Lys Asn Leu Gln Lys Gly Asp
65 70 75 80
Arg Val Val Ile Pro Phe Val Ile Ala Cys Gly Asp Cys Phe Phe Cys
85 90 95
Arg Leu Gln Gln Tyr Ala Ala Cys Glu Asn Thr Asn Ala Gly Lys Gly
100 105 110
Ala Ala Leu Asn Lys Lys Gln Ile Pro Ala Pro Ala Ala Leu Phe Gly
115 120 125
Tyr Ser His Leu Tyr Gly Gly Val Pro Gly Gly Gln Ala Glu Tyr Val
130 135 140
Arg Val Pro Lys Gly Asn Val Gly Pro Phe Lys Val Pro Pro Leu Leu
145 150 155 160
Ser Asp Asp Lys Ala Leu Phe Leu Ser Asp Ile Leu Pro Thr Ala Trp
165 170 175
Gln Ala Ala Lys Asn Ala Gln Ile Gln Gln Gly Ser Ser Val Ala Val
180 185 190
Tyr Gly Ala Gly Pro Val Gly Leu Leu Thr Ile Ala Cys Ala Arg Leu
195 200 205
Leu Gly Ala Glu Gln Ile Phe Val Val Asp His His Pro Tyr Arg Leu
210 215 220
His Phe Ala Ala Asp Arg Tyr Gly Ala Ile Pro Ile Asn Phe Asp Glu
225 230 235 240
Asp Ser Asp Pro Ala Gln Ser Ile Ile Glu Gln Thr Ala Gly His Arg
245 250 255
Gly Val Asp Ala Val Ile Asp Ala Val Gly Phe Glu Ala Lys Gly Ser
260 265 270
Thr Thr Glu Thr Val Leu Thr Asn Leu Lys Leu Glu Gly Ser Ser Gly
275 280 285
Lys Ala Leu Arg Gln Cys Ile Ala Ala Val Arg Arg Gly Gly Ile Val
290 295 300
Ser Val Pro Gly Val Tyr Ala Gly Phe Ile His Gly Phe Leu Phe Gly
305 310 315 320
Asp Ala Phe Asp Lys Gly Leu Ser Phe Lys Met Gly Gln Thr His Val
325 330 335
His Ala Trp Leu Gly Glu Leu Leu Pro Leu Ile Glu Lys Gly Leu Leu
340 345 350
Lys Pro Glu Glu Ile Val Thr His Tyr Met Pro Phe Glu Glu Ala Ala
355 360 365
Arg Gly Tyr Glu Ile Phe Glu Lys Arg Glu Glu Glu Cys Arg Lys Val
370 375 380
Ile Leu Val Pro Gly Ala Gln Ser Ala Glu Ala Ala Gln Lys Ala Val
385 390 395 400
Ser Gly Leu Val Asn Ala Met Pro Gly Gly Thr Ile
405 410
<210> 10
<211> 362
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 10
Met Pro His Asn Pro Ile Arg Val Val Val Gly Pro Ala Asn Tyr Phe
1 5 10 15
Ser His Pro Gly Ser Phe Asn His Leu His Asp Phe Phe Thr Asp Glu
20 25 30
Gln Leu Ser Arg Ala Val Trp Ile Tyr Gly Lys Arg Ala Ile Ala Ala
35 40 45
Ala Gln Thr Lys Leu Pro Pro Ala Phe Gly Leu Pro Gly Ala Lys His
50 55 60
Ile Leu Phe Arg Gly His Cys Ser Glu Ser Asp Val Gln Gln Leu Ala
65 70 75 80
Ala Glu Ser Gly Asp Asp Arg Ser Val Val Ile Gly Val Gly Gly Gly
85 90 95
Ala Leu Leu Asp Thr Ala Lys Ala Leu Ala Arg Arg Leu Gly Leu Pro
100 105 110
Phe Val Ala Val Pro Thr Ile Ala Ala Thr Cys Ala Ala Trp Thr Pro
115 120 125
Leu Ser Val Trp Tyr Asn Asp Ala Gly Gln Ala Leu His Tyr Glu Ile
130 135 140
Phe Asp Asp Ala Asn Phe Met Val Leu Val Glu Pro Glu Ile Ile Leu
145 150 155 160
Asn Ala Pro Gln Gln Tyr Leu Leu Ala Gly Ile Gly Asp Thr Leu Ala
165 170 175
Lys Trp Tyr Glu Ala Val Val Leu Ala Pro Gln Pro Glu Thr Leu Pro
180 185 190
Leu Thr Val Arg Leu Gly Ile Asn Asn Ala Gln Ala Ile Arg Asp Val
195 200 205
Leu Leu Asn Ser Ser Glu Gln Ala Leu Ser Asp Gln Gln Asn Gln Gln
210 215 220
Leu Thr Gln Ser Phe Cys Asp Val Val Asp Ala Ile Ile Ala Gly Gly
225 230 235 240
Gly Met Val Gly Gly Leu Gly Asp Arg Phe Thr Arg Val Ala Ala Ala
245 250 255
His Ala Val His Asn Gly Leu Thr Val Leu Pro Gln Thr Glu Lys Phe
260 265 270
Leu His Gly Thr Lys Val Ala Tyr Gly Ile Leu Val Gln Ser Ala Leu
275 280 285
Leu Gly Gln Asp Asp Val Leu Ala Gln Leu Thr Gly Ala Tyr Gln Arg
290 295 300
Phe His Leu Pro Thr Thr Leu Ala Glu Leu Glu Val Asp Ile Asn Asn
305 310 315 320
Gln Ala Glu Ile Asp Lys Val Ile Ala His Thr Leu Arg Pro Val Glu
325 330 335
Ser Ile His Tyr Leu Pro Val Thr Leu Thr Pro Asp Thr Leu Arg Ala
340 345 350
Ala Phe Lys Lys Val Glu Ser Phe Lys Ala
355 360
<210> 11
<211> 347
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 11
Met Lys Asn Ser Lys Ala Ile Leu Gln Val Pro Gly Thr Met Lys Ile
1 5 10 15
Ile Ser Ala Glu Ile Pro Val Pro Lys Glu Asp Glu Val Leu Ile Lys
20 25 30
Val Glu Tyr Val Gly Ile Cys Gly Ser Asp Val His Gly Phe Glu Ser
35 40 45
Gly Pro Phe Ile Pro Pro Lys Asp Pro Asn Gln Glu Ile Gly Leu Gly
50 55 60
His Glu Cys Ala Gly Thr Val Val Ala Val Gly Ser Arg Val Arg Lys
65 70 75 80
Phe Lys Pro Gly Asp Arg Val Asn Ile Glu Pro Gly Val Pro Cys Gly
85 90 95
His Cys Arg Tyr Cys Leu Glu Gly Lys Tyr Asn Ile Cys Pro Asp Val
100 105 110
Asp Phe Met Ala Thr Gln Pro Asn Tyr Arg Gly Ala Leu Thr His Tyr
115 120 125
Leu Cys His Pro Glu Ser Phe Thr Tyr Lys Leu Pro Asp Asn Met Asp
130 135 140
Thr Met Glu Gly Ala Leu Val Glu Pro Ala Ala Val Gly Met His Ala
145 150 155 160
Ala Met Leu Ala Asp Val Lys Pro Gly Lys Lys Ile Ile Ile Leu Gly
165 170 175
Ala Gly Cys Ile Gly Leu Met Thr Leu Gln Ala Cys Lys Cys Leu Gly
180 185 190
Ala Thr Glu Ile Ala Val Val Asp Val Leu Glu Lys Arg Leu Ala Met
195 200 205
Ala Glu Gln Leu Gly Ala Thr Val Val Ile Asn Gly Ala Lys Glu Asp
210 215 220
Thr Ile Ala Arg Cys Gln Gln Phe Thr Glu Asp Met Gly Ala Asp Ile
225 230 235 240
Val Phe Glu Thr Ala Gly Ser Ala Val Thr Val Lys Gln Ala Pro Tyr
245 250 255
Leu Val Met Arg Gly Gly Lys Ile Met Ile Val Gly Thr Val Pro Gly
260 265 270
Asp Ser Ala Ile Asn Phe Leu Lys Ile Asn Arg Glu Val Thr Ile Gln
275 280 285
Thr Val Phe Arg Tyr Ala Asn Arg Tyr Pro Val Thr Ile Glu Ala Ile
290 295 300
Ser Ser Gly Arg Phe Asp Val Lys Ser Met Val Thr His Ile Tyr Asp
305 310 315 320
Tyr Arg Asp Val Gln Gln Ala Phe Glu Glu Ser Val Asn Asn Lys Arg
325 330 335
Asp Ile Ile Lys Gly Val Ile Lys Ile Ser Asp
340 345
<210> 12
<211> 358
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 12
Met Lys Ala Leu Ala Arg Phe Gly Lys Ala Phe Gly Gly Tyr Lys Met
1 5 10 15
Ile Asp Val Pro Gln Pro Met Cys Gly Pro Glu Asp Val Val Ile Glu
20 25 30
Ile Lys Ala Ala Ala Ile Cys Gly Ala Asp Met Lys His Tyr Asn Val
35 40 45
Asp Ser Gly Ser Asp Glu Phe Asn Ser Ile Arg Gly His Glu Phe Ala
50 55 60
Gly Cys Ile Ala Gln Val Gly Glu Lys Val Lys Asp Trp Lys Val Gly
65 70 75 80
Gln Arg Val Val Ser Asp Asn Ser Gly His Val Cys Gly Val Cys Pro
85 90 95
Ala Cys Glu Gln Gly Asp Phe Leu Cys Cys Thr Glu Lys Val Asn Leu
100 105 110
Gly Leu Asp Asn Asn Thr Trp Gly Gly Gly Phe Ser Lys Tyr Cys Leu
115 120 125
Val Pro Gly Glu Ile Leu Lys Ile His Arg His Ala Leu Trp Glu Ile
130 135 140
Pro Asp Gly Val Asp Tyr Glu Asp Ala Ala Val Leu Asp Pro Ile Cys
145 150 155 160
Asn Ala Tyr Lys Ser Ile Ala Gln Gln Ser Lys Phe Leu Pro Gly Gln
165 170 175
Asp Val Val Val Ile Gly Thr Gly Pro Leu Gly Leu Phe Ser Val Gln
180 185 190
Met Ala Arg Ile Met Gly Ala Val Asn Ile Val Val Val Gly Leu Gln
195 200 205
Glu Asp Val Ala Val Arg Phe Pro Val Ala Lys Glu Leu Gly Ala Thr
210 215 220
Ala Val Val Asn Gly Ser Thr Glu Asp Val Val Ala Arg Cys Gln Gln
225 230 235 240
Ile Cys Gly Lys Asp Asn Leu Gly Leu Val Ile Glu Cys Ser Gly Ala
245 250 255
Asn Ile Ala Leu Lys Gln Ala Ile Glu Met Leu Arg Pro Asn Gly Glu
260 265 270
Val Val Arg Val Gly Met Gly Phe Lys Pro Leu Asp Phe Ser Ile Asn
275 280 285
Asp Ile Thr Ala Trp Asn Lys Ser Ile Ile Gly His Met Ala Tyr Asp
290 295 300
Ser Thr Ser Trp Arg Asn Ala Ile Arg Leu Leu Ala Ser Gly Ala Ile
305 310 315 320
Lys Val Lys Pro Met Ile Thr His Arg Ile Gly Leu Ser Gln Trp Arg
325 330 335
Glu Gly Phe Asp Ala Met Val Asp Lys Thr Ala Ile Lys Val Ile Met
340 345 350
Thr Tyr Asp Phe Asp Glu
355
<210> 13
<211> 390
<212> PRT
<213> 丙酮丁醇梭菌(Clostridium acetobutylicum)
<400> 13
Met Val Asp Phe Glu Tyr Ser Ile Pro Thr Arg Ile Phe Phe Gly Lys
1 5 10 15
Asp Lys Ile Asn Val Leu Gly Arg Glu Leu Lys Lys Tyr Gly Ser Lys
20 25 30
Val Leu Ile Val Tyr Gly Gly Gly Ser Ile Lys Arg Asn Gly Ile Tyr
35 40 45
Asp Lys Ala Val Ser Ile Leu Glu Lys Asn Ser Ile Lys Phe Tyr Glu
50 55 60
Leu Ala Gly Val Glu Pro Asn Pro Arg Val Thr Thr Val Glu Lys Gly
65 70 75 80
Val Lys Ile Cys Arg Glu Asn Gly Val Glu Val Val Leu Ala Ile Gly
85 90 95
Gly Gly Ser Ala Ile Asp Cys Ala Lys Val Ile Ala Ala Ala Cys Glu
100 105 110
Tyr Asp Gly Asn Pro Trp Asp Ile Val Leu Asp Gly Ser Lys Ile Lys
115 120 125
Arg Val Leu Pro Ile Ala Ser Ile Leu Thr Ile Ala Ala Thr Gly Ser
130 135 140
Glu Met Asp Thr Trp Ala Val Ile Asn Asn Met Asp Thr Asn Glu Lys
145 150 155 160
Leu Ile Ala Ala His Pro Asp Met Ala Pro Lys Phe Ser Ile Leu Asp
165 170 175
Pro Thr Tyr Thr Tyr Thr Val Pro Thr Asn Gln Thr Ala Ala Gly Thr
180 185 190
Ala Asp Ile Met Ser His Ile Phe Glu Val Tyr Phe Ser Asn Thr Lys
195 200 205
Thr Ala Tyr Leu Gln Asp Arg Met Ala Glu Ala Leu Leu Arg Thr Cys
210 215 220
Ile Lys Tyr Gly Gly Ile Ala Leu Glu Lys Pro Asp Asp Tyr Glu Ala
225 230 235 240
Arg Ala Asn Leu Met Trp Ala Ser Ser Leu Ala Ile Asn Gly Leu Leu
245 250 255
Thr Tyr Gly Lys Asp Thr Asn Trp Ser Val His Leu Met Glu His Glu
260 265 270
Leu Ser Ala Tyr Tyr Asp Ile Thr His Gly Val Gly Leu Ala Ile Leu
275 280 285
Thr Pro Asn Trp Met Glu Tyr Ile Leu Asn Asn Asp Thr Val Tyr Lys
290 295 300
Phe Val Glu Tyr Gly Val Asn Val Trp Gly Ile Asp Lys Glu Lys Asn
305 310 315 320
His Tyr Asp Ile Ala His Gln Ala Ile Gln Lys Thr Arg Asp Tyr Phe
325 330 335
Val Asn Val Leu Gly Leu Pro Ser Arg Leu Arg Asp Val Gly Ile Glu
340 345 350
Glu Glu Lys Leu Asp Ile Met Ala Lys Glu Ser Val Lys Leu Thr Gly
355 360 365
Gly Thr Ile Gly Asn Leu Arg Pro Val Asn Ala Ser Glu Val Leu Gln
370 375 380
Ile Phe Lys Lys Ser Val
385 390
<210> 14
<211> 389
<212> PRT
<213> 丙酮丁醇梭菌(Clostridium acetobutylicum)
<400> 14
Met Leu Ser Phe Asp Tyr Ser Ile Pro Thr Lys Val Phe Phe Gly Lys
1 5 10 15
Gly Lys Ile Asp Val Ile Gly Glu Glu Ile Lys Lys Tyr Gly Ser Arg
20 25 30
Val Leu Ile Val Tyr Gly Gly Gly Ser Ile Lys Arg Asn Gly Ile Tyr
35 40 45
Asp Arg Ala Thr Ala Ile Leu Lys Glu Asn Asn Ile Ala Phe Tyr Glu
50 55 60
Leu Ser Gly Val Glu Pro Asn Pro Arg Ile Thr Thr Val Lys Lys Gly
65 70 75 80
Ile Glu Ile Cys Arg Glu Asn Asn Val Asp Leu Val Leu Ala Ile Gly
85 90 95
Gly Gly Ser Ala Ile Asp Cys Ser Lys Val Ile Ala Ala Gly Val Tyr
100 105 110
Tyr Asp Gly Asp Thr Trp Asp Met Val Lys Asp Pro Ser Lys Ile Thr
115 120 125
Lys Val Leu Pro Ile Ala Ser Ile Leu Thr Leu Ser Ala Thr Gly Ser
130 135 140
Glu Met Asp Gln Ile Ala Val Ile Ser Asn Met Glu Thr Asn Glu Lys
145 150 155 160
Leu Gly Val Gly His Asp Asp Met Arg Pro Lys Phe Ser Val Leu Asp
165 170 175
Pro Thr Tyr Thr Phe Thr Val Pro Lys Asn Gln Thr Ala Ala Gly Thr
180 185 190
Ala Asp Ile Met Ser His Thr Phe Glu Ser Tyr Phe Ser Gly Val Glu
195 200 205
Gly Ala Tyr Val Gln Asp Gly Ile Ala Glu Ala Ile Leu Arg Thr Cys
210 215 220
Ile Lys Tyr Gly Lys Ile Ala Met Glu Lys Thr Asp Asp Tyr Glu Ala
225 230 235 240
Arg Ala Asn Leu Met Trp Ala Ser Ser Leu Ala Ile Asn Gly Leu Leu
245 250 255
Ser Leu Gly Lys Asp Arg Lys Trp Ser Cys His Pro Met Glu His Glu
260 265 270
Leu Ser Ala Tyr Tyr Asp Ile Thr His Gly Val Gly Leu Ala Ile Leu
275 280 285
Thr Pro Asn Trp Met Glu Tyr Ile Leu Asn Asp Asp Thr Leu His Lys
290 295 300
Phe Val Ser Tyr Gly Ile Asn Val Trp Gly Ile Asp Lys Asn Lys Asp
305 310 315 320
Asn Tyr Glu Ile Ala Arg Glu Ala Ile Lys Asn Thr Arg Glu Tyr Phe
325 330 335
Asn Ser Leu Gly Ile Pro Ser Lys Leu Arg Glu Val Gly Ile Gly Lys
340 345 350
Asp Lys Leu Glu Leu Met Ala Lys Gln Ala Val Arg Asn Ser Gly Gly
355 360 365
Thr Ile Gly Ser Leu Arg Pro Ile Asn Ala Glu Asp Val Leu Glu Ile
370 375 380
Phe Lys Lys Ser Tyr
385
<210> 15
<211> 386
<212> PRT
<213> 丙酮丁醇梭菌(Clostridium acetobutylicum)
<400> 15
Met Tyr Asn Phe Asp Phe Phe Asn Pro Thr His Ile Val Phe Gly Lys
1 5 10 15
Asp Arg Leu Asn Glu Leu Asp Asn Leu Val Pro Arg Asp Ala Lys Val
20 25 30
Leu Val Leu Tyr Gly Gly Gly Ser Val Lys Lys Phe Gly Thr Leu Glu
35 40 45
Lys Val Ile Asn Gly Leu Gly Asn Arg Gln Val Ile Glu Phe Gly Gly
50 55 60
Ile Glu Pro Asn Pro Gln Phe Thr Thr Leu Met Lys Ala Val Asp Ile
65 70 75 80
Val Lys Lys Glu Asn Ile Asp Phe Leu Leu Ala Val Gly Gly Gly Ser
85 90 95
Val Met Asp Gly Thr Lys Phe Val Ala Leu Ala Ala Tyr Tyr Glu Gly
100 105 110
Asp Asn Ala Ala Glu Ile Leu Tyr Ser Arg Glu Lys Ala Ala Ala Ile
115 120 125
Asn Lys Ala Val Pro Leu Gly Thr Val Val Thr Leu Pro Ala Thr Gly
130 135 140
Ser Glu Met Asn Asn Gly Gly Val Ile Ser Tyr Glu His Gly Lys Tyr
145 150 155 160
Gly Phe Gly Ser Lys Leu Val Phe Pro Lys Phe Ser Val Leu Asp Pro
165 170 175
Thr Leu Thr Tyr Thr Leu Pro Glu Ser Gln Val Ala Asn Gly Val Ala
180 185 190
Asp Thr Phe Val His Val Leu Glu Gln Tyr Ala Thr Phe Lys Ala Glu
195 200 205
Gly Arg Phe Gln Asp Arg Thr Ala Glu Gly Ile Leu Gln Thr Leu Ile
210 215 220
Glu Ile Gly Arg Lys Thr Ile Asp Asn Pro Thr Asp Tyr Asp Thr Arg
225 230 235 240
Ala Asn Leu Val Trp Cys Ala Thr Met Ala Leu Asn Gly Leu Ile Gly
245 250 255
Ala Gly Val Pro Gln Asp Trp Ser Thr His Met Ile Gly His Glu Leu
260 265 270
Thr Ala Met Phe Gly Ile Asp His Gly Lys Thr Leu Ala Ile Ile Leu
275 280 285
Pro Ser Ile Trp Asn Val Met Arg Glu Gln Lys Lys Gly Lys Ile Leu
290 295 300
Gln Tyr Ala Glu Arg Val Leu Gly Ile Thr Glu Gly Asp Asp Asp Ser
305 310 315 320
Arg Ile Asp Leu Ala Ile Leu Arg Thr Arg Glu Phe Phe Glu Ser Leu
325 330 335
Gly Ile Lys Thr His Leu Ser Glu Tyr Gly Val Thr Ala Asp Lys Ile
340 345 350
Asp Asp Ile Val Asn Ala Leu Asp Lys His Gly Met Lys Ala Leu Ser
355 360 365
Glu Thr Gly Ala Ile Thr Leu Glu Val Ser Arg Lys Ile Leu Glu Gly
370 375 380
Ala Met
385
<210> 16
<211> 348
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 16
Met Ser Ile Pro Glu Thr Gln Lys Gly Val Ile Phe Tyr Glu Ser His
1 5 10 15
Gly Lys Leu Glu Tyr Lys Asp Ile Pro Val Pro Lys Pro Lys Ala Asn
20 25 30
Glu Leu Leu Ile Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu
35 40 45
His Ala Trp His Gly Asp Trp Pro Leu Pro Val Lys Leu Pro Leu Val
50 55 60
Gly Gly His Glu Gly Ala Gly Val Val Val Gly Met Gly Glu Asn Val
65 70 75 80
Lys Gly Trp Lys Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly
85 90 95
Ser Cys Met Ala Cys Glu Tyr Cys Glu Leu Gly Asn Glu Ser Asn Cys
100 105 110
Pro His Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Gln
115 120 125
Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro Gln Gly Thr
130 135 140
Asp Leu Ala Gln Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr
145 150 155 160
Lys Ala Leu Lys Ser Ala Asn Leu Met Ala Gly His Trp Val Ala Ile
165 170 175
Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys
180 185 190
Ala Met Gly Tyr Arg Val Leu Gly Ile Asp Gly Gly Glu Gly Lys Glu
195 200 205
Glu Leu Phe Arg Ser Ile Gly Gly Glu Val Phe Ile Asp Phe Thr Lys
210 215 220
Glu Lys Asp Ile Val Gly Ala Val Leu Lys Ala Thr Asp Gly Gly Ala
225 230 235 240
His Gly Val Ile Asn Val Ser Val Ser Glu Ala Ala Ile Glu Ala Ser
245 250 255
Thr Arg Tyr Val Arg Ala Asn Gly Thr Thr Val Leu Val Gly Met Pro
260 265 270
Ala Gly Ala Lys Cys Cys Ser Asp Val Phe Asn Gln Val Val Lys Ser
275 280 285
Ile Ser Ile Val Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu
290 295 300
Ala Leu Asp Phe Phe Ala Arg Gly Leu Val Lys Ser Pro Ile Lys Val
305 310 315 320
Val Gly Leu Ser Thr Leu Pro Glu Ile Tyr Glu Lys Met Glu Lys Gly
325 330 335
Gln Ile Val Gly Arg Tyr Val Val Asp Thr Ser Lys
340 345
<210> 17
<211> 348
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 17
Met Ser Ile Pro Glu Thr Gln Lys Ala Ile Ile Phe Tyr Glu Ser Asn
1 5 10 15
Gly Lys Leu Glu His Lys Asp Ile Pro Val Pro Lys Pro Lys Pro Asn
20 25 30
Glu Leu Leu Ile Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu
35 40 45
His Ala Trp His Gly Asp Trp Pro Leu Pro Thr Lys Leu Pro Leu Val
50 55 60
Gly Gly His Glu Gly Ala Gly Val Val Val Gly Met Gly Glu Asn Val
65 70 75 80
Lys Gly Trp Lys Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly
85 90 95
Ser Cys Met Ala Cys Glu Tyr Cys Glu Leu Gly Asn Glu Ser Asn Cys
100 105 110
Pro His Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Glu
115 120 125
Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro Gln Gly Thr
130 135 140
Asp Leu Ala Glu Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr
145 150 155 160
Lys Ala Leu Lys Ser Ala Asn Leu Arg Ala Gly His Trp Ala Ala Ile
165 170 175
Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys
180 185 190
Ala Met Gly Tyr Arg Val Leu Gly Ile Asp Gly Gly Pro Gly Lys Glu
195 200 205
Glu Leu Phe Thr Ser Leu Gly Gly Glu Val Phe Ile Asp Phe Thr Lys
210 215 220
Glu Lys Asp Ile Val Ser Ala Val Val Lys Ala Thr Asn Gly Gly Ala
225 230 235 240
His Gly Ile Ile Asn Val Ser Val Ser Glu Ala Ala Ile Glu Ala Ser
245 250 255
Thr Arg Tyr Cys Arg Ala Asn Gly Thr Val Val Leu Val Gly Leu Pro
260 265 270
Ala Gly Ala Lys Cys Ser Ser Asp Val Phe Asn His Val Val Lys Ser
275 280 285
Ile Ser Ile Val Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu
290 295 300
Ala Leu Asp Phe Phe Ala Arg Gly Leu Val Lys Ser Pro Ile Lys Val
305 310 315 320
Val Gly Leu Ser Ser Leu Pro Glu Ile Tyr Glu Lys Met Glu Lys Gly
325 330 335
Gln Ile Ala Gly Arg Tyr Val Val Asp Thr Ser Lys
340 345
<210> 18
<211> 375
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 18
Met Leu Arg Thr Ser Thr Leu Phe Thr Arg Arg Val Gln Pro Ser Leu
1 5 10 15
Phe Ser Arg Asn Ile Leu Arg Leu Gln Ser Thr Ala Ala Ile Pro Lys
20 25 30
Thr Gln Lys Gly Val Ile Phe Tyr Glu Asn Lys Gly Lys Leu His Tyr
35 40 45
Lys Asp Ile Pro Val Pro Glu Pro Lys Pro Asn Glu Ile Leu Ile Asn
50 55 60
Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu His Ala Trp His Gly
65 70 75 80
Asp Trp Pro Leu Pro Val Lys Leu Pro Leu Val Gly Gly His Glu Gly
85 90 95
Ala Gly Val Val Val Lys Leu Gly Ser Asn Val Lys Gly Trp Lys Val
100 105 110
Gly Asp Leu Ala Gly Ile Lys Trp Leu Asn Gly Ser Cys Met Thr Cys
115 120 125
Glu Phe Cys Glu Ser Gly His Glu Ser Asn Cys Pro Asp Ala Asp Leu
130 135 140
Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Gln Phe Ala Thr Ala Asp
145 150 155 160
Ala Ile Gln Ala Ala Lys Ile Gln Gln Gly Thr Asp Leu Ala Glu Val
165 170 175
Ala Pro Ile Leu Cys Ala Gly Val Thr Val Tyr Lys Ala Leu Lys Glu
180 185 190
Ala Asp Leu Lys Ala Gly Asp Trp Val Ala Ile Ser Gly Ala Ala Gly
195 200 205
Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Thr Ala Met Gly Tyr Arg
210 215 220
Val Leu Gly Ile Asp Ala Gly Glu Glu Lys Glu Lys Leu Phe Lys Lys
225 230 235 240
Leu Gly Gly Glu Val Phe Ile Asp Phe Thr Lys Thr Lys Asn Met Val
245 250 255
Ser Asp Ile Gln Glu Ala Thr Lys Gly Gly Pro His Gly Val Ile Asn
260 265 270
Val Ser Val Ser Glu Ala Ala Ile Ser Leu Ser Thr Glu Tyr Val Arg
275 280 285
Pro Cys Gly Thr Val Val Leu Val Gly Leu Pro Ala Asn Ala Tyr Val
290 295 300
Lys Ser Glu Val Phe Ser His Val Val Lys Ser Ile Asn Ile Lys Gly
305 310 315 320
Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu Ala Leu Asp Phe Phe
325 330 335
Ser Arg Gly Leu Ile Lys Ser Pro Ile Lys Ile Val Gly Leu Ser Glu
340 345 350
Leu Pro Lys Val Tyr Asp Leu Met Glu Lys Gly Lys Ile Leu Gly Arg
355 360 365
Tyr Val Val Asp Thr Ser Lys
370 375
<210> 19
<211> 382
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 19
Met Ser Ser Val Thr Gly Phe Tyr Ile Pro Pro Ile Ser Phe Phe Gly
1 5 10 15
Glu Gly Ala Leu Glu Glu Thr Ala Asp Tyr Ile Lys Asn Lys Asp Tyr
20 25 30
Lys Lys Ala Leu Ile Val Thr Asp Pro Gly Ile Ala Ala Ile Gly Leu
35 40 45
Ser Gly Arg Val Gln Lys Met Leu Glu Glu Arg Asp Leu Asn Val Ala
50 55 60
Ile Tyr Asp Lys Thr Gln Pro Asn Pro Asn Ile Ala Asn Val Thr Ala
65 70 75 80
Gly Leu Lys Val Leu Lys Glu Gln Asn Ser Glu Ile Val Val Ser Ile
85 90 95
Gly Gly Gly Ser Ala His Asp Asn Ala Lys Ala Ile Ala Leu Leu Ala
100 105 110
Thr Asn Gly Gly Glu Ile Gly Asp Tyr Glu Gly Val Asn Gln Ser Lys
115 120 125
Lys Ala Ala Leu Pro Leu Phe Ala Ile Asn Thr Thr Ala Gly Thr Ala
130 135 140
Ser Glu Met Thr Arg Phe Thr Ile Ile Ser Asn Glu Glu Lys Lys Ile
145 150 155 160
Lys Met Ala Ile Ile Asp Asn Asn Val Thr Pro Ala Val Ala Val Asn
165 170 175
Asp Pro Ser Thr Met Phe Gly Leu Pro Pro Ala Leu Thr Ala Ala Thr
180 185 190
Gly Leu Asp Ala Leu Thr His Cys Ile Glu Ala Tyr Val Ser Thr Ala
195 200 205
Ser Asn Pro Ile Thr Asp Ala Cys Ala Leu Lys Gly Ile Asp Leu Ile
210 215 220
Asn Glu Ser Leu Val Ala Ala Tyr Lys Asp Gly Lys Asp Lys Lys Ala
225 230 235 240
Arg Thr Asp Met Cys Tyr Ala Glu Tyr Leu Ala Gly Met Ala Phe Asn
245 250 255
Asn Ala Ser Leu Gly Tyr Val His Ala Leu Ala His Gln Leu Gly Gly
260 265 270
Phe Tyr His Leu Pro His Gly Val Cys Asn Ala Val Leu Leu Pro His
275 280 285
Val Gln Glu Ala Asn Met Gln Cys Pro Lys Ala Lys Lys Arg Leu Gly
290 295 300
Glu Ile Ala Leu His Phe Gly Ala Ser Gln Glu Asp Pro Glu Glu Thr
305 310 315 320
Ile Lys Ala Leu His Val Leu Asn Arg Thr Met Asn Ile Pro Arg Asn
325 330 335
Leu Lys Glu Leu Gly Val Lys Thr Glu Asp Phe Glu Ile Leu Ala Glu
340 345 350
His Ala Met His Asp Ala Cys His Leu Thr Asn Pro Val Gln Phe Thr
355 360 365
Lys Glu Gln Val Val Ala Ile Ile Lys Lys Ala Tyr Glu Tyr
370 375 380
<210> 20
<211> 331
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 20
Met Glu Tyr Thr Ser Ile Ala Asp Thr Gly Ile Glu Ala Ser Arg Ile
1 5 10 15
Gly Leu Gly Thr Trp Ala Ile Gly Gly Thr Met Trp Gly Gly Thr Asp
20 25 30
Glu Lys Thr Ser Ile Glu Thr Ile Arg Ala Ala Leu Asp Gln Gly Ile
35 40 45
Thr Leu Ile Asp Thr Ala Pro Ala Tyr Gly Phe Gly Gln Ser Glu Glu
50 55 60
Ile Val Gly Lys Ala Ile Lys Glu Tyr Gly Lys Arg Asp Gln Val Ile
65 70 75 80
Leu Ala Thr Lys Thr Ala Leu Asp Trp Lys Asn Asn Gln Leu Phe Arg
85 90 95
His Ala Asn Arg Ala Arg Ile Val Glu Glu Val Glu Asn Ser Leu Lys
100 105 110
Arg Leu Gln Thr Asp Tyr Ile Asp Leu Tyr Gln Val His Trp Pro Asp
115 120 125
Pro Leu Val Pro Ile Glu Glu Thr Ala Glu Val Met Lys Glu Leu Tyr
130 135 140
Asp Ala Gly Lys Ile Arg Ala Ile Gly Val Ser Asn Phe Ser Ile Glu
145 150 155 160
Gln Met Asp Thr Phe Arg Ala Val Ala Pro Leu His Thr Ile Gln Pro
165 170 175
Pro Tyr Asn Leu Phe Glu Arg Glu Met Glu Glu Ser Val Leu Pro Tyr
180 185 190
Ala Lys Asp Asn Lys Ile Thr Thr Leu Leu Tyr Gly Ser Leu Cys Arg
195 200 205
Gly Leu Leu Thr Gly Lys Met Thr Glu Glu Tyr Thr Phe Glu Gly Asp
210 215 220
Asp Leu Arg Asn His Asp Pro Lys Phe Gln Lys Pro Arg Phe Lys Glu
225 230 235 240
Tyr Leu Ser Ala Val Asn Gln Leu Asp Lys Leu Ala Lys Thr Arg Tyr
245 250 255
Gly Lys Ser Val Ile His Leu Ala Val Arg Trp Ile Leu Asp Gln Pro
260 265 270
Gly Ala Asp Ile Ala Leu Trp Gly Ala Arg Lys Pro Gly Gln Leu Glu
275 280 285
Ala Leu Ser Glu Ile Thr Gly Trp Thr Leu Asn Ser Glu Asp Gln Lys
290 295 300
Asp Ile Asn Thr Ile Leu Glu Asn Thr Ile Ser Asp Pro Val Gly Pro
305 310 315 320
Glu Phe Met Ala Pro Pro Thr Arg Glu Glu Ile
325 330
<210> 21
<211> 332
<212> PRT
<213> 氧化葡糖杆菌(Gluconobacter oxydans)
<400> 21
Met Ala Ser Asp Thr Ile Arg Ile Pro Gly Ile Asp Thr Pro Leu Ser
1 5 10 15
Arg Val Ala Leu Gly Thr Trp Ala Ile Gly Gly Trp Met Trp Gly Gly
20 25 30
Pro Asp Asp Asp Asn Gly Val Arg Thr Ile His Ala Ala Leu Asp Glu
35 40 45
Gly Ile Asn Leu Ile Asp Thr Ala Pro Val Tyr Gly Phe Gly His Ser
50 55 60
Glu Glu Ile Val Gly Arg Ala Leu Ala Glu Lys Pro Asn Lys Ala His
65 70 75 80
Val Ala Thr Lys Leu Gly Leu His Trp Val Gly Glu Asp Glu Lys Asn
85 90 95
Met Lys Val Phe Arg Asp Ser Arg Pro Ala Arg Ile Arg Lys Glu Val
100 105 110
Glu Asp Ser Leu Arg Arg Leu Arg Val Glu Thr Ile Asp Leu Glu Gln
115 120 125
Ile His Trp Pro Asp Asp Lys Thr Pro Ile Asp Glu Ser Ala Arg Glu
130 135 140
Leu Gln Lys Leu His Gln Asp Gly Lys Ile Arg Ala Leu Gly Val Ser
