CN107406821B - 用于生产3-羟基丙酸的突变宿主细胞 - Google Patents

用于生产3-羟基丙酸的突变宿主细胞 Download PDF

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CN107406821B
CN107406821B CN201680011830.1A CN201680011830A CN107406821B CN 107406821 B CN107406821 B CN 107406821B CN 201680011830 A CN201680011830 A CN 201680011830A CN 107406821 B CN107406821 B CN 107406821B
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J·弗里亚斯
G·巴比尔
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Abstract

在此提供了具有主动3‑羟基丙酸(3‑HP)途径的重组宿主细胞,其中该宿主细胞包含对编码丙酮酸还原酶的内源基因进行的破坏。还描述了制造宿主细胞的方法,以及使用这些细胞来生产3‑HP和3‑HP的衍生物(例如丙烯酸)的方法。

Description

用于生产3-羟基丙酸的突变宿主细胞
相关申请的交叉引用
本申请要求于2015年2月27日提交的美国临时申请序列号62/126,377的优先权益。该申请的内容全部通过引用结合在此。
序列表的引用
本申请包括一个计算机可读形式的序列表,将其通过引用结合在此。
背景技术
3-羟基丙酸(3-HP)是一种由美国能源部鉴定为可以通过发酵制造的排名前12的高潜力结构单元化学品之一的三碳羧酸。作为乳酸(2-羟基丙酸)的异构体的3-HP的替代名称包括乙烯乳酸和3-羟基丙酸化物。3-HP是一种有吸引力的可再生平台化学品,来自葡萄糖的理论产量为100%,具有允许它参与不同种化学反应的多种官能团,以及低的毒性。可以将3-HP用作底物来形成若干日用化学品,例如1,3-丙二醇、丙二酸、丙烯酰胺、以及丙烯酸。丙烯酸是一种用来生产丙烯酸酯和超强吸收能力聚合物的大量化学品(>70亿磅/年),并且目前来源于丙烯的催化氧化。3-HP的发酵性生产将为作为这些商业上有意义的化学品的原料的石油化学产品提供可持续的替代物,从而减少能量消耗、对外国石油供应的依赖以及温室气体的产生。
不同于乳酸,3-HP不是在自然界中已知的任何途径的主要终产物,仅仅在一些细菌和真菌中发现了痕量。因此,需要更大量的遗传工程产生生产3-HP的酵母。已经描述了用来产生3-HP的若干种代谢途径(参见WO 01/16346、WO 02/042418、WO 2012/074818)。然而,在本领域中仍需要进一步以一种更成本有效的方式改进工业规模的3-HP生产。具体地,对通过将不想要的副产物的形成最小化来改进3-HP产品收得率存在需要。
发明内容
本文描述的是包含与丙酮酸还原酶基因相关的修饰的微生物菌株。
在一个方面,是具有主动3-HP途径的重组宿主细胞,其中该细胞包含对编码丙酮酸还原酶(例如,SEQ ID NO:205的丙酮酸还原酶)的内源基因的破坏。在一些实施例中,当在相同的条件下培养时,与缺乏内源基因(编码丙酮酸还原酶)的破坏的亲本菌株相比,宿主细胞产生更少的丙酮酸还原酶和/或D-乳酸盐/D-乳酸酯(D-lactate)。在一些实施例中,编码丙酮酸还原酶的内源基因是失活的。
在一些实施例中,重组细胞是细菌细胞或真菌细胞。在一些实施例中,重组细胞是酵母细胞,例如伊萨酵母属(Issatchenkia)、假丝酵母属(Candida)、克鲁维酵母属(Kluyveromyces)、毕赤酵母属(Pichia)、裂殖酵母属(Schizosaccharomyces)、有孢圆酵母属(Torulaspora)、接合酵母属(Zygosaccharomyces)或酵母属(Saccharomyces)酵母细胞。在一些实施例中,重组细胞是3-HP抗性酵母细胞。在一些实施例中,重组细胞不能发酵戊糖。
在一些实施例中,重组细胞包含一种或多种(例如两种、若干种)异源多核苷酸,这些)异源多核苷酸选自编码丙酮酸脱氢酶(PDH)的异源多核苷酸、编码乙酰辅酶A羧化酶(ACC)的异源多核苷酸、编码丙二酰辅酶A还原酶的异源多核苷酸、以及编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。
在一些实施例中,重组细胞包含一种或多种(例如两种、若干种)异源多核苷酸,这些异源多核苷酸选自编码PEP羧化酶(PPC)的异源多核苷酸、编码丙酮酸羧化酶(PYC)的异源多核苷酸、编码天冬氨酸转氨酶(AAT)的异源多核苷酸、编码天冬氨酸1-脱羧酶(ADC)的异源多核苷酸、编码β-丙氨酸氨基转移酶(BAAT)或氨基丁酸氨基转移酶(gabT)的异源多核苷酸、以及编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。在一些实施例中,重组细胞包含编码天冬氨酸1-脱羧酶(ADC)的异源多核苷酸。
还描述了用于获得重组宿主细胞的方法,该方法包括:(a)培养亲本菌株;(b)(i)用一个或多个3-HP途径基因转化该亲本菌株以在(a)的亲本菌株中提供主动3-HP途径;(b)(ii)破坏在(a)的亲本菌株中编码丙酮酸还原酶的内源基因;并且(c)分离生成自(b)(i)和(b)(ii)的突变菌株。
还描述了生产3-HP以及相关化合物的方法。在一个方面,是产生3-HP的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞以产生3-HP;并且(b)回收3-HP。在另一个方面,是产生丙烯酸或其盐的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞以产生3-HP;(b)回收该3-HP;(c)在适合的条件下,将该3-HP脱水,以产生丙烯酸或其盐;并且(d)回收该丙烯酸或其盐。
附图说明
图1示出了从葡萄糖的选择性3-HP途径的概述。
图2示出了pMHCT260b的质粒图。
图3示出了pMHCT247质粒图。
图4示出了pMHCT261的质粒图。
图5示出了菌株MhCt235、MhCt236和Ckle210在16小时和24hr的丙酮酸还原酶活性。
图6示出了菌株MhCt235、MhCt236和Ckle210经过80hr的发酵后D-乳酸盐/D-乳酸酯形成的量。
定义
3-HP:术语“3-HP”包括“3-羟基丙酸”的盐和酸形式,除非上下文中另有说明。
缩写:3-HPA,3-羟基丙醛;3-HPDH,3-羟基丙酸脱氢酶;AAM,丙氨酸2,3-氨基变位酶;AAT,天冬氨酸转氨酶;ACC,乙酰辅酶A羧化酶;ADC,天冬氨酸1-脱羧酶;AKG,α-酮戊二酸ALD,醛脱氢酶;BAAT,β-丙氨酸转氨酶;BCKA,支链α-酮酸脱羧酶;bp,碱基对;CYB2,L-(+)-乳酸-细胞色素c氧化还原酶;CYC,异-2-细胞色素c;EMS,乙烷甲基磺化酶;ENO,烯醇酶;gabT,4-氨基丁酸转氨酶;GAPDH,甘油醛-3-磷酸脱氢酶3;GPD,甘油3-磷酸脱氢酶;GPP,甘油3-磷酸磷酸酶;HIBADH,3-羟基异丁酸脱氢酶;IPDA,吲哚丙酮酸脱羧酶;KGD,α-酮戊二酸脱羧酶;LDH,乳酸脱氢酶;MAE,苹果酸酶;MDHB,苹果酸脱氢酶B;OAA,草酰乙酸PCK,磷酸烯醇丙酮酸羧激酶;PDC,丙酮酸脱羧酶;PDH,丙酮酸脱氢酶;PEP,磷酸烯醇丙酮酸;PGK,磷酸甘油酸激酶;PPC,磷酸烯醇丙酮酸羧化酶;PYC,丙酮酸羧化酶;RKI,核糖5-磷酸酮醇异构酶;TAL,转醛醇酶;TEF1,翻译延长因子-1;TEF2,翻译延长因子-2;TKL,转酮醇酶,XDH,木糖醇脱氢酶;XR,木糖还原酶;YP,酵母提取物/蛋白胨。
主动3-HP途径:如在此使用,具有“主动3-HP途径”的宿主细胞以如下量产生催化代谢途径的每个反应所必需的活性酶,该量足以从可发酵糖产生3-HP,并且因此当在至少一种可发酵糖的存在下在发酵条件下培养时,该宿主细胞能够以可测量的产量产生3-HP。具有主动3-HP途径的宿主细胞包含一个或多个3-HP途径基因。如在此使用,“3-HP途径基因”是指一种编码在主动3-HP途径中涉及的酶的基因。
催化主动3-HP途径中的每个反应所必需的活性酶可以来自内源基因表达的活性、异源基因表达的活性、或来自内源基因和异源基因表达的活性的组合,如在此更加详细地描述。
等位基因变体:术语“等位基因变体”意指占用同一染色体位点的一种基因的两个或更多个替代形式中的任一者。等位基因变异由突变天然产生,并且可以导致群体内多态性。基因突变可以是沉默的(在编码的多肽方面无变化)或可以编码具有改变的氨基酸序列的多肽。多肽的等位基因变体是由基因的等位基因变体编码的多肽。
编码序列:术语“编码序列”或“编码区”意指指定一个多肽的氨基酸序列的多核苷酸序列。编码序列的边界一般由开放阅读框架决定,该开放阅读框架通常以ATG起始密码子或替代性起始密码子(例如GTG和TTG)开始,并且以终止密码子(例如TAA、TAG、和TGA)结束。编码序列可以是基因组DNA、cDNA、合成的多核苷酸、和/或重组多核苷酸的一个序列。
控制序列:术语“控制序列”意指多肽表达所必需的核酸序列。控制序列对于编码多肽的多核苷酸可以是天然的或外源的,并且彼此可以是天然的或外源的。此类控制序列包括但不限于,前导子序列、多聚腺苷酸化序列、前肽序列、启动子序列、信号肽序列及转录终止子序列。出于引入有利于将这些控制序列与编码多肽的多核苷酸的编码区连接的特异性限制酶切位点的目的,这些控制序列可以提供有多个接头。
破坏:术语“破坏”意指参考基因的编码区和/或控制序列被部分或完全修饰(例如通过缺失、插入和/或取代一个或多个核苷酸),从而使得编码的多肽的表达不存在(失活)或降低,和/或编码的多肽的酶活性不存在或降低。可以使用本领域已知的技术测量破坏效果,如使用来自在此参考的无细胞提取物测量值检测酶活性的不存在或降低;或通过相应的mRNA的不存在或降低(例如,至少25%降低、至少50%降低、至少60%降低、至少70%降低、至少80%降低或至少90%降低);具有酶活性的相应多肽的量的不存在或降低(例如,至少25%降低、至少50%降低、至少60%降低、至少70%降低、至少80%降低或至少90%降低);或具有酶活性的相应多肽的比活性的不存在或降低(例如,至少25%降低、至少50%降低、至少60%降低、至少70%降低、至少80%降低或至少90%降低)。可以通过本领域已知的方法破坏感兴趣的具体基因,例如通过直接同源重组(参见Methods in Yeast Genetics(1997edition),Adams,Gottschling,Kaiser,and Stems,Cold Spring Harbor Press(1998)[酵母遗传学方法,1997版,Adams,Gottschling,Kaiser和Stems,冷泉港出版社(1998))。