145 150 155 160
Asn Phe Ser Pro Glu Gln Met Asp Ile Phe Arg Glu Val Ala Pro Leu
165 170 175
Ala Thr Ile Gln Pro Pro Leu Asn Leu Phe Glu Arg Thr Ile Glu Lys
180 185 190
Asp Ile Leu Pro Tyr Ala Glu Lys His Asn Ala Val Val Leu Ala Tyr
195 200 205
Gly Ala Leu Cys Arg Gly Leu Leu Thr Gly Lys Met Asn Arg Asp Thr
210 215 220
Thr Phe Pro Lys Asp Asp Leu Arg Ser Asn Asp Pro Lys Phe Gln Lys
225 230 235 240
Pro Asn Phe Glu Lys Tyr Leu Ala Ala Met Asp Glu Phe Glu Lys Leu
245 250 255
Ala Glu Lys Arg Gly Lys Ser Val Met Ala Phe Ala Val Arg Trp Val
260 265 270
Leu Asp Gln Gly Pro Val Ile Ala Leu Trp Gly Ala Arg Lys Pro Gly
275 280 285
Gln Val Ser Gly Val Lys Asp Val Phe Gly Trp Ser Leu Thr Asp Glu
290 295 300
Glu Lys Lys Ala Val Asp Asp Ile Leu Ala Arg His Val Pro Asn Pro
305 310 315 320
Ile Asp Pro Thr Phe Met Ala Pro Pro Ala Arg Asp
325 330
<210> 22
<211> 339
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 22
Met Ser Met Ile Lys Ser Tyr Ala Ala Lys Glu Ala Gly Gly Glu Leu
1 5 10 15
Glu Val Tyr Glu Tyr Asp Pro Gly Glu Leu Arg Pro Gln Asp Val Glu
20 25 30
Val Gln Val Asp Tyr Cys Gly Ile Cys His Ser Asp Leu Ser Met Ile
35 40 45
Asp Asn Glu Trp Gly Phe Ser Gln Tyr Pro Leu Val Ala Gly His Glu
50 55 60
Val Ile Gly Arg Val Val Ala Leu Gly Ser Ala Ala Gln Asp Lys Gly
65 70 75 80
Leu Gln Val Gly Gln Arg Val Gly Ile Gly Trp Thr Ala Arg Ser Cys
85 90 95
Gly His Cys Asp Ala Cys Ile Ser Gly Asn Gln Ile Asn Cys Glu Gln
100 105 110
Gly Ala Val Pro Thr Ile Met Asn Arg Gly Gly Phe Ala Glu Lys Leu
115 120 125
Arg Ala Asp Trp Gln Trp Val Ile Pro Leu Pro Glu Asn Ile Asp Ile
130 135 140
Glu Ser Ala Gly Pro Leu Leu Cys Gly Gly Ile Thr Val Phe Lys Pro
145 150 155 160
Leu Leu Met His His Ile Thr Ala Thr Ser Arg Val Gly Val Ile Gly
165 170 175
Ile Gly Gly Leu Gly His Ile Ala Ile Lys Leu Leu His Ala Met Gly
180 185 190
Cys Glu Val Thr Ala Phe Ser Ser Asn Pro Ala Lys Glu Gln Glu Val
195 200 205
Leu Ala Met Gly Ala Asp Lys Val Val Asn Ser Arg Asp Pro Gln Ala
210 215 220
Leu Lys Ala Leu Ala Gly Gln Phe Asp Leu Ile Ile Asn Thr Val Asn
225 230 235 240
Val Ser Leu Asp Trp Gln Pro Tyr Phe Glu Ala Leu Thr Tyr Gly Gly
245 250 255
Asn Phe His Thr Val Gly Ala Val Leu Thr Pro Leu Ser Val Pro Ala
260 265 270
Phe Thr Leu Ile Ala Gly Asp Arg Ser Val Ser Gly Ser Ala Thr Gly
275 280 285
Thr Pro Tyr Glu Leu Arg Lys Leu Met Arg Phe Ala Ala Arg Ser Lys
290 295 300
Val Ala Pro Thr Thr Glu Leu Phe Pro Met Ser Lys Ile Asn Asp Ala
305 310 315 320
Ile Gln His Val Arg Asp Gly Lys Ala Arg Tyr Arg Val Val Leu Lys
325 330 335
Ala Asp Phe
<210> 23
<211> 315
<212> PRT
<213> 拟南芥(Arabidopsis thaliana)
<400> 23
Met Ala Asn Ala Ile Thr Phe Phe Lys Leu Asn Thr Gly Ala Lys Phe
1 5 10 15
Pro Ser Val Gly Leu Gly Thr Trp Gln Ala Ser Pro Gly Leu Val Gly
20 25 30
Asp Ala Val Ala Ala Ala Val Lys Ile Gly Tyr Arg His Ile Asp Cys
35 40 45
Ala Gln Ile Tyr Gly Asn Glu Lys Glu Ile Gly Ala Val Leu Lys Lys
50 55 60
Leu Phe Glu Asp Arg Val Val Lys Arg Glu Asp Leu Phe Ile Thr Ser
65 70 75 80
Lys Leu Trp Cys Thr Asp His Asp Pro Gln Asp Val Pro Glu Ala Leu
85 90 95
Asn Arg Thr Leu Lys Asp Leu Gln Leu Glu Tyr Val Asp Leu Tyr Leu
100 105 110
Ile His Trp Pro Ala Arg Ile Lys Lys Gly Ser Val Gly Ile Lys Pro
115 120 125
Glu Asn Leu Leu Pro Val Asp Ile Pro Ser Thr Trp Lys Ala Met Glu
130 135 140
Ala Leu Tyr Asp Ser Gly Lys Ala Arg Ala Ile Gly Val Ser Asn Phe
145 150 155 160
Ser Thr Lys Lys Leu Ala Asp Leu Leu Glu Leu Ala Arg Val Pro Pro
165 170 175
Ala Val Asn Gln Val Glu Cys His Pro Ser Trp Arg Gln Thr Lys Leu
180 185 190
Gln Glu Phe Cys Lys Ser Lys Gly Val His Leu Ser Ala Tyr Ser Pro
195 200 205
Leu Gly Ser Pro Gly Thr Thr Trp Leu Lys Ser Asp Val Leu Lys Asn
210 215 220
Pro Ile Leu Asn Met Val Ala Glu Lys Leu Gly Lys Ser Pro Ala Gln
225 230 235 240
Val Ala Leu Arg Trp Gly Leu Gln Met Gly His Ser Val Leu Pro Lys
245 250 255
Ser Thr Asn Glu Gly Arg Ile Lys Glu Asn Phe Asn Val Phe Asp Trp
260 265 270
Ser Ile Pro Asp Tyr Met Phe Ala Lys Phe Ala Glu Ile Glu Gln Ala
275 280 285
Arg Leu Val Thr Gly Ser Phe Leu Val His Glu Thr Leu Ser Pro Tyr
290 295 300
Lys Ser Ile Glu Glu Leu Trp Asp Gly Glu Ile
305 310 315
<210> 24
<211> 284
<212> PRT
<213> 杜氏利什曼原虫(Leishmania donovani)
<400> 24
Met Ala Asp Val Gly Lys Ala Met Val Thr Leu Ser Asn Gly Val Gln
1 5 10 15
Met Pro Gln Leu Gly Leu Gly Val Trp Gln Ser Pro Ala Gly Glu Val
20 25 30
Thr Ala Asn Ala Val Lys Trp Ala Leu Cys Ala Gly Tyr Arg His Ile
35 40 45
Asp Thr Ala Ala Ile Tyr Lys Asn Glu Glu Ser Val Gly Ala Gly Leu
50 55 60
Arg Ala Ser Gly Val Pro Arg Glu Asp Val Phe Ile Thr Thr Lys Leu
65 70 75 80
Trp Asn Thr Glu Gln Gly Tyr Glu Ser Thr Leu Ala Ala Phe Glu Glu
85 90 95
Ser Arg Gln Lys Leu Gly Val Asp Tyr Ile Asp Leu Tyr Leu Ile His
100 105 110
Trp Pro Arg Gly Lys Asp Ile Val Ser Lys Glu Gly Lys Lys Tyr Leu
115 120 125
Asp Ser Trp Arg Ala Phe Glu Gln Leu Tyr Lys Asp Lys Lys Val Arg
130 135 140
Ala Ile Gly Val Ser Asn Phe His Ile His His Leu Glu Asp Val Leu
145 150 155 160
Ala Met Cys Thr Val Thr Pro Met Val Asn Gln Val Glu Leu His Pro
165 170 175
Leu Asn Asn Gln Ala Glu Leu Arg Ala Phe Cys Asp Ala Lys Gln Ile
180 185 190
Lys Val Glu Ala Trp Ser Pro Leu Gly Gln Gly Lys Leu Leu Ser Asn
195 200 205
Pro Ile Leu Ala Ala Ile Gly Ala Lys Tyr Asn Lys Thr Ala Ala Gln
210 215 220
Val Ile Leu Arg Trp Asn Ile Gln Lys Asn Leu Ile Thr Ile Pro Lys
225 230 235 240
Ser Val His Lys Glu Arg Ile Glu Glu Asn Ala Asp Val Phe Asn Phe
245 250 255
Glu Leu Asp Ala Glu Asp Val Met Ser Ile Asp Ala Leu Asn Thr Asn
260 265 270
Ser Arg Tyr Gly Pro Asp Pro Asp Glu Ala Gln Phe
275 280
<210> 25
<211> 349
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 25
Met Lys Ile Lys Ala Val Gly Ala Tyr Ser Ala Lys Gln Pro Leu Glu
1 5 10 15
Pro Met Asp Ile Thr Arg Arg Glu Pro Gly Pro Asn Asp Val Lys Ile
20 25 30
Glu Ile Ala Tyr Cys Gly Val Cys His Ser Asp Leu His Gln Val Arg
35 40 45
Ser Glu Trp Ala Gly Thr Val Tyr Pro Cys Val Pro Gly His Glu Ile
50 55 60
Val Gly Arg Val Val Ala Val Gly Asp Gln Val Glu Lys Tyr Ala Pro
65 70 75 80
Gly Asp Leu Val Gly Val Gly Cys Ile Val Asp Ser Cys Lys His Cys
85 90 95
Glu Glu Cys Glu Asp Gly Leu Glu Asn Tyr Cys Asp His Met Thr Gly
100 105 110
Thr Tyr Asn Ser Pro Thr Pro Asp Glu Pro Gly His Thr Leu Gly Gly
115 120 125
Tyr Ser Gln Gln Ile Val Val His Glu Arg Tyr Val Leu Arg Ile Arg
130 135 140
His Pro Gln Glu Gln Leu Ala Ala Val Ala Pro Leu Leu Cys Ala Gly
145 150 155 160
Ile Thr Thr Tyr Ser Pro Leu Arg His Trp Gln Ala Gly Pro Gly Lys
165 170 175
Lys Val Gly Val Val Gly Ile Gly Gly Leu Gly His Met Gly Ile Lys
180 185 190
Leu Ala His Ala Met Gly Ala His Val Val Ala Phe Thr Thr Ser Glu
195 200 205
Ala Lys Arg Glu Ala Ala Lys Ala Leu Gly Ala Asp Glu Val Val Asn
210 215 220
Ser Arg Asn Ala Asp Glu Met Ala Ala His Leu Lys Ser Phe Asp Phe
225 230 235 240
Ile Leu Asn Thr Val Ala Ala Pro His Asn Leu Asp Asp Phe Thr Thr
245 250 255
Leu Leu Lys Arg Asp Gly Thr Met Thr Leu Val Gly Ala Pro Ala Thr
260 265 270
Pro His Lys Ser Pro Glu Val Phe Asn Leu Ile Met Lys Arg Arg Ala
275 280 285
Ile Ala Gly Ser Met Ile Gly Gly Ile Pro Glu Thr Gln Glu Met Leu
290 295 300
Asp Phe Cys Ala Glu His Gly Ile Val Ala Asp Ile Glu Met Ile Arg
305 310 315 320
Ala Asp Gln Ile Asn Glu Ala Tyr Glu Arg Met Leu Arg Gly Asp Val
325 330 335
Lys Tyr Arg Phe Val Ile Asp Asn Arg Thr Leu Thr Asp
340 345
<210> 26
<211> 346
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 26
Met Gln Tyr His Arg Ile Pro His Ser Ser Leu Glu Val Ser Thr Leu
1 5 10 15
Gly Leu Gly Thr Met Thr Phe Gly Glu Gln Asn Ser Glu Ala Asp Ala
20 25 30
His Ala Gln Leu Asp Tyr Ala Val Ala Gln Gly Ile Asn Leu Ile Asp
35 40 45
Val Ala Glu Met Tyr Pro Val Pro Pro Arg Pro Glu Thr Gln Gly Leu
50 55 60
Thr Glu Thr Tyr Val Gly Asn Trp Leu Ala Lys His Gly Ser Arg Glu
65 70 75 80
Lys Leu Ile Ile Ala Ser Lys Val Ser Gly Pro Ser Arg Asn Asn Asp
85 90 95
Lys Gly Ile Arg Pro Asp Gln Ala Leu Asp Arg Lys Asn Ile Arg Glu
100 105 110
Ala Leu His Asp Ser Leu Lys Arg Leu Gln Thr Asp Tyr Leu Asp Leu
115 120 125
Tyr Gln Val His Trp Pro Gln Arg Pro Thr Asn Cys Phe Gly Lys Leu
130 135 140
Gly Tyr Ser Trp Thr Asp Ser Ala Pro Ala Val Ser Leu Leu Asp Thr
145 150 155 160
Leu Asp Ala Leu Ala Glu Tyr Gln Arg Ala Gly Lys Ile Arg Tyr Ile
165 170 175
Gly Val Ser Asn Glu Thr Ala Phe Gly Val Met Arg Tyr Leu His Leu
180 185 190
Ala Asp Lys His Asp Leu Pro Arg Ile Val Thr Ile Gln Asn Pro Tyr
195 200 205
Ser Leu Leu Asn Arg Ser Phe Glu Val Gly Leu Ala Glu Val Ser Gln
210 215 220
Tyr Glu Gly Val Glu Leu Leu Ala Tyr Ser Cys Leu Gly Phe Gly Thr
225 230 235 240
Leu Thr Gly Lys Tyr Leu Asn Gly Ala Lys Pro Ala Gly Ala Arg Asn
245 250 255
Thr Leu Phe Ser Arg Phe Thr Arg Tyr Ser Gly Glu Gln Thr Gln Lys
260 265 270
Ala Val Ala Ala Tyr Val Asp Ile Ala Arg Arg His Gly Leu Asp Pro
275 280 285
Ala Gln Met Ala Leu Ala Phe Val Arg Arg Gln Pro Phe Val Ala Ser
290 295 300
Thr Leu Leu Gly Ala Thr Thr Met Asp Gln Leu Lys Thr Asn Ile Glu
305 310 315 320
Ser Leu His Leu Glu Leu Ser Glu Asp Val Leu Ala Glu Ile Glu Ala
325 330 335
Val His Gln Val Tyr Thr Tyr Pro Ala Pro
340 345
<210> 27
<211> 399
<212> PRT
<213> 乙醇热厌氧杆菌(Thermoanaerobacter ethanolicus)
<400> 27
Met Trp Glu Thr Lys Ile Asn Pro Asn Lys Val Phe Glu Leu Arg Cys
1 5 10 15
Lys Asn Thr Thr Tyr Phe Gly Ile Gly Ser Ile Lys Lys Ile Lys Asp
20 25 30
Ile Leu Glu Val Leu Lys Asn Lys Gly Ile Asn Asn Val Ile Leu Val
35 40 45
Thr Gly Lys Gly Ser Tyr Lys Ala Ser Gly Ala Trp Asp Val Val Lys
50 55 60
Pro Ala Leu Glu Thr Leu Gly Phe Lys Tyr Ser Leu Tyr Asp Lys Val
65 70 75 80
Gly Pro Asn Pro Thr Val Asp Met Ile Asp Glu Ala Ala Lys Ile Gly
85 90 95
Arg Glu Thr Gly Ala Lys Ala Val Ile Gly Ile Gly Gly Gly Ser Pro
100 105 110
Ile Asp Thr Ala Lys Ser Val Ala Val Leu Leu Glu Tyr Thr Asp Lys
115 120 125
Asn Ala Arg Glu Leu Tyr Glu Gln Lys Phe Ile Pro Glu Lys Ala Ala
130 135 140
Pro Ile Ile Ala Ile Asn Leu Thr His Gly Thr Gly Thr Glu Val Asp
145 150 155 160
Arg Phe Ala Val Ala Thr Ile Pro Glu Lys Asn Tyr Lys Pro Ala Ile
165 170 175
Ala Tyr Asp Cys Leu Tyr Pro Met Tyr Ala Ile Asp Asp Pro Ser Leu
180 185 190
Met Thr Lys Leu Asp Lys Lys Gln Thr Ile Ala Val Thr Ile Asp Ala
195 200 205
Leu Asn His Val Thr Glu Ala Ala Thr Thr Leu Val Ala Ser Pro Tyr
210 215 220
Ser Val Leu Met Ala Lys Glu Thr Val Arg Leu Ile Val Arg Tyr Leu
225 230 235 240
Pro Ala Ala Val Asn Asp Pro Glu Asn Leu Val Ala Arg Tyr Tyr Leu
245 250 255
Leu Tyr Ala Ser Ala Leu Ala Gly Ile Ser Phe Asp Asn Gly Leu Leu
260 265 270
His Leu Thr His Ala Leu Glu His Pro Leu Ser Ala Val Lys Pro Glu
275 280 285
Ile Ala His Gly Leu Gly Leu Gly Ala Ile Leu Pro Ala Val Val Lys
290 295 300
Ala Ile Tyr Pro Ser Val Ala Glu Val Leu Ala Glu Val Tyr Ser Pro
305 310 315 320
Ile Val Pro Gly Leu Lys Gly Leu Pro Ala Glu Ala Glu Tyr Val Ala
325 330 335
Lys Lys Val Glu Glu Trp Leu Phe Lys Val Gly Cys Thr Gln Lys Leu
340 345 350
Ser Asp Phe Gly Phe Thr Lys Glu Asp Ile Pro Thr Leu Val Arg Leu
355 360 365
Ala Lys Thr Thr Pro Ser Leu Asp Gly Leu Leu Ser Asn Ala Pro Val
370 375 380
Glu Ala Thr Glu Ala Val Ile Ala Lys Ile Tyr Glu Glu Ser Phe
385 390 395
<210> 28
<211> 346
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 28
Met Lys Ala Ala Arg Trp His Asn Gln Lys Asp Ile Arg Ile Glu His
1 5 10 15
Ile Glu Glu Pro Lys Thr Glu Pro Gly Lys Val Lys Ile Lys Val Lys
20 25 30
Trp Cys Gly Ile Cys Gly Ser Asp Leu His Glu Tyr Leu Gly Gly Pro
35 40 45
Ile Phe Ile Pro Val Asp Lys Pro His Pro Leu Thr Asn Glu Thr Ala
50 55 60
Pro Val Thr Met Gly His Glu Phe Ser Gly Glu Val Val Glu Val Gly
65 70 75 80
Glu Gly Val Glu Asn Tyr Lys Val Gly Asp Arg Val Val Val Glu Pro
85 90 95
Ile Phe Ala Thr His Gly His Gln Gly Ala Tyr Asn Leu Asp Glu Gln
100 105 110
Met Gly Phe Leu Gly Leu Ala Gly Gly Gly Gly Gly Phe Ser Glu Tyr
115 120 125
Val Ser Val Asp Glu Glu Leu Leu Phe Lys Leu Pro Asp Glu Leu Ser
130 135 140
Tyr Glu Gln Gly Ala Leu Val Glu Pro Ser Ala Val Ala Leu Tyr Ala
145 150 155 160
Val Arg Ser Ser Lys Leu Lys Ala Gly Asp Lys Ala Ala Val Phe Gly
165 170 175
Cys Gly Pro Ile Gly Leu Leu Val Ile Glu Ala Leu Lys Ala Ala Gly
180 185 190
Ala Thr Asp Ile Tyr Ala Val Glu Leu Ser Pro Glu Arg Gln Gln Lys
195 200 205
Ala Glu Glu Leu Gly Ala Ile Ile Val Asp Pro Ser Lys Thr Asp Asp
210 215 220
Val Val Ala Glu Ile Ala Glu Arg Thr Gly Gly Gly Val Asp Val Ala
225 230 235 240
Phe Glu Val Thr Gly Val Pro Val Val Leu Arg Gln Ala Ile Gln Ser
245 250 255
Thr Thr Ile Ala Gly Glu Thr Val Ile Val Ser Ile Trp Glu Lys Gly
260 265 270
Ala Glu Ile His Pro Asn Asp Ile Val Ile Lys Glu Arg Thr Val Lys
275 280 285
Gly Ile Ile Gly Tyr Arg Asp Ile Phe Pro Ala Val Leu Ser Leu Met
290 295 300
Lys Glu Gly Tyr Phe Ser Ala Asp Lys Leu Val Thr Lys Lys Ile Val
305 310 315 320
Leu Asp Asp Leu Ile Glu Glu Gly Phe Gly Ala Leu Ile Lys Glu Lys
325 330 335
Ser Gln Val Lys Ile Leu Val Arg Pro Asn
340 345
<210> 29
<211> 382
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 29
Met Arg Ala Leu Ala Tyr Phe Lys Lys Gly Asp Ile His Phe Thr Asn
1 5 10 15
Asp Ile Pro Arg Pro Glu Ile Gln Thr Asp Asp Glu Val Ile Ile Asp
20 25 30
Val Ser Trp Cys Gly Ile Cys Gly Ser Asp Leu His Glu Tyr Leu Asp
35 40 45
Gly Pro Ile Phe Met Pro Lys Asp Gly Glu Cys His Lys Leu Ser Asn
50 55 60
Ala Ala Leu Pro Leu Ala Met Gly His Glu Met Ser Gly Ile Val Ser
65 70 75 80
Lys Val Gly Pro Lys Val Thr Lys Val Lys Val Gly Asp His Val Val
85 90 95
Val Asp Ala Ala Ser Ser Cys Ala Asp Leu His Cys Trp Pro His Ser
100 105 110
Lys Phe Tyr Asn Ser Lys Pro Cys Asp Ala Cys Gln Arg Gly Ser Glu
115 120 125
Asn Leu Cys Thr His Ala Gly Phe Val Gly Leu Gly Val Ile Ser Gly
130 135 140
Gly Phe Ala Glu Gln Val Val Val Ser Gln His His Ile Ile Pro Val
145 150 155 160
Pro Lys Glu Ile Pro Leu Asp Val Ala Ala Leu Val Glu Pro Leu Ser
165 170 175
Val Thr Trp His Ala Val Lys Ile Ser Gly Phe Lys Lys Gly Ser Ser
180 185 190
Ala Leu Val Leu Gly Ala Gly Pro Ile Gly Leu Cys Thr Ile Leu Val
195 200 205
Leu Lys Gly Met Gly Ala Ser Lys Ile Val Val Ser Glu Ile Ala Glu
210 215 220
Arg Arg Ile Glu Met Ala Lys Lys Leu Gly Val Glu Val Phe Asn Pro
225 230 235 240
Ser Lys His Gly His Lys Ser Ile Glu Ile Leu Arg Gly Leu Thr Lys
245 250 255
Ser His Asp Gly Phe Asp Tyr Ser Tyr Asp Cys Ser Gly Ile Gln Val
260 265 270
Thr Phe Glu Thr Ser Leu Lys Ala Leu Thr Phe Lys Gly Thr Ala Thr
275 280 285
Asn Ile Ala Val Trp Gly Pro Lys Pro Val Pro Phe Gln Pro Met Asp
290 295 300
Val Thr Leu Gln Glu Lys Val Met Thr Gly Ser Ile Gly Tyr Val Val
305 310 315 320
Glu Asp Phe Glu Glu Val Val Arg Ala Ile His Asn Gly Asp Ile Ala
325 330 335
Met Glu Asp Cys Lys Gln Leu Ile Thr Gly Lys Gln Arg Ile Glu Asp
340 345 350
Gly Trp Glu Lys Gly Phe Gln Glu Leu Met Asp His Lys Glu Ser Asn
355 360 365
Val Lys Ile Leu Leu Thr Pro Asn Asn His Gly Glu Met Lys
370 375 380
<210> 30
<211> 258
<212> PRT
<213> 谷氨酸棒状杆菌(Corynebacterium glutamicum)
<400> 30
Met Ser Lys Val Ala Met Val Thr Gly Gly Ala Gln Gly Ile Gly Arg
1 5 10 15
Gly Ile Ser Glu Lys Leu Ala Ala Asp Gly Phe Asp Ile Ala Val Ala
20 25 30
Asp Leu Pro Gln Gln Glu Glu Gln Ala Ala Glu Thr Ile Lys Leu Ile
35 40 45
Glu Ala Ala Gly Gln Lys Ala Val Phe Val Gly Leu Asp Val Thr Asp
50 55 60
Lys Ala Asn Phe Asp Ser Ala Ile Asp Glu Ala Ala Glu Lys Leu Gly
65 70 75 80
Gly Phe Asp Val Leu Val Asn Asn Ala Gly Ile Ala Gln Ile Lys Pro
85 90 95
Leu Leu Glu Val Thr Glu Glu Asp Leu Lys Gln Ile Tyr Ser Val Asn
100 105 110
Val Phe Ser Val Phe Phe Gly Ile Gln Ala Ala Ser Arg Lys Phe Asp
115 120 125
Glu Leu Gly Val Lys Gly Lys Ile Ile Asn Ala Ala Ser Ile Ala Ala
130 135 140
Ile Gln Gly Phe Pro Ile Leu Ser Ala Tyr Ser Thr Thr Lys Phe Ala
145 150 155 160
Val Arg Gly Leu Thr Gln Ala Ala Ala Gln Glu Leu Ala Pro Lys Gly
165 170 175
His Thr Val Asn Ala Tyr Ala Pro Gly Ile Val Gly Thr Gly Met Trp
180 185 190
Glu Gln Ile Asp Ala Glu Leu Ser Lys Ile Asn Gly Lys Pro Ile Gly
195 200 205
Glu Asn Phe Lys Glu Tyr Ser Ser Ser Ile Ala Leu Gly Arg Pro Ser
210 215 220
Val Pro Glu Asp Val Ala Gly Leu Val Ser Phe Leu Ala Ser Glu Asn
225 230 235 240
Ser Asn Tyr Ile Thr Gly Gln Val Met Leu Val Asp Gly Gly Met Leu
245 250 255
Tyr Asn
<210> 31
<211> 256
<212> PRT
<213> 肺炎克雷伯氏菌(Klebsiella pneumoniae)
<400> 31
Met Lys Lys Val Ala Leu Val Thr Gly Ala Gly Gln Gly Ile Gly Lys
1 5 10 15
Ala Ile Ala Leu Arg Leu Val Lys Asp Gly Phe Ala Val Ala Ile Ala
20 25 30
Asp Tyr Asn Asp Ala Thr Ala Lys Ala Val Ala Ser Glu Ile Asn Gln
35 40 45
Ala Gly Gly Arg Ala Met Ala Val Lys Val Asp Val Ser Asp Arg Asp
50 55 60
Gln Val Phe Ala Ala Val Glu Gln Ala Arg Lys Thr Leu Gly Gly Phe
65 70 75 80
Asp Val Ile Val Asn Asn Ala Gly Val Ala Pro Ser Thr Pro Ile Glu
85 90 95
Ser Ile Thr Pro Glu Ile Val Asp Lys Val Tyr Asn Ile Asn Val Lys
100 105 110
Gly Val Ile Trp Gly Ile Gln Ala Ala Val Glu Ala Phe Lys Lys Glu
115 120 125
Gly His Gly Gly Lys Ile Ile Asn Ala Cys Ser Gln Ala Gly His Val
130 135 140
Gly Asn Pro Glu Leu Ala Val Tyr Ser Ser Ser Lys Phe Ala Val Arg
145 150 155 160
Gly Leu Thr Gln Thr Ala Ala Arg Asp Leu Ala Pro Leu Gly Ile Thr
165 170 175
Val Asn Gly Tyr Cys Pro Gly Ile Val Lys Thr Pro Met Trp Ala Glu
180 185 190
Ile Asp Arg Gln Val Ser Glu Ala Ala Gly Lys Pro Leu Gly Tyr Gly
195 200 205
Thr Ala Glu Phe Ala Lys Arg Ile Thr Leu Gly Arg Leu Ser Glu Pro
210 215 220
Glu Asp Val Ala Ala Cys Val Ser Tyr Leu Ala Ser Pro Asp Ser Asp
225 230 235 240
Tyr Met Thr Gly Gln Ser Leu Leu Ile Asp Gly Gly Met Val Phe Asn
245 250 255
<210> 32
<211> 251
<212> PRT
<213> 粘质沙雷氏菌(Serratia marcescens)
<400> 32
Met Arg Phe Asp Asn Lys Val Val Val Ile Thr Gly Ala Gly Thr Gly
1 5 10 15
Met Gly Glu Ala Ala Ala Arg Arg Phe Ser Ala Glu Gly Ala Ile Val
20 25 30
Val Leu Ala Asp Trp Ala Lys Glu Ala Val Asp Lys Val Ala Ala Ser
35 40 45
Leu Pro Lys Gly Arg Ala Met Ala Val His Ile Asp Val Ser Asp His
50 55 60
Val Ala Val Glu Lys Met Met Asn Glu Val Ala Glu Lys Leu Gly Arg
65 70 75 80
Ile Asp Val Leu Leu Asn Asn Ala Gly Val His Val Ala Gly Ser Val
85 90 95
Leu Glu Thr Ser Val Asp Asp Trp Arg Arg Ile Ala Gly Val Asp Ile
100 105 110
Asp Gly Val Val Phe Cys Ser Lys Phe Ala Leu Pro His Leu Leu Lys
115 120 125
Thr Lys Gly Cys Ile Val Asn Thr Ala Ser Val Ser Gly Leu Gly Gly
130 135 140
Asp Trp Gly Ala Ala Tyr Tyr Cys Ala Ala Lys Gly Ala Val Val Asn
145 150 155 160
Leu Thr Arg Ala Met Ala Leu Asp His Gly Gly Asp Gly Val Arg Ile
165 170 175
Asn Ser Val Cys Pro Ser Leu Val Lys Thr Asn Met Thr Asn Gly Trp
180 185 190
Pro Gln Glu Ile Arg Asp Lys Phe Asn Glu Arg Ile Ala Leu Gly Arg
195 200 205
Ala Ala Glu Pro Glu Glu Val Ala Ala Val Met Ala Phe Leu Ala Ser
210 215 220
Asp Asp Ala Ser Phe Ile Asn Gly Ala Asn Ile Pro Val Asp Gly Gly
225 230 235 240
Ala Thr Ala Ser Asp Gly Gln Gln Asn Ile Val
245 250
<210> 33
<211> 367
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 33
Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp
1 5 10 15
Val Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp
20 25 30
Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val
35 40 45
Glu Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe
50 55 60
Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala
65 70 75 80
Glu Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr
85 90 95
Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala
100 105 110
Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
Val Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro
130 135 140
Asn Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala
145 150 155 160
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly
180 185 190
Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn
195 200 205
Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His
210 215 220
Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val
245 250 255
His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly
260 265 270
Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala
275 280 285
Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly
290 295 300
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala
305 310 315 320
Lys Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile
325 330 335
His Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu
340 345 350
Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
355 360 365
<210> 34
<211> 258
<212> PRT
<213> 谷氨酸棒状杆菌(Corynebacterium glutamicum)
<400> 34
Met Ser Lys Val Ala Met Val Thr Gly Gly Ala Gln Gly Ile Gly Arg
1 5 10 15
Gly Ile Ser Glu Lys Leu Ala Ala Asp Gly Phe Asp Ile Ala Val Ala
20 25 30
Asp Leu Pro Gln Gln Glu Glu Gln Ala Ala Glu Thr Ile Lys Leu Ile
35 40 45
Glu Ala Ala Asp Gln Lys Ala Val Phe Val Gly Leu Asp Val Thr Asp
50 55 60
Lys Ala Asn Phe Asp Ser Ala Ile Asp Glu Ala Ala Glu Lys Leu Gly
65 70 75 80
Gly Phe Asp Val Leu Val Asn Asn Ala Gly Ile Ala Gln Ile Lys Pro
85 90 95
Leu Leu Glu Val Thr Glu Glu Asp Leu Lys Gln Ile Tyr Ser Val Asn
100 105 110
Val Phe Ser Val Phe Phe Gly Ile Gln Ala Ala Ser Arg Lys Phe Asp
115 120 125
Glu Leu Gly Val Lys Gly Lys Ile Ile Asn Ala Ala Ser Ile Ala Ala
130 135 140
Ile Gln Gly Phe Pro Ile Leu Ser Ala Tyr Ser Thr Thr Lys Phe Ala
145 150 155 160
Val Arg Gly Leu Thr Gln Ala Ala Ala Gln Glu Leu Ala Pro Lys Gly
165 170 175
His Thr Val Asn Ala Tyr Ala Pro Gly Ile Val Gly Thr Gly Met Trp
180 185 190
Glu Gln Ile Asp Ala Glu Leu Ser Lys Ile Asn Gly Lys Pro Ile Gly
195 200 205
Glu Asn Phe Lys Glu Tyr Ser Ser Ser Ile Ala Leu Gly Arg Pro Ser
210 215 220
Val Pro Glu Asp Val Ala Gly Leu Val Ser Phe Leu Ala Ser Glu Asn
225 230 235 240
Ser Asn Tyr Val Thr Gly Gln Val Met Leu Val Asp Gly Gly Met Leu
245 250 255
Tyr Asn
<210> 35
<211> 318
<212> PRT
<213> 红褐肉座菌(Hypocrea jecorina)
<400> 35
Met Ala Ser Lys Thr Tyr Thr Leu Asn Thr Gly Ala Lys Ile Pro Ala
1 5 10 15
Val Gly Phe Gly Thr Phe Ala Asn Glu Gly Ala Lys Gly Glu Thr Tyr
20 25 30
Ala Ala Val Thr Lys Ala Leu Asp Val Gly Tyr Arg His Leu Asp Cys
35 40 45
Ala Trp Phe Tyr His Asn Glu Asp Glu Val Gly Asp Ala Val Arg Asp
50 55 60
Phe Leu Ala Arg Arg Pro Asp Val Lys Arg Glu Asp Leu Phe Ile Cys
65 70 75 80
Thr Lys Val Trp Asn His Leu His Glu Pro Glu Asp Val Lys Trp Ser
85 90 95
Ala Lys Asn Ser Cys Glu Asn Leu Lys Val Asp Tyr Ile Asp Leu Phe
100 105 110
Leu Val His Trp Pro Ile Ala Ala Glu Lys Asn Ser Asp Arg Ser Val
115 120 125
Lys Leu Gly Pro Asp Gly Lys Tyr Val Ile Asn Gln Ala Leu Thr Glu
130 135 140
Asn Pro Glu Pro Thr Trp Arg Ala Met Glu Glu Leu Val Glu Ser Gly
145 150 155 160
Leu Val Lys Ala Ile Gly Val Ser Asn Trp Thr Ile Pro Gly Leu Lys
165 170 175
Lys Leu Leu Gln Ile Ala Lys Ile Lys Pro Ala Val Asn Gln Ile Glu
180 185 190
Ile His Pro Phe Leu Pro Asn Glu Glu Leu Val Ala Phe Cys Phe Glu
195 200 205
Asn Gly Ile Leu Pro Glu Ala Tyr Ser Pro Leu Gly Ser Gln Asn Gln
210 215 220
Val Pro Ser Thr Gly Glu Arg Val Arg Asp Asn Pro Thr Leu Lys Ala
225 230 235 240
Val Ala Glu Arg Ser Gly Tyr Ser Leu Ala Gln Ile Leu Leu Ala Trp
245 250 255
Gly Leu Lys Arg Gly Tyr Val Val Leu Pro Lys Ser Ser Thr Pro Ser
260 265 270
Arg Ile Glu Ser Asn Phe Asn Ile Pro Glu Leu Ser Asp Glu Asp Phe
275 280 285
Glu Ala Ile Gln Gln Val Ala Lys Gly Arg His Thr Arg Phe Val Asn
290 295 300
Met Lys Asp Thr Phe Gly Tyr Asn Val Trp Pro Glu Glu Glu
305 310 315
<210> 36
<211> 385
<212> PRT
<213> 丁酸梭菌(Clostridium butyricum)
<400> 36
Met Arg Met Tyr Asp Tyr Leu Val Pro Ser Val Asn Phe Met Gly Ala
1 5 10 15
Asn Ser Val Ser Val Val Gly Glu Arg Cys Lys Ile Leu Gly Gly Lys
20 25 30
Lys Ala Leu Ile Val Thr Asp Lys Phe Leu Lys Asp Met Glu Gly Gly
35 40 45
Ala Val Glu Leu Thr Val Lys Tyr Leu Lys Glu Ala Gly Leu Asp Val
50 55 60
Val Tyr Tyr Asp Gly Val Glu Pro Asn Pro Lys Asp Val Asn Val Ile
65 70 75 80
Glu Gly Leu Lys Ile Phe Lys Glu Glu Asn Cys Asp Met Ile Val Thr
85 90 95
Val Gly Gly Gly Ser Ser His Asp Cys Gly Lys Gly Ile Gly Ile Ala
100 105 110
Ala Thr His Glu Gly Asp Leu Tyr Asp Tyr Ala Gly Ile Glu Thr Leu
115 120 125
Val Asn Pro Leu Pro Pro Ile Val Ala Val Asn Thr Thr Ala Gly Thr
130 135 140
Ala Ser Glu Leu Thr Arg His Cys Val Leu Thr Asn Thr Lys Lys Lys
145 150 155 160
Ile Lys Phe Val Ile Val Ser Trp Arg Asn Leu Pro Leu Val Ser Ile
165 170 175
Asn Asp Pro Met Leu Met Val Lys Lys Pro Ala Gly Leu Thr Ala Ala
180 185 190
Thr Gly Met Asp Ala Leu Thr His Ala Ile Glu Ala Tyr Val Ser Lys
195 200 205
Asp Ala Asn Pro Val Thr Asp Ala Ser Ala Ile Gln Ala Ile Lys Leu
210 215 220
Ile Ser Gln Asn Leu Arg Gln Ala Val Ala Leu Gly Glu Asn Leu Glu
225 230 235 240
Ala Arg Glu Asn Met Ala Tyr Ala Ser Leu Leu Ala Gly Met Ala Phe
245 250 255
Asn Asn Ala Asn Leu Gly Tyr Val His Ala Met Ala His Gln Leu Gly
260 265 270
Gly Leu Tyr Asp Met Pro His Gly Val Ala Asn Ala Met Leu Leu Pro
275 280 285
His Val Glu Arg Tyr Asn Met Leu Ser Asn Pro Lys Lys Phe Ala Asp
290 295 300
Ile Ala Glu Phe Met Gly Glu Asn Ile Ser Gly Leu Ser Val Met Glu
305 310 315 320
Ala Ala Glu Lys Ala Ile Asn Ala Met Phe Arg Leu Ser Glu Asp Val
325 330 335
Gly Ile Pro Lys Ser Leu Lys Glu Met Gly Val Lys Gln Glu Asp Phe
340 345 350
Glu His Met Ala Glu Leu Ala Leu Leu Asp Gly Asn Ala Phe Ser Asn
355 360 365
Pro Arg Lys Gly Asn Ala Lys Asp Ile Ile Asn Ile Phe Lys Ala Ala
370 375 380
Tyr
385
<210> 37
<211> 387
<212> PRT
<213> 弗氏柠檬酸杆菌(Citrobacter Freundii)
<400> 37
Met Ser Tyr Arg Met Phe Asp Tyr Leu Val Pro Asn Val Asn Phe Phe
1 5 10 15
Gly Pro Asn Ala Ile Ser Val Val Gly Glu Arg Cys Lys Leu Leu Gly
20 25 30
Gly Lys Lys Ala Leu Leu Val Thr Asp Lys Gly Leu Arg Ala Ile Lys
35 40 45
Asp Gly Ala Val Asp Lys Thr Leu Thr His Leu Arg Glu Ala Gly Ile
50 55 60
Asp Val Val Val Phe Asp Gly Val Glu Pro Asn Pro Lys Asp Thr Asn
65 70 75 80
Val Arg Asp Gly Leu Glu Val Phe Arg Lys Glu His Cys Asp Ile Ile
85 90 95
Val Thr Val Gly Gly Gly Ser Pro His Asp Cys Gly Lys Gly Ile Gly
100 105 110
Ile Ala Ala Thr His Glu Gly Asp Leu Tyr Ser Tyr Ala Gly Ile Glu
115 120 125
Thr Leu Thr Asn Pro Leu Pro Pro Ile Val Ala Val Asn Thr Thr Ala
130 135 140
Gly Thr Ala Ser Glu Val Thr Arg His Cys Val Leu Thr Asn Thr Lys
145 150 155 160
Thr Lys Val Lys Phe Val Ile Val Ser Trp Arg Asn Leu Pro Ser Val
165 170 175
Ser Ile Asn Asp Pro Leu Leu Met Leu Gly Lys Pro Ala Pro Leu Thr
180 185 190
Ala Ala Thr Gly Met Asp Ala Leu Thr His Ala Val Glu Ala Tyr Ile
195 200 205
Ser Lys Asp Ala Asn Pro Val Thr Asp Ala Ala Ala Ile Gln Ala Ile
210 215 220
Arg Leu Ile Ala Arg Asn Leu Arg Gln Ala Val Ala Leu Gly Ser Asn
225 230 235 240
Leu Lys Ala Arg Glu Asn Met Ala Tyr Ala Ser Leu Leu Ala Gly Met
245 250 255
Ala Phe Asn Asn Ala Asn Leu Gly Tyr Val His Ala Met Ala His Gln
260 265 270
Leu Gly Gly Leu Tyr Asp Met Pro His Gly Val Ala Asn Ala Val Leu
275 280 285
Leu Pro His Val Ala Arg Tyr Asn Leu Ile Ala Asn Pro Glu Lys Phe
290 295 300
Ala Asp Ile Ala Glu Phe Met Gly Glu Asn Thr Asp Gly Leu Ser Thr
305 310 315 320
Met Asp Ala Ala Glu Leu Ala Ile His Ala Ile Ala Arg Leu Ser Ala
325 330 335
Asp Ile Gly Ile Pro Gln His Leu Arg Asp Leu Gly Val Lys Glu Ala
340 345 350
Asp Phe Pro Tyr Met Ala Glu Met Ala Leu Lys Asp Gly Asn Ala Phe
355 360 365
Ser Asn Pro Arg Lys Gly Asn Glu Lys Glu Ile Ala Glu Ile Phe Arg
370 375 380
Gln Ala Phe
385
<210> 38
<211> 387
<212> PRT
<213> 肺炎克雷伯氏菌(Klebsiella pneumoniae)
<400> 38
Met Ser Tyr Arg Met Phe Asp Tyr Leu Val Pro Asn Val Asn Phe Phe
1 5 10 15
Gly Pro Asn Ala Ile Ser Val Val Gly Glu Arg Cys Gln Leu Leu Gly
20 25 30
Gly Lys Lys Ala Leu Leu Val Thr Asp Lys Gly Leu Arg Ala Ile Lys
35 40 45
Asp Gly Ala Val Asp Lys Thr Leu His Tyr Leu Arg Glu Ala Gly Ile
50 55 60
Glu Val Ala Ile Phe Asp Gly Val Glu Pro Asn Pro Lys Asp Thr Asn
65 70 75 80
Val Arg Asp Gly Leu Ala Val Phe Arg Arg Glu Gln Cys Asp Ile Ile
85 90 95
Val Thr Val Gly Gly Gly Ser Pro His Asp Cys Gly Lys Gly Ile Gly
100 105 110
Ile Ala Ala Thr His Glu Gly Asp Leu Tyr Gln Tyr Ala Gly Ile Glu
115 120 125
Thr Leu Thr Asn Pro Leu Pro Pro Ile Val Ala Val Asn Thr Thr Ala
130 135 140
Gly Thr Ala Ser Glu Val Thr Arg His Cys Val Leu Thr Asn Thr Glu
145 150 155 160
Thr Lys Val Lys Phe Val Ile Val Ser Trp Arg Asn Leu Pro Ser Val
165 170 175
Ser Ile Asn Asp Pro Leu Leu Met Ile Gly Lys Pro Ala Ala Leu Thr
180 185 190
Ala Ala Thr Gly Met Asp Ala Leu Thr His Ala Val Glu Ala Tyr Ile
195 200 205
Ser Lys Asp Ala Asn Pro Val Thr Asp Ala Ala Ala Met Gln Ala Ile
210 215 220
Arg Leu Ile Ala Arg Asn Leu Arg Gln Ala Val Ala Leu Gly Ser Asn
225 230 235 240
Leu Gln Ala Arg Glu Asn Met Ala Tyr Ala Ser Leu Leu Ala Gly Met
245 250 255
Ala Phe Asn Asn Ala Asn Leu Gly Tyr Val His Ala Met Ala His Gln
260 265 270
Leu Gly Gly Leu Tyr Asp Met Pro His Gly Val Ala Asn Ala Val Leu
275 280 285
Leu Pro His Val Ala Arg Tyr Asn Leu Ile Ala Asn Pro Glu Lys Phe
290 295 300
Ala Asp Ile Ala Glu Leu Met Gly Glu Asn Ile Thr Gly Leu Ser Thr
305 310 315 320
Leu Asp Ala Ala Glu Lys Ala Ile Ala Ala Ile Thr Arg Leu Ser Met
325 330 335
Asp Ile Gly Ile Pro Gln His Leu Arg Asp Leu Gly Val Lys Glu Ala
340 345 350
Asp Phe Pro Tyr Met Ala Glu Met Ala Leu Lys Asp Gly Asn Ala Phe
355 360 365
Ser Asn Pro Arg Lys Gly Asn Glu Gln Glu Ile Ala Ala Ile Phe Arg
370 375 380
Gln Ala Phe
385
<210> 39
<211> 22
<212> PRT
<213> 红串红球菌(Rhodoccus erythropolis)
<400> 39
Met Lys Thr Lys Ala Ala Val Leu Phe Glu Thr His Lys Pro Phe Glu
1 5 10 15
Ile Val Glu Leu Glu Leu
20
<210> 40
<211> 371
<212> PRT
<213> 甲醇拟无枝酸菌(Amycolatopsis methanolica)
<400> 40
Met Lys Thr Lys Ala Ala Val Leu His Ser Ala Gly Lys Pro Phe Glu
1 5 10 15
Ile Glu Glu Leu Glu Leu Asp Gly Pro Arg Glu Gly Glu Val Leu Ile
20 25 30
Lys Tyr Thr Ala Ala Gly Leu Cys His Ser Asp Leu His Leu Ile Asp
35 40 45
Asn Asp Leu Val Pro Arg Phe Pro Ile Val Gly Gly His Glu Gly Ala
50 55 60
Gly Val Ile Glu Asp Val Gly Pro Gly Val Thr Lys Val Lys Pro Gly
65 70 75 80
Asp His Val Val Cys Ser Phe Ile Pro Asn Cys Gly Thr Cys Arg Tyr
85 90 95
Cys Ala Thr Gly Arg Ser Asn Leu Cys Asp Met Gly Ala Thr Ile Leu
100 105 110
Asp Gly Gly Met Pro Asp Gly Ser Phe Arg Phe His Arg Gly Gly Thr
115 120 125
Asp Tyr Gly Ala Met Cys Met Leu Gly Thr Phe Ser Glu Arg Ala Thr
130 135 140
Ile Ser Gln His Ser Val Val Lys Val Asp Asp Trp Leu Pro Leu Glu
145 150 155 160
Thr Ala Val Leu Val Gly Cys Gly Val Pro Thr Gly Trp Ala Ser Ala
165 170 175
Asn Tyr Ala Gly Gly Val Arg Ala Gly Asp Thr Cys Val Val Tyr Gly
180 185 190
Ile Gly Gly Ile Gly Ile Asn Ala Val Gln Gly Ala Ala His Ala Gly
195 200 205
Ala Ala Asn Val Ile Ala Val Asp Pro Val Ala Phe Lys Arg Glu Lys
210 215 220
Ala Leu Glu Leu Gly Ala Thr His Ala Phe Ala Ser Ala Asp Glu Ala
225 230 235 240
Ala Ala Lys Val Ala Glu Leu Thr Trp Gly Gln Met Ala Asp Gln Ala
245 250 255
Leu Ile Thr Val Gly Thr Val Val Glu Gln Val Val Thr Asp Ala Phe
260 265 270
Asn Val Ile Gly Lys Gly Gly Thr Val Val Ile Thr Gly Leu Ala Asn
275 280 285
Pro Glu Lys Leu Thr Val His Leu Ser Gly Gly Val Met Thr Leu Phe
290 295 300
Glu Lys Thr Val Lys Gly Thr Leu Phe Gly Ser Ala Asn Pro Gln Tyr
305 310 315 320
Asp Ile Val Arg Leu Leu Arg Leu Tyr Gln Ala Gly His Val Lys Leu
325 330 335
Asp Glu Leu Val Thr Lys Arg Tyr Ser Leu Glu Glu Val Asn Glu Gly
340 345 350
Tyr Gln Asp Leu Arg Asp Gly Lys Asn Ile Arg Gly Val Ile Met His
355 360 365
Ser Ala Asp
370
<210> 41
<211> 382
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 41
Met Ala Asn Arg Met Ile Leu Asn Glu Thr Ala Trp Phe Gly Arg Gly
1 5 10 15
Ala Val Gly Ala Leu Thr Asp Glu Val Lys Arg Arg Gly Tyr Gln Lys
20 25 30
Ala Leu Ile Val Thr Asp Lys Thr Leu Val Gln Cys Gly Val Val Ala
35 40 45
Lys Val Thr Asp Lys Met Asp Ala Ala Gly Leu Ala Trp Ala Ile Tyr
50 55 60
Asp Gly Val Val Pro Asn Pro Thr Ile Thr Val Val Lys Glu Gly Leu
65 70 75 80
Gly Val Phe Gln Asn Ser Gly Ala Asp Tyr Leu Ile Ala Ile Gly Gly
85 90 95
Gly Ser Pro Gln Asp Thr Cys Lys Ala Ile Gly Ile Ile Ser Asn Asn
100 105 110
Pro Glu Phe Ala Asp Val Arg Ser Leu Glu Gly Leu Ser Pro Thr Asn
115 120 125
Lys Pro Ser Val Pro Ile Leu Ala Ile Pro Thr Thr Ala Gly Thr Ala
130 135 140
Ala Glu Val Thr Ile Asn Tyr Val Ile Thr Asp Glu Glu Lys Arg Arg
145 150 155 160
Lys Phe Val Cys Val Asp Pro His Asp Ile Pro Gln Val Ala Phe Ile
165 170 175
Asp Ala Asp Met Met Asp Gly Met Pro Pro Ala Leu Lys Ala Ala Thr
180 185 190
Gly Val Asp Ala Leu Thr His Ala Ile Glu Gly Tyr Ile Thr Arg Gly
195 200 205
Ala Trp Ala Leu Thr Asp Ala Leu His Ile Lys Ala Ile Glu Ile Ile
210 215 220
Ala Gly Ala Leu Arg Gly Ser Val Ala Gly Asp Lys Asp Ala Gly Glu
225 230 235 240
Glu Met Ala Leu Gly Gln Tyr Val Ala Gly Met Gly Phe Ser Asn Val
245 250 255
Gly Leu Gly Leu Val His Gly Met Ala His Pro Leu Gly Ala Phe Tyr
260 265 270
Asn Thr Pro His Gly Val Ala Asn Ala Ile Leu Leu Pro His Val Met
275 280 285
Arg Tyr Asn Ala Asp Phe Thr Gly Glu Lys Tyr Arg Asp Ile Ala Arg
290 295 300
Val Met Gly Val Lys Val Glu Gly Met Ser Leu Glu Glu Ala Arg Asn
305 310 315 320
Ala Ala Val Glu Ala Val Phe Ala Leu Asn Arg Asp Val Gly Ile Pro
325 330 335
Pro His Leu Arg Asp Val Gly Val Arg Lys Glu Asp Ile Pro Ala Leu
340 345 350
Ala Gln Ala Ala Leu Asp Asp Val Cys Thr Gly Gly Asn Pro Arg Glu
355 360 365
Ala Thr Leu Glu Asp Ile Val Glu Leu Tyr His Thr Ala Trp
370 375 380
<210> 42
<211> 312
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 42
Met Asp Ile Ile Phe Tyr His Pro Thr Phe Asp Thr Gln Trp Trp Ile
1 5 10 15
Glu Ala Leu Arg Lys Ala Ile Pro Gln Ala Arg Val Arg Ala Trp Lys
20 25 30
Ser Gly Asp Asn Asp Ser Ala Asp Tyr Ala Leu Val Trp His Pro Pro
35 40 45
Val Glu Met Leu Ala Gly Arg Asp Leu Lys Ala Val Phe Ala Leu Gly
50 55 60
Ala Gly Val Asp Ser Ile Leu Ser Lys Leu Gln Ala His Pro Glu Met
65 70 75 80
Leu Asn Pro Ser Val Pro Leu Phe Arg Leu Glu Asp Thr Gly Met Gly
85 90 95
Glu Gln Met Gln Glu Tyr Ala Val Ser Gln Val Leu His Trp Phe Arg
100 105 110
Arg Phe Asp Asp Tyr Arg Ile Gln Gln Asn Ser Ser His Trp Gln Pro
115 120 125
Leu Pro Glu Tyr His Arg Glu Asp Phe Thr Ile Gly Ile Leu Gly Ala
130 135 140
Gly Val Leu Gly Ser Lys Val Ala Gln Ser Leu Gln Thr Trp Arg Phe
145 150 155 160
Pro Leu Arg Cys Trp Ser Arg Thr Arg Lys Ser Trp Pro Gly Val Gln
165 170 175
Ser Phe Ala Gly Arg Glu Glu Leu Ser Ala Phe Leu Ser Gln Cys Arg
180 185 190
Val Leu Ile Asn Leu Leu Pro Asn Thr Pro Glu Thr Val Gly Ile Ile
195 200 205
Asn Gln Gln Leu Leu Glu Lys Leu Pro Asp Gly Ala Tyr Leu Leu Asn
210 215 220
Leu Ala Arg Gly Val His Val Val Glu Asp Asp Leu Leu Ala Ala Leu
225 230 235 240
Asp Ser Gly Lys Val Lys Gly Ala Met Leu Asp Val Phe Asn Arg Glu
245 250 255
Pro Leu Pro Pro Glu Ser Pro Leu Trp Gln His Pro Arg Val Thr Ile
260 265 270
Thr Pro His Val Ala Ala Ile Thr Arg Pro Ala Glu Ala Val Glu Tyr
275 280 285
Ile Ser Arg Thr Ile Ala Gln Leu Glu Lys Gly Glu Arg Val Cys Gly
290 295 300
Gln Val Asp Arg Ala Arg Gly Tyr
305 310
<210> 43
<211> 324
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 43
Met Lys Pro Ser Val Ile Leu Tyr Lys Ala Leu Pro Asp Asp Leu Leu
1 5 10 15
Gln Arg Leu Gln Glu His Phe Thr Val His Gln Val Ala Asn Leu Ser
20 25 30
Pro Gln Thr Val Glu Gln Asn Ala Ala Ile Phe Ala Glu Ala Glu Gly
35 40 45
Leu Leu Gly Ser Asn Glu Asn Val Asn Ala Ala Leu Leu Glu Lys Met
50 55 60
Pro Lys Leu Arg Ala Thr Ser Thr Ile Ser Val Gly Tyr Asp Asn Phe
65 70 75 80
Asp Val Asp Ala Leu Thr Ala Arg Lys Ile Leu Leu Met His Thr Pro
85 90 95
Thr Val Leu Thr Glu Thr Val Ala Asp Thr Leu Met Ala Leu Val Leu
100 105 110
Ser Thr Ala Arg Arg Val Val Glu Val Ala Glu Arg Val Lys Ala Gly
115 120 125
Glu Trp Thr Ala Ser Ile Gly Pro Asp Trp Tyr Gly Thr Asp Val His
130 135 140
His Lys Thr Leu Gly Ile Val Gly Met Gly Arg Ile Gly Met Ala Leu
145 150 155 160
Ala Gln Arg Ala His Phe Gly Phe Asn Met Pro Ile Leu Tyr Asn Ala
165 170 175
Arg Arg His His Lys Glu Ala Glu Glu Arg Phe Asn Ala Arg Tyr Cys
180 185 190
Asp Leu Asp Thr Leu Leu Gln Glu Ser Asp Phe Val Cys Leu Ile Leu
195 200 205
Pro Leu Thr Asp Glu Thr His His Leu Phe Gly Ala Glu Gln Phe Ala
210 215 220
Lys Met Lys Ser Ser Ala Ile Phe Ile Asn Ala Gly Arg Gly Pro Val
225 230 235 240
Val Asp Glu Asn Ala Leu Ile Ala Ala Leu Gln Lys Gly Glu Ile His
245 250 255
Ala Ala Gly Leu Asp Val Phe Glu Gln Glu Pro Leu Ser Val Asp Ser
260 265 270
Pro Leu Leu Ser Met Ala Asn Val Val Ala Val Pro His Ile Gly Ser
275 280 285
Ala Thr His Glu Thr Arg Tyr Gly Met Ala Ala Cys Ala Val Asp Asn
290 295 300
Leu Ile Asp Ala Leu Gln Gly Lys Val Glu Lys Asn Cys Val Asn Pro
305 310 315 320
His Val Ala Asp
<210> 44
<211> 275
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 44
Met Ala Asn Pro Thr Val Ile Lys Leu Gln Asp Gly Asn Val Met Pro
1 5 10 15
Gln Leu Gly Leu Gly Val Trp Gln Ala Ser Asn Glu Glu Val Ile Thr
20 25 30
Ala Ile Gln Lys Ala Leu Glu Val Gly Tyr Arg Ser Ile Asp Thr Ala
35 40 45
Ala Ala Tyr Lys Asn Glu Glu Gly Val Gly Lys Ala Leu Lys Asn Ala
50 55 60
Ser Val Asn Arg Glu Glu Leu Phe Ile Thr Thr Lys Leu Trp Asn Asp
65 70 75 80
Asp His Lys Arg Pro Arg Glu Ala Leu Leu Asp Ser Leu Lys Lys Leu
85 90 95
Gln Leu Asp Tyr Ile Asp Leu Tyr Leu Met His Trp Pro Val Pro Ala
100 105 110
Ile Asp His Tyr Val Glu Ala Trp Lys Gly Met Ile Glu Leu Gln Lys
115 120 125
Glu Gly Leu Ile Lys Ser Ile Gly Val Cys Asn Phe Gln Ile His His
130 135 140
Leu Gln Arg Leu Ile Asp Glu Thr Gly Val Thr Pro Val Ile Asn Gln
145 150 155 160
Ile Glu Leu His Pro Leu Met Gln Gln Arg Gln Leu His Ala Trp Asn
165 170 175
Ala Thr His Lys Ile Gln Thr Glu Ser Trp Ser Pro Leu Ala Gln Gly
180 185 190
Gly Lys Gly Val Phe Asp Gln Lys Val Ile Arg Asp Leu Ala Asp Lys
195 200 205
Tyr Gly Lys Thr Pro Ala Gln Ile Val Ile Arg Trp His Leu Asp Ser
210 215 220
Gly Leu Val Val Ile Pro Lys Ser Val Thr Pro Ser Arg Ile Ala Glu
225 230 235 240
Asn Phe Asp Val Trp Asp Phe Arg Leu Asp Lys Asp Glu Leu Gly Glu
245 250 255
Ile Ala Lys Leu Asp Gln Gly Lys Arg Leu Gly Pro Asp Pro Asp Gln
260 265 270
Phe Gly Gly
275
<210> 45
<211> 267
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 45
Met Ala Ile Pro Ala Phe Gly Leu Gly Thr Phe Arg Leu Lys Asp Asp
1 5 10 15
Val Val Ile Ser Ser Val Ile Thr Ala Leu Glu Leu Gly Tyr Arg Ala
20 25 30
Ile Asp Thr Ala Gln Ile Tyr Asp Asn Glu Ala Ala Val Gly Gln Ala
35 40 45
Ile Ala Glu Ser Gly Val Pro Arg His Glu Leu Tyr Ile Thr Thr Lys
50 55 60
Ile Trp Ile Glu Asn Leu Ser Lys Asp Lys Leu Ile Pro Ser Leu Lys
65 70 75 80
Glu Ser Leu Gln Lys Leu Arg Thr Asp Tyr Val Asp Leu Thr Leu Ile
85 90 95
His Trp Pro Ser Pro Asn Asp Glu Val Ser Val Glu Glu Phe Met Gln
100 105 110
Ala Leu Leu Glu Ala Lys Lys Gln Gly Leu Thr Arg Glu Ile Gly Ile
115 120 125
Ser Asn Phe Thr Ile Pro Leu Met Glu Lys Ala Ile Ala Ala Val Gly
130 135 140
Ala Glu Asn Ile Ala Thr Asn Gln Ile Glu Leu Ser Pro Tyr Leu Gln
145 150 155 160
Asn Arg Lys Val Val Ala Trp Ala Lys Gln His Gly Ile His Ile Thr
165 170 175
Ser Tyr Met Thr Leu Ala Tyr Gly Lys Ala Leu Lys Asp Glu Val Ile
180 185 190
Ala Arg Ile Ala Ala Lys His Asn Ala Thr Pro Ala Gln Val Ile Leu
195 200 205
Ala Trp Ala Met Gly Glu Gly Tyr Ser Val Ile Pro Ser Ser Thr Lys
210 215 220
Arg Lys Asn Leu Glu Ser Asn Leu Lys Ala Gln Asn Leu Gln Leu Asp
225 230 235 240
Ala Glu Asp Lys Lys Ala Ile Ala Ala Leu Asp Cys Asn Asp Arg Leu
245 250 255
Val Ser Pro Glu Gly Leu Ala Pro Glu Trp Asp
260 265
<210> 46
<211> 495
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 46
Met Asn Phe His His Leu Ala Tyr Trp Gln Asp Lys Ala Leu Ser Leu
1 5 10 15
Ala Ile Glu Asn Arg Leu Phe Ile Asn Gly Glu Tyr Thr Ala Ala Ala
20 25 30
Glu Asn Glu Thr Phe Glu Thr Val Asp Pro Val Thr Gln Ala Pro Leu
35 40 45
Ala Lys Ile Ala Arg Gly Lys Ser Val Asp Ile Asp Arg Ala Met Ser
50 55 60
Ala Ala Arg Gly Val Phe Glu Arg Gly Asp Trp Ser Leu Ser Ser Pro
65 70 75 80
Ala Lys Arg Lys Ala Val Leu Asn Lys Leu Ala Asp Leu Met Glu Ala
85 90 95
His Ala Glu Glu Leu Ala Leu Leu Glu Thr Leu Asp Thr Gly Lys Pro
100 105 110
Ile Arg His Ser Leu Arg Asp Asp Ile Pro Gly Ala Ala Arg Ala Ile
115 120 125
Arg Trp Tyr Ala Glu Ala Ile Asp Lys Val Tyr Gly Glu Val Ala Thr
130 135 140
Thr Ser Ser His Glu Leu Ala Met Ile Val Arg Glu Pro Val Gly Val
145 150 155 160
Ile Ala Ala Ile Val Pro Trp Asn Phe Pro Leu Leu Leu Thr Cys Trp
165 170 175
Lys Leu Gly Pro Ala Leu Ala Ala Gly Asn Ser Val Ile Leu Lys Pro
180 185 190
Ser Glu Lys Ser Pro Leu Ser Ala Ile Arg Leu Ala Gly Leu Ala Lys
195 200 205
Glu Ala Gly Leu Pro Asp Gly Val Leu Asn Val Val Thr Gly Phe Gly
210 215 220
His Glu Ala Gly Gln Ala Leu Ser Arg His Asn Asp Ile Asp Ala Ile
225 230 235 240
Ala Phe Thr Gly Ser Thr Arg Thr Gly Lys Gln Leu Leu Lys Asp Ala
245 250 255
Gly Asp Ser Asn Met Lys Arg Val Trp Leu Glu Ala Gly Gly Lys Ser
260 265 270
Ala Asn Ile Val Phe Ala Asp Cys Pro Asp Leu Gln Gln Ala Ala Ser
275 280 285
Ala Thr Ala Ala Gly Ile Phe Tyr Asn Gln Gly Gln Val Cys Ile Ala
290 295 300
Gly Thr Arg Leu Leu Leu Glu Glu Ser Ile Ala Asp Glu Phe Leu Ala
305 310 315 320
Leu Leu Lys Gln Gln Ala Gln Asn Trp Gln Pro Gly His Pro Leu Asp
325 330 335
Pro Ala Thr Thr Met Gly Thr Leu Ile Asp Cys Ala His Ala Asp Ser
340 345 350
Val His Ser Phe Ile Arg Glu Gly Glu Ser Lys Gly Gln Leu Leu Leu
355 360 365
Asp Gly Arg Asn Ala Gly Leu Ala Ala Ala Ile Gly Pro Thr Ile Phe
370 375 380
Val Asp Val Asp Pro Asn Ala Ser Leu Ser Arg Glu Glu Ile Phe Gly
385 390 395 400
Pro Val Leu Val Val Thr Arg Phe Thr Ser Glu Glu Gln Ala Leu Gln
405 410 415
Leu Ala Asn Asp Ser Gln Tyr Gly Leu Gly Ala Ala Val Trp Thr Arg
420 425 430
Asp Leu Ser Arg Ala His Arg Met Ser Arg Arg Leu Lys Ala Gly Ser
435 440 445
Val Phe Val Asn Asn Tyr Asn Asp Gly Asp Met Thr Val Pro Phe Gly
450 455 460
Gly Tyr Lys Gln Ser Gly Asn Gly Arg Asp Lys Ser Leu His Ala Leu
465 470 475 480
Glu Lys Phe Thr Glu Leu Lys Thr Ile Trp Ile Ser Leu Glu Ala
485 490 495
<210> 47
<211> 496
<212> PRT
<213> 肺炎克雷伯氏菌(Klebsiella pneumoniae)
<400> 47
Met Met Asn Phe Gln His Leu Ala Tyr Trp Gln Glu Lys Ala Lys Asn
1 5 10 15
Leu Ala Ile Glu Thr Arg Leu Phe Ile Asn Gly Glu Tyr Cys Ala Ala
20 25 30
Ala Asp Asn Thr Thr Phe Glu Thr Ile Asp Pro Ala Ala Gln Gln Thr
35 40 45
Leu Ala Gln Val Ala Arg Gly Lys Lys Ala Asp Val Glu Arg Ala Val
50 55 60
Lys Ala Ala Arg Gln Ala Phe Asp Asn Gly Asp Trp Ser Gln Ala Ser
65 70 75 80
Pro Ala Gln Arg Lys Ala Ile Leu Thr Arg Phe Ala Asn Leu Met Glu
85 90 95
Ala His Arg Glu Glu Leu Ala Leu Leu Glu Thr Leu Asp Thr Gly Lys
100 105 110
Pro Ile Arg His Ser Leu Arg Asp Asp Ile Pro Gly Ala Ala Arg Ala
115 120 125
Ile Arg Trp Tyr Ala Glu Ala Leu Asp Lys Val Tyr Gly Glu Val Ala
130 135 140
Pro Thr Gly Ser Asn Glu Leu Ala Met Ile Val Arg Glu Pro Ile Gly
145 150 155 160
Val Ile Ala Ala Val Val Pro Trp Asn Phe Pro Leu Leu Leu Ala Cys
165 170 175
Trp Lys Leu Gly Pro Ala Leu Ala Ala Gly Asn Ser Val Ile Leu Lys
180 185 190
Pro Ser Glu Lys Ser Pro Leu Thr Ala Leu Arg Leu Ala Gly Leu Ala
195 200 205
Lys Glu Ala Gly Leu Pro Asp Gly Val Leu Asn Val Val Ser Gly Phe
210 215 220
Gly His Glu Ala Gly Gln Ala Leu Ala Leu His Pro Asp Val Glu Val
225 230 235 240
Ile Thr Phe Thr Gly Ser Thr Arg Thr Gly Lys Gln Leu Leu Lys Asp
245 250 255
Ala Gly Asp Ser Asn Met Lys Arg Val Trp Leu Glu Ala Gly Gly Lys
260 265 270
Ser Ala Asn Ile Val Phe Ala Asp Cys Pro Asp Leu Gln Gln Ala Val
275 280 285
Arg Ala Thr Ala Gly Gly Ile Phe Tyr Asn Gln Gly Gln Val Cys Ile
290 295 300
Ala Gly Thr Arg Leu Leu Leu Glu Glu Ser Ile Ala Asp Glu Phe Leu
305 310 315 320
Ala Arg Leu Lys Ala Glu Ala Gln His Trp Gln Pro Gly Asn Pro Leu
325 330 335
Asp Pro Asp Thr Thr Met Gly Met Leu Ile Asp Asn Thr His Ala Asp
340 345 350
Asn Val His Ser Phe Ile Arg Gly Gly Glu Ser Gln Ser Thr Leu Phe
355 360 365
Leu Asp Gly Arg Lys Asn Pro Trp Pro Ala Ala Val Gly Pro Thr Ile
370 375 380
Phe Val Asp Val Asp Pro Ala Ser Thr Leu Ser Arg Glu Glu Ile Phe
385 390 395 400
Gly Pro Val Leu Val Val Thr Arg Phe Lys Ser Glu Glu Glu Ala Leu
405 410 415
Lys Leu Ala Asn Asp Ser Asp Tyr Gly Leu Gly Ala Ala Val Trp Thr
420 425 430
Arg Asp Leu Ser Arg Ala His Arg Met Ser Arg Arg Leu Lys Ala Gly
435 440 445
Ser Val Phe Val Asn Asn Tyr Asn Asp Gly Asp Met Thr Val Pro Phe
450 455 460
Gly Gly Tyr Lys Gln Ser Gly Asn Gly Arg Asp Lys Ser Leu His Ala
465 470 475 480
Leu Glu Lys Phe Thr Glu Leu Lys Thr Ile Trp Ile Ala Leu Glu Ser
485 490 495
<210> 48
<211> 519
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 48
Met Phe Ser Arg Ser Thr Leu Cys Leu Lys Thr Ser Ala Ser Ser Ile
1 5 10 15
Gly Arg Leu Gln Leu Arg Tyr Phe Ser His Leu Pro Met Thr Val Pro
20 25 30
Ile Lys Leu Pro Asn Gly Leu Glu Tyr Glu Gln Pro Thr Gly Leu Phe
35 40 45
Ile Asn Asn Lys Phe Val Pro Ser Lys Gln Asn Lys Thr Phe Glu Val
50 55 60
Ile Asn Pro Ser Thr Glu Glu Glu Ile Cys His Ile Tyr Glu Gly Arg
65 70 75 80
Glu Asp Asp Val Glu Glu Ala Val Gln Ala Ala Asp Arg Ala Phe Ser
85 90 95
Asn Gly Ser Trp Asn Gly Ile Asp Pro Ile Asp Arg Gly Lys Ala Leu
100 105 110
Tyr Arg Leu Ala Glu Leu Ile Glu Gln Asp Lys Asp Val Ile Ala Ser
115 120 125
Ile Glu Thr Leu Asp Asn Gly Lys Ala Ile Ser Ser Ser Arg Gly Asp
130 135 140
Val Asp Leu Val Ile Asn Tyr Leu Lys Ser Ser Ala Gly Phe Ala Asp
145 150 155 160
Lys Ile Asp Gly Arg Met Ile Asp Thr Gly Arg Thr His Phe Ser Tyr
165 170 175
Thr Lys Arg Gln Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn
180 185 190
Phe Pro Leu Leu Met Trp Ala Trp Lys Ile Ala Pro Ala Leu Val Thr
195 200 205
Gly Asn Thr Val Val Leu Lys Thr Ala Glu Ser Thr Pro Leu Ser Ala
210 215 220
Leu Tyr Val Ser Lys Tyr Ile Pro Gln Ala Gly Ile Pro Pro Gly Val
225 230 235 240
Ile Asn Ile Val Ser Gly Phe Gly Lys Ile Val Gly Glu Ala Ile Thr
245 250 255
Asn His Pro Lys Ile Lys Lys Val Ala Phe Thr Gly Ser Thr Ala Thr
260 265 270
Gly Arg His Ile Tyr Gln Ser Ala Ala Ala Gly Leu Lys Lys Val Thr
275 280 285
Leu Glu Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Glu
290 295 300
Leu Lys Lys Ala Val Gln Asn Ile Ile Leu Gly Ile Tyr Tyr Asn Ser
305 310 315 320
Gly Glu Val Cys Cys Ala Gly Ser Arg Val Tyr Val Glu Glu Ser Ile