内源基因:术语“内源基因”意指对参考宿主细胞而言天然的基因。“内源基因表达”意指内源基因的表达。
表达:术语“表达”包括涉及多肽产生的任何步骤,包括但不限于转录、转录后修饰、翻译、翻译后修饰以及分泌。可以对表达进行测量-例如,来检测增加的表达-通过本领域已知的技术,例如测量mRNA和/或翻译的多肽的水平。
表达载体:术语“表达载体”意指一种线性或环形的DNA分子,该分子包括编码一种多肽的多核苷酸并且被可操作地连接至控制序列,其中这些控制序列提供编码该多肽的多核苷酸的表达。最低限度上,该表达载体包括启动子序列,以及转录和翻译终止信号序列。
可发酵的培养基:术语“可发酵的培养基”或“发酵培养基”是指包括一种或多种(例如,两种、若干种)糖的培养基,例如葡萄糖、果糖、蔗糖、纤维二糖、木糖、木酮糖、阿拉伯糖、甘露糖、半乳糖和/或可溶性低聚糖,其中该培养基能够部分地被宿主细胞转化(发酵)为希望的产物,例如3-HP。在一些情况下,这种发酵培养基来源于天然来源,例如甘蔗、淀粉、或纤维素;并且可以来自这种来源的酶水解(糖化作用)的预处理。
异源多核苷酸:在此将术语“异源多核苷酸”定义为以下多核苷酸:对该宿主细胞而言不是天然的多核苷酸;已经对编码区做出结构修饰的天然多核苷酸;由于通过重组DNA技术(例如,一种不同的(外源的)启动子)对DNA的操作而定量改变其表达的天然多核苷酸;或具有该多核苷酸的一个或多个额外的拷贝以定量改变表达的宿主细胞中的天然多核苷酸。“异源基因”是包括异源多核苷酸的基因。
宿主细胞:术语“宿主细胞”意指对用核酸构建体或表达载体进行的转化、转染、转导等是易感的任何细胞类型。术语“宿主细胞”涵盖由于复制期间发生的突变而与亲本细胞不同的亲本细胞的任何后代。在此将术语“重组细胞”定义为包括一种或多种(例如,两种、若干种)异源多核苷酸的非天然存在的宿主细胞。宿主细胞可以是在此描述的突变菌株,或被进一步破坏以提供在此描述的突变菌株。
杂交条件:语“非常低严格条件”是指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和25%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在45℃下洗涤三次,每次15分钟。
术语“低严格条件”意指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和25%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在50℃下洗涤三次,每次15分钟。
术语“中严格条件”意指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和35%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在55℃下洗涤三次,每次15分钟。
术语“中-高严格条件”意指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和35%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在60℃下洗涤三次,每次15分钟。
术语“高严格条件”意指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和50%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在65℃下洗涤三次,每次15分钟。
术语“非常高严格条件”意指对于长度为至少100个核苷酸的探针而言,遵循标准DNA印迹程序,在42℃下在5X SSPE、0.3%SDS、200微克/ml剪切并变性的鲑鱼精子DNA和50%甲酰胺中预杂交和杂交12至24小时。载体材料最终使用0.2X SSC、0.2%SDS,在70℃下洗涤三次,每次15分钟。
分离的:术语“分离的”意指处于自然界中不存在的形式或环境中的物质。分离的物质的非限制性实例包括(1)任何非天然存在的物质,(2)任何物质,包括(但不限于)从与其性质上相关的一种或多种或所有天然存在的组分至少部分除去的任何宿主细胞、酶、变体、核酸、蛋白质、肽或辅因子;(3)相对于自然中发现的物质,由人工修饰的任何物质;或(4)通过相对于与其天然相关的其他组分,增加物质的量而修饰的任何物质。
突变体:术语“突变体”意指对亲本菌株进行一个或多个破坏后的所得菌株。
核酸构建体:术语“核酸构建体”意指一种包括一个或多个(例如,两个、若干个)控制序列的多核苷酸。多核苷酸可以是单链的或双链的,并且可以分离自天然存在的基因、可以被修饰成以另外的不会在自然界中存在的方式包含核酸的区段,或可以是合成的。
可操作地连接:术语“可操作地连接”意指如下的构造,其中,控制序列相对于多核苷酸的编码序列安置在适当位置,从而使得该控制序列指导该编码序列的表达。
亲本:术语“亲本”或“亲本菌株”意指对其进行破坏以产生在此描述的突变菌株的菌株。亲本可以是天然存在的(野生型)或预先修饰的菌株(例如,经修饰以包含主动3-HP途径的菌株)。
丙酮酸还原酶:术语“丙酮酸还原酶”意指具有参考序列的多肽,其可以在表达该多肽的宿主细胞中有助于增加水平的D-乳酸盐/D-乳酸酯,例如通过在NAD(H)或NADP(H)辅因子的存在下,催化丙酮酸还原成D-乳酸盐/D-乳酸酯。在此描述的任何方面,参考丙酮酸还原酶可以具有丙酮酸还原酶活性。丙酮酸还原酶活性可根据实例中描述的丙酮酸还原酶测定来确定。在一个方面,本发明的丙酮酸还原酶具有至少20%,例如,至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少95%或至少100%的SEQ ID NO:205的丙酮酸还原酶。
序列一致性:两个氨基酸序列之间或者两个核苷酸序列之间的关联度通过参数“序列一致性”来描述。
出于在此所述的目的,两个氨基酸序列之间的序列一致性的程度使用Needleman-Wunsch算法(Needleman和Wunsch,J.Mol.Biol.[分子生物学杂志],1970,48:443-453)确定,如在EMBOSS包的Needle程序(EMBOSS:The European Molecular Biology OpenSoftware Suite[欧洲分子生物学开放软件包],Rice等人,Trends Genet.[遗传学趋势],2000,16:276-277),优选地3.0.0版或更高版本中所实施。所用的任选参数是空位开放罚分10、空位延伸罚分0.5,和EBLOSUM62(BLOSUM62的EMBOSS版)替代矩阵。Needle标注的“最长的一致性”的输出(使用-非简化选项获得)被用作百分比一致性,并且如下计算:
(一致的残基X 100)/(参考序列的长度-比对中的空位总数)
出于在此所述的目的,两个脱氧核糖核苷酸序列之间的序列一致性的程度使用Needleman-Wunsch算法(Needleman和Wunsch,1970,同上)确定,如在EMBOSS包的Needle程序(EMBOSS:The European Molecular Biology Open Software Suite[欧洲分子生物学开放软件包],Rice等人,2000,同上),优选地3.0.0版或更高版本中所实施。所用的任选参数是空位开放罚分10、空位延伸罚分0.5,和EDNAFULL(NCBI NUC4.4的EMBOSS版)替代矩阵。Needle标注的“最长的一致性”的输出(使用-非简化选项获得)被用作百分比一致性,并且如下计算:
(一致的脱氧核糖核苷酸X 100)/(参考序列的长度-比对中的空位总数)
体积生产力:术语“体积生产力”是指每单位时间使用的每体积系统(例如,培养基和其中的内容物的总体积)产生的参考产物的量(例如,产生的3-HP的量)。
在此提及“约”一个数值或参数包括指向那个数值或参数本身的方面。例如,提及“约X”的描述包括方面“X”。当与测量值组合使用时,“约”包括涵盖至少与测量该具体数值的方法相关的不确定性的范围,并且可以包括在给定的数值附近正或负两个标准差的范围。
如在此和所附权利要求书中所使用,单数形式“一种/个”、“或”以及“该”包括复数指示物,除非上下文以另外的方式清楚表明。应该理解的是在此描述的这些方面包括“由……方面组成”和/或“基本由……方面组成”。
除非由上下文以另外的方式定义或清楚表明,在此使用的所有技术术语和科学术语具有如本领域普通技术人员所通常理解的相同的含义。
详细说明
在此尤其描述了包含与丙酮酸还原酶基因相关的修饰的微生物菌株。本申请人已经发现了包含SEQ ID NO:204的序列(编码SEQ ID NO:205的丙酮酸还原酶)的内源基因,并且当在相同条件下培养时,与亲本菌株相比,该基因的破坏导致突变体中D-乳酸盐/D-乳酸酯的减少的产生。因此,向技术人员提供了产生D-乳酸盐/D-乳酸酯(通过丙酮酸还原酶的过表达),或减少不希望的D-乳酸盐/D-乳酸酯污染物以改进发酵宿主细胞中的碳产率(通过破坏丙酮酸还原酶基因)的手段。
丙酮酸还原酶表达
在一个方面,是包含编码丙酮酸还原酶的异源多核苷酸的重组宿主细胞。丙酮酸还原酶可以是适合宿主细胞及其使用方法的在此描述的任何丙酮酸还原酶,例如SEQ IDNO:205的丙酮酸还原酶,或保留了丙酮酸还原酶活性的其变体,例如人造变体或来自其他物种的天然变体。
在一个实施例中,异源多核苷酸编码包含SEQ ID NO:205的氨基酸序列或由其组成的丙酮酸还原酶。在一些方面,丙酮酸还原酶在相同条件下具有至少20%,例如,至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%的SEQ ID NO:205的丙酮酸还原酶活性。