325 330 335
Tyr Asp Lys Phe Ile Glu Glu Phe Lys Ala Ala Ser Glu Ser Ile Lys
340 345 350
Val Gly Asp Pro Phe Asp Glu Ser Thr Phe Gln Gly Ala Gln Thr Ser
355 360 365
Gln Met Gln Leu Asn Lys Ile Leu Lys Tyr Val Asp Ile Gly Lys Asn
370 375 380
Glu Gly Ala Thr Leu Ile Thr Gly Gly Glu Arg Leu Gly Ser Lys Gly
385 390 395 400
Tyr Phe Ile Lys Pro Thr Val Phe Gly Asp Val Lys Glu Asp Met Arg
405 410 415
Ile Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Thr Lys Phe
420 425 430
Lys Ser Ala Asp Glu Val Ile Asn Met Ala Asn Asp Ser Glu Tyr Gly
435 440 445
Leu Ala Ala Gly Ile His Thr Ser Asn Ile Asn Thr Ala Leu Lys Val
450 455 460
Ala Asp Arg Val Asn Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp
465 470 475 480
Phe His His Ala Val Pro Phe Gly Gly Phe Asn Ala Ser Gly Leu Gly
485 490 495
Arg Glu Met Ser Val Asp Ala Leu Gln Asn Tyr Leu Gln Val Lys Ala
500 505 510
Val Arg Ala Lys Leu Asp Glu
515
<210> 49
<211> 520
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 49
Met Leu Ser Arg Thr Arg Ala Ala Ala Pro Asn Ser Arg Ile Phe Thr
1 5 10 15
Arg Ser Leu Leu Arg Leu Tyr Ser Gln Ala Pro Leu Arg Val Pro Ile
20 25 30
Thr Leu Pro Asn Gly Phe Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile
35 40 45
Asn Gly Glu Phe Val Ala Ser Lys Gln Lys Lys Thr Phe Asp Val Ile
50 55 60
Asn Pro Ser Asn Glu Glu Lys Ile Thr Thr Val Tyr Lys Ala Met Glu
65 70 75 80
Asp Asp Val Asp Glu Ala Val Ala Ala Ala Lys Lys Ala Phe Glu Thr
85 90 95
Lys Trp Ser Ile Val Glu Pro Glu Val Arg Ala Lys Ala Leu Phe Asn
100 105 110
Leu Ala Asp Leu Val Glu Lys His Gln Glu Thr Leu Ala Ala Ile Glu
115 120 125
Ser Met Asp Asn Gly Lys Ser Leu Phe Cys Ala Arg Gly Asp Val Ala
130 135 140
Leu Val Ser Lys Tyr Leu Arg Ser Cys Gly Gly Trp Ala Asp Lys Ile
145 150 155 160
Tyr Gly Asn Val Ile Asp Thr Gly Lys Asn His Phe Thr Tyr Ser Ile
165 170 175
Lys Glu Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro
180 185 190
Leu Leu Met Trp Ser Trp Lys Ile Gly Pro Ala Leu Ala Thr Gly Asn
195 200 205
Thr Val Val Leu Lys Pro Ala Glu Thr Thr Pro Leu Ser Ala Leu Phe
210 215 220
Ala Ser Gln Leu Cys Gln Glu Ala Gly Ile Pro Ala Gly Val Val Asn
225 230 235 240
Ile Leu Pro Gly Ser Gly Arg Val Val Gly Glu Arg Leu Ser Ala His
245 250 255
Pro Asp Val Lys Lys Ile Ala Phe Thr Gly Ser Thr Ala Thr Gly Arg
260 265 270
His Ile Met Lys Val Ala Ala Asp Thr Val Lys Lys Val Thr Leu Glu
275 280 285
Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Asp Leu Asp
290 295 300
Lys Ala Val Lys Asn Ile Ala Phe Gly Ile Phe Tyr Asn Ser Gly Glu
305 310 315 320
Val Cys Cys Ala Gly Ser Arg Ile Tyr Ile Gln Asp Thr Val Tyr Glu
325 330 335
Glu Val Leu Gln Lys Leu Lys Asp Tyr Thr Glu Ser Leu Lys Val Gly
340 345 350
Asp Pro Phe Asp Glu Glu Val Phe Gln Gly Ala Gln Thr Ser Asp Lys
355 360 365
Gln Leu His Lys Ile Leu Asp Tyr Val Asp Val Ala Lys Ser Glu Gly
370 375 380
Ala Arg Leu Val Thr Gly Gly Ala Arg His Gly Ser Lys Gly Tyr Phe
385 390 395 400
Val Lys Pro Thr Val Phe Ala Asp Val Lys Glu Asp Met Arg Ile Val
405 410 415
Lys Glu Glu Val Phe Gly Pro Ile Val Thr Val Ser Lys Phe Ser Thr
420 425 430
Val Asp Glu Val Ile Ala Met Ala Asn Asp Ser Gln Tyr Gly Leu Ala
435 440 445
Ala Gly Ile His Thr Asn Asp Ile Asn Lys Ala Val Asp Val Ser Lys
450 455 460
Arg Val Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asn Phe His
465 470 475 480
Gln Asn Val Pro Phe Gly Gly Phe Gly Gln Ser Gly Ile Gly Arg Glu
485 490 495
Met Gly Glu Ala Ala Leu Ser Asn Tyr Thr Gln Thr Lys Ser Val Arg
500 505 510
Ile Ala Ile Asp Lys Pro Ile Arg
515 520
<210> 50
<211> 462
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 50
Met Thr Ile Thr Pro Ala Thr His Ala Ile Ser Ile Asn Pro Ala Thr
1 5 10 15
Gly Glu Gln Leu Ser Val Leu Pro Trp Ala Gly Ala Asp Asp Ile Glu
20 25 30
Asn Ala Leu Gln Leu Ala Ala Ala Gly Phe Arg Asp Trp Arg Glu Thr
35 40 45
Asn Ile Asp Tyr Arg Ala Glu Lys Leu Arg Asp Ile Gly Lys Ala Leu
50 55 60
Arg Ala Arg Ser Glu Glu Met Ala Gln Met Ile Thr Arg Glu Met Gly
65 70 75 80
Lys Pro Ile Asn Gln Ala Arg Ala Glu Val Ala Lys Ser Ala Asn Leu
85 90 95
Cys Asp Trp Tyr Ala Glu His Gly Pro Ala Met Leu Lys Ala Glu Pro
100 105 110
Thr Leu Val Glu Asn Gln Gln Ala Val Ile Glu Tyr Arg Pro Leu Gly
115 120 125
Thr Ile Leu Ala Ile Met Pro Trp Asn Phe Pro Leu Trp Gln Val Met
130 135 140
Arg Gly Ala Val Pro Ile Ile Leu Ala Gly Asn Gly Tyr Leu Leu Lys
145 150 155 160
His Ala Pro Asn Val Met Gly Cys Ala Gln Leu Ile Ala Gln Val Phe
165 170 175
Lys Asp Ala Gly Ile Pro Gln Gly Val Tyr Gly Trp Leu Asn Ala Asp
180 185 190
Asn Asp Gly Val Ser Gln Met Ile Lys Asp Ser Arg Ile Ala Ala Val
195 200 205
Thr Val Thr Gly Ser Val Arg Ala Gly Ala Ala Ile Gly Ala Gln Ala
210 215 220
Gly Ala Ala Leu Lys Lys Cys Val Leu Glu Leu Gly Gly Ser Asp Pro
225 230 235 240
Phe Ile Val Leu Asn Asp Ala Asp Leu Glu Leu Ala Val Lys Ala Ala
245 250 255
Val Ala Gly Arg Tyr Gln Asn Thr Gly Gln Val Cys Ala Ala Ala Lys
260 265 270
Arg Phe Ile Ile Glu Glu Gly Ile Ala Ser Ala Phe Thr Glu Arg Phe
275 280 285
Val Ala Ala Ala Ala Ala Leu Lys Met Gly Asp Pro Arg Asp Glu Glu
290 295 300
Asn Ala Leu Gly Pro Met Ala Arg Phe Asp Leu Arg Asp Glu Leu His
305 310 315 320
His Gln Val Glu Lys Thr Leu Ala Gln Gly Ala Arg Leu Leu Leu Gly
325 330 335
Gly Glu Lys Met Ala Gly Ala Gly Asn Tyr Tyr Pro Pro Thr Val Leu
340 345 350
Ala Asn Val Thr Pro Glu Met Thr Ala Phe Arg Glu Glu Met Phe Gly
355 360 365
Pro Val Ala Ala Ile Thr Ile Ala Lys Asp Ala Glu His Ala Leu Glu
370 375 380
Leu Ala Asn Asp Ser Glu Phe Gly Leu Ser Ala Thr Ile Phe Thr Thr
385 390 395 400
Asp Glu Thr Gln Ala Arg Gln Met Ala Ala Arg Leu Glu Cys Gly Gly
405 410 415
Val Phe Ile Asn Gly Tyr Cys Ala Ser Asp Ala Arg Val Ala Phe Gly
420 425 430
Gly Val Lys Lys Ser Gly Phe Gly Arg Glu Leu Ser His Phe Gly Leu
435 440 445
His Glu Phe Cys Asn Ile Gln Thr Val Trp Lys Asp Arg Ile
450 455 460
<210> 51
<211> 482
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 51
Met Lys Leu Asn Asp Ser Asn Leu Phe Arg Gln Gln Ala Leu Ile Asn
1 5 10 15
Gly Glu Trp Leu Asp Ala Asn Asn Gly Glu Ala Ile Asp Val Thr Asn
20 25 30
Pro Ala Asn Gly Asp Lys Leu Gly Ser Val Pro Lys Met Gly Ala Asp
35 40 45
Glu Thr Arg Ala Ala Ile Asp Ala Ala Asn Arg Ala Leu Pro Ala Trp
50 55 60
Arg Ala Leu Thr Ala Lys Glu Arg Ala Thr Ile Leu Arg Asn Trp Phe
65 70 75 80
Asn Leu Met Met Glu His Gln Asp Asp Leu Ala Arg Leu Met Thr Leu
85 90 95
Glu Gln Gly Lys Pro Leu Ala Glu Ala Lys Gly Glu Ile Ser Tyr Ala
100 105 110
Ala Ser Phe Ile Glu Trp Phe Ala Glu Glu Gly Lys Arg Ile Tyr Gly
115 120 125
Asp Thr Ile Pro Gly His Gln Ala Asp Lys Arg Leu Ile Val Ile Lys
130 135 140
Gln Pro Ile Gly Val Thr Ala Ala Ile Thr Pro Trp Asn Phe Pro Ala
145 150 155 160
Ala Met Ile Thr Arg Lys Ala Gly Pro Ala Leu Ala Ala Gly Cys Thr
165 170 175
Met Val Leu Lys Pro Ala Ser Gln Thr Pro Phe Ser Ala Leu Ala Leu
180 185 190
Ala Glu Leu Ala Ile Arg Ala Gly Val Pro Ala Gly Val Phe Asn Val
195 200 205
Val Thr Gly Ser Ala Gly Ala Val Gly Asn Glu Leu Thr Ser Asn Pro
210 215 220
Leu Val Arg Lys Leu Ser Phe Thr Gly Ser Thr Glu Ile Gly Arg Gln
225 230 235 240
Leu Met Glu Gln Cys Ala Lys Asp Ile Lys Lys Val Ser Leu Glu Leu
245 250 255
Gly Gly Asn Ala Pro Phe Ile Val Phe Asp Asp Ala Asp Leu Asp Lys
260 265 270
Ala Val Glu Gly Ala Leu Ala Ser Lys Phe Arg Asn Ala Gly Gln Thr
275 280 285
Cys Val Cys Ala Asn Arg Leu Tyr Val Gln Asp Gly Val Tyr Asp Arg
290 295 300
Phe Ala Glu Lys Leu Gln Gln Ala Val Ser Lys Leu His Ile Gly Asp
305 310 315 320
Gly Leu Asp Asn Gly Val Thr Ile Gly Pro Leu Ile Asp Glu Lys Ala
325 330 335
Val Ala Lys Val Glu Glu His Ile Ala Asp Ala Leu Glu Lys Gly Ala
340 345 350
Arg Val Val Cys Gly Gly Lys Ala His Glu Arg Gly Gly Asn Phe Phe
355 360 365
Gln Pro Thr Ile Leu Val Asp Val Pro Ala Asn Ala Lys Val Ser Lys
370 375 380
Glu Glu Thr Phe Gly Pro Leu Ala Pro Leu Phe Arg Phe Lys Asp Glu
385 390 395 400
Ala Asp Val Ile Ala Gln Ala Asn Asp Thr Glu Phe Gly Leu Ala Ala
405 410 415
Tyr Phe Tyr Ala Arg Asp Leu Ser Arg Val Phe Arg Val Gly Glu Ala
420 425 430
Leu Glu Tyr Gly Ile Val Gly Ile Asn Thr Gly Ile Ile Ser Asn Glu
435 440 445
Val Ala Pro Phe Gly Gly Ile Lys Ala Ser Gly Leu Gly Arg Glu Gly
450 455 460
Ser Lys Tyr Gly Ile Glu Asp Tyr Leu Glu Ile Lys Tyr Met Cys Ile
465 470 475 480
Gly Leu
<210> 52
<211> 474
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 52
Met Gln His Lys Leu Leu Ile Asn Gly Glu Leu Val Ser Gly Glu Gly
1 5 10 15
Glu Lys Gln Pro Val Tyr Asn Pro Ala Thr Gly Asp Val Leu Leu Glu
20 25 30
Ile Ala Glu Ala Ser Ala Glu Gln Val Asp Ala Ala Val Arg Ala Ala
35 40 45
Asp Ala Ala Phe Ala Glu Trp Gly Gln Thr Thr Pro Lys Val Arg Ala
50 55 60
Glu Cys Leu Leu Lys Leu Ala Asp Val Ile Glu Glu Asn Gly Gln Val
65 70 75 80
Phe Ala Glu Leu Glu Ser Arg Asn Cys Gly Lys Pro Leu His Ser Ala
85 90 95
Phe Asn Asp Glu Ile Pro Ala Ile Val Asp Val Phe Arg Phe Phe Ala
100 105 110
Gly Ala Ala Arg Cys Leu Asn Gly Leu Ala Ala Gly Glu Tyr Leu Glu
115 120 125
Gly His Thr Ser Met Ile Arg Arg Asp Pro Leu Gly Val Val Ala Ser
130 135 140
Ile Ala Pro Trp Asn Tyr Pro Leu Met Met Ala Ala Trp Lys Leu Ala
145 150 155 160
Pro Ala Leu Ala Ala Gly Asn Cys Val Val Leu Lys Pro Ser Glu Ile
165 170 175
Thr Pro Leu Thr Ala Leu Lys Leu Ala Glu Leu Ala Lys Asp Ile Phe
180 185 190
Pro Ala Gly Val Ile Asn Ile Leu Phe Gly Arg Gly Lys Thr Val Gly
195 200 205
Asp Pro Leu Thr Gly His Pro Lys Val Arg Met Val Ser Leu Thr Gly
210 215 220
Ser Ile Ala Thr Gly Glu His Ile Ile Ser His Thr Ala Ser Ser Ile
225 230 235 240
Lys Arg Thr His Met Glu Leu Gly Gly Lys Ala Pro Val Ile Val Phe
245 250 255
Asp Asp Ala Asp Ile Glu Ala Val Val Glu Gly Val Arg Thr Phe Gly
260 265 270
Tyr Tyr Asn Ala Gly Gln Asp Cys Thr Ala Ala Cys Arg Ile Tyr Ala
275 280 285
Gln Lys Gly Ile Tyr Asp Thr Leu Val Glu Lys Leu Gly Ala Ala Val
290 295 300
Ala Thr Leu Lys Ser Gly Ala Pro Asp Asp Glu Ser Thr Glu Leu Gly
305 310 315 320
Pro Leu Ser Ser Leu Ala His Leu Glu Arg Val Gly Lys Ala Val Glu
325 330 335
Glu Ala Lys Ala Thr Gly His Ile Lys Val Ile Thr Gly Gly Glu Lys
340 345 350
Arg Lys Gly Asn Gly Tyr Tyr Tyr Ala Pro Thr Leu Leu Ala Gly Ala
355 360 365
Leu Gln Asp Asp Ala Ile Val Gln Lys Glu Val Phe Gly Pro Val Val
370 375 380
Ser Val Thr Pro Phe Asp Asn Glu Glu Gln Val Val Asn Trp Ala Asn
385 390 395 400
Asp Ser Gln Tyr Gly Leu Ala Ser Ser Val Trp Thr Lys Asp Val Gly
405 410 415
Arg Ala His Arg Val Ser Ala Arg Leu Gln Tyr Gly Cys Thr Trp Val
420 425 430
Asn Thr His Phe Met Leu Val Ser Glu Met Pro His Gly Gly Gln Lys
435 440 445
Leu Ser Gly Tyr Gly Lys Asp Met Ser Leu Tyr Gly Leu Glu Asp Tyr
450 455 460
Thr Val Val Arg His Val Met Val Lys His
465 470
<210> 53
<211> 480
<212> PRT
<213> 恶臭假单胞菌(Pseudomonas putida)
<400> 53
Met Gln Leu Lys Asp Ala Gln Leu Phe Arg Gln Gln Ala Tyr Ile Asn
1 5 10 15
Gly Glu Trp Leu Asp Ala Asp Asn Gly Gln Thr Ile Lys Val Thr Asn
20 25 30
Pro Ala Thr Gly Glu Val Ile Gly Thr Val Pro Lys Met Gly Thr Ala
35 40 45
Glu Thr Arg Arg Ala Ile Glu Ala Ala Asp Lys Ala Leu Pro Ala Trp
50 55 60
Arg Ala Leu Thr Ala Lys Glu Arg Ser Ala Lys Leu Arg Arg Trp Phe
65 70 75 80
Glu Leu Met Ile Glu Asn Gln Asp Asp Leu Ala Arg Leu Met Thr Thr
85 90 95
Glu Gln Gly Lys Pro Leu Ala Glu Ala Lys Gly Glu Ile Ala Tyr Ala
100 105 110
Ala Ser Phe Ile Glu Trp Phe Ala Glu Glu Ala Lys Arg Ile Tyr Gly
115 120 125
Asp Thr Ile Pro Gly His Gln Pro Asp Lys Arg Leu Ile Val Ile Lys
130 135 140
Gln Pro Ile Gly Val Thr Ala Ala Ile Thr Pro Trp Asn Phe Pro Ala
145 150 155 160
Ala Met Ile Thr Arg Lys Ala Gly Pro Ala Leu Ala Ala Gly Cys Thr
165 170 175
Met Val Leu Lys Pro Ala Ser Gln Thr Pro Tyr Ser Ala Leu Ala Leu
180 185 190
Val Glu Leu Ala His Arg Ala Gly Ile Pro Ala Gly Val Leu Ser Val
195 200 205
Val Thr Gly Ser Ala Gly Glu Val Gly Gly Glu Leu Thr Gly Asn Ser
210 215 220
Leu Val Arg Lys Leu Ser Phe Thr Gly Ser Thr Glu Ile Gly Arg Gln
225 230 235 240
Leu Met Glu Glu Cys Ala Lys Asp Ile Lys Lys Val Ser Leu Glu Leu
245 250 255
Gly Gly Asn Ala Pro Phe Ile Val Phe Asp Asp Ala Asp Leu Asp Lys
260 265 270
Ala Val Glu Gly Ala Ile Ile Ser Lys Tyr Arg Asn Asn Gly Gln Thr
275 280 285
Cys Val Cys Ala Asn Arg Ile Tyr Val Gln Asp Gly Val Tyr Asp Ala
290 295 300
Phe Ala Glu Lys Leu Ala Ala Ala Val Ala Lys Leu Lys Ile Gly Asn
305 310 315 320
Gly Leu Glu Glu Gly Thr Thr Thr Gly Pro Leu Ile Asp Gly Lys Ala
325 330 335
Val Ala Lys Val Gln Glu His Ile Glu Asp Ala Val Ser Lys Gly Ala
340 345 350
Lys Val Leu Ser Gly Gly Lys Leu Ile Glu Gly Asn Phe Phe Glu Pro
355 360 365
Thr Ile Leu Val Asp Val Pro Lys Thr Ala Ala Val Ala Lys Glu Glu
370 375 380
Thr Phe Gly Pro Leu Ala Pro Leu Phe Arg Phe Lys Asp Glu Ala Glu
385 390 395 400
Val Ile Ala Met Ser Asn Asp Thr Glu Phe Gly Leu Ala Ser Tyr Phe
405 410 415
Tyr Ala Arg Asp Met Ser Arg Val Phe Arg Val Ala Glu Ala Leu Glu
420 425 430
Tyr Gly Met Val Gly Ile Asn Thr Gly Leu Ile Ser Asn Glu Val Ala
435 440 445
Pro Phe Gly Gly Ile Lys Ala Ser Gly Leu Gly Arg Glu Gly Ser Lys
450 455 460
Tyr Gly Ile Glu Asp Tyr Leu Glu Ile Lys Tyr Leu Cys Ile Ser Val
465 470 475 480
<210> 54
<211> 479
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 54
Met Ser Val Pro Val Gln His Pro Met Tyr Ile Asp Gly Gln Phe Val
1 5 10 15
Thr Trp Arg Gly Asp Ala Trp Ile Asp Val Val Asn Pro Ala Thr Glu
20 25 30
Ala Val Ile Ser Arg Ile Pro Asp Gly Gln Ala Glu Asp Ala Arg Lys
35 40 45
Ala Ile Asp Ala Ala Glu Arg Ala Gln Pro Glu Trp Glu Ala Leu Pro
50 55 60
Ala Ile Glu Arg Ala Ser Trp Leu Arg Lys Ile Ser Ala Gly Ile Arg
65 70 75 80
Glu Arg Ala Ser Glu Ile Ser Ala Leu Ile Val Glu Glu Gly Gly Lys
85 90 95
Ile Gln Gln Leu Ala Glu Val Glu Val Ala Phe Thr Ala Asp Tyr Ile
100 105 110
Asp Tyr Met Ala Glu Trp Ala Arg Arg Tyr Glu Gly Glu Ile Ile Gln
115 120 125
Ser Asp Arg Pro Gly Glu Asn Ile Leu Leu Phe Lys Arg Ala Leu Gly
130 135 140
Val Thr Thr Gly Ile Leu Pro Trp Asn Phe Pro Phe Phe Leu Ile Ala
145 150 155 160
Arg Lys Met Ala Pro Ala Leu Leu Thr Gly Asn Thr Ile Val Ile Lys
165 170 175
Pro Ser Glu Phe Thr Pro Asn Asn Ala Ile Ala Phe Ala Lys Ile Val
180 185 190
Asp Glu Ile Gly Leu Pro Arg Gly Val Phe Asn Leu Val Leu Gly Arg
195 200 205
Gly Glu Thr Val Gly Gln Glu Leu Ala Gly Asn Pro Lys Val Ala Met
210 215 220
Val Ser Met Thr Gly Ser Val Ser Ala Gly Glu Lys Ile Met Ala Thr
225 230 235 240
Ala Ala Lys Asn Ile Thr Lys Val Cys Leu Glu Leu Gly Gly Lys Ala
245 250 255
Pro Ala Ile Val Met Asp Asp Ala Asp Leu Glu Leu Ala Val Lys Ala
260 265 270
Ile Val Asp Ser Arg Val Ile Asn Ser Gly Gln Val Cys Asn Cys Ala
275 280 285
Glu Arg Val Tyr Val Gln Lys Gly Ile Tyr Asp Gln Phe Val Asn Arg
290 295 300
Leu Gly Glu Ala Met Gln Ala Val Gln Phe Gly Asn Pro Ala Glu Arg
305 310 315 320
Asn Asp Ile Ala Met Gly Pro Leu Ile Asn Ala Ala Ala Leu Glu Arg
325 330 335
Val Glu Gln Lys Val Ala Arg Ala Val Glu Glu Gly Ala Arg Val Ala
340 345 350
Phe Gly Gly Lys Ala Val Glu Gly Lys Gly Tyr Tyr Tyr Pro Pro Thr
355 360 365
Leu Leu Leu Asp Val Arg Gln Glu Met Ser Ile Met His Glu Glu Thr
370 375 380
Phe Gly Pro Val Leu Pro Val Val Ala Phe Asp Thr Leu Glu Asp Ala
385 390 395 400
Ile Ser Met Ala Asn Asp Ser Asp Tyr Gly Leu Thr Ser Ser Ile Tyr
405 410 415
Thr Gln Asn Leu Asn Val Ala Met Lys Ala Ile Lys Gly Leu Lys Phe
420 425 430
Gly Glu Thr Tyr Ile Asn Arg Glu Asn Phe Glu Ala Met Gln Gly Phe
435 440 445
His Ala Gly Trp Arg Lys Ser Gly Ile Gly Gly Ala Asp Gly Lys His
450 455 460
Gly Leu His Glu Tyr Leu Gln Thr Gln Val Val Tyr Leu Gln Ser
465 470 475
<210> 55
<211> 512
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 55
Met Thr Asn Asn Pro Pro Ser Ala Gln Ile Lys Pro Gly Glu Tyr Gly
1 5 10 15
Phe Pro Leu Lys Leu Lys Ala Arg Tyr Asp Asn Phe Ile Gly Gly Glu
20 25 30
Trp Val Ala Pro Ala Asp Gly Glu Tyr Tyr Gln Asn Leu Thr Pro Val
35 40 45
Thr Gly Gln Leu Leu Cys Glu Val Ala Ser Ser Gly Lys Arg Asp Ile
50 55 60
Asp Leu Ala Leu Asp Ala Ala His Lys Val Lys Asp Lys Trp Ala His
65 70 75 80
Thr Ser Val Gln Asp Arg Ala Ala Ile Leu Phe Lys Ile Ala Asp Arg
85 90 95
Met Glu Gln Asn Leu Glu Leu Leu Ala Thr Ala Glu Thr Trp Asp Asn
100 105 110
Gly Lys Pro Ile Arg Glu Thr Ser Ala Ala Asp Val Pro Leu Ala Ile
115 120 125
Asp His Phe Arg Tyr Phe Ala Ser Cys Ile Arg Ala Gln Glu Gly Gly
130 135 140
Ile Ser Glu Val Asp Ser Glu Thr Val Ala Tyr His Phe His Glu Pro
145 150 155 160
Leu Gly Val Val Gly Gln Ile Ile Pro Trp Asn Phe Pro Leu Leu Met
165 170 175
Ala Ser Trp Lys Met Ala Pro Ala Leu Ala Ala Gly Asn Cys Val Val
180 185 190
Leu Lys Pro Ala Arg Leu Thr Pro Leu Ser Val Leu Leu Leu Met Glu
195 200 205
Ile Val Gly Asp Leu Leu Pro Pro Gly Val Val Asn Val Val Asn Gly
210 215 220
Ala Gly Gly Val Ile Gly Glu Tyr Leu Ala Thr Ser Lys Arg Ile Ala
225 230 235 240
Lys Val Ala Phe Thr Gly Ser Thr Glu Val Gly Gln Gln Ile Met Gln
245 250 255
Tyr Ala Thr Gln Asn Ile Ile Pro Val Thr Leu Glu Leu Gly Gly Lys
260 265 270
Ser Pro Asn Ile Phe Phe Ala Asp Val Met Asp Glu Glu Asp Ala Phe
275 280 285
Phe Asp Lys Ala Leu Glu Gly Phe Ala Leu Phe Ala Phe Asn Gln Gly
290 295 300
Glu Val Cys Thr Cys Pro Ser Arg Ala Leu Val Gln Glu Ser Ile Tyr
305 310 315 320
Glu Arg Phe Met Glu Arg Ala Ile Arg Arg Val Glu Ser Ile Arg Ser
325 330 335
Gly Asn Pro Leu Asp Ser Val Thr Gln Met Gly Ala Gln Val Ser His
340 345 350
Gly Gln Leu Glu Thr Ile Leu Asn Tyr Ile Asp Ile Gly Lys Lys Glu
355 360 365
Gly Ala Asp Val Leu Thr Gly Gly Arg Arg Lys Leu Leu Glu Gly Glu
370 375 380
Leu Lys Asp Gly Tyr Tyr Leu Glu Pro Thr Ile Leu Phe Gly Gln Asn
385 390 395 400
Asn Met Arg Val Phe Gln Glu Glu Ile Phe Gly Pro Val Leu Ala Val
405 410 415
Thr Thr Phe Lys Thr Met Glu Glu Ala Leu Glu Leu Ala Asn Asp Thr
420 425 430
Gln Tyr Gly Leu Gly Ala Gly Val Trp Ser Arg Asn Gly Asn Leu Ala
435 440 445
Tyr Lys Met Gly Arg Gly Ile Gln Ala Gly Arg Val Trp Thr Asn Cys
450 455 460
Tyr His Ala Tyr Pro Ala His Ala Ala Phe Gly Gly Tyr Lys Gln Ser
465 470 475 480
Gly Ile Gly Arg Glu Thr His Lys Met Met Leu Glu His Tyr Gln Gln
485 490 495
Thr Lys Cys Leu Leu Val Ser Tyr Ser Asp Lys Pro Leu Gly Leu Phe
500 505 510
<210> 56
<211> 463
<212> PRT
<213> 詹氏甲烷球菌(Methanocaldococcus jannaschii)
<400> 56
Met Phe Ile Asp Gly Lys Trp Ile Asn Arg Glu Asp Met Asp Val Ile
1 5 10 15
Asn Pro Tyr Ser Leu Glu Val Ile Lys Lys Ile Pro Ala Leu Ser Arg
20 25 30
Glu Glu Ala Lys Glu Ala Ile Asp Thr Ala Glu Lys Tyr Lys Glu Val
35 40 45
Met Lys Asn Leu Pro Ile Thr Lys Arg Tyr Asn Ile Leu Met Asn Ile
50 55 60
Ala Lys Gln Ile Lys Glu Lys Lys Glu Glu Leu Ala Lys Ile Leu Ala
65 70 75 80
Ile Asp Ala Gly Lys Pro Ile Lys Gln Ala Arg Val Glu Val Glu Arg
85 90 95
Ser Ile Gly Thr Phe Lys Leu Ala Ala Phe Tyr Val Lys Glu His Arg
100 105 110
Asp Glu Val Ile Pro Ser Asp Asp Arg Leu Ile Phe Thr Arg Arg Glu
115 120 125
Pro Val Gly Ile Val Gly Ala Ile Thr Pro Phe Asn Phe Pro Leu Asn
130 135 140
Leu Ser Ala His Lys Ile Ala Pro Ala Ile Ala Thr Gly Asn Val Ile
145 150 155 160
Val His His Pro Ser Ser Lys Ala Pro Leu Val Cys Ile Glu Leu Ala
165 170 175
Lys Ile Ile Glu Asn Ala Leu Lys Lys Tyr Asn Val Pro Leu Gly Val
180 185 190
Tyr Asn Leu Leu Thr Gly Ala Gly Glu Val Val Gly Asp Glu Ile Val
195 200 205
Val Asn Glu Lys Val Asn Met Ile Ser Phe Thr Gly Ser Ser Lys Val
210 215 220
Gly Glu Leu Ile Thr Lys Lys Ala Gly Phe Lys Lys Ile Ala Leu Glu
225 230 235 240
Leu Gly Gly Val Asn Pro Asn Ile Val Leu Lys Asp Ala Asp Leu Asn
245 250 255
Lys Ala Val Asn Ala Leu Ile Lys Gly Ser Phe Ile Tyr Ala Gly Gln
260 265 270
Val Cys Ile Ser Val Gly Met Ile Leu Val Asp Glu Ser Ile Ala Asp
275 280 285
Lys Phe Ile Glu Met Phe Val Asn Lys Ala Lys Val Leu Asn Val Gly
290 295 300
Asn Pro Leu Asp Glu Lys Thr Asp Val Gly Pro Leu Ile Ser Val Glu
305 310 315 320
His Ala Glu Trp Val Glu Lys Val Val Glu Lys Ala Ile Asp Glu Gly
325 330 335
Gly Lys Leu Leu Leu Gly Gly Lys Arg Asp Lys Ala Leu Phe Tyr Pro
340 345 350
Thr Ile Leu Glu Val Asp Arg Asp Asn Ile Leu Cys Lys Thr Glu Thr
355 360 365
Phe Ala Pro Val Ile Pro Ile Ile Arg Thr Asn Glu Glu Glu Met Ile
370 375 380
Asp Ile Ala Asn Ser Thr Glu Tyr Gly Leu His Ser Ala Ile Phe Thr
385 390 395 400
Asn Asp Ile Asn Lys Ser Leu Lys Phe Ala Glu Asn Leu Glu Phe Gly
405 410 415
Gly Val Val Ile Asn Asp Ser Ser Leu Phe Arg Gln Asp Asn Met Pro
420 425 430
Phe Gly Gly Val Lys Lys Ser Gly Leu Gly Arg Glu Gly Val Lys Tyr
435 440 445
Ala Met Glu Glu Met Ser Asn Ile Lys Thr Ile Ile Ile Ser Lys
450 455 460
<210> 57
<211> 488
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 57
Met Ser Val Ala Ala Glu Ser Lys Thr Tyr Phe Asn Phe Ile Asn Gly
1 5 10 15
Arg Trp Val Lys Ala Glu Ser Gly Gly Met Glu Gln Ser Leu Asn Pro
20 25 30
Ala Asp Thr Arg Asp Ile Val Gly Leu Val Gln Lys Ser Ser Ile Glu
35 40 45
Asp Val Asp Arg Ala Val Glu Ala Ala Lys Gln Ala Lys Lys Ala Trp
50 55 60
Arg Lys Leu Ser Gly Ala Glu Arg Gly Gln Phe Leu Tyr Lys Ala Ala
65 70 75 80
Asp Ile Met Glu Gln Arg Leu Asp Glu Ile Ala Glu Cys Ala Thr Arg
85 90 95
Glu Met Gly Lys Thr Leu Pro Glu Ala Lys Gly Glu Thr Ala Arg Gly
100 105 110
Ile Ala Ile Leu Arg Tyr Tyr Ala Gly Glu Gly Leu Arg Lys Thr Gly
115 120 125
Asp Val Ile Pro Ser Thr Asp Ser Ser Ala Phe Met Tyr Thr Asp Arg
130 135 140
Val Pro Leu Gly Val Val Gly Val Ile Ser Pro Trp Asn Phe Pro Val
145 150 155 160
Ala Ile Pro Ile Trp Lys Met Ala Pro Ala Leu Ile Tyr Gly Asn Thr
165 170 175
Val Val Ile Lys Pro Ala Thr Glu Thr Ala Val Thr Cys Leu Lys Val
180 185 190
Ile Ser Cys Phe Glu Glu Ala Gly Ile Pro Ser Gly Val Val Asn Ala
195 200 205
Val Thr Gly Pro Gly Ser Ser Ala Gly Gln Arg Leu Ala Glu His Pro
210 215 220
Asp Val Asn Gly Ile Thr Phe Thr Gly Ser Asn Gln Thr Gly Lys Ile
225 230 235 240
Ile Gly Arg Thr Ala Phe Glu Arg Gly Ala Lys Tyr Gln Leu Glu Met
245 250 255
Gly Gly Lys Asn Pro Val Ile Val Ala Asp Asp Ala Asp Leu Asp Ile
260 265 270
Ala Val Glu Ala Val Ile Ser Gly Ala Phe Arg Ser Thr Gly Gln Lys
275 280 285
Cys Thr Ala Thr Ser Arg Val Ile Val Leu Asn Gly Val Tyr Asp Arg
290 295 300
Phe Lys Glu Lys Leu Leu Gln Gln Thr Lys Glu Ile Thr Ile Gly Asp
305 310 315 320
Ser Leu Lys Glu Asp Val Trp Met Gly Pro Ile Ala Asn Lys Gln Gln
325 330 335
Leu Asp Asn Cys Leu Ser Tyr Ile Ala Lys Gly Lys Gln Glu Gly Ala
340 345 350
Asp Leu Ile Phe Gly Gly Glu Arg Leu Ala Asp Gly Lys Tyr Glu Asn
355 360 365
Gly Tyr Tyr Ile Arg Pro Ala Ile Phe Asp Asn Val Thr Ser Gly Met
370 375 380
Thr Ile Ala Gln Glu Glu Ile Phe Gly Pro Val Ile Ala Leu Ile Lys
385 390 395 400
Ala Asp Thr Leu Glu Glu Ala Leu Glu Thr Ala Asn Asp Val Lys Phe
405 410 415
Gly Leu Ser Ala Ser Ile Phe Thr Gln Asn Ile Arg Arg Met Leu Ser
420 425 430
Phe Thr Asp Glu Ile Glu Ala Gly Leu Ile Arg Val Asn Ala Glu Ser
435 440 445
Ala Gly Val Glu Leu Gln Ala Pro Phe Gly Gly Val Lys Gln Ser Ser
450 455 460
Ser His Ser Arg Glu Gln Gly Glu Ala Ala Lys Glu Phe Phe Thr Ala
465 470 475 480
Val Lys Thr Val Phe Val Lys Pro
485
<210> 58
<211> 491
<212> PRT
<213> 假单胞菌属菌种(Pseudomonas sp.)
<400> 58
Met Lys Gln Tyr Arg Asn Phe Val Asp Gly Lys Trp Val Glu Ser Ser
1 5 10 15
Lys Thr Phe Gln Asp Val Thr Pro Ile Asp Gly Ser Val Val Ala Val
20 25 30
Val His Glu Ala Asp Arg Asp Leu Val Asp Ala Ala Val Lys Ala Gly
35 40 45
His Arg Ala Leu Glu Gly Glu Trp Gly Arg Thr Thr Ala Ala Gln Arg
50 55 60
Val Asp Trp Leu Arg Arg Ile Ala Asn Glu Met Glu Arg Arg Gln Gln
65 70 75 80
Asp Phe Leu Asp Ala Glu Met Ala Asp Thr Gly Lys Pro Leu Ser Met
85 90 95
Ala Ala Thr Ile Asp Ile Pro Arg Gly Ile Ala Asn Phe Arg Asn Phe
100 105 110
Ala Asp Ile Leu Ala Thr Ala Pro Val Asp Ser His Arg Leu Asp Leu
115 120 125
Pro Asp Gly Ala Tyr Ala Leu Asn Tyr Ala Ala Arg Lys Pro Leu Gly
130 135 140
Val Val Gly Val Ile Ser Pro Trp Asn Leu Pro Leu Leu Leu Leu Thr
145 150 155 160
Trp Lys Val Ala Pro Ala Leu Ala Cys Gly Asn Ala Val Val Val Lys
165 170 175
Pro Ser Glu Asp Thr Pro Gly Thr Ala Thr Leu Leu Ala Glu Val Met
180 185 190
Glu Ala Val Gly Ile Pro Pro Gly Val Phe Asn Leu Val His Gly Phe
195 200 205
Gly Pro Asn Ser Ala Gly Glu Phe Ile Ser Gln His Pro Asp Ile Ser
210 215 220
Ala Ile Thr Phe Thr Gly Glu Ser Lys Thr Gly Ser Thr Ile Met Arg
225 230 235 240
Ala Ala Ala Glu Gly Val Lys Pro Val Ser Phe Glu Leu Gly Gly Lys
245 250 255
Asn Ala Ala Val Ile Phe Ala Asp Cys Asp Phe Glu Lys Met Leu Asp
260 265 270
Gly Met Met Arg Ala Leu Phe Leu Asn Ser Gly Gln Val Cys Leu Cys
275 280 285
Ser Glu Arg Val Tyr Val Glu Arg Pro Ile Phe Asp Arg Phe Cys Val
290 295 300
Ala Leu Ala Glu Arg Ile Lys Ala Leu Lys Val Asp Trp Pro His Glu
305 310 315 320
Thr Asp Thr Gln Met Gly Pro Leu Ile Ser Ser Lys His Arg Asp Lys
325 330 335
Val Leu Ser Tyr Phe Glu Leu Ala Arg Gln Glu Gly Ala Thr Phe Leu
340 345 350
Ala Gly Gly Gly Val Pro Arg Phe Gly Asp Glu Arg Asp Asn Gly Ala
355 360 365
Trp Val Glu Pro Thr Val Ile Ala Gly Leu Ser Asp Asp Ala Arg Val
370 375 380
Val Arg Glu Glu Ile Phe Gly Pro Ile Cys His Val Thr Pro Phe Asp
385 390 395 400
Ser Glu Ser Glu Val Ile Arg Arg Ala Asn Asp Thr Arg Tyr Gly Leu
405 410 415
Ala Ala Thr Ile Trp Thr Thr Asn Leu Ser Arg Ala His Arg Val Ser
420 425 430
Glu Leu Met Arg Val Gly Ile Ser Trp Val Asn Thr Trp Phe Leu Arg
435 440 445
Asp Leu Arg Thr Pro Phe Gly Gly Ala Gly Leu Ser Gly Ile Gly Arg
450 455 460
Glu Gly Gly Met His Ser Leu Asn Phe Tyr Ser Glu Leu Thr Asn Val
465 470 475 480
Cys Val Arg Ile Asp Lys Glu Ser Pro Asp Val
485 490
<210> 59
<211> 499
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 59
Met Thr Glu Pro His Val Ala Val Leu Ser Gln Val Gln Gln Phe Leu
1 5 10 15
Asp Arg Gln His Gly Leu Tyr Ile Asp Gly Arg Pro Gly Pro Ala Gln
20 25 30
Ser Glu Lys Arg Leu Ala Ile Phe Asp Pro Ala Thr Gly Gln Glu Ile
35 40 45
Ala Ser Thr Ala Asp Ala Asn Glu Ala Asp Val Asp Asn Ala Val Met
50 55 60
Ser Ala Trp Arg Ala Phe Val Ser Arg Arg Trp Ala Gly Arg Leu Pro
65 70 75 80
Ala Glu Arg Glu Arg Ile Leu Leu Arg Phe Ala Asp Leu Val Glu Gln
85 90 95
His Ser Glu Glu Leu Ala Gln Leu Glu Thr Leu Glu Gln Gly Lys Ser
100 105 110
Ile Ala Ile Ser Arg Ala Phe Glu Val Gly Cys Thr Leu Asn Trp Met
115 120 125
Arg Tyr Thr Ala Gly Leu Thr Thr Lys Ile Ala Gly Lys Thr Leu Asp
130 135 140
Leu Ser Ile Pro Leu Pro Gln Gly Ala Arg Tyr Gln Ala Trp Thr Arg
145 150 155 160
Lys Glu Pro Val Gly Val Val Ala Gly Ile Val Pro Trp Asn Phe Pro
165 170 175
Leu Met Ile Gly Met Trp Lys Val Met Pro Ala Leu Ala Ala Gly Cys
180 185 190
Ser Ile Val Ile Lys Pro Ser Glu Thr Thr Pro Leu Thr Met Leu Arg
195 200 205
Val Ala Glu Leu Ala Ser Glu Ala Gly Ile Pro Asp Gly Val Phe Asn
210 215 220
Val Val Thr Gly Ser Gly Ala Val Cys Gly Ala Ala Leu Thr Ser His
225 230 235 240
Pro His Val Ala Lys Ile Ser Phe Thr Gly Ser Thr Ala Thr Gly Lys
245 250 255
Gly Ile Ala Arg Thr Ala Ala Asp His Leu Thr Arg Val Thr Leu Glu
260 265 270
Leu Gly Gly Lys Asn Pro Ala Ile Val Leu Lys Asp Ala Asp Pro Gln
275 280 285
Trp Val Ile Glu Gly Leu Met Thr Gly Ser Phe Leu Asn Gln Gly Gln
290 295 300
Val Cys Ala Ala Ser Ser Arg Ile Tyr Ile Glu Ala Pro Leu Phe Asp
305 310 315 320
Thr Leu Val Ser Gly Phe Glu Gln Ala Val Lys Ser Leu Gln Val Gly
325 330 335
Pro Gly Met Ser Pro Val Ala Gln Ile Asn Pro Leu Val Ser Arg Ala
340 345 350
His Cys Asp Lys Val Cys Ser Phe Leu Asp Asp Ala Gln Ala Gln Gln
355 360 365
Ala Glu Leu Ile Arg Gly Ser Asn Gly Pro Ala Gly Glu Gly Tyr Tyr
370 375 380
Val Ala Pro Thr Leu Val Val Asn Pro Asp Ala Lys Leu Arg Leu Thr
385 390 395 400
Arg Glu Glu Val Phe Gly Pro Val Val Asn Leu Val Arg Val Ala Asp
405 410 415
Gly Glu Glu Ala Leu Gln Leu Ala Asn Asp Thr Glu Tyr Gly Leu Thr
420 425 430
Ala Ser Val Trp Thr Gln Asn Leu Ser Gln Ala Leu Glu Tyr Ser Asp
435 440 445
Arg Leu Gln Ala Gly Thr Val Trp Val Asn Ser His Thr Leu Ile Asp
450 455 460
Ala Asn Leu Pro Phe Gly Gly Met Lys Gln Ser Gly Thr Gly Arg Asp
465 470 475 480
Phe Gly Pro Asp Trp Leu Asp Gly Trp Cys Glu Thr Lys Ser Val Cys
485 490 495
Val Arg Tyr
<210> 60
<211> 492
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 60
Met Thr Leu Trp Ile Asn Gly Asp Trp Ile Thr Gly Gln Gly Ala Ser
1 5 10 15
Arg Val Lys Arg Asn Pro Val Ser Gly Glu Val Leu Trp Gln Gly Asn
20 25 30
Asp Ala Asp Ala Ala Gln Val Glu Gln Ala Cys Arg Ala Ala Arg Ala
35 40 45
Ala Phe Pro Arg Trp Ala Arg Leu Ser Phe Ala Glu Arg His Ala Val
50 55 60
Val Glu Arg Phe Ala Ala Leu Leu Glu Ser Asn Lys Ala Glu Leu Thr
65 70 75 80
Ala Ile Ile Ala Arg Glu Thr Gly Lys Pro Arg Trp Glu Ala Ala Thr
85 90 95
Glu Val Thr Ala Met Ile Asn Lys Ile Ala Ile Ser Ile Lys Ala Tyr
100 105 110
His Val Arg Thr Gly Glu Gln Arg Ser Glu Met Pro Asp Gly Ala Ala
115 120 125
Ser Leu Arg His Arg Pro His Gly Val Leu Ala Val Phe Gly Pro Tyr
130 135 140
Asn Phe Pro Gly His Leu Pro Asn Gly His Ile Val Pro Ala Leu Leu
145 150 155 160
Ala Gly Asn Thr Ile Ile Phe Lys Pro Ser Glu Leu Thr Pro Trp Ser
165 170 175
Gly Glu Ala Val Met Arg Leu Trp Gln Gln Ala Gly Leu Pro Pro Gly
180 185 190
Val Leu Asn Leu Val Gln Gly Gly Arg Glu Thr Gly Gln Ala Leu Ser
195 200 205
Ala Leu Glu Asp Leu Asp Gly Leu Leu Phe Thr Gly Ser Ala Asn Thr
210 215 220
Gly Tyr Gln Leu His Arg Gln Leu Ser Gly Gln Pro Glu Lys Ile Leu
225 230 235 240
Ala Leu Glu Met Gly Gly Asn Asn Pro Leu Ile Ile Asp Glu Val Ala
245 250 255
Asp Ile Asp Ala Ala Val His Leu Thr Ile Gln Ser Ala Phe Val Thr
260 265 270
Ala Gly Gln Arg Cys Thr Cys Ala Arg Arg Leu Leu Leu Lys Ser Gly
275 280 285
Ala Gln Gly Asp Ala Phe Leu Ala Arg Leu Val Ala Val Ser Gln Arg
290 295 300
Leu Thr Pro Gly Asn Trp Asp Asp Glu Pro Gln Pro Phe Ile Gly Gly
305 310 315 320
Leu Ile Ser Glu Gln Ala Ala Gln Gln Val Val Thr Ala Trp Gln Gln
325 330 335
Leu Glu Ala Met Gly Gly Arg Pro Leu Leu Ala Pro Arg Leu Leu Gln
340 345 350
Ala Gly Thr Ser Leu Leu Thr Pro Gly Ile Ile Glu Met Thr Gly Val
355 360 365
Ala Gly Val Pro Asp Glu Glu Val Phe Gly Pro Leu Leu Arg Val Trp
370 375 380
Arg Tyr Asp Thr Phe Asp Glu Ala Ile Arg Met Ala Asn Asn Thr Arg
385 390 395 400
Phe Gly Leu Ser Cys Gly Leu Val Ser Pro Glu Arg Glu Lys Phe Asp
405 410 415
Gln Leu Leu Leu Glu Ala Arg Ala Gly Ile Val Asn Trp Asn Lys Pro
420 425 430
Leu Thr Gly Ala Ala Ser Thr Ala Pro Phe Gly Gly Ile Gly Ala Ser
435 440 445
Gly Asn His Arg Pro Ser Ala Trp Tyr Ala Ala Asp Tyr Cys Ala Trp
450 455 460
Pro Met Ala Ser Leu Glu Ser Asp Ser Leu Thr Leu Pro Ala Thr Leu
465 470 475 480
Asn Pro Gly Leu Asp Phe Ser Asp Glu Val Val Arg
485 490
<210> 61
<211> 1358
<212> PRT
<213> 玉米(Zea mays) (玉蜀黍(Maize))
<400> 61
Met Gly Lys Glu Ala Gly Ala Ala Glu Ser Ser Thr Val Val Leu Ala
1 5 10 15
Val Asn Gly Lys Arg Tyr Glu Ala Ala Gly Val Ala Pro Ser Thr Ser
20 25 30
Leu Leu Glu Phe Leu Arg Thr Gln Thr Pro Val Arg Gly Pro Lys Leu
35 40 45
Gly Cys Gly Glu Gly Gly Cys Gly Ala Cys Val Val Leu Val Ser Lys
50 55 60
Tyr Asp Pro Ala Thr Asp Glu Val Thr Glu Phe Ser Ala Ser Ser Cys
65 70 75 80
Leu Thr Leu Leu His Ser Val Asp Arg Cys Ser Val Thr Thr Ser Glu
85 90 95
Gly Ile Gly Asn Thr Arg Asp Gly Tyr His Pro Val Gln Gln Arg Leu
100 105 110
Ser Gly Phe His Ala Ser Gln Cys Gly Phe Cys Thr Pro Gly Met Cys
115 120 125
Met Ser Ile Phe Ser Ala Leu Val Lys Ala Asp Asn Lys Ser Asp Arg
130 135 140
Pro Asp Pro Pro Ala Gly Phe Ser Lys Ile Thr Thr Ser Glu Ala Glu
145 150 155 160
Lys Ala Val Ser Gly Asn Leu Cys Arg Cys Thr Gly Tyr Arg Pro Ile
165 170 175
Val Asp Thr Cys Lys Ser Phe Ala Ser Asp Val Asp Leu Glu Asp Leu
180 185 190
Gly Leu Asn Cys Phe Trp Lys Lys Gly Glu Glu Pro Ala Glu Val Ser
195 200 205
Arg Leu Pro Gly Tyr Asn Ser Gly Ala Val Cys Thr Phe Pro Glu Phe
210 215 220
Leu Lys Ser Glu Ile Lys Ser Thr Met Lys Gln Val Asn Asp Val Pro
225 230 235 240
Ile Ala Ala Ser Gly Asp Gly Trp Tyr His Pro Lys Ser Ile Glu Glu
245 250 255
Leu His Arg Leu Phe Asp Ser Ser Trp Phe Asp Asp Ser Ser Val Lys
260 265 270
Ile Val Ala Ser Asn Thr Gly Ser Gly Val Tyr Lys Asp Gln Asp Leu
275 280 285
Tyr Asp Lys Tyr Ile Asp Ile Lys Gly Ile Pro Glu Leu Ser Val Ile
290 295 300
Asn Lys Asn Asp Lys Ala Ile Glu Leu Gly Ser Val Val Ser Ile Ser
305 310 315 320
Lys Ala Ile Glu Val Leu Ser Asp Gly Asn Leu Val Phe Arg Lys Ile
325 330 335
Ala Asp His Leu Asn Lys Val Ala Ser Pro Phe Val Arg Asn Thr Ala
340 345 350
Thr Ile Gly Gly Asn Ile Met Met Ala Gln Arg Leu Pro Phe Glu Ser
355 360 365
Asp Val Ala Thr Val Leu Leu Ala Ala Gly Ser Thr Val Thr Val Gln
370 375 380
Val Ala Ser Lys Arg Leu Cys Phe Thr Leu Glu Glu Phe Leu Glu Gln
385 390 395 400
Pro Pro Cys Asp Ser Arg Thr Leu Leu Leu Ser Ile Phe Ile Pro Glu
405 410 415
Trp Gly Ser Asp Tyr Val Thr Phe Glu Thr Phe Arg Ala Ala Pro Arg
420 425 430
Pro Phe Gly Asn Ala Val Ser Tyr Val Asn Ser Ala Phe Leu Ala Arg
435 440 445
Thr Ser Gly Ser Leu Leu Ile Glu Asp Ile Cys Leu Ala Phe Gly Ala
450 455 460
Tyr Gly Val Asp His Ala Ile Arg Ala Lys Lys Val Glu Asp Phe Leu
465 470 475 480
Lys Gly Lys Ser Leu Ser Ser Phe Val Ile Leu Glu Ala Ile Lys Leu
485 490 495
Leu Lys Asp Thr Val Ser Pro Ser Glu Gly Thr Thr His His Glu Tyr
500 505 510
Arg Val Ser Leu Ala Val Ser Phe Leu Phe Ser Phe Leu Ser Ser Leu
515 520 525
Ala Asn Ser Ser Ser Ala Pro Ser Asn Ile Asp Thr Pro Asn Gly Ser
530 535 540
Tyr Thr His Glu Thr Gly Ser Asn Val Asp Ser Pro Glu Arg His Ile
545 550 555 560
Lys Val Asp Ser Asn Asp Leu Pro Ile Arg Ser Arg Gln Glu Met Val
565 570 575
Phe Ser Asp Glu Tyr Lys Pro Val Gly Lys Pro Ile Lys Lys Val Gly
580 585 590
Ala Glu Ile Gln Ala Ser Gly Glu Ala Val Tyr Val Asp Asp Ile Pro
595 600 605
Ala Pro Lys Asp Cys Leu Tyr Gly Ala Phe Ile Tyr Ser Thr His Pro
610 615 620
His Ala His Val Arg Ser Ile Asn Phe Lys Ser Ser Leu Ala Ser Gln
625 630 635 640
Lys Val Ile Thr Val Ile Thr Ala Lys Asp Ile Pro Ser Gly Gly Glu
645 650 655
Asn Ile Gly Ser Ser Phe Leu Met Gln Gly Glu Ala Leu Phe Ala Asp
660 665 670
Pro Ile Ala Glu Phe Ala Gly Gln Asn Ile Gly Val Val Ile Ala Glu
675 680 685
Thr Gln Arg Tyr Ala Asn Met Ala Ala Lys Gln Ala Val Val Glu Tyr
690 695 700
Ser Thr Glu Asn Leu Gln Pro Pro Ile Leu Thr Ile Glu Asp Ala Ile
705 710 715 720
Gln Arg Asn Ser Tyr Ile Gln Ile Pro Pro Phe Leu Ala Pro Lys Pro
725 730 735
Val Gly Asp Tyr Asn Lys Gly Met Ala Glu Ala Asp His Lys Ile Leu
740 745 750
Ser Ala Glu Val Lys Leu Glu Ser Gln Tyr Tyr Phe Tyr Met Glu Thr
755 760 765
Gln Ala Ala Leu Ala Ile Pro Asp Glu Asp Asn Cys Ile Thr Ile Tyr
770 775 780
Ser Ser Thr Gln Met Pro Glu Leu Thr Gln Asn Leu Ile Ala Arg Cys
785 790 795 800
Leu Gly Ile Pro Phe His Asn Val Arg Val Ile Ser Arg Arg Val Gly
805 810 815
Gly Gly Phe Gly Gly Lys Ala Met Lys Ala Thr His Thr Ala Cys Ala
820 825 830
Cys Ala Leu Ala Ala Phe Lys Leu Arg Arg Pro Val Arg Met Tyr Leu
835 840 845
Asp Arg Lys Thr Asp Met Ile Met Ala Gly Gly Arg His Pro Met Lys
850 855 860
Ala Lys Tyr Ser Val Gly Phe Lys Ser Asp Gly Lys Ile Thr Ala Leu
865 870 875 880
His Leu Asp Leu Gly Ile Asn Ala Gly Ile Ser Pro Asp Val Ser Pro
885 890 895
Leu Met Pro Arg Ala Ile Ile Gly Ala Leu Lys Lys Tyr Asn Trp Gly
900 905 910
Thr Leu Glu Phe Asp Thr Lys Val Cys Lys Thr Asn Val Ser Ser Lys
915 920 925
Ser Ala Met Arg Ala Pro Gly Asp Val Gln Gly Ser Phe Ile Ala Glu
930 935 940
Ala Ile Ile Glu His Val Ala Ser Ala Leu Ala Leu Asp Thr Asn Thr
945 950 955 960
Val Arg Arg Lys Asn Leu His Asp Phe Glu Ser Leu Glu Val Phe Tyr
965 970 975
Gly Glu Ser Ala Gly Glu Ala Ser Thr Tyr Ser Leu Val Ser Met Phe
980 985 990
Asp Lys Leu Ala Leu Ser Pro Glu Tyr Gln His Arg Ala Ala Met Ile
995 1000 1005
Glu Gln Phe Asn Ser Ser Asn Lys Trp Lys Lys Arg Gly Ile Ser
1010 1015 1020
Cys Val Pro Ala Thr Tyr Glu Val Asn Leu Arg Pro Thr Pro Gly
1025 1030 1035
Lys Val Ser Ile Met Asn Asp Gly Ser Ile Ala Val Glu Val Gly
1040 1045 1050
Gly Ile Glu Ile Gly Gln Gly Leu Trp Thr Lys Val Lys Gln Met
1055 1060 1065
Thr Ala Phe Gly Leu Gly Gln Leu Cys Pro Asp Gly Gly Glu Cys
1070 1075 1080
Leu Leu Asp Lys Val Arg Val Ile Gln Ala Asp Thr Leu Ser Leu
1085 1090 1095
Ile Gln Gly Gly Met Thr Ala Gly Ser Thr Thr Ser Glu Thr Ser
1100 1105 1110
Cys Glu Thr Val Arg Gln Ser Cys Val Ala Leu Val Glu Lys Leu
1115 1120 1125
Asn Pro Ile Lys Glu Ser Leu Glu Ala Lys Ser Asn Thr Val Glu
1130 1135 1140
Trp Ser Ala Leu Ile Ala Gln Ala Ser Met Ala Ser Val Asn Leu
1145 1150 1155
Ser Ala Gln Pro Tyr Trp Thr Pro Asp Pro Ser Phe Lys Ser Tyr
1160 1165 1170
Leu Asn Tyr Gly Ala Gly Thr Ser Glu Val Glu Val Asp Ile Leu
1175 1180 1185
Thr Gly Ala Thr Thr Ile Leu Arg Ser Asp Leu Val Tyr Asp Cys
1190 1195 1200
Gly Gln Ser Leu Asn Pro Ala Val Asp Leu Gly Gln Ile Glu Gly
1205 1210 1215
Cys Phe Val Gln Gly Ile Gly Phe Phe Thr Asn Glu Asp Tyr Lys
1220 1225 1230
Thr Asn Ser Asp Gly Leu Val Ile His Asp Gly Thr Trp Thr Tyr
1235 1240 1245
Lys Ile Pro Thr Val Asp Asn Ile Pro Lys Glu Phe Asn Val Glu
1250 1255 1260
Met Phe Asn Ser Ala Pro Asp Lys Lys Arg Val Leu Ser Ser Lys
1265 1270 1275
Ala Ser Gly Glu Pro Pro Leu Val Leu Ala Thr Ser Val His Cys
1280 1285 1290
Ala Met Arg Glu Ala Ile Arg Ala Ala Arg Lys Glu Phe Ser Val
1295 1300 1305
Ser Thr Ser Pro Ala Lys Ser Ala Val Thr Phe Gln Met Asp Val
1310 1315 1320
Pro Ala Thr Met Pro Val Val Lys Glu Leu Cys Gly Leu Asp Val
1325 1330 1335
Val Glu Arg Tyr Leu Glu Asn Val Ser Ala Ala Ser Ala Gly Pro
1340 1345 1350
Asn Thr Ala Lys Ala
1355
<210> 62
<211> 747
<212> DNA
<213> 新月柄杆菌(Caulobacter crescentus)
<400> 62
atgtcctcag ccatctatcc cagcctgaag ggcaagcgcg tcgtcatcac cggcggcggc 60
tcgggcatcg gggccggcct caccgccggc ttcgcccgtc agggcgcgga ggtgatcttc 120
ctcgacatcg ccgacgagga ctccagggct cttgaggccg agctggccgg ctcgccgatc 180
ccgccggtct acaagcgctg cgacctgatg aacctcgagg cgatcaaggc ggtcttcgcc 240
gagatcggcg acgtcgacgt gctggtcaac aacgccggca atgacgaccg ccacaagctg 300
gccgacgtga ccggcgccta ttgggacgag cggatcaacg tcaacctgcg ccacatgctg 360
ttctgcaccc aggccgtcgc gccgggcatg aagaagcgtg gcggcggggc ggtgatcaac 420
ttcggttcga tcagctggca cctggggctt gaggacctcg tcctctacga aaccgccaag 480
gccggcatcg aaggcatgac ccgcgcgctg gcccgggagc tgggtcccga cgacatccgc 540
gtcacctgcg tggtgccggg caacgtcaag accaagcgcc aggagaagtg gtacacgccc 600
gaaggcgagg cccagatcgt ggcggcccaa tgcctgaagg gccgcatcgt cccggagaac 660
gtcgccgcgc tggtgctgtt cctggcctcg gatgacgcgt cgctctgcac cggccacgaa 720
tactggatcg acgccggctg gcgttga 747
<210> 63
<211> 870
<212> DNA
<213> 新月柄杆菌(Caulobacter crescentus)
<400> 63
atgaccgctc aagtcacttg cgtatgggat ctgaaggcca cgttgggcga aggcccgatc 60
tggcatggcg acaccctgtg gttcgtcgac atcaagcagc gtaaaatcca caactaccac 120
cccgccaccg gcgagcgctt cagcttcgac gcgccggatc aggtgacctt cctcgcgccg 180
atcgtcggcg cgaccggctt tgtcgtcggt ctgaagaccg ggattcaccg cttccacccg 240
gccacgggct tcagcctgct gctcgaggtc gaggacgcgg cgctgaacaa ccgccccaac 300
gacgccacgg tcgacgcgca aggccgtctg tggttcggca ccatgcacga cggggaagag 360
aacaatagcg gctcgctcta tcggatggac ctcaccggcg tcgcccggat ggaccgcgac 420
atctgcatca ccaacggccc gtgcgtctcg cccgacggca agaccttcta ccacaccgac 480
accctggaaa agacgatcta cgccttcgac ctggccgagg acggcctgct gtcgaacaag 540
cgcgtcttcg tgcagttcgc cctgggcgac gatgtctatc cggacggttc ggtcgtcgat 600
tccgaaggct atctgtggac cgccctgtgg ggcggtttcg gcgcggtccg cttctcgccg 660
caaggcgacg ccgtgacgcg catcgaactg cccgccccca acgtcaccaa gccctgcttc 720
ggcgggcctg acctgaagac cctctatttc accaccgccc gcaagggcct gagcgacgag 780
accctggccc agtacccgct ggccggcggt gtgttcgccg ttccggtcga tgtggccggc 840
caaccccagc atgaggtccg ccttgtctaa 870
<210> 64
<211> 1968
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 64
atgtctgttc gcaatatttt tgctgacgag agccacgata tttacaccgt cagaacgcac 60
gccgatggcc cggacggcga actcccatta accgcagaga tgcttatcaa ccgcccgagc 120
ggggatctgt tcggtatgac catgaatgcc ggaatgggtt ggtctccgga cgagctggat 180
cgggacggta ttttactgct cagtacactc ggtggcttac gcggcgcaga cggtaaaccc 240
gtggcgctgg cgttgcacca ggggcattac gaactggaca tccagatgaa agcggcggcc 300
gaggttatta aagccaacca tgccctgccc tatgccgtgt acgtctccga tccttgtgac 360
gggcgtactc agggtacaac ggggatgttt gattcgctac cataccgaaa tgacgcatcg 420
atggtaatgc gccgccttat tcgctctctg cccgacgcga aagcagttat tggtgtggcg 480
agttgcgata aggggcttcc ggccaccatg atggcactcg ccgcgcagca caacatcgca 540
accgtgctgg tccccggcgg cgcgacgctg cccgcaaagg atggagaaga caacggcaag 600
gtgcaaacca ttggcgcacg cttcgccaat ggcgaattat ctctacagga cgcacgccgt 660
gcgggctgta aagcctgtgc ctcttccggc ggcggctgtc aatttttggg cactgccggg 720
acatctcagg tggtggccga aggattggga ctggcaatcc cacattcagc cctggcccct 780
tccggtgagc ctgtgtggcg ggagatcgcc agagcttccg cgcgagctgc gctgaacctg 840
agtcaaaaag gcatcaccac ccgggaaatt ctcaccgata aagcgataga gaatgcgatg 900
acggtccatg ccgcgttcgg tggttcaaca aacctgctgt tacacatccc ggcaattgct 960
caccaggcag gttgccatat cccgaccgtt gatgactgga tccgcatcaa caagcgcgtg 1020
ccccgactgg tgagcgtact gcctaatggc ccggtttatc atccaacggt caatgccttt 1080
atggcaggtg gtgtgccgga agtcatgttg catctgcgca gcctcggatt gttgcatgaa 1140
gacgttatga cggttaccgg cagcacgctg aaagaaaacc tcgactggtg ggagcactcc 1200
gaacggcgtc agcggttcaa gcaactcctg ctcgatcagg aacaaatcaa cgctgacgaa 1260
gtgatcatgt ctccgcagca agcaaaagcg cgcggattaa cctcaactat caccttcccg 1320
gtgggcaata ttgcgccaga aggttcggtg atcaaatcca ccgccattga cccctcgatg 1380
attgatgagc aaggtatcta ttaccataaa ggtgtggcga aggtttatct gtccgagaaa 1440
agtgcgattt acgatatcaa acatgacaag atcaaggcgg gcgatattct ggtcattatt 1500
ggcgttggac cttcaggtac agggatggaa gaaacctacc aggttaccag tgccctgaag 1560
catctgtcat acggtaagca tgtttcgtta atcaccgatg cacgtttctc gggcgtttct 1620
actggcgcgt gcatcggcca tgtggggcca gaagcgctgg ccggaggccc catcggtaaa 1680
ttacgcaccg gggatttaat tgaaattaaa attgattgtc gcgagcttca cggcgaagtc 1740
aatttcctcg gaacccgtag cgatgaacaa ttaccttcac aggaggaggc aactgcaata 1800
ttaaatgcca gacccagcca tcaggattta cttcccgatc ctgaattgcc agatgatacc 1860
cggctatggg caatgcttca ggccgtgagt ggtgggacat ggaccggttg tatttatgat 1920
gtaaacaaaa ttggcgcggc tttgcgcgat tttatgaata aaaactga 1968
<210> 65
<211> 1383
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 65
atgacgcaat taaccatgaa agacaaaatt ggctacgggc tgggagacac cgcctgcggc 60
ttcgtctggc aggccacgat gttcctgctg gcctatttct acaccgacgt cttcggcctg 120
tcggcgggga ttatgggcac gctgtttttg gtctcccgcg tgctcgacgc cgtcaccgac 180
ccgctgatgg ggctgctggt agaccgcacc cgcacgcggc acggccagtt ccgcccgttc 240
ctgctgtggg gggccatccc gttcggcatc gtctgcgtgc tgaccttcta cacgccggac 300
ttctccgcac agggcaagat catctacgcc tgcgtgacct acattctcct gaccctggtc 360
tacaccttcg ttaacgtgcc gtactgcgcc atgccgggcg tcatcaccgc cgacccgaaa 420
gagcgtcacg ccctgcagtc ctggcgcttc ttcctggcgg cggcgggctc gctcgctatc 480
agcggcatcg cgctgccgct ggtgagcatc atcggcaaag gggacgagca ggtgggctac 540
ttcggcgcca tgtgcgtgct ggggctgagc ggcgtggtgc tgctctacgt ctgcttcttc 600
acgaccaaag agcgctacac ctttgaggtg cagccgggct cgtcggtggc gaaagacctt 660
aagctgctgc tgggcaacag ccagtggcgc atcatgtgcg cgttcaagat gatggcgacc 720
tgctccaacg tggtgcgcgg cggggcgacg ctctacttcg tgaaatacgt gatggatcac 780
ccggagttgg cgacccagtt tttactttac ggcagcctcg ccaccatgtt cggctcgctt 840
tgctcctcac gcctgctggg ccgcttcgac cgcgtcaccg ccttcaagtg gatcatcgtc 900
gcctactcgc tgatcagcct gctgattttc gtcaccccgg cggagcacat cgcgctcatt 960
tttgccctca acatcctgtt cctgttcgtc tttaatacca ccacgccgct gcagtggctg 1020
atggcttctg acgtggtgga ctacgaggag agccgcagcg gtcgccgcct cgacgggctg 1080
gtgttctcca cctacctgtt cagcctgaag attggcctgg cgattggcgg ggcggtggtg 1140
ggctggatcc tggcgtacgt caactattcc gccagcagca gcgtgcagcc ggttgaggtg 1200
ctcaccacca tcaaaattct gttctgcgtg gtgccggtgg tgctctacgc gggcatgttc 1260
atcatgctgt cgctctacaa gctcaccgat gcccgcgtgg aggccatcag ccggcagctg 1320
attaagcacc gcgcggcgca gggcgaggcc gttcccgacg ccgcgacagc cgcatcccat 1380
taa 1383
<210> 66
<211> 1041
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 66
atgcagaaca tcatccgaaa aggaggaact atgaaggctg cagttgttac gaaggatcat 60
catgttgacg ttacgtataa aacactgcgc tcactgaaac atggcgaagc cctgctgaaa 120
atggagtgtt gtggtgtatg tcataccgat cttcatgtta agaatggcga ttttggtgac 180
aaaaccggcg taattctggg ccatgaaggc atcggtgtgg tggcagaagt gggtccaggt 240
gtcacctcat taaaaccagg cgatcgtgcc agcgtggcgt ggttctacga aggatgcggt 300
cattgcgaat actgtaacag tggtaacgaa acgctctgcc gttcagttaa aaatgccgga 360
tacagcgttg atggcgggat ggcggaagag tgcatcgtgg tcgccgatta cgcggtaaaa 420
gtgccagatg gtctggactc ggcggcggcc agcagcatta cctgtgcggg agtcaccacc 480
tacaaagccg ttaagctgtc aaaaattcgt ccagggcagt ggattgctat ctacggtctt 540
ggcggtctgg gtaacctcgc cctgcaatac gcgaagaatg tctttaacgc caaagtgatc 600
gccattgatg tcaatgatga gcagttaaaa ctggcaaccg aaatgggcgc agatttagcg 660
attaactcac acaccgaaga cgccgccaaa attgtgcagg agaaaactgg tggcgctcac 720
gctgcggtgg taacagcggt agctaaagct gcgtttaact cggcagttga tgctgtccgt 780
gcaggcggtc gtgttgtggc tgtcggtcta ccgccggagt ctatgagcct ggatatccca 840
cgtcttgtgc tggatggtat tgaagtggtc ggttcgctgg tcggcacgcg ccaggattta 900
actgaagcct tccagtttgc cgccgaaggt aaagtggtgc cgaaagtcgc cctgcgtccg 960
ttagcggaca tcaacaccat ctttactgag atggaagaag gcaaaatccg tggccgcatg 1020
gtgattgatt tccgtcacta a 1041
<210> 67
<211> 1164
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 67
atgaacaact ttaatctgca caccccaacc cgcattctgt ttggtaaagg cgcaatcgct 60
ggtttacgcg aacaaattcc tcacgatgct cgcgtattga ttacctacgg cggcggcagc 120
gtgaaaaaaa ccggcgttct cgatcaagtt ctggatgccc tgaaaggcat ggacgtgctg 180
gaatttggcg gtattgagcc aaacccggct tatgaaacgc tgatgaacgc cgtgaaactg 240
gttcgcgaac agaaagtgac tttcctgctg gcggttggcg gcggttctgt actggacggc 300
accaaattta tcgccgcagc ggctaactat ccggaaaata tcgatccgtg gcacattctg 360
caaacgggcg gtaaagagat taaaagcgcc atcccgatgg gctgtgtgct gacgctgcca 420
gcaaccggtt cagaatccaa cgcaggcgcg gtgatctccc gtaaaaccac aggcgacaag 480
caggcgttcc attctgccca tgttcagccg gtatttgccg tgctcgatcc ggtttatacc 540
tacaccctgc cgccgcgtca ggtggctaac ggcgtagtgg acgcctttgt acacaccgtg 600
gaacagtatg ttaccaaacc ggttgatgcc aaaattcagg accgtttcgc agaaggcatt 660
ttgctgacgc taatcgaaga tggtccgaaa gccctgaaag agccagaaaa ctacgatgtg 720
cgcgccaacg tcatgtgggc ggcgactcag gcgctgaacg gtttgattgg cgctggcgta 780
ccgcaggact gggcaacgca tatgctgggc cacgaactga ctgcgatgca cggtctggat 840
cacgcgcaaa cactggctat cgtcctgcct gcactgtgga atgaaaaacg cgataccaag 900
cgcgctaagc tgctgcaata tgctgaacgc gtctggaaca tcactgaagg ttccgatgat 960
gagcgtattg acgccgcgat tgccgcaacc cgcaatttct ttgagcaatt aggcgtgccg 1020
acccacctct ccgactacgg tctggacggc agctccatcc cggctttgct gaaaaaactg 1080
gaagagcacg gcatgaccca actgggcgaa aatcatgaca ttacgttgga tgtcagccgc 1140
cgtatatacg aagccgcccg ctaa 1164
<210> 68
<211> 1587
<212> DNA
<213> 恶臭假单胞菌(Pseudomonas putida)
<400> 68
atggcttcgg tacacggcac cacatacgaa ctcttgcgac gtcaaggcat cgatacggtc 60
ttcggcaatc ctggctcgaa cgagctcccg tttttgaagg actttccaga ggactttcga 120
tacatcctgg ctttgcagga agcgtgtgtg gtgggcattg cagacggcta tgcgcaagcc 180
agtcggaagc cggctttcat taacctgcat tctgctgctg gtaccggcaa tgctatgggt 240
gcactcagta acgcctggaa ctcacattcc ccgctgatcg tcactgccgg ccagcagacc 300
agggcgatga ttggcgttga agctctgctg accaacgtcg atgccgccaa cctgccacga 360
ccacttgtca aatggagcta cgagcccgca agcgcagcag aagtccctca tgcgatgagc 420
agggctatcc atatggcaag catggcgcca caaggccctg tctatctttc ggtgccatat 480
gacgattggg ataaggatgc tgatcctcag tcccaccacc tttttgatcg ccatgtcagt 540
tcatcagtac gcctgaacga ccaggatctc gatattctgg tgaaagctct caacagcgca 600
tccaacccgg cgatcgtcct gggcccggac gtcgacgcag caaatgcgaa cgcagactgc 660
gtcatgttgg ccgaacgcct caaagctccg gtttgggttg cgccatccgc tccacgctgc 720
ccattcccta cccgtcatcc ttgcttccgt ggattgatgc cagctggcat cgcagcgatt 780
tctcagctgc tcgaaggtca cgatgtggtt ttggtaatcg gcgctccagt gttccgttac 840
caccaatacg acccaggtca atatctcaaa cctggcacgc gattgatttc ggtgacctgc 900
gacccgctcg aagctgcacg cgcgccaatg ggcgatgcga tcgtggcaga cattggtgcg 960
atggctagcg ctcttgccaa cttggttgaa gagagcagcc gccagctccc aactgcagct 1020
ccggaacccg cgaaggttga ccaagacgct ggccgacttc acccagagac agtgttcgac 1080
acactgaacg acatggcccc ggagaatgcg atttacctga acgagtcgac ttcaacgacc 1140
gcccaaatgt ggcagcgcct gaacatgcgc aaccctggta gctactactt ctgtgcagct 1200
ggcggactgg gcttcgccct gcctgcagca attggcgttc aactcgcaga acccgagcga 1260
caagtcatcg ccgtcattgg cgacggatcg gcgaactaca gcattagtgc gttgtggact 1320
gcagctcagt acaacatccc cactatcttc gtgatcatga acaacggcac ctacggtgcg 1380
ttgcgatggt ttgccggcgt tctcgaagca gaaaacgttc ctgggctgga tgtgccaggg 1440
atcgacttcc gcgcactcgc caagggctat ggtgtccaag cgctgaaagc cgacaacctt 1500
gagcagctca agggttcgct acaagaagcg ctttctgcca aaggcccggt acttatcgaa 1560
gtaagcaccg taagcccggt gaagtga 1587
<210> 69
<211> 1323
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 69
atgcaagcct attttgacca gctcgatcgc gttcgttatg aaggctcaaa atcctcaaac 60
ccgttagcat tccgtcacta caatcccgac gaactggtgt tgggtaagcg tatggaagag 120
cacttgcgtt ttgccgcctg ctactggcac accttctgct ggaacggggc ggatatgttt 180
ggtgtggggg cgtttaatcg tccgtggcag cagcctggtg aggcactggc gttggcgaag 240