在另一个实施例中,异源多核苷酸编码SEQ ID NO:205的多肽的片段,其中该片段具有丙酮酸还原酶活性。在一个实施例中,片段中的氨基酸残基的数目为SEQ ID NO:205的至少75%,例如至少80%、85%、90%或95%。
异源多核苷酸也可以编码SEQ ID NO:205的丙酮酸还原酶的变体。在一个实施例中,丙酮酸还原酶与SEQ ID NO:205具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一个实施例中,丙酮酸还原酶序列与SEQ ID NO:205相差不超过十个氨基酸,例如不超过五个氨基酸,不超过四个氨基酸,不超过三个氨基酸,不超过两个氨基酸或一个氨基酸。在一个实施例中,丙酮酸还原酶包含SEQ ID NO:205的氨基酸序列,或等位基因变体或具有丙酮酸还原酶活性的其片段或由它们组成。在一个实施例中,丙酮酸还原酶具有SEQ ID NO:205的一个或多个(例如两个,若干个)氨基酸的氨基酸取代、缺失和/或插入。在一些实施例中,氨基酸取代、缺失和/或插入的总数不超过10,例如不超过9、8、7、6、5、4、3、2或1。
氨基酸改变的性质通常较小,也就是说不会显著影响蛋白质折叠和/或活性的保守氨基酸取代或插入;典型地为一个至约30个氨基酸的小缺失;小的氨基末端或羧基末端延伸,例如氨基末端的甲硫氨酸残基;多至约20-25个残基的小接头肽;或便于通过改变净电荷或另一种功能来纯化的小延伸,如聚组氨酸段(tract)、抗原表位或结合结构域。
保守取代的实例在下组之内:碱性氨基酸(精氨酸、赖氨酸和组氨酸)、酸性氨基酸(谷氨酸和天冬氨酸)、极性氨基酸(谷氨酰胺和天冬酰胺)、疏水氨基酸(亮氨酸、异亮氨酸和缬氨酸)、芳族氨基酸(苯丙氨酸、色氨酸和酪氨酸)、以及小氨基酸(甘氨酸、丙氨酸、丝氨酸、苏氨酸以及甲硫氨酸)。一般不会改变比活性的氨基酸取代是本领域已知的并且例如由H.Neurath,R.L.Hill,1979,在The Proteins,Academic Press,New York[蛋白质,学术出版社,纽约]中描述。最常发生的交换是Ala/Ser、Val/Ile、Asp/Glu、Thr/Ser、Ala/Gly、Ala/Thr、Ser/Asn、Ala/Val、Ser/Gly、Tyr/Phe、Ala/Pro、Lys/Arg、Asp/Asn、Leu/Ile、Leu/Val、Ala/Glu、和Asp/Gly。
可替代地,这些氨基酸改变具有如下此种性质:改变多肽的物理化学特性。例如,氨基酸改变可以提高丙酮酸还原酶的热稳定性、改变底物特异性、改变最适pH,等。
可以根据本领域已知的程序来鉴定必需氨基酸,例如定点诱变或丙氨酸扫描诱变(Cunningham和Wells,1989,Science[科学]244:1081-1085)。在后一项技术中,在该分子中的每个残基处引入单个丙氨酸突变,并且对所得突变体分子的丙酮酸还原酶活性进行测试以鉴定对于该分子的活性至关重要的氨基酸残基。还参见Hilton等人,1996,J.Biol.Chem.[生物化学杂志]271:4699-4708。丙酮酸还原酶或其他生物学相互作用的活性位点还可通过对结构的物理分析来确定,如由下述技术确定:核磁共振、晶体学、电子衍射、或光亲和标记,连同对推定的接触位点氨基酸进行突变。参见例如de Vos等人,1992,Science[科学]255:306-312;Smith等人,1992,J.Mol.Biol.[分子生物学杂志]224:899-904;Wlodaver等人,1992,FEBS Lett.[欧洲生物化学学会联盟简讯]309:59-64。还可以从与参考丙酮酸还原酶相关的其他丙酮酸还原酶的一致性的分析推断必需氨基酸的一致性。
可以使用本领域熟知的多重序列比对(MSA)技术来确定有关在此的丙酮酸还原酶的结构-活性关系的额外指导。基于在此的教导,技术人员可以与本领域已知的许多丙酮酸还原酶做相似的比对。此类比对帮助技术人员确定潜在的相关结构域(例如,结合域或催化结构域),并且在不同的丙酮酸还原酶序列中确定哪些氨基酸残基是保守的和不是保守的。本领域中应理解的是,改变在所披露的丙酮酸还原酶与本领域已知的那些丙酮酸还原酶之间特定位置处是保守的氨基酸将更有可能导致生物活性的改变(Bowie等人,1990,Science[科学]247:1306-1310:“Residues that are directly involved in protein functionssuch as binding or catalysis will certainly be among the most conserved”[直接涉及到蛋白功能如结合或催化的残基将一定是在最保守的残基中])。相比之下,取代这些丙酮酸还原酶中不是高度保守的氨基酸将不可能或不显著地改变生物活性。
可以做出单个或多个氨基酸取代、缺失和/或插入并且使用诱变、重组和/或改组的已知方法进行测试,随后进行相关筛选程序,如由Reidhaar-Olson和Sauer,1988,Science[科学]241:53-57;Bowie和Sauer,1989,Proc.Natl.Acad.Sci.USA[美国科学院院刊]86:2152-2156;WO 95/17413;或WO 95/22625所披露的那些。可以使用的其他方法包括易错PCR、噬菌体展示(例如,Lowman等人,1991,Biochemistry[生物化学]30:10832-10837;美国专利号5,223,409;WO 92/06204)以及区域定向诱变(Derbyshire等人,1986,Gene[基因]46:145;Ner等人,1988,DNA 7:127)。
可以结合诱变/改组方法与高通量自动化筛选方法来检测由宿主细胞表达的克隆的、诱变的多肽的活性(Ness等人,1999,Nature Biotechnology[自然生物技术]17:893-896)。可以从宿主细胞回收编码活性丙酮酸还原酶的诱变的DNA分子,并且使用本领域的标准方法快速测序。这些方法允许迅速确定多肽中单个氨基酸残基的重要性。
在另一个实施例中,编码丙酮酸还原酶的异源多核苷酸包含与SEQ ID NO:204具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性的编码序列。
在一个实施例中,编码丙酮酸还原酶的异源多核苷酸包括SEQ ID NO:204的编码序列。在另一个实施例中,编码丙酮酸还原酶的异源多核苷酸包括SEQ ID NO:204的编码序列的子序列,其中该子序列编码具有丙酮酸还原酶活性的多肽。在另一个实施例中,编码子序列中的核苷酸残基的数目是SEQ ID NO:204的至少75%,例如,至少80%、85%、90%或95%。
在此描述的任何相关方面或实施例的参考编码序列可以是天然编码序列或简并序列,例如为特定宿主细胞设计的密码子优化的编码序列。
SEQ ID NO:204的多核苷酸编码序列或其子序列,连同SEQ ID NO:205的多肽或其片段,可用于设计核酸探针,以根据本领域熟知的方法从不同属或种的菌株中鉴定和克隆编码丙酮酸还原酶的DNA。具体而言,此类探针可以用于按照标准DNA印迹程序与感兴趣的细胞的基因组DNA或cDNA杂交,以便鉴定和分离其中相应的基因。这类探针可以明显短于完整序列,但是长度应为至少15,例如至少25、至少35、或至少70个核苷酸。优选地,该核酸探针的长度为至少100个核苷酸,例如长度为至少200个核苷酸、至少300个核苷酸、至少400个核苷酸、至少500个核苷酸、至少600个核苷酸、至少700个核苷酸、至少800个核苷酸、或至少900个核苷酸。DNA和RNA探针两者都可使用。典型地将探针进行标记,用于检测相应的基因(例如,用32P、3H、35S、生物素或抗生物素蛋白)。
可以针对与上文所述的探针杂交并编码亲本的DNA来筛选由这类其他菌株制备的基因组DNA或cDNA文库。来自这类其他菌株的基因组DNA或其他DNA可以通过琼脂糖或聚丙烯酰胺凝胶电泳,或其他分离技术来分离。可以将来自文库的DNA或分离的DNA转移至硝化纤维素(nitrocellulose)或其他适合的载体材料并且固定于其上。为了鉴定与SEQ ID NO:204或其子序列杂交的克隆或DNA,在DNA印迹中使用载体材料。
在一个方面,核酸探针是包含SEQ ID NO:204或其子序列的多核苷酸。在另一个方面,核酸探针是编码SEQ ID NO:205的多肽或其片段的多核苷酸。
出于上述探针的目的,杂交是指,在非常低至非常高严格条件下,多核苷酸杂交至标记的核酸探针或其全长互补链或前述各项的子序列。在这些条件下,核酸探针杂交的分子可以使用例如X射线胶片而进行检测。严格性和洗涤条件如所上述所定义。
在一个实施例中,丙酮酸还原酶由多核苷酸编码,其在至少低严格条件下,例如中严格条件下、中-高严格条件下、高严格条件下、或非常高严格条件下与SEQ ID NO:204的全长互补链杂交。(Sambrook et al.,1989,Molecular Cloning,A Laboratory Manual,2dedition,Cold Spring Harbor,New York[Sambrook等人,1989,分子克隆:实验室手册,第2版,冷泉港,纽约])。
这些丙酮酸还原酶可以获得自任何适合属的微生物,这些微生物包括在UniProtKB数据库(www.uniprot.org)内可容易获得的那些。
丙酮酸还原酶可以是细菌丙酮酸还原酶。例如,该丙酮酸还原酶可以是革兰氏阳性细菌多肽,如芽胞杆菌属(Bacillus)、梭状芽胞杆菌属(Clostridium)、肠球菌属(Enterococcus)、土芽孢杆菌属(Geobacillus)、乳杆菌属(Lactobacillus)、乳球菌属(Lactococcus)、大洋芽孢杆菌属(Oceanobacillus)、葡萄球菌属(Staphylococcus)、链球菌属(Streptococcus)或链霉菌属(Streptomyces)丙酮酸还原酶,或者革兰氏阴性细菌多肽,如弯曲杆菌属(Campylobacter)、大肠杆菌(E.coli)、黄杆菌属(Flavobacterium)、梭杆菌属(Fusobacterium)、螺杆菌属(Helicobacter)、泥杆菌属(Ilyobacter)、奈瑟氏菌属(Neisseria)、假单胞菌属(Pseudomonas)、沙门氏菌属(Salmonella)、或脲原体属(Ureaplasma)丙酮酸还原酶。