cgtaaagcag atgtcgcatt tgagtttttc cacaagttac atgtgccatt ttattgcttc 300
cacgatgtgg atgtttcccc tgagggcgcg tcgttaaaag agtacatcaa taattttgcg 360
caaatggttg atgtcctggc aggcaagcaa gaagagagcg gcgtgaagct gctgtgggga 420
acggccaact gctttacaaa ccctcgctac ggcgcgggtg cggcgacgaa cccagatcct 480
gaagtcttca gctgggcggc aacgcaagtt gttacagcga tggaagcaac ccataaattg 540
ggcggtgaaa actatgtcct gtggggcggt cgtgaaggtt acgaaacgct gttaaatacc 600
gacttgcgtc aggagcgtga acaactgggc cgctttatgc agatggtggt tgagcataaa 660
cataaaatcg gtttccaggg cacgttgctt atcgaaccga aaccgcaaga accgaccaaa 720
catcaatatg attacgatgc cgcgacggtc tatggcttcc tgaaacagtt tggtctggaa 780
aaagagatta aactgaacat tgaagctaac cacgcgacgc tggcaggtca ctctttccat 840
catgaaatag ccaccgccat tgcgcttggc ctgttcggtt ctgtcgacgc caaccgtggc 900
gatgcgcaac tgggctggga caccgaccag ttcccgaaca gtgtggaaga gaatgcgctg 960
gtgatgtatg aaattctcaa agcaggcggt ttcaccaccg gtggtctgaa cttcgatgcc 1020
aaagtacgtc gtcaaagtac tgataaatat gatctgtttt acggtcatat cggcgcgatg 1080
gatacgatgg cactggcgct gaaaattgca gcgcgcatga ttgaagatgg cgagctggat 1140
aaacgcatcg cgcagcgtta ttccggctgg aatagcgaat tgggccagca aatcctgaaa 1200
ggccaaatgt cactggcaga tttagccaaa tatgctcagg aacatcattt gtctccggtg 1260
catcagagtg gtcgccagga acaactggaa aatctggtaa accattatct gttcgacaaa 1320
taa 1323
<210> 70
<211> 1455
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 70
atgtatatcg ggatagatct tggcacctcg ggcgtaaaag ttattttgct caacgagcag 60
ggtgaggtgg ttgctgcgca aacggaaaag ctgaccgttt cgcgcccgca tccactctgg 120
tcggaacaag acccggaaca gtggtggcag gcaactgatc gcgcaatgaa agctctgggc 180
gatcagcatt ctctgcagga cgttaaagca ttgggtattg ccggccagat gcacggagca 240
accttgctgg atgctcagca acgggtgtta cgccctgcca ttttgtggaa cgacgggcgc 300
tgtgcgcaag agtgcacttt gctggaagcg cgagttccgc aatcgcgggt gattaccggc 360
aacctgatga tgcccggatt tactgcgcct aaattgctat gggttcagcg gcatgagccg 420
gagatattcc gtcaaatcga caaagtatta ttaccgaaag attacttgcg tctgcgtatg 480
acgggggagt ttgccagcga tatgtctgac gcagctggca ccatgtggct ggatgtcgca 540
aagcgtgact ggagtgacgt catgctgcag gcttgcgact tatctcgtga ccagatgccc 600
gcattatacg aaggcagcga aattactggt gctttgttac ctgaagttgc gaaagcgtgg 660
ggtatggcga cggtgccagt tgtcgcaggc ggtggcgaca atgcagctgg tgcagttggt 720
gtgggaatgg ttgatgctaa tcaggcaatg ttatcgctgg ggacgtcggg ggtctatttt 780
gctgtcagcg aagggttctt aagcaagcca gaaagcgccg tacatagctt ttgccatgcg 840
ctaccgcaac gttggcattt aatgtctgtg atgctgagtg cagcgtcgtg tctggattgg 900
gccgcgaaat taaccggcct gagcaatgtc ccagctttaa tcgctgcagc tcaacaggct 960
gatgaaagtg ccgagccagt ttggtttctg ccttatcttt ccggcgagcg tacgccacac 1020
aataatcccc aggcgaaggg ggttttcttt ggtttgactc atcaacatgg ccccaatgaa 1080
ctggcgcgag cagtgctgga aggcgtgggt tatgcgctgg cagatggcat ggatgtcgtg 1140
catgcctgcg gtattaaacc gcaaagtgtt acgttgattg ggggcggggc gcgtagtgag 1200
tactggcgtc agatgctggc ggatatcagc ggtcagcagc tcgattaccg tacggggggg 1260
gatgtggggc cagcactggg cgcagcaagg ctggcgcaga tcgcggcgaa tccagagaaa 1320
tcgctcattg aattgttgcc gcaactaccg ttagaacagt cgcatctacc agatgcgcag 1380
cgttatgccg cttatcagcc acgacgagaa acgttccgtc gcctctatca gcaacttctg 1440
ccattaatgg cgtaa 1455
<210> 71
<211> 906
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 71
atgaaaaaat tcagcggcat tattccaccg gtatccagca cgtttcatcg tgacggaacc 60
cttgataaaa aggcaatgcg cgaagttgcc gacttcctga ttaataaagg ggtcgacggg 120
ctgttttatc tgggtaccgg tggtgaattt agccaaatga atacagccca gcgcatggca 180
ctcgccgaag aagctgtaac cattgtcgac gggcgagtgc cggtattgat tggcgtcggt 240
tccccttcca ctgacgaagc ggtcaaactg gcgcagcatg cgcaagccta cggcgctgat 300
ggtatcgtcg ccatcaaccc ctactactgg aaagtcgcac cacgaaatct tgacgactat 360
taccagcaga tcgcccgtag cgtcacccta ccggtgatcc tgtacaactt tccggatctg 420
acgggtcagg acttaacccc ggaaaccgtg acgcgtctgg ctctgcaaaa cgagaatatc 480
gttggcatca aagacaccat cgacagcgtt ggtcacttgc gtacgatgat caacacagtt 540
aagtcggtac gcccgtcgtt ttcggtattc tgcggttacg atgatcattt gctgaatacg 600
atgctgctgg gcggcgacgg tgcgataacc gccagcgcta actttgctcc ggaactctcc 660
gtcggcatct accgcgcctg gcgtgaaggc gatctggcga ccgctgcgac gctgaataaa 720
aaactactac aactgcccgc tatttacgcc ctcgaaacac cgtttgtctc actgatcaaa 780
tacagcatgc agtgtgtcgg gctgcctgta gagacatatt gcttaccacc gattcttgaa 840
gcatctgaag aagcaaaaga taaagtccac gtgctgctta ccgcgcaggg cattttacca 900
gtctga 906
<210> 72
<211> 909
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 72
atgccgcagt ccgcgttgtt cacgggaatc attccccctg tctccaccat ttttaccgcc 60
gacggccagc tcgataagcc gggcaccgcc gcgctgatcg acgatctgat caaagcaggc 120
gttgacggcc tgttcttcct gggcagcggt ggcgagttct cccagctcgg cgccgaagag 180
cgtaaagcca ttgcccgctt tgctatcgat catgtcgatc gtcgcgtgcc ggtgctgatc 240
ggcaccggcg gcaccaacgc ccgggaaacc atcgaactca gccagcacgc gcagcaggcg 300
ggcgcggacg gcatcgtggt gatcaacccc tactactgga aagtgtcgga agcgaacctg 360
atccgctatt tcgagcaggt ggccgacagc gtcacgctgc cggtgatgct ctataacttc 420
ccggcgctga ccgggcagga tctgactccg gcgctggtga aaaccctcgc cgactcgcgc 480
agcaatatta tcggcatcaa agacaccatc gactccgtcg cccacctgcg cagcatgatc 540
cataccgtca aaggtgccca tccgcacttc accgtgctct gcggctacga cgatcatctg 600
ttcaataccc tgctgctcgg cggcgacggg gcgatatcgg cgagcggcaa ctttgccccg 660
caggtgtcgg tgaatcttct gaaagcctgg cgcgacgggg acgtggcgaa agcggccggg 720
tatcatcaga ccttgctgca aattccgcag atgtatcagc tggatacgcc gtttgtgaac 780
gtgattaaag aggcgatcgt gctctgcggt cgtcctgtct ccacgcacgt gctgccgccc 840
gcctcgccgc tggacgagcc gcgcaaggcg cagctgaaaa ccctgctgca acagctcaag 900
ctttgctga 909
<210> 73
<211> 975
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 73
atgaagccgt ccgttatcct ctacaaagcc ttacctgatg atttactgca acgcctgcaa 60
gagcatttca ccgttcacca ggtggcaaac ctcagcccac aaaccgtcga acaaaatgca 120
gcaatttttg ccgaagctga aggtttactg ggttcaaacg agaatgtaaa tgccgcattg 180
ctggaaaaaa tgccgaaact gcgtgccaca tcaacgatct ccgtcggcta tgacaatttt 240
gatgtcgatg cgcttaccgc ccgaaaaatt ctgctgatgc acacgccaac cgtattaaca 300
gaaaccgtcg ccgatacgct gatggcgctg gtgttgtcta ccgctcgtcg ggttgtggag 360
gtagcagaac gggtaaaagc aggcgaatgg accgcgagca taggcccgga ctggtacggc 420
actgacgttc accataaaac actgggcatt gtcgggatgg gacggatcgg catggcgctg 480
gcacaacgtg cgcactttgg cttcaacatg cccatcctct ataacgcgcg ccgccaccat 540
aaagaagcag aagaacgctt caacgcccgc tactgcgatt tggatactct gttacaagag 600
tcagatttcg tttgcctgat cctgccgtta actgatgaga cgcatcatct gtttggcgca 660
gaacaattcg ccaaaatgaa atcctccgcc attttcatta atgccggacg tggcccggtg 720
gttgacgaaa atgcactgat cgcagcattg cagaaaggcg aaattcacgc tgccgggctg 780
gatgtcttcg aacaagagcc actgtccgta gattcgccgt tgctctcaat ggccaacgtc 840
gtcgcagtac cgcatattgg atctgccacc catgagacgc gttatggcat ggccgcctgt 900
gccgtggata atttgattga tgcgttacaa ggaaaggttg agaagaactg tgtgaatccg 960
cacgtcgcgg actaa 975
<210> 74
<211> 939
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 74
atggatatca tcttttatca cccaacgttc gatacccaat ggtggattga ggcactgcgc 60
aaagctattc ctcaggcaag agtcagagca tggaaaagcg gagataatga ctctgctgat 120
tatgctttag tctggcatcc tcctgttgaa atgctggcag ggcgcgatct taaagcggtg 180
ttcgcactcg gggccggtgt tgattctatt ttgagcaagc tacaggcaca ccctgaaatg 240
ctgaaccctt ctgttccact ttttcgcctg gaagataccg gtatgggcga gcaaatgcag 300
gaatatgctg tcagtcaggt gctgcattgg tttcgacgtt ttgacgatta tcgcatccag 360
caaaatagtt cgcattggca accgctgcct gaatatcatc gggaagattt taccatcggc 420
attttgggcg caggcgtact gggcagtaaa gttgctcaga gtctgcaaac ctggcgcttt 480
ccgctgcgtt gctggagtcg aacccgtaaa tcgtggcctg gcgtgcaaag ctttgccgga 540
cgggaagaac tgtctgcatt tctgagccaa tgtcgggtat tgattaattt gttaccgaat 600
acccctgaaa ccgtcggcat tattaatcaa caattactcg aaaaattacc ggatggcgcg 660
tatctcctca acctggcgcg tggtgttcat gttgtggaag atgacctgct cgcggcgctg 720
gatagcggca aagttaaagg cgcaatgttg gatgttttta atcgtgaacc cttaccgcct 780
gaaagtccgc tctggcaaca tccacgcgtg acgataacac cacatgtcgc cgcgattacc 840
cgtcccgctg aagctgtgga gtacatttct cgcaccattg cccagctcga aaaaggggag 900
agggtctgcg ggcaagtcga ccgcgcacgc ggctactaa 939
<210> 75
<211> 1434
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 75
atgtttaaga atgcatttgc taacctgcaa aaggtcggta aatcgctgat gctgccggta 60
tccgtactgc ctatcgcagg tattctgctg ggcgtcggtt ccgcgaattt cagctggctg 120
cccgccgttg tatcgcatgt tatggcagaa gcaggcggtt ccgtctttgc aaacatgcca 180
ctgatttttg cgatcggtgt cgccctcggc tttaccaata acgatggcgt atccgcgctg 240
gccgcagttg ttgcctatgg catcatggtt aaaaccatgg ccgtggttgc gccactggta 300
ctgcatttac ctgctgaaga aatcgcctct aaacacctgg cggatactgg cgtactcgga 360
gggattatct ccggtgcgat cgcagcgtac atgtttaacc gtttctaccg tattaagctg 420
cctgagtatc ttggcttctt tgccggtaaa cgctttgtgc cgatcatttc tggcctggct 480
gccatcttta ctggcgttgt gctgtccttc atttggccgc cgattggttc tgcaatccag 540
accttctctc agtgggctgc ttaccagaac ccggtagttg cgtttggcat ttacggtttc 600
atcgaacgtt gcctggtacc gtttggtctg caccacatct ggaacgtacc tttccagatg 660
cagattggtg aatacaccaa cgcagcaggt caggttttcc acggcgacat tccgcgttat 720
atggcgggtg acccgactgc gggtaaactg tctggtggct tcctgttcaa aatgtacggt 780
ctgccagctg ccgcaattgc tatctggcac tctgctaaac cagaaaaccg cgcgaaagtg 840
ggcggtatta tgatctccgc ggcgctgacc tcgttcctga ccggtatcac cgagccgatc 900
gagttctcct tcatgttcgt tgcgccgatc ctgtacatca tccacgcgat tctggcaggc 960
ctggcattcc caatctgtat tcttctgggg atgcgtgacg gtacgtcgtt ctcgcacggt 1020
ctgatcgact tcatcgttct gtctggtaac agcagcaaac tgtggctgtt cccgatcgtc 1080
ggtatcggtt atgcgattgt ttactacacc atcttccgcg tgctgattaa agcactggat 1140
ctgaaaacgc cgggtcgtga agacgcgact gaagatgcaa aagcgacagg taccagcgaa 1200
atggcaccgg ctctggttgc tgcatttggt ggtaaagaaa acattactaa cctcgacgca 1260
tgtattaccc gtctgcgcgt cagcgttgct gatgtgtcta aagtggatca ggccggcctg 1320
aagaaactgg gcgcagcggg cgtagtggtt gctggttctg gtgttcaggc gattttcggt 1380
actaaatccg ataacctgaa aaccgagatg gatgagtaca tccgtaacca ctaa 1434
<210> 76
<211> 510
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 76
atgggtttgt tcgataaact gaaatctctg gtttccgacg acaagaagga taccggaact 60
attgagatca ttgctccgct ctctggcgag atcgtcaata tcgaagacgt gccggatgtc 120
gtttttgcgg aaaaaatcgt tggtgatggt attgctatca aaccaacggg taacaaaatg 180
gtcgcgccag tagacggcac cattggtaaa atctttgaaa ccaaccacgc attctctatc 240
gaatctgata gcggcgttga actgttcgtc cacttcggta tcgacaccgt tgaactgaaa 300
ggcgaaggct tcaagcgtat tgctgaagaa ggtcagcgcg tgaaagttgg cgatactgtc 360
attgaatttg atctgccgct gctggaagag aaagccaagt ctaccctgac tccggttgtt 420
atctccaaca tggacgaaat caaagaactg atcaaactgt ccggtagcgt aaccgtgggt 480
gaaaccccgg ttatccgcat caagaagtaa 510
<210> 77
<211> 258
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 77
atgttccagc aagaagttac cattaccgct ccgaacggtc tgcacacccg ccctgctgcc 60
cagtttgtaa aagaagctaa gggcttcact tctgaaatta ctgtgacttc caacggcaaa 120
agcgccagcg cgaaaagcct gtttaaactg cagactctgg gcctgactca aggtaccgtt 180
gtgactatct ccgcagaagg cgaagacgag cagaaagcgg ttgaacatct ggttaaactg 240
atggcggaac tcgagtaa 258
<210> 78
<211> 1254
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 78
atgtactatt taaaaaacac aaacttttgg atgttcggtt tattcttttt cttttacttt 60
tttatcatgg gagcctactt cccgtttttc ccgatttggc tacatgacat caaccatatc 120
agcaaaagtg atacgggtat tatttttgcc gctatttctc tgttctcgct attattccaa 180
ccgctgtttg gtctgctttc tgacaaactc gggctgcgca aatacctgct gtggattatt 240
accggcatgt tagtgatgtt tgcgccgttc tttattttta tcttcgggcc actgttacaa 300
tacaacattt tagtaggatc gattgttggt ggtatttatc taggcttttg ttttaacgcc 360
ggtgcgccag cagtagaggc atttattgag aaagtcagcc gtcgcagtaa tttcgaattt 420
ggtcgcgcgc ggatgtttgg ctgtgttggc tgggcgctgt gtgcctcgat tgtcggcatc 480
atgttcacca tcaataatca gtttgttttc tggctgggct ctggctgtgc actcatcctc 540
gccgttttac tctttttcgc caaaacggat gcgccctctt ctgccacggt tgccaatgcg 600
gtaggtgcca accattcggc atttagcctt aagctggcac tggaactgtt cagacagcca 660
aaactgtggt ttttgtcact gtatgttatt ggcgtttcct gcacctacga tgtttttgac 720
caacagtttg ctaatttctt tacttcgttc tttgctaccg gtgaacaggg tacgcgggta 780
tttggctacg taacgacaat gggcgaatta cttaacgcct cgattatgtt ctttgcgcca 840
ctgatcatta atcgcatcgg tgggaaaaac gccctgctgc tggctggcac tattatgtct 900
gtacgtatta ttggctcatc gttcgccacc tcagcgctgg aagtggttat tctgaaaacg 960
ctgcatatgt ttgaagtacc gttcctgctg gtgggctgct ttaaatatat taccagccag 1020
tttgaagtgc gtttttcagc gacgatttat ctggtctgtt tctgcttctt taagcaactg 1080
gcgatgattt ttatgtctgt actggcgggc aatatgtatg aaagcatcgg tttccagggc 1140
gcttatctgg tgctgggtct ggtggcgctg ggcttcacct taatttccgt gttcacgctt 1200
agcggccccg gcccgctttc cctgctgcgt cgtcaggtga atgaagtcgc ttaa 1254
<210> 79
<211> 1545
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 79
atggatgtca ttaaaaagaa acattggtgg caaagcgacg cgctgaaatg gtcagtgcta 60
ggtctgctcg gcctgctggt gggttacctt gttgttttaa tgtacgcaca aggggaatac 120
ctgttcgcca ttaccacgct gatattgagt tcagcggggc tgtatatttt cgccaatcgt 180
aaagcctacg cctggcgcta tgtttacccg ggaatggctg gaatgggatt attcgtcctc 240
ttccctctgg tctgcaccat cgccattgcc ttcaccaact acagcagcac taaccagctg 300
acttttgaac gtgcgcagga agtgttgtta gatcgctcct ggcaagcagg caaaacctat 360
aactttggtc tttacccggc gggcgatgag tggcaactgg cgctcagcga cggcgaaacc 420
ggcaaaaatt acctctccga cgcttttaaa tttggcggcg agcaaaaact gcaactgaaa 480
gaaacgaccg cccagcccga aggcgaacgc gcgaatctgc gcgtgattac ccagaatcgt 540
caggcgctga gtgacattac cgccattctg ccggatggca acaaagtgat gatgagctcc 600
ctgcgccagt tttctggcac gcagccgctc tacacactcg acggtgacgg cacgttgacg 660
aataatcaga gcggcgtgaa atatcgtccg aataaccaaa ttggctttta ccagtccatt 720
accgccgacg gcaactgggg tgatgaaaag ctaagccccg gttacaccgt gaccaccggc 780
tggaaaaact ttacccgcgt ctttaccgac gaaggcattc agaaaccgtt cctcgccatt 840
ttcgtctgga ccgtggtgtt ctcgctgatc actgtctttt taacggtggc ggtcggcatg 900
gttctggcgt gtctggtgca gtgggaagcg ttgcgcggca aagcggtcta tcgcgtcctg 960
ctgattctgc cctacgcggt gccatcgttc atttcaatct tgattttcaa agggttgttt 1020
aaccagagct tcggtgaaat caacatgatg ttgagcgcgc tgtttggcgt gaagcccgcc 1080
tggttcagcg atccgaccac cgcccgcacg atgctaatta tcgtcaatac ctggctgggt 1140
tatccgtaca tgatgatcct ctgcatgggc ttgctgaaag cgattccgga cgatttgtat 1200
gaagcctcag caatggatgg cgcaggtccg ttccagaact tctttaagat tacgctgccg 1260
ctgctgatta aaccgctgac gccgctgatg atcgccagct tcgcctttaa ctttaacaac 1320
ttcgtgctga ttcaactgtt aaccaacggc ggcccggatc gtcttggcac gaccacgcca 1380
gccggttata ccgacctgct tgttaactac acctaccgca tcgcttttga aggcggcggg 1440
ggtcaggact tcggtctggc ggcagcaatt gccacgctga tcttcctgct ggtgggtgcg 1500
ctggcgatag tgaacctgaa agccacgcga atgaagtttg attaa 1545
<210> 80
<211> 891
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 80
atggcaatgg tccaaccgaa atcgcaaaaa gctcgtttat ttattactca cctgctactg 60
ctacttttta tcgcagcgat tatgttcccg ctgctgatgg tcgtcgctat ctcgctgcgt 120
cagggaaact ttgcgaccgg cagcctgatc ccggagcaaa tctcctggga tcactggaaa 180
ctggcgttag gttttagcgt tgaacaggct gatggtcgca ttacgccacc gccattcccg 240
gtactgctgt ggctgtggaa ctcggtaaag gtcgccggga tttccgcgat tggcattgtg 300
gcgctctcca ccacctgcgc ctacgctttc gcccgtatgc gctttccagg caaagcgacg 360
ctgctgaaag gaatgctgat tttccagatg ttcccggcag tactttcact ggtcgcgttg 420
tatgcgttgt ttgatcgtct gggtgagtac attccattca ttggcctgaa tactcacggc 480
ggcgtaattt tcgcgtatct gggtgggatt gcgctgcatg tctggaccat caaaggctat 540
ttcgaaacca tcgacagttc gctggaagaa gctgctgcgc tggatggtgc gacaccgtgg 600
caggccttcc gccttgtcct gttgccgctg tcagtaccga ttctggcggt ggtattcatc 660
ctgtcgttta tcgctgccat tactgaagtt ccggtcgcgt cgctgttact gcgtgacgta 720
aacagctaca ccctggccgt ggggatgcag caatacctca acccgcaaaa ctacctgtgg 780
ggtgactttg ccgccgctgc cgtgatgtct gcattaccga tcaccatcgt cttcttgctg 840
gctcaacgct ggctggtcaa cggcctgacg gcaggtggtg tgaaaggtta a 891
<210> 81
<211> 6546
<212> DNA
<213> 鼠伤寒沙门氏菌(Salmonella typhimurium)
<400> 81
atgaatgcaa aagtttgggt tctgggcgac gcggtggtgg acctgctgcc ggagagcgaa 60
gggcgcctgc tgcagtgccc tggaggcgcg ccggctaacg tggcggtagg ggttgcccgc 120
cttggcggca acagcggatt tatcggcgcc gtcggcggtg acccgtttgg ccgctacatg 180
cgtcataccc tgcaacagga gcaggtcgac gtcagccata tgtatctcga cgatcagcac 240
cgcacgtcca ctgtggtcgt cgaccttgac gaccaggggg aacgcacctt tacctttatg 300
gtacgcccca gcgcggacct gttcctggtt gaagaagacc tgccacagtt tgccgccgga 360
cagtggttgc acgtctgctc catcgcgctc agcgccgagc ccagccgtag cactaccttc 420
gcggcgatgg agagcatcag gtctgccggc ggtcgggtca gctttgaccc taatattcgt 480
cccgatctct ggcaggatca ggctttgctg ctagcctgcc tcgatcgcgc tttgcacatg 540
gccaacgtgg taaagctatc ggaagaggag ctggtcttca tcagcagcag taatgattta 600
gcatacggaa tcgccagcgt aacggagcgc tatcagccag aattgctact ggtgacccgg 660
ggcaaagcgg gggtgcttgc cgcgtttcag cagaagttta cccatttcaa cgcccggcct 720
gtggccagcg tggacaccac cggcgcggga gacgcatttg tcgccggact gctcgccagc 780
cttgcggcta acgggatgcc aacggacatg accgcactgg aaccgacact cacgcttgca 840
cagacctgcg gcgccctggc caccacagcc aaaggtgcga tgaccgcctt gccttatcag 900
cgcgatctca accgtcagtt ttaatcctta aagccgcttt gcgcggctca ctttgttgca 960
tgcatcacat ttattaaacc ggtttagcat atttgtttta agaaaaacaa aggtcgggct 1020
taacatagcg cctaaaccgg tttagcaaaa attataattt tccattttta cttttgggat 1080
gccaacagca tgtacagaaa aagcacactt gcgatgctta tcgctttgct aaccagcgct 1140
gcctcagccc atgcgcaaac ggatataagc accattgaag cccgactcaa cgcgctggaa 1200
aaacgcctgc aggaggcaga aaacagggcg caaacggcgg aaaaccgcgc cggggcggcg 1260
gagaaaaaag ttcagcaact caccgcgcag cagcaaaaaa accagaactc gactcaggaa 1320
gtggctcagc gtaccgccag acttgagaaa aaagccgatg acaaaagcgg atttgagttt 1380
cacggttacg cccgctccgg cgtgataatg aatgattccg gcgccagcac caaatccgga 1440
gcctacataa cgccggcagg tgaaaccggc ggagctatcg gccgtctggg aaaccaggcc 1500
gatacctatg ttgaaatgaa tcttgaacat aagcagaccc tggataatgg ggccacgacc 1560
cgctttaagg tgatggtcgc cgacgggcaa acctcttata acgactggac tgcaagcacc 1620
agcgatctga acgttcgtca ggcctttgtc gaattgggta acctgccgac gttcgctggg 1680
ccatttaagg gctccaccct gtgggccggg aaacgtttcg accgcgacaa tttcgatatt 1740
cactggattg actctgatgt cgtgttcctc gccggtaccg gtggtggtat ctatgacgtg 1800
aagtggaacg acggcctgcg gagtaatttc tccctgtacg ggcgtaactt cggcgacatt 1860
gatgattcca gcaacagcgt gcagaactat atcctcacca tgaatcactt cgcaggtccg 1920
ctgcagatga tggtcagcgg tctgcgggcg aaggataacg acgagcgtaa agatagcaac 1980
ggcaatctgg caaaaggcga tgcggcaaac accggcgtgc atgcgctgct cggcctgcat 2040
aacgacagtt tctacggcct gcgcgacggt agcagtaaaa ccgctctgct ttatggtcat 2100
ggtctgggcg cagaggttaa aggtatcgga tctgatggcg cacttcgtcc gggagccgac 2160
acatggcgca ttgccagtta cggcaccacg ccgctcagcg aaaactggtc tgttgccccg 2220
gcaatgctgg cgcaacgcag taaagaccgc tatgccgatg gcgacagcta tcagtgggca 2280
acattcaacc tgcgtctgat tcaggcaatc aatcagaatt tcgctctcgc ctacgaaggc 2340
agctaccagt acatggatct taaacccgaa ggttataacg atcgtcaggc ggtgaacggt 2400
agcttctaca agctcacctt cgccccgaca tttaaggtcg gcagtatcgg tgatttcttc 2460
agtcgcccgg agattcgttt ctatacctcc tggatggact ggagcaaaaa actgaataat 2520
tacgccagcg acgacgccct gggcagtgac ggttttaact cgggcggcga atggtctttc 2580
ggtgtgcaga tggaaacctg gttctgacgc ttacgcctga tgacaggaat agccgggggt 2640
cagagcatct ttgtcacccc ggactcaact aagacgcaga aaaagcgctc ccgtgaacgc 2700
gggacgacaa cataaaaatg tttaagcctt aagagggtac tatggatttt gaacagattt 2760
cctgctcgct gcttccgctt cttggaggca aagaaaatat cgccagcgcc gcgcactgcg 2820
ccacgcgcct gcgcctggtg ctggtcgatg attcgctggc cgaccagcag gccatcggca 2880
aagttgaagg ggtgaagggc tgttttcgta atgccggaca gatgcagatt attttcggca 2940
ccggggtggt aaataaggtc tacgctgcct ttactcaggc ggcgggtatt agcgaatcca 3000
gcaaatcgga agccgccgac atcgcggcaa aaaagctcaa tccgttccag cgcatcgccc 3060
gcctgctatc aaacatcttc gtgccgataa tccctgccat cgtcgcctct ggtctgctga 3120
tgggcctgct gggaatggtc aaaacatacg gctgggttga cccgggcaac gccatctaca 3180
tcatgctgga tatgtgcagc tcggcggcat ttatcattct gccgattctg attggcttta 3240
ccgccgcccg cgaattcggc ggtaatcctt atctcggcgc gacgcttggc ggcattctga 3300
ctcatccagc gctgactaac gcctggggcg tggccgcggg tttccacacc atgaactttt 3360
tcggcttcga aattgccatg atcggctatc agggtacggt gttcccggta ctgctggcag 3420
tatggtttat gagcatcgtt gagaagcagt tgcgtcgcgc aatccccgat gccctggatt 3480
tgatcctgac gccgttcctg acggtgatta tatccggttt tatcgccctg ttgattatcg 3540
gcccggccgg tcgcgcactg ggcgacggta tctcgtttgt cctcagcacc ctgattagcc 3600
acgccggctg gctcgccggg ttactgtttg gcggtctcta ttcagttatc gtcattaccg 3660
gtattcatca cagcttccat gcggttgaag ccgggttgct gggcaatccc tccatcggcg 3720
tcaacttcct gctgccgatt tgggcgatgg ccaacgtcgc tcagggcgga gcctgtctgg 3780
cggtgtggtt caaaaccaaa gatgcaaaaa ttaaagccat tactctgccc tcggcgtttt 3840
ccgccatgct gggcatcacc gaggcggcga tttttggtat taacctgcgc tttgtgaagc 3900
catttattgc ggcgctgatt ggtggtgcgg cgggcggcgc atgggtggta tctgtacacg 3960
tctacatgac cgcggtcggc ttgacagcga tccccggcat ggccatcgtg caggccagtt 4020
cgctgttgaa ctacattatc gggatggtta tcgcctttgg cgtcgccttt acggtctccc 4080
tggttttgaa atacaaaacg gacgctgaat aatgtctctt ccatcacgac tgcctgcgat 4140
tttgcaggcc gtaatgcagg gccagccgcg cgcgctggcc gatagccact atccgcgctg 4200
gcaccatgcg ccggtcaccg ggctgatgaa cgaccccaac ggctttatcg aatttgccgg 4260
acgctatcat ctgttttatc agtggaaccc gctcgcctgc gatcatacgt ttaagtgctg 4320
ggcgcactgg agttccatcg atctgctgca ctggcagcat gagcccattg cgctgatgcc 4380
ggacgaagag tatgaccgta acggctgcta ctccggcagc gcggtggata acaacggtac 4440
gcttaccctg tgctataccg gcaacgtgaa gtttgccgag ggagggcgaa ccgcctggca 4500
atgcctggca acggaaaacg ctgacggcac cttccgcaaa atcggtccgg tcctgccgct 4560
gccggagggc tacaccggcc acgtgcgcga cccaaaagtc tggcgacacg aagacctgtg 4620
gtacatggtg ctgggcgcgc aggatcggca aaagcgcggc aaggtgctgc tgttcagctc 4680
tgcggatctc catcagtgga cgagtatggg tgaaatcgcc ggccacggca tcaatggcct 4740
cgacgacgtc ggctatatgt gggagtgccc ggatcttttt ccactcggcg accagcatat 4800
tctaatctgc tgtccgcagg ggattgcccg tgaggaagag tgctacctga acacctaccc 4860
ggcagtatgg atggcgggcg agtttgatta cgctgctggc gctttcagac acggcgaact 4920
gcacgaactg gacgccgggt ttgagttcta cgccccgcaa accatgctta ccagtgatgg 4980
ccgtcgtctg ctggtcggct ggatgggcgt gccggagggc gaagagatgc ttcagccgac 5040
cctgaacaac ggctggatcc atcagatgac ctgcctgcgt gagctggagt ttatcaacgg 5100
tcagctctat cagcgtccgc tacgggaact gagcgccctg cgcggtgaag cgaacggctg 5160
gtcggggaac gccctgccgc tggcaccgat ggaaatcgat ttgcaaaccc gcgggggcga 5220
tatgttgagc ctcgattttg gcggcgtatt aacccttgag tgcgatgcca gcggactccg 5280
cctggcccga cgcagtctcg ccagtgacga gatgcattat cgttactggc gcggaaacgt 5340
ccgctcgctg cgtgttttca tcgaccagtc gagcgtggag attttcataa acggcggtga 5400
aggggtgatg agcagccgct acttcccggc ctgctccggt cagctaacat tctccggcat 5460
cacgccggac gcattctgct actggccgct gcgaacttgc atggtagaat aagcgttttg 5520
cttcaggctc atggcgtcgt aatgaaaacc aaacgcgtaa ccattaaaga tatagccgaa 5580
caggctggcg tctccaaagc gaccgccagc ttggtactga atggtcgtgg caaggagctg 5640
cgcgtggcgc aggaaacgcg tgagcgcgta ctgtcgattg cccgtaagca tcactatcag 5700
ccaagcattc atgcccgctc gctgcgcaac aaccgcagcc acaccatcgg gctggtggtg 5760
ccggagatca ccaaccacgg ctttgcggtc tttgcccatg agctggagat gctgtgccgc 5820
gaggcgggcg tccagctgtt gatctcttgt actgatgaaa accccggtca ggagagcgtg 5880
gtggtcaata atatgattgc ccgccaggtc gacgggatga tcgtcgcttc ctgtatgcac 5940
aacgatgccg actatctcaa actcagccaa cagctgccag tggtgctgtt tgaccggtgc 6000
cccaatgaaa gcgcgctgcc gctggtaatg accgattcga ttaccccaac ggcggaactg 6060
atttcccgca tcgcgcctca gcatagcgat gagttctggt ttttaggcgg tcaggcgcgt 6120
ctgtcgccct cccgcgatcg tctgaccggg ttcacgcagg gtttggctca ggcgggtatt 6180
gccctgcgcc cggaatgggt gatcaacggc aattaccacc ccagctccgg ctatgagatg 6240
tttgccgcac tctgcgcgcg ccttgggcgg ccgcctaagg cgctattcac cgccgcctgc 6300
gggctgctcg aaggggttct gcgctatatg agccagcacc atttactcga ttccgatatt 6360
catctgacga gctttgacga tcactatctt tatgattcgc tgtcgctgcg tatcgacact 6420
gtccagcagg ataatcgcca gctggcctgg cactgctacg atctgataag ccagctgatc 6480
gagggcgata cgcccgaaac gctacaacgc tacctgcccg caaccctgca gtttcggcat 6540
cagtaa 6546
<210> 82
<211> 4885
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 82
ctatattgct gaaggtacag gcgtttccat aactatttgc tcgcgttttt tactcaggaa 60
gaaaatgcca aatagcaaca tcaggcagac aatacccgaa attgcgaaga aaactgtctg 120
gtagcctgcg tggtcaaaga gtatcccagt cggcgttgaa agcagcacaa tcccaagcga 180
actggcaatt tgaaaaccaa tcagaaagat cgtcgacgac aggcgcttat caaagtttgc 240
cacgctgtat ttgaagacgg atatgacaca aagtggaacc tcaatggcat gtaacagctt 300
cactaatgaa ataatccagg ggttaacgaa caacgcgcag gaaaggatac gcaacgccat 360
aatcacaaca ccgataagta atgcattttt tggccctacc cgattcacaa agaaaggaat 420
aatcgccatg cacagcgctt cgagtaccac ctggaatgag ttgagataac catacaggcg 480
cgttcctaca tcgtgtgatt cgaataaacc tgcataaaag acaggaaaga gttgttgatc 540
aaaaatgtta tagaaagacc acgtccccac aataaatatg acgaaaaccc agaagtttcg 600
atccttgaaa actgcgataa aatcctcttt ttttacccct cccgcatccg ccgctatgca 660
ctggtgatcc ttatctttaa aacgcatgtt gatcatcata aatacagcgc caaatagcga 720
gaccaaccag aagttgatat ggggactgat actaaaaaat ataccggcaa agaacgcgcc 780
aatagcatag ccaaaagatc cccaggcgcg cgctgttcca tattcgaaat gaaaatttcg 840
cgccattttt tcggtgaagc tgtcaagcaa accgcatccc gccagatacc ccaggccaaa 900
aaagagcgcc cccagaatta gacctacaga aaaattgctt tgcagtaacg gttcataaac 960
gtaaatcata aacggtccgg tcaagaccag aatgaaactc atacaccaga tgagcggttt 1020
cttcagaccg agtttatcct gaacgatgcc gtagaacatc ataaatagaa tgctggtaaa 1080
ctggttgacc gaataaagtg tacctaattc cgtccctgtt aatcctagat gtcctttcag 1140
ccaaatagcg tataacgacc accacagcga ccaggaaata aaaaagagaa atgagtaact 1200
ggatgcaaaa cgatagtacg catttctgaa tggaatattc agtgccataa ttacctgcct 1260
gtcgttaaaa aattcatgtc ctatttagag ataagagcgg cctcgccgtt tacttctcac 1320
tttccagttc ttgtcgacat ggcagcgctg tcattgcccc tttcgctgtt actgcaagcg 1380
ctccgcaacg ttgagcgaga tcgataattc gtcgcatttc tctctcatct gtagataatc 1440
ccgtagagga cagacctgtg agtaacccgg caacgaacgc atctcccgcc ccagtgctat 1500
cgacacaatt cacagacatt ccagcaaaat ggtggacttg tcctcgataa cagaccacca 1560
ccccttctgc acctttagtc accaacagca tggcgatctc atactctttt gccagggcgc 1620
atatatcccg atcgttctgt gtttttccac tgataagtcg ccattcttct tccgagagct 1680
tgacgacatc cgccagttgt agcgcctgcc gcaaacacaa gcggagcaaa tgctcgtctt 1740
gccatagatc ttcacgaata ttgggatcga agctgacaaa acctccggca tgccggatcg 1800