在一个实施例中,该丙酮酸还原酶是嗜碱芽孢杆菌(Bacillus alkalophilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、短芽孢杆菌(Bacillus brevis)、环状芽孢杆菌(Bacillus circulans)、克劳氏芽孢杆菌(Bacillus clausii)、凝结芽孢杆菌(Bacillus coagulans)、坚强芽孢杆菌(Bacillus firmus)、灿烂芽孢杆菌(Bacilluslautus)、迟缓芽孢杆菌(Bacillus lentus)、地衣芽孢杆菌(Bacillus licheniformis)、巨大芽孢杆菌(Bacillus megaterium)、短小芽孢杆菌(Bacillus pumilus)、嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)、枯草芽孢杆菌(Bacillus subtilis)或苏云金芽孢杆菌(Bacillus thuringiensis)丙酮酸还原酶。
在另一个实施例中,该丙酮酸还原酶是似马链球菌(Streptococcusequisimilis)、酿脓链球菌(Streptococcus pyogenes)、乳房链球菌(StreptococcusUberis)、或马链球菌兽瘟亚种(Streptococcus equi subsp.Zooepidemicus)丙酮酸还原酶。
在另一个实施例中,该丙酮酸还原酶是不产色链霉菌(Streptomycesachromogenes)、除虫链霉菌(Streptomyces avermitilis)、天蓝链霉菌(Streptomycescoelicolor)、灰色链霉菌(Streptomyces griseus)或浅青紫链霉菌(Streptomyceslividans)丙酮酸还原酶。
丙酮酸还原酶可以是真菌丙酮酸还原酶。例如,该丙酮酸还原酶可以是酵母丙酮酸还原酶,例如假丝酵母属、克鲁维酵母属、毕赤酵母属、酵母属、裂殖酵母属、亚罗酵母属或伊萨酵母属丙酮酸还原酶;或丝状真菌丙酮酸还原酶,例如支顶孢属(Acremonium)、伞菌属(Agaricus)、支链孢属(Alternaria)、曲霉属(Aspergillus)、短梗霉属(Aureobasidium)、葡萄座腔菌属(Botryospaeria)、拟蜡菌属(Ceriporiopsis)、毛喙壳属(Chaetomidium)、金孢子菌属(Chrysosporium)、麦角菌属(Claviceps)、旋孢腔菌属(Cochliobolus)、拟鬼伞属(Coprinopsis)、家白蚁属(Coptotermes)、棒囊孢壳菌属(Corynascus)、隐丛壳属(Cryphonectria)、隐球酵母属(Cryptococcus)、壳色单隔孢属(Diplodia)、黑耳属(Exidia)、网孢菌属(Filibasidium)、镰孢属(Fusarium)、赤霉菌属(Gibberella)、全鞭毛虫属(Holomastigotoides)、腐质霉属(Humicola)、耙齿菌属(Irpex)、香菇属(Lentinula)、小球腔菌属(Leptospaeria)、大毁壳属(Magnaporthe)、马兰诺菌属(Melanocarpus)、亚灰树花菌属(Meripilus)、毛霉菌属(Mucor)、蚀丝霉属(Myceliophthora)、新美鞭菌属(Neocallimastix)、脉孢菌属(Neurospora)、拟青霉属(Paecilomyces)、青霉属(Penicillium)、平革菌属(Phanerochaete)、梨囊鞭菌属、Poitrasia、假黑盘菌属(Pseudoplectania)、假披发虫属(Pseudotrichonympha)、根毛霉属(Rhizomucor)、裂褶菌属(Schizophyllum)、柱顶孢霉属(Scytalidium)、踝节菌属(Talaromyces)、嗜热子囊菌属(Thermoascus)、梭孢壳菌属(Thielavia)、弯颈霉属(Tolypocladium)、木霉属(Trichoderma)、长毛盘菌属(Trichophaea)、轮枝孢属(Verticillium)、小包脚菇属(Volvariella)、或炭角菌属(Xylaria)丙酮酸还原酶。
在另一个实施例中,该丙酮酸还原酶是卡氏酵母(Saccharomycescarlsbergensis)、酿酒酵母(Saccharomyces cerevisiae)、糖化酵母(Saccharomycesdiastaticus)、道格拉氏酵母(Saccharomyces douglasii)、克鲁维酵母(Saccharomyceskluyveri)、诺地酵母(Saccharomyces norbensis)、或卵形酵母(Saccharomycesoviformis)丙酮酸还原酶。
在另一个实施例中,该丙酮酸还原酶是解纤维枝顶孢霉(Acremoniumcellulolyticus)、棘孢曲霉(Aspergillus aculeatus)、泡盛曲霉(Aspergillus awamori)、臭曲霉(Aspergillus foetidus)、烟曲霉(Aspergillusfumigatus)、日本曲霉(Aspergillus japonicus)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)、狭边金孢子菌(Chrysosporiuminops)、嗜角质金孢子菌(Chrysosporium keratinophilum)、Chrysosporiumlucknowense、Chrysosporium merdarium、租金孢子菌(Chrysosporium pannicola)、昆士兰金抱子菌(Chrysosporium queenslandicum)、热带金孢子菌(Chrysosporiumtropicum)、带纹金孢子菌(Chrysosporium zonatum)、杆孢状镰孢(Fusariumbactridioides)、谷类镰孢(Fusarium cerealis)、库威镰孢(Fusarium crookwellense)、大刀镰孢(Fusarium culmorum)、禾谷镰孢(Fusarium graminearum)、禾赤镰孢(Fusariumgraminum)、异孢镰孢(Fusarium heterosporum)、合欢木镰孢(Fusarium negundi,)、尖镰孢(Fusarium oxysporum)、多枝镰孢(Fusarium reticulatum)、粉红镰孢(Fusariumroseum)、接骨木镰孢(Fusarium sambucinum)、肤色镰孢(Fusarium sarcochroum)、拟分枝孢镰孢(Fusarium sporotrichioides)、硫色镰孢(Fusarium sulphureum)、圆镰孢(Fusarium torulosum)、拟丝孢镰孢(Fusarium trichothecioides)、镶片镰孢(Fusariumvenenatum)、灰腐质霉(Humicola grisea)、特异腐质霉(Humicola insolens)、疏棉状腐质霉(Humicola lanuginosa)、白耙齿菌(Irpex lacteus)、米黑毛霉(Mucor miehei)、嗜热毁丝霉(Myceliophthora thermophila)、粗糙链孢菌(Neurospora crassa)、绳状青霉菌(Penicillium funiculosum)、产紫青霉菌(Penicillium purpurogenum)、黄孢原毛平革菌(Phanerochaete chrysosporium)、无色梭孢壳霉(Thielavia achromatica)、成层梭抱壳菌(Thielavia albomyces)、白毛梭孢壳霉(Thielavia albopilosa)、澳洲梭孢壳霉(Thielavia australeinsis)、菲美蒂梭抱壳菌(Thielavia fimeti)、小孢梭孢壳霉(Thielavia microspora)、卵孢梭孢壳霉(Thielavia ovispora)、秘鲁梭孢壳霉(Thielavia peruviana)、毛梭孢壳霉(Thielavia setosa)、瘤孢梭孢壳霉(Thielaviaspededonium)、耐热梭孢壳(Thielavia subthermophila)、土生梭孢壳(Thielaviaterrestris)、哈茨木霉(Trichoderma harzianum)、康宁木霉(Trichoderma koningii)、长枝木霉(Trichoderma longibrachiatum)、里氏木霉(Trichoderma reesei)、或绿色木霉(Trichoderma viride)丙酮酸还原酶。
在另一个方面,丙酮酸还原酶来自伊萨酵母属,如SEQ ID NO:205的东方伊萨酵母(Issatchenkia orientalis)丙酮酸还原酶。
应理解的是对于前述的种,本发明涵盖完全和不完全阶段(perfect andimperfect states),和其他分类学的等同物(equivalent),例如无性型(anamorph),而与它们已知的种名无关。本领域普通技术人员将容易地识别适当等效物的身份。
这些物种的菌株可以容易地在许多培养物保藏中心为公众所获得,如美国典型培养物保藏中心(ATCC)、德国微生物菌种保藏中心(Deutsche Sammlung vonMikroorganismen und Zellkulturen GmbH,DSMZ)、荷兰菌种保藏中心(CentraalbureauVoor Schimmelcultures,CBS)以及美国农业研究服务专利培养物保藏中心北方地区研究中心(NRRL)。
这些物种的菌株可以容易地在许多培养物保藏中心为公众所获得,如美国典型培养物保藏中心(ATCC)、德国微生物菌种保藏中心(Deutsche Sammlung vonMikroorganismen und Zellkulturen GmbH,DSM)、荷兰菌种保藏中心(CentraalbureauVoor Schimmelcultures,CBS)以及美国农业研究服务专利培养物保藏中心北方地区研究中心(NRRL)。
也可以使用以上提到的探针从其他来源,包括从自然界(例如,土壤、堆肥、水、青贮等)分离的微生物或直接从天然材料(例如,土壤、堆肥、水、青贮等)获得的DNA样品鉴定并获得丙酮酸还原酶。直接从天然栖息地分离微生物和DNA的技术是本领域熟知的。随后可以通过类似地筛选另一种微生物的基因组或cDNA文库或混合DNA样品衍生编码丙酮酸还原酶的多核苷酸。
一旦已经用如在此描述的适合的探针检测到编码丙酮酸还原酶的多核苷酸,就可以通过使用本领域普通技术人员已知的技术来分离或克隆序列(参见例如,Sambrook等人,1989,同上)。用于分离或克隆编码丙酮酸还原酶的多核苷酸的技术包括从基因组DNA分离、从cDNA制备或其组合。可以例如通过使用熟知的聚合酶链反应(PCR)或表达文库的抗体筛选来检测具有共有结构特征的克隆DNA片段,实现从这样的基因组DNA克隆多核苷酸。参见例如,Innis et al.,1990,PCR:A Guide to Methods and Application,Academic Press,New York[Innis等人,1990,PCR:方法和应用指南,学术出版社,纽约]。还可以使用其他核酸扩增程序,诸如连接酶链式反应(LCR)、连接激活转录(LAT)和基于核苷酸序列的扩增(NASBA)。
丙酮酸还原酶可以是融合的多肽或可切割的融合多肽,其中另一个多肽在丙酮酸还原酶的N-末端或C-末端处融合。可以通过将编码另一多肽的多核苷酸融合于编码丙酮酸还原酶的多核苷酸来产生融合的多肽。用于产生融合多肽的技术是本领域已知的,并包括连接编码多肽的编码序列,这样使得它们在阅读框中,并且使所述融合的多肽的表达在相同的一个或多个启动子和终止子的控制下。融合蛋白还可以使用内含肽技术构建,其中融合在翻译后产生(Cooper等人,1993,EMBO J.[欧洲分子生物学学会杂志]12:2575-2583;Dawson等人,1994,Science[科学]266:776-779)。
在一些实施例中,当在相同条件下培养时,与不含编码丙酮酸还原酶的异源多核苷酸的宿主细胞相比,上述包含编码丙酮酸还原酶的异源多核苷酸的重组细胞产生(具有)增加的水平的丙酮酸还原酶活性(例如,至少5%、至少10%、至少15%、至少20%、至少25%、至少50%、至少100%、至少150%、至少200%、至少300%、或500%以上)。
在一些实施例中,包含编码丙酮酸还原酶的异源多核苷酸的上述宿主细胞能够产生D-乳酸盐/D-乳酸酯。因此,在一个方面,是生产D-乳酸盐/D-乳酸酯或其盐的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养在此描述的包含编码丙酮酸还原酶(例如SEQ ID NO:205的丙酮酸还原酶)的异源多核苷酸的重组细胞以产生D-乳酸盐/D-乳酸酯或其盐;并且(b)回收D-乳酸盐/D-乳酸酯。适合的发酵条件是本领域已知的,并且在本文中有更详细地描述。
在一些实施例中,当在相同条件下培养时,与没有编码丙酮酸还原酶的异源多核苷酸的细胞相比,包含编码丙酮酸还原酶的异源多核苷酸的宿主细胞产生(和/或能够产生)更大量的D-乳酸盐/D-乳酸酯(例如,多至少15%、至少20%、至少25%、至少30%、至少35%、至少40%、至少45%、至少50%、至少75%、至少100%、至少200%、至少300%,或500%以上)。
丙酮酸还原酶基因破坏
在一个方面,是包含对编码丙酮酸还原酶内源基因的破坏的突变菌株。菌株可以是任何适合的微生物细胞,例如下述任何宿主细胞。
在突变体和相关方法的一些实施例中,内源基因编码包含SEQ ID NO:205的氨基酸序列(或由其组成)的丙酮酸还原酶。在一些方面,内源基因编码丙酮酸还原酶,该丙酮酸还原酶在相同的条件下具有至少20%,例如,至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%的SEQ IDNO:205的丙酮酸还原酶活性。
在突变体及相关方法的一些实施例中,内源基因编码丙酮酸还原酶,该丙酮酸还原酶与SEQ ID NO:205具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%序列一致性。在一个实施例中,丙酮酸还原酶序列与SEQ IDNO:205相差不超过十个氨基酸,例如不超过五个氨基酸,不超过四个氨基酸,不超过三个氨基酸,不超过两个氨基酸或一个氨基酸。在一个实施例中,丙酮酸还原酶包含SEQ ID NO:205的氨基酸序列,或等位基因变体或具有丙酮酸还原酶活性的其片段或由它们组成。在一个实施例中,丙酮酸还原酶具有SEQ ID NO:205的一个或多个(例如两个,若干个)氨基酸的氨基酸取代、缺失和/或插入。在一些实施例中,氨基酸取代、缺失和/或插入的总数不超过10,例如不超过9、8、7、6、5、4、3、2或1。
在突变体和相关方法的一些实施例中,编码丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一些实施例中,内源基因的编码序列包括SEQ ID NO:204或由其组成。
在突变体和相关方法的一些实施例中,编码丙酮酸还原酶内源基因的编码序列在至少低严格条件下,例如中严格条件下、中-高严格条件下、高严格条件下、或非常高严格条件下与SEQ ID NO:204的全长互补链杂交。(Sambrook et al.,1989,Molecular Cloning,ALaboratory Manual,2d edition,Cold Spring Harbor,New York[Sambrook等人,1989,分子克隆:实验室手册,第2版,冷泉港,纽约])。
在突变体和相关方法的一些实施例中,当在相同条件下培养时,与缺乏对编码丙酮酸还原酶内源基因的破坏的亲本菌株相比,突变体产生更少的D-乳酸盐/D-乳酸酯(例如,减少至少25%、减少50%、减少至少60%、减少至少70%、减少至少80%、减少至少90%、或减少100%)。
在突变体和相关方法的一些实施例中,当在相同条件下培养时,与缺乏对编码丙酮酸还原酶内源基因的破坏的亲本菌株相比,突变体产生更少的丙酮酸还原酶(例如,减少至少25%、减少至少50%、减少至少60%、减少至少70%、减少至少80%、减少至少90%、或减少100%)。在一些实施例中,编码丙酮酸还原酶的内源基因是失活的。
在此描述的突变菌株可以通过使用本领域熟知的方法(包括在此描述的那些方法)破坏参考内源性丙酮酸还原酶来构建。可以破坏该操纵子的一部分,例如一个或多个(例如,两个、若干个)编码区或为这些编码区的表达所需的控制序列。该操纵子的这样一种控制序列可以是启动子序列或其功能部分,即足以影响该操纵子的表达的部分。例如,可以将启动子序列失活从而无表达或可以将天然启动子替换为更弱的启动子以减少编码序列的表达。可修饰的其他控制序列包括但不限于前导子、前肽序列、信号序列、转录终止子以及转录激活因子。
突变菌株可以通过基因缺失技术构建,以消除或减少丙酮酸还原酶编码序列的表达。基因缺失技术使得可以部分或完全去除该操纵子,从而消除表达。在此类方法中,使用已经构建为邻接地包含侧翼于这些基因的5’和3’区的一种或多种质粒,通过同源重组完成该操纵子的缺失。
还可以通过引入、取代和/或去除丙酮酸还原酶编码序列或其转录或翻译所需的其控制序列中的一个或多个(例如,两个、若干个)核苷酸来构建突变菌株。例如,可以插入或去除核苷酸,用于引入终止密码子、去除起始密码子或移码开放阅读框。可以根据本领域已知的方法,通过定点诱变或PCR产生的诱变完成这样的修饰。参见,例如Botstein和Shortle,1985,Science[科学]229:4719;Lo等人,1985,Proc.Natl.Acad.Sci.USA[美国国家科学院院刊]81:2285;Higuchi等人,1988,Nucleic Acids Res[核酸研究]16:7351;Shimada,1996,Meth.Mol.Biol.[分子生物学方法]57:157;Ho等人,1989,Gene[基因]77:51;Horton等人,1989,Gene[基因]77:61;以及Sarkar和Sommer,1990,BioTechniques[生物技术]8:404。
还可以通过基因破坏技术,通过将破坏性核酸构建体插入进该丙酮酸还原酶基因中而构建突变菌株,该破坏性核酸构建体包括与该编码序列同源的核酸片段,该片段将产生具有同源性的区域的重复并且在重复的区域之间掺入构建体DNA。这样的一种基因破坏可以消除表达,如果插入的构建体将该基因的启动子与编码区分离或打断编码序列,这样使得产生无功能性或功能性降低的编码序列。破坏构建体可以简单地是伴有与该基因同源的5’和3’区的选择性标记基因。该选择性标记使得可以鉴定包含破坏的基因的转化株。
还可以通过基因转化过程构建突变菌株(参见例如,Iglesias和Trautner,1985,Molecular General Genetics[分子普通遗传学]189:73-76)。例如,在基因转化方法中,将相应于该基因中的序列的核苷酸序列体外诱变,以产生缺陷核苷酸序列,然后将其转化进亲本菌株中以产生缺陷基因。通过同源重组,该缺陷核苷酸序列替换该内源序列。该缺陷核苷酸序列还包括一种用于选择包含缺陷序列的转化体的标记可以是令人希望的。
可以使用本领域熟知的方法(包括但不限于化学诱变),通过随机或特异诱变进一步构建突变菌株(参见例如,Hopwood,The Isolation of Mutants in Methods inMicrobiology(J.R.Norris and D.W.Ribbons,eds.)pp.363-433,Academic Press,NewYork,1970[Hopwood,微生物学方法中的突变体分离,J.R.Norris和D.W.Ribbon编辑,第363-433页,学术出版社,纽约,1970])。可以通过使亲本菌株经受诱变并筛选其中该基因的表达已经被减少或失活的突变菌株来修饰该基因。诱变可以是特异的或随机的,例如通过使用适合的物理或化学诱变剂、使用适合的寡核苷酸或使DNA序列经受PCR产生的诱变来进行。此外,诱变可以通过使用这些诱变方法的任何组合来进行。