ccgtcatcgc agtaaatgcg ctggtacgcg aaggctcggc agacaacgca attgaacaga 1860
gatgtaacca ttcgccatgt cgccagcagg gcaagtctgt cgtctctaaa aaaagatcgg 1920
cactggggcg gaccataaac gtaaatgaac gttctccttg atcgttcaga tcgacaagca 1980
ccgtggatgt ccggtgccat tcatcttgct tcagatacgt gatatcgaca ccctcagtta 2040
gcagcgttct ttgcattaac gcaccaaaag gatcatcacc gacccgacct ataaacccac 2100
ttgttccgcc taatctggcg attcccaccg caacgttagc tggcgcgccg ccaggacaag 2160
gcagtagccg cccgtctgat tctggcaaga gatctacgac cgcatcccct aaaacccata 2220
ctttggctga catttttttc ccttaaattc atctgactta cgcatagtga taaacctctt 2280
tttcgcaaaa tcgtcatgga tttactaaaa catgcatatt cgatcacaaa acgtcatagt 2340
taacgttaac atttgtgata ttcatcgcat ttatgaaagt aagggacttt atttttataa 2400
aagttaacgt taacaattca ccaaatttgc ttaaccagga tgattaaaat gacgcaatct 2460
cgattgcatg cggcgcaaaa cgcactagca aaacttcacg agcgccgagg taacactttc 2520
tatccccatt ttcacctcgc gcctcctgcc gggtggatga acgatccaaa cggcctgatc 2580
tggtttaacg atcgttatca cgcgttttat caacatcacc cgatgagcga acactggggg 2640
ccaatgcact ggggacatgc caccagcgac gatatgatcc actggcagca tgagcctatt 2700
gcgctagcgc caggagacga gaatgacaaa gacgggtgtt tttcaggtag tgctgtcgat 2760
gacaatggtg tcctctcact tatctacacc ggacacgtct ggctcgatgg tgcaggtaat 2820
gacgatgcaa ttcgcgaagt acaatgtctg gctaccagtc gggatggtat tcatttcgag 2880
aaacagggtg tgatcctcac tccaccagaa ggcatcatgc acttccgcga tcctaaagtg 2940
tggcgtgaag ccgacacatg gtggatggta gtcggggcga aagacccagg caacacgggg 3000
cagatcctgc tttatcgcgg cagttcattg cgtgaatgga ctttcgatcg cgtactggcc 3060
cacgctgatg cgggtgaaag ctatatgtgg gaatgtccgg actttttcag ccttggcgat 3120
cagcattatc tgatgttttc cccgcaggga atgaatgccg agggatacag ttatcgaaat 3180
cgctttcaaa gtggcgtaat acccggaatg tggtcgccag gacgactttt tgcacaatcc 3240
gggcatttta ctgaacttga taacgggcat gacttttatg caccacaaag ctttgtagcg 3300
aaggatggtc ggcgtattgt tatcggctgg atggatatgt gggaatcgcc aatgccctca 3360
aaacgtgaag gctgggcagg ctgcatgacg ctggcgcgcg agctatcaga gagcaatggc 3420
aaactcctac aacgcccggt acacgaagct gagtcgttac gccagcagca tcaatctatc 3480
tctccccgca caatcagcaa taaatatgtt ttgcaggaaa acgcgcaagc agttgagatt 3540
cagttgcagt gggcgctgaa gaacagtgat gccgaacatt acggattaca gctcggcgct 3600
ggaatgcggc tgtatattga taaccaatct gagcgacttg ttttgtggcg gtattaccca 3660
cacgagaatt tagatggcta ccgtagtatt cccctcccgc agggtgacat gctcgcccta 3720
aggatattta tcgatacatc atccgtggaa gtatttatta acgacgggga ggcggtgatg 3780
agtagccgaa tatatccgca gccagaagaa cgggaactgt cgctctatgc ctcccacgga 3840
gtggctgtgc tgcaacatgg agcactctgg caactgggtt aacataatat caggtggaac 3900
aacggatcaa cagcgggcaa gggatccgcg tcactcttcc cccttcacga ccttcaataa 3960
tatgcaatgc agcttcccgc ccgataatgt catgtggaag ctgaattgtg gtcagcggcg 4020
gtaaaaacag atgcccgacg ccaaccagat tatcaaagcc cattacggcg acatcctgcg 4080
ggatacgtac ccccttcgcc aaaagaacct gataagccac aaaggctgcg cgatcgttac 4140
cacatatcag aacatcaaaa tctggtttgc ccgatttgaa gtgggcattg agtaaacttg 4200
cgagatcggt gtagtgatca tcacctgttg ccatgtgaaa ttgtttcacc tcagccagat 4260
ctcgtccagc atcacgccag gcctgctcaa atccctgccg acgataccct gttgccaacg 4320
cactttccgg tagccagaag cataacggtt gacgatagcc cgccgcgagc aaatgctgtg 4380
ttgattcata ttgtgcagtg taatcatcag ggatataact gggtaacgct gggtcatccg 4440
ccacacagtt cgccaataca atattttcac catacagaga ctcaggcagc gtgatatgtc 4500
gcagccccat tgtagtatag ataatgccat ccggacggtg ggcaagcagc tgacgtgccg 4560
cgcgggcagc gtcatcttca gaaaaaatat tgattaaaaa actattccag ccgaactcgc 4620
tggcggtttg ctcaatggca agcagaatat caacagagaa aggagtggta gcagtgtcct 4680
gcgccagcac ggcgagagtc gacggcttac gtccttgagc gcgcatctta cgggcggaaa 4740
gatcaggaac ataattcagg gtctggattg cctgcaatac gcggtcacgc gttgcaggac 4800
gcacagattc tgcattatgc atcacccggg agactgtcat catcgacact cccgccaggc 4860
gtgcgacatc ctttaatgaa gccat 4885
<210> 83
<211> 3771
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 83
atgaaaataa agaacattct actcaccctt tgcacctcac tcctgcttac caacgttgct 60
gcacacgcca aagaagtcaa aataggtatg gcgattgatg atctccgtct tgaacgctgg 120
caaaaagatc gagatatctt tgtgaaaaag gcagaatctc tcggcgcgaa agtatttgta 180
cagtctgcaa atggcaatga agaaacacaa atgtcgcaga ttgaaaacat gataaaccgg 240
ggtgtcgatg ttcttgtcat tattccgtat aacggtcagg tattaagtaa cgttgtaaaa 300
gaagccaaac aagaaggcat taaagtatta gcttacgacc gtatgattaa cgatgcggat 360
atcgattttt atatttcttt cgataacgaa aaagtcggtg aactgcaggc aaaagccctg 420
gtcgatattg ttccgcaagg taattacttc ctgatgggcg gctcgccggt agataacaac 480
gccaagctgt tccgcgccgg acaaatgaaa gtgttaaaac cttacgttga ttccggaaaa 540
attaaagtcg ttggtgacca atgggttgat ggctggttac cggaaaacgc attgaaaatt 600
atggaaaacg cgctaaccgc caataataac aaaattgatg ctgtagttgc ctcaaacgat 660
gccaccgcag gtggggcaat tcaggcatta agcgcgcaag gtttatcagg gaaagtagca 720
atctccggcc aggatgcgga tctcgcaggt attaaacgta ttgctgccgg tacgcaaact 780
atgacggtgt ataaacctat tacgttgttg gcaaatactg ccgcagaaat tgccgttgag 840
ttgggcaatg gtcaggaacc aaaagcagat accacactga ataatggcct gaaagatgtc 900
ccctcccgcc tcctgacacc gatcgatgtg aataaaaaca acatcaaaga tacggtaatt 960
aaagacggat tccacaaaga gagcgagctg taagcgttac gccccagcgc ggagcggggg 1020
cgtgatttct ctccatgccg cgtgaatgaa ttggcttagg tggagtcgtt atgccttatc 1080
tacttgaaat gaagaacatt accaaaacct tcggcagtgt gaaggcgatt gataacgtct 1140
gcttgcggtt gaatgctggc gaaatcgtct cactttgtgg ggaaaatggg tctggtaaat 1200
caacgctgat gaaagtgctg tgtggtattt atccccatgg ctcctacgaa ggcgaaatta 1260
tttttgcggg agaagagatt caggcgagtc acatccgcga taccgaacgc aaaggtatcg 1320
ccatcattca tcaggaattg gccctggtga aagaattgac cgtgctggaa aatatcttcc 1380
tgggtaacga aataacccac aatggcatta tggattatga cctgatgacg ctacgctgtc 1440
agaagctgct cgcacaggtc agtttatcca tttcacctga tacccgcgtt ggcgatttag 1500
ggcttgggca acaacaactg gttgaaattg ccaaggcact taataaacag gtgcgcttgt 1560
taattctcga tgaaccgaca gcctcattaa ctgagcagga aacgtcgatt ttactggata 1620
ttattcgcga tctacaacag cacggtatcg cctgtattta tatttcgcac aaactcaacg 1680
aagtcaaagc gatttccgat acgatttgcg ttattcgcga cggacagcac attggtacgc 1740
gtgatgctgc cggaatgagt gaagacgata ttatcaccat gatggtcggg cgagagttaa 1800
ccgcgcttta ccctaatgaa ccacatacca ccggagatga aatattacgt attgaacatc 1860
tgacggcatg gcatccggtt aatcgtcata ttaaacgagt taatgatgtc tcgttttccc 1920
tgaaacgtgg cgaaatattg ggtattgccg gactcgttgg tgccggacgt accgagacca 1980
ttcagtgcct gtttggtgtg tggcccggac aatgggaagg aaaaatttat attgatggca 2040
aacaggtaga tattcgtaac tgtcagcaag ccatcgccca ggggattgcg atggtccccg 2100
aagacagaaa gcgcgacggc atcgttccgg taatggcggt tggtaaaaat attaccctcg 2160
ccgcactcaa taaatttacc ggtggcatta gccagcttga tgacgcggca gagcaaaaat 2220
gtattctgga atcaatccag caactcaaag ttaaaacgtc gtcccccgac cttgctattg 2280
gacgtttgag cggcggcaat cagcaaaaag cgatcctcgc tcgctgtctg ttacttaacc 2340
cgcgcattct cattcttgat gaacccacca ggggtatcga tattggcgcg aaatacgaga 2400
tctacaaatt aattaaccaa ctcgtccagc agggtattgc cgttattgtc atctcttccg 2460
aattacctga agtgctcggc cttagcgatc gtgtactggt gatgcatgaa gggaaactaa 2520
aagccaacct gataaatcat aacctgactc aggagcaggt gatggaagcc gcattgagga 2580
gcgaacatca tgtcgaaaag caatccgtct gaagtgaaat tggccgtacc gacatccggt 2640
ggcttctccg ggctgaaatc actgaatttg caggtcttcg tgatgattgc agctatcatc 2700
gcaatcatgc tgttctttac ctggaccacc gatggtgcct acttaagcgc ccgtaacgtc 2760
tccaacctgt tacgccagac cgcgattacc ggcatcctcg cggtaggaat ggtgttcgtc 2820
ataatttctg ctgaaatcga cctttccgtc ggctcaatga tggggctgtt aggtggcgtc 2880
gcggcgattt gtgacgtctg gttaggctgg cctttgccac ttaccatcat tgtgacgctg 2940
gttctgggac tgcttctcgg tgcctggaac ggatggtggg tcgcgtaccg taaagtccct 3000
tcatttattg tcaccctcgc gggcatgttg gcatttcgcg gcatactcat tggcatcacc 3060
aacggcacga ctgtatcccc caccagcgcc gcgatgtcac aaattgggca aagctatctc 3120
cccgccagta ccggcttcat cattggcgcg cttggcttaa tggcttttgt tggttggcaa 3180
tggcgcggaa gaatgcgccg tcaggctttg ggtttacagt ctccggcctc taccgcagta 3240
gtcggtcgcc aggctttaac cgctatcatc gtattaggcg caatctggct gttgaatgat 3300
taccgtggcg ttcccactcc tgttctgctg ctgacgttgc tgttactcgg cggaatgttt 3360
atggcaacgc ggacggcatt tggacgacgc atttatgcca tcggcggcaa tctggaagca 3420
gcacgtctct ccgggattaa cgttgaacgc accaaacttg ccgtgttcgc gattaacgga 3480
ttaatggtag ccatcgccgg attaatcctt agttctcgac ttggcgctgg ttcaccttct 3540
gcgggaaata tcgccgaact ggacgcaatt gcagcatgcg tgattggcgg caccagcctg 3600
gctggcggtg tgggaagcgt tgccggagca gtaatggggg catttatcat ggcttcactg 3660
gataacggca tgagtatgat ggatgtaccg accttctggc agtatatcgt taaaggtgcg 3720
attctgttgc tggcagtatg gatggactcc gcaaccaaac gccgttcttg a 3771
<210> 84
<211> 1395
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 84
atgcctgacg ctaaaaaaca ggggcggtca aacaaggcaa tgacgttttt cgtctgcttc 60
cttgccgctc tggcgggatt actctttggc ctggatatcg gtgtaattgc tggcgcactg 120
ccgtttattg cagatgaatt ccagattact tcgcacacgc aagaatgggt cgtaagctcc 180
atgatgttcg gtgcggcagt cggtgcggtg ggcagcggct ggctctcctt taaactcggg 240
cgcaaaaaga gcctgatgat cggcgcaatt ttgtttgttg ccggttcgct gttctctgcg 300
gctgcgccaa acgttgaagt actgattctt tcccgcgttc tactggggct ggcggtgggt 360
gtggcctctt ataccgcacc gctgtacctc tctgaaattg cgccggaaaa aattcgtggc 420
agtatgatct cgatgtatca gttgatgatc actatcggga tcctcggtgc ttatctttct 480
gataccgcct tcagctacac cggtgcatgg cgctggatgc tgggtgtgat tatcatcccg 540
gcaattttgc tgctgattgg tgtcttcttc ctgccagaca gcccacgttg gtttgccgcc 600
aaacgccgtt ttgttgatgc cgaacgcgtg ctgctacgcc tgcgtgacac cagcgcggaa 660
gcgaaacgcg aactggatga aatccgtgaa agtttgcagg ttaaacagag tggctgggcg 720
ctgtttaaag agaacagcaa cttccgccgc gcggtgttcc ttggcgtact gttgcaggta 780
atgcagcaat tcaccgggat gaacgtcatc atgtattacg cgccgaaaat cttcgaactg 840
gcgggttata ccaacactac cgagcaaatg tgggggaccg tgattgtcgg cctgaccaac 900
gtacttgcca cctttatcgc aatcggcctt gttgaccgct ggggacgtaa accaacgcta 960
acgctgggct tcctggtgat ggctgctggc atgggcgtac tcggtacaat gatgcatatc 1020
ggtattcact ctccgtcggc gcagtatttc gccatcgcca tgctgctgat gtttattgtc 1080
ggttttgcca tgagtgccgg tccgctgatt tgggtactgt gctccgaaat tcagccgctg 1140
aaaggccgcg attttggcat cacctgctcc actgccacca actggattgc caacatgatc 1200
gttggcgcaa cgttcctgac catgctcaac acgctgggta acgccaacac cttctgggtg 1260
tatgcggctc tgaacgtact gtttatcctg ctgacattgt ggctggtacc ggaaaccaaa 1320
cacgtttcgc tggaacatat tgaacgtaat ctgatgaaag gtcgtaaact gcgcgaaata 1380
ggcgctcacg attaa 1395
<210> 85
<211> 1422
<212> DNA
<213> 运动发酵单胞菌(Zymomonas mobilis)
<400> 85
atgagttctg aaagtagtca gggtctagtc acgcgactag ccctaatcgc tgctataggc 60
ggcttgcttt tcggttacga ttcagcggtt atcgctgcaa tcggtacacc ggttgatatc 120
cattttattg cccctcgtca cctgtctgct acggctgcgg cttccctttc tgggatggtc 180
gttgttgctg ttttggtcgg ttgtgttacc ggttctttgc tgtctggctg gattggtatt 240
cgcttcggtc gtcgcggcgg attgttgatg agttccattt gtttcgtcgc cgccggtttt 300
ggtgctgcgt taaccgaaaa attatttgga accggtggtt cggctttaca aattttttgc 360
tttttccggt ttcttgccgg tttaggtatc ggtgtcgttt caaccttgac cccaacctat 420
attgctgaaa ttcgtccgcc agacaaacgt ggtcagatgg tttctggtca gcagatggcc 480
attgtgacgg gtgctttaac cggttatatc tttacctggt tactggctca tttcggttct 540
atcgattggg ttaatgccag tggttggtgc tggtctccgg cttcagaagg cctgatcggt 600
attgccttct tattgctgct gttaaccgca ccggatacgc cgcattggtt ggtgatgaag 660
ggacgtcatt ccgaggctag caaaatcctt gctcgtctgg aaccgcaagc cgatcctaat 720
ctgacgattc aaaagattaa agctggcttt gataaagcca tggacaaaag cagcgcaggt 780
ttgtttgctt ttggtatcac cgttgttttt gccggtgtat ccgttgctgc cttccagcag 840
ttagtcggta ttaacgccgt gctgtattat gcaccgcaga tgttccagaa tttaggtttt 900
ggagctgata cggcattatt gcagaccatc tctatcggtg ttgtgaactt catcttcacc 960
atgattgctt cccgtgttgt tgaccgcttc ggccgtaaac ctctgcttat ttggggtgct 1020
ctcggtatgg ctgcaatgat ggctgtttta ggctgctgtt tctggttcaa agtcggtggt 1080
gttttgcctt tggcttctgt gcttctttat attgcagtct ttggtatgtc atggggccct 1140
gtctgctggg ttgttctgtc agaaatgttc ccgagttcca tcaagggcgc agctatgcct 1200
atcgctgtta ccggacaatg gttagctaat atcttggtta acttcctgtt taaggttgcc 1260
gatggttctc cagcattgaa tcagactttc aaccacggtt tctcctatct cgttttcgca 1320
gcattaagta tcttaggtgg cttgattgtt gctcgcttcg tgccggaaac caaaggtcgg 1380
agcctggatg aaatcgagga gatgtggcgc tcccagaagt ag 1422
<210> 86
<211> 984
<212> DNA
<213> 运动发酵单胞菌(Zymomonas mobilis)
<400> 86
atggaaattg ttgcgattga catcggtgga acgcatgcgc gtttctctat tgcggaagta 60
agcaatggtc gggttctttc tcttggagaa gaaacaactt ttaaaacggc agaacatgct 120
agcttgcagt tagcttggga acgtttcggt gaaaaactgg gtcgtcctct gccacgtgcc 180
gcagctattg catgggctgg cccggttcat ggtgaagttt taaaacttac caataaccct 240
tgggtattaa gaccagctac tctgaatgaa aagctggaca tcgatacgca tgttctgatc 300
aatgacttcg gcgcggttgc ccacgcggtt gcgcatatgg attcttctta tctggatcat 360
atttgtggtc ctgatgaagc gcttcctagc gatggtgtta tcactattct tggtccggga 420
acgggcttgg gtgttgccca tctgttgcgg actgaaggcc gttatttcgt catcgaaact 480
gaaggcggtc atatcgactt tgctccgctt gacagacttg aagacaaaat tctggcacgt 540
ttacgtgaac gtttccgccg cgtttctatc gaacgcatta tttctggccc gggtcttggt 600
aatatctacg aagcactggc tgccattgaa ggcgttccgt tcagcttgct ggatgatatt 660
aaattatggc agatggcttt ggaaggtaaa gacaaccttg ctgaagccgc tttggatcgc 720
ttctgcttga gccttggcgc tatcgctggt gatcttgctt tggcacaggg tcgaaccagt 780
gttgttattg gcggtggtgt cggtcttcgt atcgcttccc atttgccaga atctggtttc 840
cgtcagcgct ttgtttcaaa aggacgcttt gaacgcgtca tgtccaagat tccggttaag 900
ttgattactt atccgcagcc tggactgttg ggtgcgcagc tgcctatgcc aacaaatatt 960
ctgaagttga ataatatttt ttaa 984
<210> 87
<211> 966
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 87
atgacaaagt atgcattagt cggtgatgtg ggcggcacca acgcacgtct tgctctgtgt 60
gatattgcca gtggtgaaat ctcgcaggct aagacctatt cagggcttga ttaccccagc 120
ctcgaagcgg tcattcgcgt ttatcttgaa gaacataagg tcgaggtgaa agacggctgt 180
attgccatcg cttgcccaat taccggtgac tgggtggcga tgaccaacca tacctgggcg 240
ttctcaattg ccgaaatgaa aaagaatctc ggttttagcc atctggaaat tattaacgat 300
tttaccgctg tatcgatggc gatcccgatg ctgaaaaaag agcatctgat tcagtttggt 360
ggcgcagaac cggtcgaagg taagcctatt gcggtttacg gtgccggaac ggggcttggg 420
gttgcgcatc tggtccatgt cgataagcgt tgggtaagct tgccaggcga aggcggtcac 480
gttgattttg cgccgaatag tgaagaagag gccattatcc tcgaaatatt gcgtgcggaa 540
attggtcatg tttcggcgga gcgcgtgctt tctggccctg ggctggtgaa tttgtatcgc 600
gcaattgtga aagctgacaa ccgcctgcca gaaaatctca agccaaaaga tattaccgaa 660
cgcgcgctgg ctgacagctg caccgattgc cgccgcgcat tgtcgctgtt ttgcgtcatt 720
atgggccgtt ttggcggcaa tctggcgctc aatctcggga catttggcgg cgtgtttatt 780
gcgggcggta tcgtgccgcg cttccttgag ttcttcaaag cctccggttt ccgtgccgca 840
tttgaagata aagggcgctt taaagaatat gtccatgata ttccggtgta tctcatcgtc 900
catgacaatc cgggccttct cggttccggt gcacatttac gccagacctt aggtcacatt 960
ctgtaa 966
<210> 88
<211> 2547
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 88
ttgtacctct atattgagac tctgaaacag agactggatg ccataaatca attgcgtgtg 60
gatcgcgcgc ttgctgctat ggggcctgca ttccaacagg tctacagtct actgccgaca 120
ttgttgcact atcaccatcc gctaatgccg ggttaccttg atggtaacgt tcccaaaggc 180
atttgccttt acacgcctga tgaaactcaa cgccactacc tgaacgagct tgaactgtat 240
cgtggaatgt cagtacagga tccgccgaaa ggtgagcttc caattactgg tgtatacacc 300
atgggcagca cctcgtccgt agggcaaagt tgttcctctg acctggatat ctgggtctgt 360
catcaatcct ggctcgatag cgaagagcgc caattgctac aacgtaaatg tagcctgctg 420
gaaaactggg ccgcctcgct gggtgtggaa gtcagcttct tcctgattga tgaaaaccgc 480
ttccgtcata atgaaagcgg cagcctgggg ggcgaagatt gtggctccac ccagcatata 540
ctgctgcttg acgaatttta tcgtaccgcc gtgcgtctcg ccggtaagcg tattctgtgg 600
aatatggtgc cgtgcgacga agaagagcat tacgacgact atgtgatgac gctttacgcg 660
cagggcgtgc tgacgccaaa tgaatggctg gatctcggtg gcttaagctc gctttctgct 720
gaagagtact ttggtgccag cctttggcag ctctacaaga gtatcgattc cccatacaaa 780
gcggtactga aaacactgct gctggaagcc tattcctggg aatacccgaa cccacgtctg 840
ctggcgaaag atatcaaaca gcgtttgcac gacggcgaga ttgtatcgtt tggtctcgat 900
ccatactgca tgatgctgga gcgtgttact gaatacctga cggcgattga agattttacc 960
cgtctggatt tagtacgtcg ctgcttctat ttaaaagtgt gcgaaaagct cagccgtgaa 1020
cgcgcctgcg taggctggcg tcgcgcagtg ttgagccagt tagtgagcga gtggggttgg 1080
gacgaagctc gtctggcaat gctcgataac cgcgctaact ggaagattga tcaggtgcgt 1140
gaggcgcaca acgagttgct cgacgcgatg atgcagagct accgtaatct gatccgcttt 1200
gcgcgtcgca ataaccttag cgtctccgcc agtccgcagg atatcggcgt gctgacgcgt 1260
aagctgtatg ccgcgtttga agcattacca ggtaaagtga cgctggtaaa cccgcagatt 1320
tcacccgatc tctcggaacc gaatctgacc tttatttatg tgccgccggg ccgggctaac 1380
cgttcaggtt ggtatctgta taaccgcgcg ccaaatattg agtcgatcat cagccatcag 1440
ccgctggaat ataaccgtta cctgaataaa ctggtggcgt gggcatggtt taacggcctg 1500
ctgacctcgc gcacccgttt gtatattaaa ggtaacggca ttgtcgattt gcctaagttg 1560
caggagatgg tcgccgacgt gtcgcaccat ttcccgctgc gcttacctgc accgacaccg 1620
aaggcgctct acagcccgtg tgagatccgc catctggcga ttatcgttaa cctggaatat 1680
gacccgacag cggcgttccg caatcaggtg gtgcatttcg atttccgtaa gctggatgtc 1740
ttcagctttg gcgagaatca aaattgcctg gtaggtagcg ttgacctgct gtaccgcaac 1800
tcgtggaacg aagtgcgtac gctgcacttc aacggcgagc aatcgatgat cgaagccctg 1860
aaaactattc tcggcaaaat gcatcaggac gccgcaccgc cagatagcgt ggaagtcttc 1920
tgttatagcc agcatctgcg cggcttaatt cgtactcgcg tgcagcaact ggtttctgag 1980
tgtattgaat tgcgtctttc cagcacccgc caggaaaccg ggcgtttcaa ggcgctgcgc 2040
gtttctggtc aaacctgggg gttgttcttc gaacgcctga atgtatcggt acagaaactg 2100
gaaaacgcca tcgagtttta tggcgcgatt tcgcataaca aactgcacgg cctgtcagtg 2160
caggttgaaa ccaatcacgt caaattaccg gcggtggtgg acggctttgc cagcgaaggg 2220
atcatccagt tctttttcga agaaacgcaa gacgagaatg gctttaatat ctacattctc 2280
gacgaaagca accgggttga ggtatatcac cactgcgaag gcagcaaaga ggagctggta 2340
cgtgacgtca gtcgcttcta ctcgtcatcg catgaccgtt ttacctacgg ctcaagcttc 2400
atcaacttca acctgccgca gttctatcag attgtgaagg ttgatggtcg tgaacaggtg 2460
attccgttcc gcacaaaatc tatcggtaac atgccgcctg ccaatcagga tcacgatacg 2520
ccgctattac agcaatattt ttcgtga 2547
<210> 89
<211> 210
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 89
Met Val Leu Gly Lys Pro Gln Thr Asp Pro Thr Leu Glu Trp Phe Leu
1 5 10 15
Ser His Cys His Ile His Lys Tyr Pro Ser Lys Ser Thr Leu Ile His
20 25 30
Gln Gly Glu Lys Ala Glu Thr Leu Tyr Tyr Ile Val Lys Gly Ser Val
35 40 45
Ala Val Leu Ile Lys Asp Glu Glu Gly Lys Glu Met Ile Leu Ser Tyr
50 55 60
Leu Asn Gln Gly Asp Phe Ile Gly Glu Leu Gly Leu Phe Glu Glu Gly
65 70 75 80
Gln Glu Arg Ser Ala Trp Val Arg Ala Lys Thr Ala Cys Glu Val Ala
85 90 95
Glu Ile Ser Tyr Lys Lys Phe Arg Gln Leu Ile Gln Val Asn Pro Asp
100 105 110
Ile Leu Met Arg Leu Ser Ala Gln Met Ala Arg Arg Leu Gln Val Thr
115 120 125
Ser Glu Lys Val Gly Asn Leu Ala Phe Leu Asp Val Thr Gly Arg Ile
130 135 140
Ala Gln Thr Leu Leu Asn Leu Ala Lys Gln Pro Asp Ala Met Thr His
145 150 155 160
Pro Asp Gly Met Gln Ile Lys Ile Thr Arg Gln Glu Ile Gly Gln Ile
165 170 175
Val Gly Cys Ser Arg Glu Thr Val Gly Arg Ile Leu Lys Met Leu Glu
180 185 190
Asp Gln Asn Leu Ile Ser Ala His Gly Lys Thr Ile Val Val Tyr Gly
195 200 205
Thr Arg
210
<210> 90
<211> 210
<212> PRT
<213> 人工的
<220>
<223> Crp*突变体
<400> 90
Met Val Leu Gly Lys Pro Gln Thr Asp Pro Thr Leu Glu Trp Phe Leu
1 5 10 15
Ser His Cys His Ile His Lys Tyr Pro Ser Lys Ser Thr Leu Ile His
20 25 30
Gln Gly Glu Lys Ala Glu Thr Leu Tyr Tyr Ile Val Lys Gly Ser Val
35 40 45
Ala Val Leu Ile Lys His Glu Glu Gly Lys Glu Met Ile Leu Ser Tyr
50 55 60
Leu Asn Gln Gly Asp Phe Ile Gly Glu Leu Gly Leu Phe Glu Glu Gly
65 70 75 80
Gln Glu Arg Ser Ala Trp Val Arg Ala Lys Thr Ala Cys Glu Val Ala
85 90 95
Glu Ile Ser Tyr Lys Lys Phe Arg Gln Leu Ile Gln Val Asn Pro Asp
100 105 110
Ile Leu Met Arg Leu Ser Ala Gln Met Ala Arg Arg Leu Gln Val Thr
115 120 125
Ser Glu Lys Val Gly Asn Leu Ala Phe Leu Asp Val Thr Gly Arg Ile
130 135 140
Ala Gln Thr Leu Leu Asn Leu Ala Lys Gln Pro Asp Ala Met Thr His
145 150 155 160
Pro Asp Gly Met Gln Ile Lys Ile Thr Arg Gln Glu Ile Gly Gln Ile
165 170 175
Val Gly Cys Ser Arg Glu Thr Val Gly Arg Ile Leu Lys Met Leu Glu
180 185 190
Asp Gln Asn Leu Ile Ser Ala His Gly Lys Thr Ile Val Val Tyr Gly
195 200 205
Thr Arg
210
<210> 91
<211> 210
<212> PRT
<213> 人工的
<220>
<223> Crp*突变体
<400> 91
Met Val Leu Gly Lys Pro Gln Thr Asp Pro Thr Leu Glu Trp Phe Leu
1 5 10 15
Ser His Cys His Ile His Lys Tyr Pro Ser Lys Ser Thr Leu Ile His
20 25 30
Gln Gly Glu Lys Ala Glu Thr Leu Tyr Tyr Ile Val Lys Gly Ser Val
35 40 45
Ala Val Leu Ile Lys Asp Glu Glu Gly Lys Glu Met Ile Leu Phe Tyr
50 55 60
Leu Asn Gln Gly Asp Phe Ile Gly Glu Leu Gly Leu Phe Glu Glu Gly
65 70 75 80
Gln Glu Arg Ser Ala Trp Val Arg Ala Lys Thr Ala Cys Glu Val Ala
85 90 95
Glu Ile Ser Tyr Lys Lys Phe Arg Gln Leu Ile Gln Val Asn Pro Asp
100 105 110
Ile Leu Met Arg Leu Ser Ala Gln Met Ala Arg Arg Leu Gln Val Thr
115 120 125
Ser Glu Lys Val Gly Asn Leu Ala Phe Leu Asp Val Thr Gly Arg Ile
130 135 140
Ala Gln Thr Leu Leu Asn Leu Ala Lys Gln Pro Asp Ala Met Thr His
145 150 155 160
Pro Asp Gly Met Gln Ile Lys Ile Thr Arg Gln Glu Ile Gly Gln Ile
165 170 175
Val Gly Cys Ser Arg Glu Thr Val Gly Arg Ile Leu Lys Met Leu Glu
180 185 190
Asp Gln Asn Leu Ile Ser Ala His Gly Lys Thr Ile Val Val Tyr Gly
195 200 205
Thr Arg
210
<210> 92
<211> 210
<212> PRT
<213> 人工的
<220>
<223> Crp*突变体
<400> 92
Met Val Leu Gly Lys Pro Gln Thr Asp Pro Thr Leu Glu Trp Phe Leu
1 5 10 15
Ser His Cys His Ile His Lys Tyr Pro Ser Lys Ser Thr Leu Ile His
20 25 30
Gln Gly Glu Lys Ala Glu Thr Leu Tyr Tyr Ile Val Lys Gly Ser Val
35 40 45
Ala Val Leu Ile Lys Asp Glu Glu Gly Lys Glu Met Ile Leu Ser Tyr
50 55 60
Leu Asn Gln Gly Asp Phe Ile Gly Glu Leu Gly Leu Phe Glu Glu Gly
65 70 75 80
Gln Glu Arg Ser Ala Trp Val Arg Ala Lys Thr Ala Cys Glu Val Ala
85 90 95
Glu Ile Ser Tyr Lys Lys Phe Arg Gln Leu Ile Gln Val Asn Pro Asp
100 105 110
Ile Leu Met Arg Leu Ser Ala Gln Met Ala Arg Arg Leu Gln Val Thr
115 120 125
Ser Glu Lys Val Gly Asn Leu Ala Phe Leu Asp Val Thr Asp Arg Ile
130 135 140
Ala Gln Thr Leu Leu Asn Leu Ala Lys Gln Pro Asp Ala Met Thr His
145 150 155 160
Pro Asp Gly Met Gln Ile Lys Ile Thr Arg Gln Glu Ile Gly Gln Ile
165 170 175
Val Gly Cys Ser Arg Glu Thr Val Gly Arg Ile Leu Lys Met Leu Glu
180 185 190
Asp Gln Asn Leu Ile Ser Ala His Gly Lys Thr Ile Val Val Tyr Gly
195 200 205
Thr Arg
210
<210> 93
<211> 210
<212> PRT
<213> 人工的
<220>
<223> Crp*突变体
<400> 93
Met Val Leu Gly Lys Pro Gln Thr Asp Pro Thr Leu Glu Trp Phe Leu
1 5 10 15
Ser His Cys His Ile His Lys Tyr Pro Ser Lys Ser Thr Leu Ile His
20 25 30
Gln Gly Glu Lys Ala Glu Thr Leu Tyr Tyr Ile Val Lys Gly Ser Val
35 40 45
Ala Val Leu Ile Lys Asp Glu Glu Gly Lys Glu Met Ile Leu Ser Tyr
50 55 60
Leu Asn Gln Gly Asp Phe Ile Gly Glu Leu Gly Leu Phe Glu Glu Gly
65 70 75 80
Gln Glu Arg Ser Ala Trp Val Arg Ala Lys Thr Ala Cys Glu Val Ala
85 90 95
Glu Ile Ser Tyr Lys Lys Phe Arg Gln Leu Ile Gln Val Asn Pro Asp
100 105 110
Ile Leu Met Arg Leu Ser Ala Gln Met Ala Arg Arg Leu Gln Val Thr
115 120 125
Ser Glu Lys Val Gly Asn Leu Ala Phe Leu Asp Val Thr Gly Asp Ile
130 135 140
Ala Gln Thr Leu Leu Asn Leu Ala Lys Gln Pro Asp Ala Met Thr His
145 150 155 160
Pro Asp Gly Met Gln Ile Lys Ile Thr Arg Gln Glu Ile Gly Gln Ile
165 170 175
Val Gly Cys Ser Arg Glu Thr Val Gly Arg Ile Leu Lys Met Leu Glu
180 185 190
Asp Gln Asn Leu Ile Ser Ala His Gly Lys Thr Ile Val Val Tyr Gly
195 200 205
Thr Arg
210
<210> 94
<211> 2932
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 94
atgcgaattg gcataccaag agaacggtta accaatgaaa cccgtgttgc agcaacgcca 60
aaaacagtgg aacagctgct gaaactgggt tttaccgtcg cggtagagag cggcgcgggt 120
caactggcaa gttttgacga taaagcgttt gtgcaagcgg gcgctgaaat tgtagaaggg 180
aatagcgtct ggcagtcaga gatcattctg aaggtcaatg cgccgttaga tgatgaaatt 240
gcgttactga atcctgggac aacgctggtg agttttatct ggcctgcgca gaatccggaa 300
ttaatgcaaa aacttgcgga acgtaacgtg accgtgatgg cgatggactc tgtgccgcgt 360
atctcacgcg cacaatcgct ggacgcacta agctcgatgg cgaacatcgc cggttatcgc 420
gccattgttg aagcggcaca tgaatttggg cgcttcttta ccgggcaaat tactgcggcc 480
gggaaagtgc caccggcaaa agtgatggtg attggtgcgg gtgttgcagg tctggccgcc 540
attggcgcag caaacagtct cggcgcgatt gtgcgtgcat tcgacacccg cccggaagtg 600
aaagaacaag ttcaaagtat gggcgcggaa ttcctcgagc tggattttaa agaggaagct 660
ggcagcggcg atggctatgc caaagtgatg tcggacgcgt tcatcaaagc ggaaatggaa 720
ctctttgccg cccaggcaaa agaggtcgat atcattgtca ccaccgcgct tattccaggc 780
aaaccagcgc cgaagctaat tacccgtgaa atggttgact ccatgaaggc gggcagtgtg 840
attgtcgacc tggcagccca aaacggcggc aactgtgaat acaccgtgcc gggtgaaatc 900
ttcactacgg aaaatggtgt caaagtgatt ggttataccg atcttccggg ccgtctgccg 960
acgcaatcct cacagcttta cggcacaaac ctcgttaatc tgctgaaact gttgtgcaaa 1020
gagaaagacg gcaatatcac tgttgatttt gatgatgtgg tgattcgcgg cgtgaccgtg 1080
atccgtgcgg gcgaaattac ctggccggca ccgccgattc aggtatcagc tcagccgcag 1140
gcggcacaaa aagcggcacc ggaagtgaaa actgaggaaa aatgtacctg ctcaccgtgg 1200
cgtaaatacg cgttgatggc gctggcaatc attctttttg gctggatggc aagcgttgcg 1260
ccgaaagaat tccttgggca cttcaccgtt ttcgcgctgg cctgcgttgt cggttattac 1320
gtggtgtgga atgtatcgca cgcgctgcat acaccgttga tgtcggtcac caacgcgatt 1380
tcagggatta ttgttgtcgg agcactgttg cagattggcc agggcggctg ggttagcttc 1440
cttagtttta tcgcggtgct tatagccagc attaatattt tcggtggctt caccgtgact 1500
cagcgcatgc tgaaaatgtt ccgcaaaaat taaggggtaa catatgtctg gaggattagt 1560
tacagctgca tacattgttg ccgcgatcct gtttatcttc agtctggccg gtctttcgaa 1620
acatgaaacg tctcgccagg gtaacaactt cggtatcgcc gggatggcga ttgcgttaat 1680
cgcaaccatt tttggaccgg atacgggtaa tgttggctgg atcttgctgg cgatggtcat 1740
tggtggggca attggtatcc gtctggcgaa gaaagttgaa atgaccgaaa tgccagaact 1800
ggtggcgatc ctgcatagct tcgtgggtct ggcggcagtg ctggttggct ttaacagcta 1860
tctgcatcat gacgcgggaa tggcaccgat tctggtcaat attcacctga cggaagtgtt 1920
cctcggtatc ttcatcgggg cggtaacgtt cacgggttcg gtggtggcgt tcggcaaact 1980
gtgtggcaag atttcgtcta aaccattgat gctgccaaac cgtcacaaaa tgaacctggc 2040
ggctctggtc gtttccttcc tgctgctgat tgtatttgtt cgcacggaca gcgtcggcct 2100
gcaagtgctg gcattgctga taatgaccgc aattgcgctg gtattcggct ggcatttagt 2160