适合本发明目的的物理或化学诱变剂的实例包括紫外线(UV)照射,羟胺,N-甲基-N’-硝基-N-亚硝基胍(MNNG),N-甲基-N’-亚硝基胍(NTG)邻甲基羟胺,亚硝酸,乙基甲磺酸(EMS),亚硫酸氢钠,甲酸和核苷酸类似物。当使用此类试剂时,诱变典型地是在适合条件下在所选的诱变剂的存在下通过孵育有待诱变的亲本菌株并选择展现出该基因的减少表达或无表达的突变体来进行的。
可以使用来自其他微生物来源的与在此描述的序列同源或互补的核苷酸序列来破坏所选菌株中的相应序列。
在一个方面,突变体中基因的破坏未用选择性标记加以标记。可以通过将突变体在反向选择培养基中进行培养来去除选择性标记基因。在该选择性标记基因包含侧翼于其5'和3'端的重复序列的情况下,当该突变菌株经受反向选择时,这些重复序列将有助于该选择性标记基因通过同源重组而环出。还可以通过向该突变菌株中引入一个核酸片段,该核酸片段包括缺陷基因的5'和3'区但是缺乏该选择性标记基因,随后在反向选择培养基上进行选择,通过同源重组来去除该选择性标记基因。通过同源重组,包含该选择性标记基因的缺陷基因被缺乏该选择性标记基因的核酸片段替换。还可以使用本领域已知的其他方法。
还描述了产生在此描述的突变体的方法。在一个方面,是用于获得在此描述的突变体的方法,该方法包括在亲本菌株中破坏编码丙酮酸还原酶的内源基因。在另一个方面,是一种用于获得在此描述的突变体的方法,该方法包括:(a)培养亲本菌株;(b)在(a)的亲本菌株中破坏编码丙酮酸还原酶的内源基因;并且(c)分离生成自(b)的突变菌株。
宿主、表达载体与核酸构建体
在此描述的重组宿主细胞和/或突变菌株可以选自任何适合的微生物细胞(例如下述能够具有主动3-HP途径的宿主细胞)。本领域普通技术人员应该理解,遗传改变(包括在此示例的代谢修饰)可以参考适合的宿主生物及其相应的代谢反应或用于希望的遗传材料(例如希望的代谢途径的基因)的适合的来源生物加以描述。然而,考虑到多种多样的生物的全基因组测序以及基因组学领域中的较高水平的技能,本领域普通技术人员可以将在此提供的教导和指导应用于其他生物中。例如,可以通过掺入相同的或来自不同于参考物种的物种的类似编码核酸而容易地将在此示例的代谢改变应用于其他物种中。在一些实施例中,分离宿主细胞和/或突变菌株。
该宿主细胞可以是任何革兰氏阳性或革兰氏阴性细菌。革兰氏阳性细菌包括,但不限于芽孢杆菌属(Bacillus)、梭菌属(Clostridium)、肠球菌属(Enterococcus)、土芽孢杆菌属(Geobacillus)、乳杆菌属(Lactobacillus)、乳球菌属(Lactococcus)、大洋芽孢杆菌属(Oceanobacillus)、葡萄球菌属(Staphylococcus)、链球菌属(Streptococcus)、和链霉菌属(Streptomyces)。革兰氏阴性细菌包括,但不限于弯曲杆菌属(Campylobacter)、大肠杆菌(E.coli)、黄杆菌属(Flavobacterium)、梭杆菌属(Fusobacterium)、螺杆菌属(Helicobacter,)、泥杆菌属(Ilyobacter)、奈瑟球菌属(Neisseria)、假单胞菌属(Pseudomonas)、沙门菌属(Salmonella)、和脲原体属(Ureaplasma)。
细菌宿主细胞可以是任何芽孢杆菌(Bacillus)细胞,包括但不限于嗜碱芽孢杆菌(Bacillus alkalophilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、短杆菌(Bacillus brevis)、环状芽孢杆菌(Bacillus circulans)、克劳氏芽孢杆菌(Bacillusclausii)、凝结芽孢杆菌(Bacillus coagulans)、坚硬芽孢杆菌(Bacillus firmus)、灿烂芽孢杆菌(Bacillus lautus)、迟缓芽孢杆菌(Bacillus lentus)、地衣芽孢杆菌(Bacilluslicheniformis)、巨大芽孢杆菌(Bacillus megaterium)、短小芽孢杆菌(Bacilluspumilus)、嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)、枯草芽孢杆菌(Bacillussubtilis)、以及苏云金芽孢杆菌(Bacillus thuringiensis)的细胞。
细菌宿主细胞还可以是任何链球菌属(Streptococcus)细胞,包括但不限于似马链球菌(Streptococcus equisimilis)、化酿脓链球菌(Streptococcus pyogenes)、乳房链球菌(Streptococcus uberis)和马链球菌兽瘟亚种(Streptococcus equisubsp.Zooepidemicus)的细胞。
细菌宿主细胞还可以是任何链霉菌(Streptomyces)细胞,包括但不限于不产色链霉菌(Streptomyces achromogenes)、除虫链霉菌(Streptomyces avermitilis)、天蓝链霉菌(Streptomyces coelicolor)、灰色链霉菌(Streptomyces griseus)以及浅青紫链霉菌(Streptomyces lividans)细胞。
将DNA引入芽孢杆菌属细胞中可通过以下来实现:原生质体转化(参见,例如,Chang和Cohen,1979,Mol.Gen.Genet.[分子遗传学与基因组学]168:111-115)、感受态细胞转化(参见,例如,Young和Spizizen,1961,J.Bacteriol.[细菌学杂志]81:823-829;或Dubnau和Davidoff-Abelson,1971,J.Mol.Biol.[分子生物学杂志]56:209-221)、电穿孔(参见,例如,Shigekawa和Dower,1988,Biotechniques[生物技术]6:742-751)、或者接合(参见,例如,Koehler和Thorne,1987,J.Bacteriol.[细菌学杂志]169:5271-5278)。将DNA引入大肠杆菌细胞中可通过以下来实现:原生质体转化(参见,例如,Hanahan,1983,J.Mol.Biol.[分子生物学杂志]166:557-580)或电穿孔(参见,例如,Dower等人,1988,Nucleic Acids Res.[核酸研究]16:6127-6145)。将DNA引入链霉菌属细胞中可通过以下来实现:原生质体转化、电穿孔(参见,例如,Gong等人,2004,Folia Microbiol.(Praha)[叶线形微生物学(布拉格)]49:399-405)、接合(参见,例如,Mazodier等人,1989,J.Bacteriol.[细菌学杂志]171:3583-3585)、或转导(参见,例如,Burke等人,2001,Proc.Natl.Acad.Sci.USA[美国科学院院刊]98:6289-6294)。将DNA引入假单孢菌属细胞中可通过以下来实现:电穿孔(参见,例如,Choi等人,2006,J.Microbiol.Methods[微生物学方法杂志]64:391-397)或接合(参见,例如,Pinedo和Smets,2005,[Appl.Environ.Microbiol.[应用与环境微生物学]71:51-57)。将DNA引入链球菌属细胞中可通过以下来实现:天然感受态(参见例如,Perry和Kuramitsu,1981,Infect.Immun.[感染与免疫]32:1295-1297)、原生质体转化(参见例如,Catt和Jollick,1991,Microbios[微生物学]68:189-207)、电穿孔(参见例如,Buckley等人,1999,Appl.Environ.Microbiol.[应用与环境微生物学]65:3800-3804)、或者接合(参见例如,Clewell,1981,Microbiol.Rev.[微生物学评论]45:409-436)。然而,可以使用本领域已知的将DNA引入宿主细胞的任何方法。
宿主细胞还可以是真核生物,如哺乳动物、昆虫、植物或真菌细胞。
如在此使用的“真菌”包括子囊菌门(Ascomycota)、担子菌门(Basidiomycota)、壶菌门(Chytridiomycota)、以及接合菌门(Zygomycota)、连同卵菌门(Oomycota)和全部有丝分裂孢子真菌(如由Hawksworth等人在Ainsworth and Bisby’s Dictionary of TheFungi[安斯沃思和拜斯比真菌词典],第8版,1995,国际应用生物科学中心(CABInternational),大学出版社(University Press),英国剑桥(Cambridge,UK)中所定义)。
在一个实施例中,该宿主细胞是酵母细胞。如在此使用的“酵母”包括产子嚢酵母(内孢霉目(Endomycetales))、产担子酵母和属于半知菌类(Fungi Imperfecti)(芽孢纲(Blastomycetes))的酵母。由于酵母的分类在将来可能改变,出于在此描述的目的,酵母应如在Biology and Activities ofYeast[酵母生物学与活性](Skinner,F.A.,Passmore,S.M.和Davenport,R.R.编辑,Soc.App.Bacteriol.Symposium Series[应用细菌学研讨会系列]第9期,1980)中所述进行定义。
酵母宿主细胞可以是假丝酵母属、汉逊酵母属(Hansenula)、伊萨酵母属、克鲁维酵母属、毕赤酵母属、酵母属、裂殖酵母属或亚罗酵母属(Yarrowia)的细胞,例如萨纳瑞西斯假丝酵母(Candida sonorensis)、甲醇山梨糖假丝酵母(Candida methanosorbosa)、嗜酒假丝酵母(Candida ethanolica)、东方伊萨酵母(Issatchenkia orientalis)、乳酸克鲁维酵母(Kluyveromyces lactis)、马克思克鲁维酵母(Kluyveromyces marxianus)、发酵毕赤酵母(Pichia fermentans)、盔形毕赤酵母(Pichia galeiformis)、膜醭毕赤酵母(Pichia membranifaciens)、沙生毕赤酵母(Pichia deserticola)、卡尔斯伯酵母(Saccharomyces carlsbergensis)、酿酒酵母(Saccharomyces cerevisiae)、糖化酵母(Saccharomyces diastaticus)、布拉迪酵母(Saccharomyces bulderi)、道格拉斯酵母(Saccharomyces douglasii)、克鲁维酵母(Saccharomyces kluyveri)、诺地酵母(Saccharomyces norbensis)、卵形酵母(Saccharomyces oviformis)、或解脂耶氏酵母(Yarrowia lipolytica)的细胞。