cgcctccatc ggtggtgcag atatgccagt ggtggtgtcg atgctgaact cgtactccgg 2220
ctgggcggct gcggctgcgg gctttatgct cagcaacgac ctgctgattg tgaccggtgc 2280
gctggtcggt tcttcggggg ctatcctttc ttacattatg tgtaaggcga tgaaccgttc 2340
ctttatcagc gttattgcgg gtggtttcgg caccgacggc tcttctactg gcgatgatca 2400
ggaagtgggt gagcaccgcg aaatcaccgc agaagagaca gcggaactgc tgaaaaactc 2460
ccattcagtg atcattactc cggggtacgg catggcagtc gcgcaggcgc aatatcctgt 2520
cgctgaaatt actgagaaat tgcgcgctcg tggtattaat gtgcgtttcg gtatccaccc 2580
ggtcgcgggg cgtttgcctg gacatatgaa cgtattgctg gctgaagcaa aagtaccgta 2640
tgacatcgtg ctggaaatgg acgagatcaa tgatgacttt gctgataccg ataccgtact 2700
ggtgattggt gctaacgata cggttaaccc ggcggcgcag gatgatccga agagtccgat 2760
tgctggtatg cctgtgctgg aagtgtggaa agcgcagaac gtgattgtct ttaaacgttc 2820
gatgaacact ggctatgctg gtgtgcaaaa cccgctgttc ttcaaggaaa acacccacat 2880
gctgtttggt gacgccaaag ccagcgtgga tgcaatcctg aaagctctgt aa 2932
<210> 95
<211> 1401
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 95
atgccacatt cctacgatta cgatgccata gtaataggtt ccggccccgg cggcgaaggc 60
gctgcaatgg gcctggttaa gcaaggtgcg cgcgtcgcag ttatcgagcg ttatcaaaat 120
gttggcggcg gttgcaccca ctggggcacc atcccgtcga aagctctccg tcacgccgtc 180
agccgcatta tagaattcaa tcaaaaccca ctttacagcg accattcccg actgctccgc 240
tcttcttttg ccgatatcct taaccatgcc gataacgtga ttaatcaaca aacgcgcatg 300
cgtcagggat tttacgaacg taatcactgt gaaatattgc agggaaacgc tcgctttgtt 360
gacgagcata cgttggcgct ggattgcccg gacggcagcg ttgaaacact aaccgctgaa 420
aaatttgtta ttgcctgcgg ctctcgtcca tatcatccaa cagatgttga tttcacccat 480
ccacgcattt acgacagcga ctcaattctc agcatgcacc acgaaccgcg ccatgtactt 540
atctatggtg ctggagtgat cggctgtgaa tatgcgtcga tcttccgcgg tatggatgta 600
aaagtggatc tgatcaacac ccgcgatcgc ctgctggcat ttctcgatca agagatgtca 660
gattctctct cctatcactt ctggaacagt ggcgtagtga ttcgtcacaa cgaagagtac 720
gagaagatcg aaggctgtga cgatggtgtg atcatgcatc tgaagtcggg taaaaaactg 780
aaagctgact gcctgctcta tgccaacggt cgcaccggta ataccgattc gctggcgtta 840
cagaacattg ggctagaaac tgacagccgc ggacagctga aggtcaacag catgtatcag 900
accgcacagc cacacgttta cgcggtgggc gacgtgattg gttatccgag cctggcgtcg 960
gcggcctatg accaggggcg cattgccgcg caggcgctgg taaaaggcga agccaccgca 1020
catctgattg aagatatccc taccggtatt tacaccatcc cggaaatcag ctctgtgggc 1080
aaaaccgaac agcagctgac cgcaatgaaa gtgccatatg aagtgggccg cgcccagttt 1140
aaacatctgg cacgcgcaca aatcgtcggc atgaacgtgg gcacgctgaa aattttgttc 1200
catcgggaaa caaaagagat tctgggtatt cactgctttg gcgagcgcgc tgccgaaatt 1260
attcatatcg gtcaggcgat tatggaacag aaaggtggcg gcaacactat tgagtacttc 1320
gtcaacacca cctttaacta cccgacgatg gcggaagcct atcgggtagc tgcgttaaac 1380
ggtttaaacc gcctgtttta a 1401
<210> 96
<211> 1434
<212> DNA
<213> 变形链球菌(Streptococcus mutans)
<400> 96
atggcaatga caaaacaata taaaaattat gtcaatggcg agtggaagct ttcagaaaat 60
gaaattaaaa tctacgaacc agccagtgga gctgaattgg gttcagttcc agcaatgagt 120
actgaagaag tagattatgt ttatgcttca gccaagaaag ctcaaccagc ttggcgagca 180
ctttcataca tagaacgtgc tgcctacctt cataaggtag cagatatttt gatgcgtgat 240
aaagaaaaaa taggtgctat tctttccaaa gaggttgcta aaggttataa atcagcagtc 300
agcgaagttg ttcgtactgc agaaatcatt aattatgcag ctgaagaagg tcttcgtatg 360
gaaggtgaag tccttgaagg cggcagtttt gaagcagcca gcaagaaaaa aattgccgtt 420
gttcgtcgtg aaccagtagg tcttgtatta gctatttcac catttaacta ccctgttaac 480
ttggcaggtt cgaaaattgc accggctctt attgcgggaa atgttattgc ttttaaacca 540
ccgacgcaag gatcaatctc agggctctta cttgctgaag catttgctga agctggactt 600
cctgcaggtg tctttaatac cattacaggt cgtggttctg aaattggaga ctatattgta 660
gaacatcaag ccgttaactt tatcaatttc actggttcaa caggaattgg cgaacgtatt 720
ggcaaaatgg ctggtatgcg tccgattatg cttgaactcg gtggaaaaga ttcagccatc 780
gttcttgaag atgcggacct tgaattgact gctaaaaata ttattgcagg tgcttttggt 840
tattcaggtc aacgctgtac agcagttaaa cgtgttcttg tgatggaaag tgttgctgat 900
gaactggtcg aaaaaatccg tgaaaaagtt cttgcattaa caattggtaa tccagaagac 960
gatgcagata ttacaccgtt gattgataca aaatcagctg attatgtaga aggtcttatt 1020
aatgatgcca atgataaagg agccactgcc cttactgaaa tcaaacgtga aggtaatctt 1080
atctgtccaa tcctctttga taaggtaacg acagatatgc gtcttgcttg ggaagaacca 1140
tttggtcctg ttcttccgat cattcgtgtg acatctgtag aagaagccat tgaaatttct 1200
aacaaatcgg aatatggact tcaggcttct atctttacaa atgatttccc acgcgctttt 1260
ggtattgctg agcagcttga agttggtaca gttcatatca ataataagac acagcgcggc 1320
acggacaact tcccattctt aggggctaaa aaatcaggtg caggtattca aggggtaaaa 1380
tattctattg aagctatgac aactgttaaa tccgtcgtat ttgatatcaa ataa 1434
<210> 97
<211> 996
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 97
atgactatca aagtaggtat caacggtttt ggccgtatcg gtcgcattgt tttccgtgct 60
gctcagaaac gttctgacat cgagatcgtt gcaatcaacg acctgttaga cgctgattac 120
atggcataca tgctgaaata tgactccact cacggccgtt tcgacggtac cgttgaagtg 180
aaagacggtc atctgatcgt taacggtaaa aaaatccgtg ttaccgctga acgtgatccg 240
gctaacctga aatgggacga agttggtgtt gacgttgtcg ctgaagcaac tggtctgttc 300
ctgactgacg aaactgctcg taaacacatc accgctggtg cgaagaaagt ggttatgact 360
ggtccgtcta aagacaacac tccgatgttc gttaaaggcg ctaacttcga caaatatgct 420
ggccaggaca tcgtttccaa cgcttcctgc accaccaact gcctggctcc gctggctaaa 480
gttatcaacg ataacttcgg catcatcgaa ggtctgatga ccaccgttca cgctactacc 540
gctactcaga aaaccgttga tggcccgtct cacaaagact ggcgcggcgg ccgcggcgct 600
tcccagaaca tcatcccgtc ctctaccggt gctgctaaag ctgtaggtaa agtactgcca 660
gaactgaatg gcaaactgac tggtatggcg ttccgcgttc cgaccccgaa cgtatctgta 720
gttgacctga ccgttcgtct ggaaaaagct gcaacttacg agcagatcaa agctgccgtt 780
aaagctgctg ctgaaggcga aatgaaaggc gttctgggct acaccgaaga tgacgtagta 840
tctaccgatt tcaacggcga agtttgcact tccgtgttcg atgctaaagc tggtatcgct 900
ctgaacgaca acttcgtgaa actggtatcc tggtacgaca acgaaaccgg ttactccaac 960
aaagttctgg acctgatcgc tcacatctcc aaataa 996
<210> 98
<211> 1650
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 98
atgaaaaaca tcaatccaac gcagaccgct gcctggcagg cactacagaa acacttcgat 60
gaaatgaaag acgttacgat cgccgatctt tttgctaaag acggcgatcg tttttctaag 120
ttctccgcaa ccttcgacga tcagatgctg gtggattact ccaaaaaccg catcactgaa 180
gagacgctgg cgaaattaca ggatctggcg aaagagtgcg atctggcggg cgcgattaag 240
tcgatgttct ctggcgagaa gatcaaccgc actgaaaacc gcgccgtgct gcacgtagcg 300
ctgcgtaacc gtagcaatac cccgattttg gttgatggca aagacgtaat gccggaagtc 360
aacgcggtgc tggagaagat gaaaaccttc tcagaagcga ttatttccgg tgagtggaaa 420
ggttataccg gcaaagcaat cactgacgta gtgaacatcg ggatcggcgg ttctgacctc 480
ggcccataca tggtgaccga agctctgcgt ccgtacaaaa accacctgaa catgcacttt 540
gtttctaacg tcgatgggac tcacatcgcg gaagtgctga aaaaagtaaa cccggaaacc 600
acgctgttct tggtagcatc taaaaccttc accactcagg aaactatgac caacgcccat 660
agcgcgcgtg actggttcct gaaagcggca ggtgatgaaa aacacgttgc aaaacacttt 720
gcggcgcttt ccaccaatgc caaagccgtt ggcgagtttg gtattgatac tgccaacatg 780
ttcgagttct gggactgggt tggcggccgt tactctttgt ggtcagcgat tggcctgtcg 840
attgttctct ccatcggctt tgataacttc gttgaactgc tttccggcgc acacgcgatg 900
gacaagcatt tctccaccac gcctgccgag aaaaacctgc ctgtactgct ggcgctgatt 960
ggcatctggt acaacaattt ctttggtgcg gaaactgaag cgattctgcc gtatgaccag 1020
tatatgcacc gtttcgcggc gtacttccag cagggcaata tggagtccaa cggtaagtat 1080
gttgaccgta acggtaacgt tgtggattac cagactggcc cgattatctg gggtgaacca 1140
ggcactaacg gtcagcacgc gttctaccag ctgatccacc agggaaccaa aatggtaccg 1200
tgcgatttca tcgctccggc tatcacccat aacccgctct ctgatcatca ccagaaactg 1260
ctgtctaact tcttcgccca gaccgaagcg ctggcgtttg gtaaatcccg cgaagtggtt 1320
gagcaggaat atcgtgatca gggtaaagat ccggcaacgc ttgactacgt ggtgccgttc 1380
aaagtattcg aaggtaaccg cccgaccaac tccatcctgc tgcgtgaaat cactccgttc 1440
agcctgggtg cgttgattgc gctgtatgag cacaaaatct ttactcaggg cgtgatcctg 1500
aacatcttca ccttcgacca gtggggcgtg gaactgggta aacagctggc gaaccgtatt 1560
ctgccagagc tgaaagatga taaagaaatc agcagccacg atagctcgac caatggtctg 1620
attaaccgct ataaagcgtg gcgcggttaa 1650
<210> 99
<211> 963
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 99
atgattaaga aaatcggtgt gttgacaagc ggcggtgatg cgccaggcat gaacgccgca 60
attcgcgggg ttgttcgttc tgcgctgaca gaaggtctgg aagtaatggg tatttatgac 120
ggctatctgg gtctgtatga agaccgtatg gtacagctag accgttacag cgtgtctgac 180
atgatcaacc gtggcggtac gttcctcggt tctgcgcgtt tcccggaatt ccgcgacgag 240
aacatccgcg ccgtggctat cgaaaacctg aaaaaacgtg gtatcgacgc gctggtggtt 300
atcggcggtg acggttccta catgggtgca atgcgtctga ccgaaatggg cttcccgtgc 360
atcggtctgc cgggcactat cgacaacgac atcaaaggca ctgactacac tatcggtttc 420
ttcactgcgc tgagcaccgt tgtagaagcg atcgaccgtc tgcgtgacac ctcttcttct 480
caccagcgta tttccgtggt ggaagtgatg ggccgttatt gtggagatct gacgttggct 540
gcggccattg ccggtggctg tgaattcgtt gtggttccgg aagttgaatt cagccgtgaa 600
gacctggtaa acgaaatcaa agcgggtatc gcgaaaggta aaaaacacgc gatcgtggcg 660
attaccgaac atatgtgtga tgttgacgaa ctggcgcatt tcatcgagaa agaaaccggt 720
cgtgaaaccc gcgcaactgt gctgggccac atccagcgcg gtggttctcc ggtgccttac 780
gaccgtattc tggcttcccg tatgggcgct tacgctatcg atctgctgct ggcaggttac 840
ggcggtcgtt gtgtaggtat ccagaacgaa cagctggttc accacgacat catcgacgct 900
atcgaaaaca tgaagcgtcc gttcaaaggt gactggctgg actgcgcgaa aaaactgtat 960
taa 963
<210> 100
<211> 1476
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 100
atggcggtaa cgcaaacagc ccaggcctgt gacctggtca ttttcggcgc gaaaggcgac 60
cttgcgcgtc gtaaattgct gccttccctg tatcaactgg aaaaagccgg tcagctcaac 120
ccggacaccc ggattatcgg cgtagggcgt gctgactggg ataaagcggc atataccaaa 180
gttgtccgcg aggcgctcga aactttcatg aaagaaacca ttgatgaagg tttatgggac 240
accctgagtg cacgtctgga tttttgtaat ctcgatgtca atgacactgc tgcattcagc 300
cgtctcggcg cgatgctgga tcaaaaaaat cgtatcacca ttaactactt tgccatgccg 360
cccagcactt ttggcgcaat ttgcaaaggg cttggcgagg caaaactgaa tgctaaaccg 420
gcacgcgtag tcatggagaa accgctgggg acgtcgctgg cgacctcgca ggaaatcaat 480
gatcaggttg gcgaatactt cgaggagtgc caggtttacc gtatcgacca ctatcttggt 540
aaagaaacgg tgctgaacct gttggcgctg cgttttgcta actccctgtt tgtgaataac 600
tgggacaatc gcaccattga tcatgttgag attaccgtgg cagaagaagt ggggatcgaa 660
gggcgctggg gctattttga taaagccggt cagatgcgcg acatgatcca gaaccacctg 720
ctgcaaattc tttgcatgat tgcgatgtct ccgccgtctg acctgagcgc agacagcatc 780
cgcgatgaaa aagtgaaagt actgaagtct ctgcgccgca tcgaccgctc caacgtacgc 840
gaaaaaaccg tacgcgggca atatactgcg ggcttcgccc agggcaaaaa agtgccggga 900
tatctggaag aagagggcgc gaacaagagc agcaatacag aaactttcgt ggcgatccgc 960
gtcgacattg ataactggcg ctgggccggt gtgccattct acctgcgtac tggtaaacgt 1020
ctgccgacca aatgttctga agtcgtggtc tatttcaaaa cacctgaact gaatctgttt 1080
aaagaatcgt ggcaggatct gccgcagaat aaactgacta tccgtctgca acctgatgaa 1140
ggcgtggata tccaggtact gaataaagtt cctggccttg accacaaaca taacctgcaa 1200
atcaccaagc tggatctgag ctattcagaa acctttaatc agacgcatct ggcggatgcc 1260
tatgaacgtt tgctgctgga aaccatgcgt ggtattcagg cactgtttgt acgtcgcgac 1320
gaagtggaag aagcctggaa atgggtagac tccattactg aggcgtgggc gatggacaat 1380
gatgcgccga aaccgtatca ggccggaacc tggggacccg ttgcctcggt ggcgatgatt 1440
acccgtgatg gtcgttcctg gaatgagttt gagtaa 1476
<210> 101
<211> 474
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 101
Met Ser Thr Glu Ile Lys Thr Gln Val Val Val Leu Gly Ala Gly Pro
1 5 10 15
Ala Gly Tyr Ser Ala Ala Phe Arg Cys Ala Asp Leu Gly Leu Glu Thr
20 25 30
Val Ile Val Glu Arg Tyr Asn Thr Leu Gly Gly Val Cys Leu Asn Val
35 40 45
Gly Cys Ile Pro Ser Lys Ala Leu Leu His Val Ala Lys Val Ile Glu
50 55 60
Glu Ala Lys Ala Leu Ala Glu His Gly Ile Val Phe Gly Glu Pro Lys
65 70 75 80
Thr Asp Ile Asp Lys Ile Arg Thr Trp Lys Glu Lys Val Ile Asn Gln
85 90 95
Leu Thr Gly Gly Leu Ala Gly Met Ala Lys Gly Arg Lys Val Lys Val
100 105 110
Val Asn Gly Leu Gly Lys Phe Thr Gly Ala Asn Thr Leu Glu Val Glu
115 120 125
Gly Glu Asn Gly Lys Thr Val Ile Asn Phe Asp Asn Ala Ile Ile Ala
130 135 140
Ala Gly Ser Arg Pro Ile Gln Leu Pro Phe Ile Pro His Glu Asp Pro
145 150 155 160
Arg Ile Trp Asp Ser Thr Asp Ala Leu Glu Leu Lys Glu Val Pro Glu
165 170 175
Arg Leu Leu Val Met Gly Gly Gly Ile Ile Gly Leu Glu Met Gly Thr
180 185 190
Val Tyr His Ala Leu Gly Ser Gln Ile Asp Val Val Glu Met Phe Asp
195 200 205
Gln Val Ile Pro Ala Ala Asp Lys Asp Ile Val Lys Val Phe Thr Lys
210 215 220
Arg Ile Ser Lys Lys Phe Asn Leu Met Leu Glu Thr Lys Val Thr Ala
225 230 235 240
Val Glu Ala Lys Glu Asp Gly Ile Tyr Val Thr Met Glu Gly Lys Lys
245 250 255
Ala Pro Ala Glu Pro Gln Arg Tyr Asp Ala Val Leu Val Ala Ile Gly
260 265 270
Arg Val Pro Asn Gly Lys Asn Leu Asp Ala Gly Lys Ala Gly Val Glu
275 280 285
Val Asp Asp Arg Gly Phe Ile Arg Val Asp Lys Gln Leu Arg Thr Asn
290 295 300
Val Pro His Ile Phe Ala Ile Gly Asp Ile Val Gly Gln Pro Met Leu
305 310 315 320
Ala His Lys Gly Val His Glu Gly His Val Ala Ala Glu Val Ile Ala
325 330 335
Gly Lys Lys His Tyr Phe Asp Pro Lys Val Ile Pro Ser Ile Ala Tyr
340 345 350
Thr Glu Pro Glu Val Ala Trp Val Gly Leu Thr Glu Lys Glu Ala Lys
355 360 365
Glu Lys Gly Ile Ser Tyr Glu Thr Ala Thr Phe Pro Trp Ala Ala Ser
370 375 380
Gly Arg Ala Ile Ala Ser Asp Cys Ala Asp Gly Met Thr Lys Leu Ile
385 390 395 400
Phe Asp Lys Glu Ser His Arg Val Ile Gly Gly Ala Ile Val Gly Thr
405 410 415
Asn Gly Gly Glu Leu Leu Gly Glu Ile Gly Leu Ala Ile Glu Met Gly
420 425 430
Cys Asp Ala Glu Asp Ile Ala Leu Thr Ile His Ala His Pro Thr Leu
435 440 445
His Glu Ser Val Gly Leu Ala Ala Glu Val Phe Glu Gly Ser Ile Thr
450 455 460
Asp Leu Pro Asn Pro Lys Ala Lys Lys Lys
465 470
<210> 102
<211> 474
<212> PRT
<213> 人工的
<220>
<223> Lpd突变体
<400> 102
Met Ser Thr Glu Ile Lys Thr Gln Val Val Val Leu Gly Ala Gly Pro
1 5 10 15
Ala Gly Tyr Ser Ala Ala Phe Arg Cys Ala Asp Leu Gly Leu Glu Thr
20 25 30
Val Ile Val Glu Arg Tyr Asn Thr Leu Gly Gly Val Cys Leu Asn Val
35 40 45
Gly Cys Ile Pro Ser Lys Ala Leu Leu His Val Ala Lys Val Ile Glu
50 55 60
Glu Ala Lys Ala Leu Ala Glu His Gly Ile Val Phe Gly Glu Pro Lys
65 70 75 80
Thr Asp Ile Asp Lys Ile Arg Thr Trp Lys Glu Lys Val Ile Asn Gln
85 90 95
Leu Thr Gly Gly Leu Ala Gly Met Ala Lys Gly Arg Lys Val Lys Val
100 105 110
Val Asn Gly Leu Gly Lys Phe Thr Gly Ala Asn Thr Leu Glu Val Glu
115 120 125
Gly Glu Asn Gly Lys Thr Val Ile Asn Phe Asp Asn Ala Ile Ile Ala
130 135 140
Ala Gly Ser Arg Pro Ile Gln Leu Pro Phe Ile Pro His Glu Asp Pro
145 150 155 160
Arg Ile Trp Asp Ser Thr Asp Ala Leu Glu Leu Lys Glu Val Pro Glu
165 170 175
Arg Leu Leu Val Met Gly Gly Gly Ile Ile Ala Leu Glu Met Ala Thr
180 185 190
Val Tyr His Ala Leu Gly Ser Gln Ile Asp Val Val Val Arg Lys His
195 200 205
Gln Val Ile Arg Ala Ala Asp Lys Asp Ile Val Lys Val Phe Thr Lys
210 215 220
Arg Ile Ser Lys Lys Phe Asn Leu Met Leu Glu Thr Lys Val Thr Ala
225 230 235 240
Val Glu Ala Lys Glu Asp Gly Ile Tyr Val Thr Met Glu Gly Lys Lys
245 250 255
Ala Pro Ala Glu Pro Gln Arg Tyr Asp Ala Val Leu Val Ala Ile Gly
260 265 270
Arg Val Pro Asn Gly Lys Asn Leu Asp Ala Gly Lys Ala Gly Val Glu
275 280 285
Val Asp Asp Arg Gly Phe Ile Arg Val Asp Lys Gln Leu Arg Thr Asn
290 295 300
Val Pro His Ile Phe Ala Ile Gly Asp Ile Val Gly Gln Pro Met Leu
305 310 315 320
Ala His Lys Gly Val His Glu Gly His Val Ala Ala Glu Val Ile Ala
325 330 335
Gly Lys Lys His Tyr Phe Asp Pro Lys Val Ile Pro Ser Ile Ala Tyr
340 345 350
Thr Glu Pro Glu Val Ala Trp Val Gly Leu Thr Glu Lys Glu Ala Lys
355 360 365
Glu Lys Gly Ile Ser Tyr Glu Thr Ala Thr Phe Pro Trp Ala Ala Ser
370 375 380
Gly Arg Ala Ile Ala Ser Asp Cys Ala Asp Gly Met Thr Lys Leu Ile
385 390 395 400
Phe Asp Lys Glu Ser His Arg Val Ile Gly Gly Ala Ile Val Gly Thr
405 410 415
Asn Gly Gly Glu Leu Leu Gly Glu Ile Gly Leu Ala Ile Glu Met Gly
420 425 430
Cys Asp Ala Glu Asp Ile Ala Leu Thr Ile His Ala His Pro Thr Leu
435 440 445
His Glu Ser Val Gly Leu Ala Ala Glu Val Phe Glu Gly Ser Ile Thr
450 455 460
Asp Leu Pro Asn Pro Lys Ala Lys Lys Lys
465 470
<210> 103
<211> 1548
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 103
atgacggacc atacaatgaa gaaaaacccc gtaagtatac cacacaccgt ctggtacgcc 60
gacgatatcc gccgcggaga acgcgaggcg gcagatgtgc tggggctcac actctatgag 120
ctgatgcttc gcgctggcga ggccgcattc caggtgtgtc gttcggcgta tcctgacgcc 180
cgccactggc tggtgctgtg cggtcatggt aataacggcg gcgatggcta cgtggtcgcg 240
cgactggcca aagcggtcgg cattgaggtc acgttgttgg cccaggagag cgacaaaccg 300
ttgccggaag aggccgcgct ggcacgcgaa gcatggttaa acgcgggtgg cgagatccat 360
gcttcgaata ttgtctggcc cgaatcggta gatctgattg ttgatgcgct gctcggtacc 420
ggtttgcggc aagcgccccg cgaatccatt agccagttaa tcgaccacgc taattcccat 480
cctgcgccga ttgtggcggt tgatatccct tccggcctgc tggctgaaac tggcgctacg 540
ccaggcgcgg tgatcaacgc cgatcacacc atcactttta ttgcgctgaa accaggcttg 600
ctcactggaa aagcgcggga tgttaccgga caactgcatt ttgactcact ggggctggat 660
agttggctgg caggtcagga gacgaaaatt cagcggtttt cagcagaaca actttctcac 720
tggctaaaac cgcgtcgccc gacttcgcat aaaggcgatc acgggcggct ggtaattatc 780
ggtggcgatc acggcacggc gggggctatt cgtatgacgg gggaagcggc gctgcgtgct 840
ggtgctggtt tagtccgagt actgacccgc agtgaaaaca ttgcgccgct gctgactgca 900
cgaccggaat tgatggtgca tgaactgacg atggactctc ttaccgaaag cctggaatgg 960
gccgatgtgg tggtgattgg tcccggtctg ggccagcaag agtgggggaa aaaagcactg 1020
caaaaagttg agaattttcg caaaccgatg ttgtgggatg ccgatgcatt gaacctgctg 1080
gcaatcaatc ccgataagcg tcacaatcgc gtgatcacgc cgcatcctgg cgaggccgca 1140
cggttgttag gctgttccgt cgctgaaatt gaaagtgacc gcttacattg cgccaaacgt 1200
ctggtacaac gttatggcgg cgtagcggtg ctgaaaggtg ccggaaccgt ggtcgccgcc 1260
catcctgacg ctttaggcat tattgatgcc ggaaatgcag gcatggcgag cggcggcatg 1320
ggcgatgtgc tctctggtat tattggcgca ttgcttgggc aaaaactgtc gccgtatgat 1380
gcagcctgtg caggctgtgt cgcgcacggt gcggcagctg acgtactggc ggcgcgtttt 1440
ggaacgcgcg ggatgctggc aaccgatctc ttttccacgc tacagcgtat tgttaacccg 1500
gaagtgactg ataaaaacca tgatgaatcg agtaattccg ctccctga 1548
<210> 104
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失xylAB 1
<400> 104
acgacatcat ccatcacccg cggcattacc tgattatgga gttcaatatg 50
<210> 105
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失xylAB 2
<400> 105
cccccacccg gtcaggcagg ggataacgtt tacgccatta atggcagaag 50
<210> 106
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yjhH 1
<400> 106
aatgcgcgaa gttgccgact tcctgattaa taaaggggtc gacgggctgt 50
<210> 107
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yfhH 2
<400> 107
gtaccgactt aactgtgttg atcatcgtac gcaagtgacc aacgctgtcg 50
<210> 108
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yagE 1
<400> 108
ggcggcacca acgcccggga aaccatcgaa ctcagccagc acgcgcagca 50
<210> 109
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yagE 2
<400> 109
agcacggtga agtgcggatg ggcacctttg acggtatgga tcatgctgcg 50
<210> 110
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yiaE 1
<400> 110
catatttcag gctaaggtga tcgccttatc agtgaatgga gagaagcatg 50
<210> 111
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失yiaE 2
<400> 111
tatcgggctt tactctacgc agtcgcggct tagtccgcga cgtgcggatt 50
<210> 112
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失ycdW 1
<400> 112
aacgataagt gcgaataaat ttcgcacaac gcttttcggg agtcagtatg 50
<210> 113
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物缺失ycdW 2
<400> 113
ccaaggatag caggaatcct gatgctttat tagtagccgc gtgcgcggtc 50
<210> 114
<211> 100
<212> DNA
<213> 人工的
<220>
<223> 引物缺失ptsG 1
<400> 114
atgtttaaga atgcatttgc taacctgcaa aaggtcggta aatcgctgat gctgccggta 60
tccgtactgc ctatcgcagg tgtaggctgg agctgcttcg 100
<210> 115
<211> 100
<212> DNA
<213> 人工的
<220>
<223> 引物缺失ptsG 2
<400> 115
ttagtggtta cggatgtact catccatctc ggttttcagg ttatcggatt tagtaccgaa 60
aatcgcctga acaccagaac catatgaata tcctccttag 100
<210> 116
<211> 21
<212> DNA
<213> 人工的
<220>
<223> RBS120
<400> 116
atccggtata ggaggtatag a 21
<210> 117
<211> 100
<212> DNA
<213> 人工的
<220>
<223> 引物缺失udhA 1
<400> 117
ggtgcgcgcg tcgcagttat cgagcgttat caaaatgttg gcggcggttg cacccactgg 60
ggcaccatcc cgtcgaaagc catatgaata tcctccttag 100
<210> 118
<211> 100
<212> DNA
<213> 人工的
<220>
<223> 引物缺失udhA 2
<400> 118
cccagaatct cttttgtttc ccgatggaac aaaattttca gcgtgcccac gttcatgccg 60
acgatttgtg cgcgtgccag tgtaggctgg agctgcttcg 100
<210> 119
<211> 101
<212> DNA
<213> 人工的
<220>
<223> 引物缺失pfkA 1
<400> 119
gttcctcggt tctgcgcgtt tcccggaatt ccgcgacgag aacatccgcg ccgtggctat 60
cgaaaacctg aaaaaacgtg gtgtaggctg gagctgcttc g 101
<210> 120
<211> 102
<212> DNA
<213> 人工的
<220>
<223> 引物缺失pfkA 2
<400> 120
ggcctgataa gcgaagcgca tcaggcattt ttgcttctgt catcggtttc agggtaaagg 60
aatctgcctt tttccgaaat cacatatgaa tatcctcctt ag 102
<210> 121
<211> 83
<212> DNA
<213> 人工的
<220>
<223> PR01
<400> 121
acgttaaatc tatcaccgca agggataaat atctaacacc gtgcgtgttg acaattttac 60
ctctggcggt gataatggtt gca 83
<210> 122
<211> 714
<212> DNA
<213> 人工的
<220>
<223> CI857
<400> 122
atgagcacaa aaaagaaacc attaacacaa gagcagcttg aggacgcacg tcgccttaaa 60
gcaatttatg aaaaaaagaa aaatgaactt ggcttatccc aggaatctgt cgcagacaag 120
atggggatgg ggcagtcagg cgttggtgct ttatttaatg gcatcaatgc attaaatgct 180
tataacgccg cattgcttac aaaaattctc aaagttagcg ttgaagaatt tagcccttca 240
atcgccagag aaatctacga gatgtatgaa gcggttagta tgcagccgtc acttagaagt 300
gagtatgagt accctgtttt ttctcatgtt caggcaggga tgttctcacc taagcttaga 360
acctttacca aaggtgatgc ggagagatgg gtaagcacaa ccaaaaaagc cagtgattct 420
gcattctggc ttgaggttga aggtaattcc atgaccgcac caacaggctc caagccaagc 480
tttcctgacg gaatgttaat tctcgttgac cctgagcagg ctgttgagcc aggtgatttc 540
tgcatagcca gacttggggg tgatgagttt accttcaaga aactgatcag ggatagcggt 600
caggtgtttt tacaaccact aaacccacag tacccaatga tcccatgcaa tgagagttgt 660
tccgttgtgg ggaaagttat cgctagtcag tggcctgaag agacgtttgg ctga 714
<210> 123
<211> 34
<212> DNA
<213> 人工的
<220>
<223> RBS150
<400> 123
taccaactaa cgcacgttta agtaggaacc gtat 34
<210> 124
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 由gapN代替gapA
<400> 124
acgtgactga ttctaacaaa acattaacac caactggcaa aattttgtcc 50
<210> 125
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 由gapN代替gapA
<400> 125
aaaaaagagc gaccgaagtc gctcttttta gatcacagtg tcatctcaac 50
<210> 126
<211> 43
<212> DNA
<213> 人工的
<220>
<223> ptrc
<400> 126
gagctgttga caattaatca tccggctcgt ataatgtgtg gaa 43
<210> 127
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物代替xylFGH启动子1
<400> 127
ctaaaaattg gttacgttta tcgcggtgat tgttacttat taaaactgtc 50
<210> 128
<211> 50
<212> DNA
<213> 人工的
<220>
<223> 引物代替xylFGH启动子2
<400> 128
tgagtagaat gttctttatt ttcatggtgt agggccttct gtagttagag 50

Claims (18)

1.一种大肠杆菌(E.coli)微生物，所述微生物经遗传修饰用于通过将木糖转化成1,2,4-丁三醇而生产2,4-二羟基丁酸，其中所述用于将木糖转化成1,2,4-丁三醇的遗传修饰是以下基因中的至少两个的过表达：
- 来自新月柄杆菌（Caulobacter crescentus）的xdh基因，
- 来自新月柄杆菌的xylC基因，
- 来自大肠杆菌的yjhG和/或yagF基因，
- 来自恶臭假单胞菌(Pseudomonas putida)的mdlC基因，
-大肠杆菌的adhP和/或yqhD基因，
其中所述至少两个基因包括来自新月柄杆菌的xdh基因和来自恶臭假单胞菌的mdlC基因，并且
其中所述微生物经进一步遗传修饰而用于：
i) 将1,2,4-丁三醇氧化成2,4-二羟基丁醛；以及
ii) 将2,4-二羟基丁醛氧化成2,4-二羟基丁酸，
其中所述遗传修饰i)是选自由以下组成的组的至少一个编码酶的基因的过表达：来自丁酸梭菌(Clostridium butyricum)、弗氏柠檬酸杆菌(Citrobacter freudii)或肺炎克雷伯氏菌(Klebsiella pneumoniae)的编码醇脱氢酶的基因dhaT和来自大肠杆菌的编码乳醛还原酶的基因fucO，
并且其中所述遗传修饰ii)是选自由以下组成的组的至少一个编码酶的基因的过表达：来自大肠杆菌的编码醛脱氢酶的基因puuC和基因aldB。
2.根据权利要求1所述的微生物，其中所述醇脱氢酶具有SEQ ID NO: 36、37或38中的任一个的序列；和/或所述乳醛还原酶具有SEQ ID NO：41的序列。
3.根据权利要求1所述的微生物，其中所述醛脱氢酶具有SEQ ID NO: 46或55的序列。
4.根据权利要求1至3中任一项所述的微生物，其中用于将木糖转化成1,2,4-丁三醇的遗传修饰是以下三个基因的过表达：来自新月柄杆菌的xdh和xylC基因以及来自恶臭假单胞菌的mdlC基因。
5.根据权利要求1至3中任一项所述的微生物，其中所述微生物经进一步遗传修饰而用于：
iii) 从除木糖外的碳源中提供还原能力和/或能量，用于2,4-二羟基丁酸生产和微生物生长，和/或
iv) 至少部分抑制碳分解代谢物阻抑，
其中所述遗传修饰iii)是至少一个以下基因的缺失和/或弱化：
- 编码木糖异构酶的基因，
- 编码木酮糖激酶的基因，
- 编码3-脱氧-D-甘油-戊酮糖酸醛缩酶的基因，
- 编码酮酸脱氢酶的基因，和
- 其任何组合，
其中所述遗传修饰iv)选自以下至少一个：
- 缺失编码磷酸转移酶系统的葡萄糖通透酶的基因，
- 缺失编码磷酸载体Hpr蛋白的基因，
- 表达糖输入蛋白系统中牵涉的基因和/或操纵子，其中所述糖是除木糖外的碳源，
- 表达编码木糖转运蛋白的基因，
- 过表达编码葡萄糖同向转运蛋白的基因，
- 过表达编码葡萄糖易化蛋白的基因，
- 过表达编码葡糖激酶的基因，
-表达编码腺苷酸环化酶的基因，
- 表达编码CRP和/或CRP样蛋白的基因，
- 表达编码不依赖于cAMP的CRP蛋白的基因，和
- 其任何组合。
6.根据权利要求5所述的微生物，其中所述遗传修饰iii)是至少一个以下基因的缺失和/或弱化：
- 来自大肠杆菌的xylA基因，
- 来自大肠杆菌的xylB基因，
- 来自大肠杆菌的yjhH和/或yagE基因，
- 来自大肠杆菌的yiaE和/或ycdW基因，和
- 其任何组合。
7.根据权利要求5所述的微生物，其中所述遗传修饰iv)选自以下至少一个：
- 来自大肠杆菌的pstG基因的缺失，
- 来自大肠杆菌的ptsH基因的缺失，
- 来自大肠杆菌的lacY基因、来自大肠杆菌的malFG基因、来自鼠伤寒沙门氏菌(Salmonella typhimurium)的scrKYABR操纵子和/或来自大肠杆菌的cscBKAR操纵子的表达，
- 来自大肠杆菌的xylFGH操纵子、来自大肠杆菌的xylE基因或来自大肠杆菌的araE基因的表达，
- 来自大肠杆菌的galP基因的过表达，
- 来自运动发酵单胞菌(Zymomonas mobilis)的glf基因的过表达，
- 来自大肠杆菌的glk基因的过表达，
- 来自大肠杆菌的cyaA基因的表达，
- 来自大肠杆菌的CRP基因或来自枯草芽孢杆菌(Bacillus subtilis)的ccpA基因的表达，
- 编码不依赖于cAMP的CRP蛋白的基因的表达，其中所述蛋白在包含氨基酸残基53至62的区域内被突变和/或在包含氨基酸残基141至148的区域内被突变，和
- 其任何组合。
8.根据权利要求7所述的微生物，其中所述不依赖于cAMP的CRP蛋白包含至少一个以下突变：D53H、S62F、G141D、R142D和L148R。
9.根据权利要求4所述的微生物，其中遗传修饰i)和ii)由以下两个基因的过表达组成：
- 来自大肠杆菌的fucO和puuC，或
- 来自大肠杆菌的fucO和aldB，或
- 来自丁酸梭菌的dhaT和来自大肠杆菌的puuC，或
- 来自丁酸梭菌的dhaT和来自大肠杆菌的aldB，
并且其中遗传修饰iii)是以下基因的缺失：
- 编码木糖异构酶的xylA和xylB，
- 编码3-脱氧-D-甘油-戊酮糖酸醛缩酶的基因yjhH和yagE，和
- 编码酮酸脱氢酶的基因yiaE和ycdW，
并且其中遗传修饰iv)是编码葡萄糖磷酸转移酶的ptsG基因的缺失。
10.根据权利要求1至3中任一项所述的微生物，其包含作为还原能力来源的NADPH的产生中牵涉的至少一个基因的进一步遗传修饰，所述修饰选自：
- 过表达编码膜结合的转氢酶的基因或操纵子，
- 缺失或弱化编码可溶性转氢酶的基因，
- 过表达编码产生NADPH的甘油醛-3-磷酸脱氢酶的基因，
- 缺失或弱化编码磷酸葡糖异构酶的基因，
- 缺失或弱化编码磷酸果糖激酶的基因，
- 过表达编码葡萄糖-6-磷酸脱氢酶的基因，
- 过表达编码具有序列SEQ ID NO: 102的能够产生NADPH的硫辛酰胺脱氢酶的突变体lpd基因，
- 过表达编码双功能NAD(P)H水合物修复酶的基因，和
- 其任何组合。
11.根据权利要求10所述的微生物，其包含参与产生作为还原力来源的NADPH的至少一个基因的进一步遗传修饰，所述修饰选自：
- 来自大肠杆菌的pntAB操纵子的过表达，
- 来自大肠杆菌的udhA基因的缺失或弱化，
- 通过用来自变异链球菌(Streptococcus mutans)的gapN基因取代来自大肠杆菌的gapA，编码甘油醛-3-磷酸脱氢酶的基因的过表达，所述甘油醛-3-磷酸脱氢酶产生NADPH，
- 来自大肠杆菌的pgi基因的缺失或弱化，
- 来自大肠杆菌的pfkA基因的缺失或弱化，
- 来自大肠杆菌的zwf基因的过表达，
- 来自大肠杆菌的yjeF基因的过表达，和
- 其任何组合。
12.一种生产2,4-二羟基丁酸的方法，其包括：
a) 在允许将木糖转化成2,4-二羟基丁酸的发酵条件下，在包含木糖的培养基中培养如权利要求1至11中任一项所述的经遗传修饰的微生物，和
b) 从所述培养基中回收2,4-二羟基丁酸。
13.根据权利要求12所述的方法，其中所述培养基包含除木糖外的碳源。
14.根据权利要求13所述的方法，其中所述除木糖外的碳源是包含3、6或12个碳原子的碳水化合物。
15.根据权利要求14所述的方法，其中所述除木糖外的碳源选自下组：甘油、葡萄糖、半乳糖、果糖、乳糖、麦芽糖、蔗糖、及其任何组合。
16.根据权利要求13至15中任一项所述的方法，其中所述碳源源自植物生物质。
17.根据权利要求12至15中任一项所述的方法，其中所述方法还包括纯化步骤b)的所述2,4-二羟基丁酸的步骤c)。
18.根据权利要求17所述的方法，其中通过离子交换层析纯化所述2,4-二羟基丁酸。
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