该酵母宿主细胞可以是克拉布特里(crabtree)阳性表型或克拉布特里阴性表型。克拉布特里阴性生物由被诱导为增加的发酵性状态的能力表征。天然存在的生物和重组生物两者都可以被表征为克拉布特里阴性的。克拉布特里效应被定义为:当在高浓度的葡萄糖(例如>5mM葡萄糖)的存在下在需氧条件下培养微生物时,该微生物中的耗氧抑制。不论氧利用率如何,克拉布特里阳性生物在葡萄糖的存在下继续发酵(而非呼吸),而克拉布特里阴性生物未展示出葡萄糖介导的耗氧抑制。这一特征对于有机产品合成而言是有用的,因为它允许细胞在较高的底物浓度下生长,但保留氧化磷酸化的有益的高能作用。在一个方面,该酵母具有克拉布特里阴性表型。
在某些实施例中,该宿主酵母细胞属于伊萨酵母属(Issatchenkia)、假丝酵母属(Candida)或酵母属(Saccharomyces),并且在这些实施例的某些实施例中,该宿主细胞属于东方伊萨酵母(I.orientalis)/发酵毕赤酵母(P.fermentans)或酵母属(Saccharomyces)进化枝。在这些实施例的某些实施例中,该宿主细胞是东方伊萨酵母(I.orientalis)或郎比可假丝酵母(C.lambica)或布拉迪酵母(S.bulderi)。在某些实施例中,该酵母细胞是CNB1酵母细胞(如WO 2012/074818中所述,将其内容通过引用结合在此)和/或CB1酵母细胞。CB1细胞包括WO 2012/074818中描述的CNB1细胞以及与在此描述的CNB1细胞相关的或者从其衍生的任何细胞。
该酵母宿主细胞可以来自一个细胞,或者被工程化使得细胞已受到遗传修饰以产生高的乳酸或3-HP滴度,表现出对酸性pH的耐受性增加,表现出对发酵产物(例如乳酸盐/乳酸酯、3-HP、乙醇或丙醇)的耐受性增加,以及/或者显示出发酵戊糖能力的增加(然而,在一些实施例中,该酵母细胞不能发酵戊糖)。示例性遗传修饰的酵母细胞描述于WO 00/71738、WO 03/049525、WO 03/102201、WO 03/102152、WO 02/42471、WO 2007/032792、WO2007/106524、WO 2007/117282中,将其内容相对于所述细胞通过引用结合在此。将前述申请中所描述的任何酵母细胞的修饰与如下所述的主动3-HP途径一起考虑。
如上所述,宿主细胞可以具有对乳酸盐/乳酸酯和/或3-HP的改进的抗性。在某些实施例中,如US 2012/0135481中所描述,宿主细胞是“3-HP抗性的酵母细胞”。在这些实施例的某些中,这些宿主细胞能以其天然形式展示出3-HP抗性。在其他实施例中,在引入与主动3-HP途径相关的遗传修饰前、中或后,这些细胞可以已经经历突变和/或选择(例如,恒化器选择或重复的系列传代培养),这样使得这些突变的和/或选择的细胞具有比同一物种的野生型细胞对3-HP的更高程度的抗性。例如,在一些实施例中,在用一个或多个异源3-HP途径基因遗传修饰之前或之后,这些细胞已经在3-HP或乳酸的存在下经历了突变和/或选择。在某些实施例中,可以在以其天然形式展示出3-HP抗性的细胞上进行突变和/或选择。可以在不同水平的3-HP的存在下,针对糖消耗以及其他特征对经历突变和/或选择的细胞进行测试,以便确定它们作为用于生产3-HP的工业化宿主的可能性。除了3-HP抗性以外,宿主细胞可以已经经历用于抵抗一种或多种另外的有机酸(例如,乳酸)或抵抗其他发酵产物、副产物或培养基组分的突变和/或选择。可以使用本领域熟知的方法完成选择,例如对3-HP或对其他化合物的抗性的选择(例如,如在US 2012/0135481中描述的)。
在一个实施例中,该宿主细胞是丝状真菌细胞。“丝状真菌”包括所有丝状形式的细分真菌亚门(Eumycota)和卵菌亚门(Oomycota)(如由上文的Hawksworth等人,1995所定义的)。丝状真菌通常的特征在于由几丁质、纤维素、葡聚糖、壳多糖、甘露聚糖、以及其他复杂多糖构成的菌丝体壁。营养生长是通过菌丝延伸,而碳分解代谢是专性需氧的。相反,酵母(如酿酒酵母(Saccharomyces cerevisiae))的营养生长是通过单细胞菌体的出芽(budding),而碳分解代谢可以是发酵性的。
丝状真菌宿主细胞可以是枝顶孢霉属(Acremonium)、曲霉属(Aspergillus)、短梗霉属(Aureobasidium)、烟管菌属(Bjerkandera)、拟蜡菌属(Ceriporiopsis)、金孢子菌属(Chrysosporium)、鬼伞菌属(Coprinus)、革盖菌属(Coriolus)、隐球菌属(Cryptococcus)、线黑粉酵母属(Filibasidium)、镰孢属(Fusarium)、腐质霉属(Humicola)、梨孢菌属(Magnaporthe)、毛霉属(Mucor)、毁丝霉属(Myceliophthora)、新美鞭菌属(Neocallimastix)、脉孢菌属(Neurospora)、拟青霉属(Paecilomyces)、青霉菌属(Penicillium)、平革菌属(Phanerochaete)、白腐菌属(Phlebia)、瘤胃壶菌属(Piromyces)、侧耳属(Pleurotus)、裂褶菌属(Schizophyllum)、踝节菌属(Talaromyces)、嗜热子囊菌属(Thermoascus)、梭孢壳属(Thielavia)、弯颈霉属(Tolypocladium)、栓菌属(Trametes)、或木霉属(Trichoderma)的细胞。
例如,丝状真菌宿主细胞可以是泡盛曲霉(Aspergillus awamori)、臭曲霉(Aspergillus foetidus)、烟曲霉(Aspergillus fumigatus)、日本曲霉(Aspergillusjaponicus)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)、黑刺烟管菌(Bjerkandera adusta)、干拟蜡菌(Ceriporiopsisaneirina)、卡内基拟蜡菌(Ceriporiopsis caregiea)、浅黄拟蜡孔菌(Ceriporiopsisgilvescens)、潘诺希塔拟蜡菌(Ceriporiopsis pannocinta)、环带拟蜡菌(Ceriporiopsisrivulosa)、微红拟蜡菌(Ceriporiopsis subrufa)、虫拟蜡菌(Ceriporiopsissubvermispora)、狭边金孢子菌(Chrysosporium inops)、嗜角质金孢子菌(Chrysosporiumkeratinophilum)、卢克诺文思金孢子菌(Chrysosporium lucknowense)、粪状金孢子菌(Chrysosporium merdarium)、租金孢子菌(Chrysosporium pannicola)、昆士兰金孢子菌(Chrysosporium queenslandicum)、热带金孢子菌(Chrysosporium tropicum)、褐薄金孢子菌(Chrysosporium zonatum)、灰盖鬼伞(Coprinus cinereus)、毛革盖菌(Coriolushirsutus)、杆孢状镰孢(Fusarium bactridioides)、谷类镰孢(Fusarium cerealis)、库威镰孢(Fusarium crookwellense)、大刀镰孢(Fusarium culmorum)、禾谷镰孢(Fusariumgraminearum)、禾赤镰孢(Fusarium graminum)、异孢镰孢(Fusarium heterosporum)、合欢木镰孢(Fusarium negundi)、尖镰孢(Fusarium oxysporum)、多枝镰孢(Fusariumreticulatum)、粉红镰孢(Fusarium roseum)、接骨木镰孢(Fusarium sambucinum)、肤色镰孢(Fusarium sarcochroum)、拟分枝孢镰孢(Fusarium sporotrichioides)、硫色镰孢(Fusarium sulphureum)、圆镰孢(Fusarium torulosum)、拟丝孢镰孢(Fusariumtrichothecioides)、镶片镰孢(Fusarium venenatum)、特异腐质霉(Humicola insolens)、柔毛腐质霉(Humicola lanuginosa)、米黑毛霉(Mucor miehei)、嗜热毁丝霉(Myceliophthora thermophila)、粗糙链孢菌(Neurospora crassa)、产紫青霉菌(Penicillium purpurogenum)、黄孢原毛平革菌(Phanerochaete chrysosporium)、射脉菌(Phlebia radiata)、刺芹侧耳(Pleurotus eryngii)、土生梭孢壳(Thielaviaterrestris)、长域毛栓菌(Trametes villosa)、变色栓菌(Trametes versicolor)、哈茨木霉(Trichoderma harzianum)、康宁木霉(Trichoderma koningii)、长枝木霉(Trichodermalongibrachiatum)、里氏木霉(Trichoderma reesei)或绿色木霉(Trichoderma viride)细胞。
可以将真菌细胞通过涉及原生质体形成、原生质体转化、以及细胞壁再生的方法以本身已知的方式转化。用于转化曲霉属和木霉属宿主细胞的适合程序在EP 238023,Yelton等人,1984,Proc.Natl.Acad.Sci.USA[美国国家科学院院刊]81:1470-1474,和Christensen等人,1988,Bio/Technology[生物/技术]6:1419-1422中描述。用于转化镰孢属(Fusarium)物种的适合方法由Malardier等人,1989,Gene[基因]78:147-156以及WO 96/00787描述。可以使用由如以下文献描述的程序转化酵母:Becker和Guarente,在Abelson,J.N.和Simon,M.I.,编辑,Guide to Yeast Genetics and Molecular Biology[酵母遗传学与分子生物学指南],Methods in Enzymology[酶学方法],第194卷,第182-187页,学术出版社有限公司(Academic Press,Inc.),纽约中;Ito等人,1983,J.Bacteriol.[细菌学杂志]153:163;以及Hinnen等人,1978,Proc.Natl.Acad.Sci.USA[美国国家科学院院刊]75:1920,或如在此所述的。
对有机酸(如乳酸或3-HP)而言,理想细胞能够在低pH值水平下生长。在低的pH下进行发酵的能力降低了下游的回收成本,从而使得生产更经济。因此,在某些实施例中,该宿主细胞能够在低的pH水平下(例如,在低于7、6、5、4或3的pH水平下)生长。
除有机酸抗性和/或低pH生长能力之外,适合的宿主细胞可以拥有一种或多种有利的特征。例如,可以基于糖分解速度、比生长速率、耐热性、对生物质水解产物抑制剂的耐受性、整体流程稳健性等对展现出酸抗性的潜在宿主细胞进行进一步选择。可以在进行与代谢途径相关的任何遗传修饰之前评估这些标准,或可以在已经发生一种或多种此类修饰之后对其进行评估。
在这些方面的任一项中,重组细胞产生(和/或能够产生)发酵产物,例如D-乳酸盐/D-乳酸酯或3-HP,产率至少为理论值的10%,例如至少20%,在至少30%、至少40%、至少50%、至少60%、至少70%、至少80%,或至少90%。
在这些方面的任一项中,该重组细胞具有大于约0.1g/L/小时的体积生产力(例如,D-乳酸盐/D-乳酸酯或3-HP体积生产力),例如大于约0.2g/L/小时、0.5g/L/小时、0.6g/L/小时、0.7g/L/小时、0.8g/L/小时、0.9g/L/小时、1.0g/L/小时、1.1g/L/小时、1.2g/L/小时、1.3g/L/小时、1.5g/L/小时、1.75g/L/小时、2.0g/L/小时、2.25g/L/小时、2.5g/L/小时、2.75g/L/小时、3.0g/L/小时、3.25g/L/小时、3.5g/L/小时、3.75g/L/小时、或4.0g/L/小时;或在约0.1g/L/小时和约2.0g/L/小时之间,例如在约0.3g/L/小时和约1.7g/L/小时之间、约0.5g/L/小时和约1.5g/L/小时之间、约0.7g/L/小时和约1.3g/L/小时之间、约0.8g/L/小时和约1.2g/L/小时之间、或约0.9g/L/小时和约1.1g/L/小时之间;或在约0.1g/L/小时和约4.0g/L/小时之间,例如在约0.5g/L/小时和约3.75g/L/小时之间、约1.0g/L/小时之和约3.5g/L/小时之间、约2.0g/L/小时和约3.25g/L/小时之间。
重组细胞可以使用本领域熟知的方法培养于适合的营养培养基中。例如,可以通过在适合的培养基中和在允许表达和/或分离所希望的多肽的条件下,进行摇瓶培养,以及在实验室或工业发酵罐中进行小规模或大规模发酵(包括连续,分批,分批补料,或固态发酵)来培养细胞。如在此所述,该培养是使用本领域中已知的程序,在一种适合营养培养基中发生,该培养基包括碳源和氮源及无机盐。适合的培养基从商业供应商处可获得或者可根据公开的组成(例如,在美国典型培养物保藏中心目录中)来制备,或可以从可商购的成分制备。
在此描述的重组细胞还可以经受适应进化,以进一步增加产物生物合成,包括在接近理论最大生长的条件下。
在此描述的重组细胞可以利用以下表达载体,这些表达载体包括一个或多个(例如,两个、若干个)异源3-HP途径基因的编码序列,这些编码序列被连接至一个或多个控制序列,这一个或多个控制序列指导在与这一个或多个控制序列相容的条件下在适合的细胞中的表达。可以在任何于此描述的细胞和方法中使用此类表达载体。在此描述的多核苷酸可以按多种方式操纵,以提供希望的多肽的表达。取决于表达载体,在其插入载体以前操纵多核苷酸可以是希望的或必需的。用于利用重组DNA方法修饰多核苷酸的技术是本领域熟知的。
可以将一个构建体或载体(或多个构建体或载体)引入细胞中,这样使得该构建体或载体被维持作为染色体整合体或作为自主复制的染色体外载体,如早前所述;该构建体或载体(或这些构建体或载体)包括一个或多个(例如,两个、若干个)异源3-HP途径基因。
各种核苷酸和控制序列可以连接在一起以产生重组表达载体,该重组表达载体可以包括一个或多个(例如,两个、若干个)合宜的限制性位点以允许在此类位点插入或取代该多核苷酸。可替代地,可以通过将这种或这些多核苷酸或者包括该序列的核酸构建体插入用于表达的适当载体中而表达这种或这些多核苷酸。在产生该表达载体时,该编码序列位于该载体中,这样使得该编码序列与该用于表达的适当控制序列可操作地连接。
重组表达载体可以是可便利地经受重组DNA程序并且可引起多核苷酸表达的任何载体(例如,质粒或病毒)。载体的选择将典型地取决于该载体与有待引入该载体的宿主细胞的相容性。该载体可以是一种线性的或闭合的环形质粒。
在一个方面中,该重组细胞包含在独立载体中包含的异源多核苷酸。在一个方面,该重组细胞包含多个各自包含在独立载体中的异源多核苷酸。在一个方面,该重组细胞包含至少两个包含在单个载体中的异源多核苷酸。在一个方面,该重组细胞包含至少三个包含在单个载体上的异源多核苷酸。在一个方面,该重组细胞包含至少四个包含在单个载体上的异源多核苷酸。在一个方面,该重组细胞的所有异源多核苷酸都包含在单个载体上。编码蛋白复合体的异聚亚基的多核苷酸可以被包含在单一载体的单一异源多核苷酸中或可替代地,被包含在分开的载体的分开的异源多核苷酸中。
该载体可以是自主复制载体,即,作为染色体外实体存在的载体,其复制独立于染色体复制,例如,质粒、染色体外元件、微染色体或人工染色体。该载体可以包含用于确保自我复制的任何装置。可替代地,该载体可以是这样载体,当它被引入该宿主细胞中时,被整合到基因组中并且与其中已整合了它的一个或多个染色体一起复制。此外,可以使用单一载体或质粒或两个或更多个载体或质粒(这些载体或质粒共同包含待引入到细胞的基因组中的总DNA)或转座子。
该表达载体可以包含任何适合的启动子序列,该启动子序列可被细胞识别,以表达丙酮酸还原酶基因或在此描述的任何3-HP途径基因。启动子序列包含转录控制序列,其介导多肽的表达。该启动子可以是在选择的细胞中显示出转录活性的任何多核苷酸,包括突变型、截短型及杂合型启动子,并且可以是由编码与该细胞同源或异源的细胞外或细胞内多肽的基因获得。
在此描述的每种异源多核苷酸都可以被可操作地连接至对于该多核苷酸而言外源的启动子上。例如,在一个方面,编码丙酮酸还原酶的异源多核苷酸被可操作地连接至对于该多核苷酸而言外源的启动子上。在另一个方面,编码在此描述的3-HP途径的多肽(例如,PPC、PYC、AAT、ADC、BAAT、gabT或3-HPDH)的异源多核苷酸被可操作地连接至对于该多核苷酸而言外源的启动子上。这些启动子可以与所选的天然启动子相同或与其具有高水平的序列一致性(例如,至少约80%、至少约85%、至少约90%、至少约95%、或至少约99%)。
在细菌宿主细胞中,适合指导本发明的核酸构建体转录的启动子的实例是获得自以下各项的启动子:解淀粉芽孢杆菌(Bacillus amyloliquefaciens)α-淀粉酶基因(amyQ)、地衣芽孢杆菌(Bacillus licheniformis)α-淀粉酶基因(amyL)、地衣芽孢杆菌青霉素酶基因(penP)、嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)麦芽淀粉酶基因(amyM)、枯草芽孢杆菌(Bacillus subtilis)果聚糖蔗糖酶基因(sacB)、枯草芽孢杆菌xylA和xylB基因、苏云金芽孢杆菌(Bacillus thuringiensis)cryIIIA基因(Agaisse和Lereclus,1994,Molecular Microbiology[分子微生物学]13:97-107)、大肠杆菌(E.coli)lac操纵子、大肠杆菌trc启动子(Egon等人,1988,Gene[基因]69:301-315),天蓝链霉菌(Streptomyces coelicolor)琼脂酶基因(dagA)、和原核β-内酰胺酶基因(Villa-Kamaroff等人,1978,Proc.Natl.Acad.Sci.USA[美国国家科学院院刊]75:3727-3731)、以及tac启动子(DeBoer等人,1983,Proc.Natl.Acad.Sci.USA[美国国家科学院院刊]80:21-25)。另外的启动子描述于“Useful proteins from recombinant bacteria[来自重组细菌的有用蛋白]”,Gilbert等人,1980,Scientific American[科学美国人])242:74-94;以及在Sambrook等人,1989,同上中。串联启动子的实例披露在WO 99/43835中。
用于指导核酸构建体在酵母细胞中的转录的适合的启动子的实例包括但不限于获得自以下各项的基因的启动子:烯醇酶(例如,酿酒酵母(S.cerevisiae)烯醇酶或东方伊萨酵母烯醇酶(ENO1))、半乳糖激酶(例如,酿酒酵母半乳糖激酶或东方伊萨酵母半乳糖激酶(GAL1))、醇脱氢酶/甘油醛-3磷酸脱氢酶(例如,酿酒酵母醇脱氢酶/甘油醛-3磷酸脱氢酶或东方伊萨酵母醇脱氢酶/甘油醛-3磷酸脱氢酶(ADH1、ADH2/GAP))、磷酸甘油醛异构酶(例如,酿酒酵母磷酸甘油醛异构酶或东方伊萨酵母磷酸甘油醛异构酶(TPI))、金属硫蛋白(例如,酿酒酵母金属硫蛋白或东方伊萨酵母金属硫蛋白(CUP1))、3-磷酸甘油酸激酶(例如,酿酒酵母3磷酸甘油酸激酶或东方伊萨酵母3-磷酸甘油酸激酶(PGK))、PDC1、木糖还原酶(XR)、木糖醇脱氢酶(XDH)、L-(+)-乳酸-细胞色素C氧化还原酶(CYB2)、翻译延长因子-1(TEF1)、翻译延长因子-2(TEF2)、甘油醛-3-磷酸脱氢酶(GAPDH)、和乳清酸核苷5'-磷酸脱羧酶(URA3)基因。Romanos等人,1992,Yeast[酵母]8:423-488描述了酵母宿主细胞的其他有用的启动子。
用于指导本发明的核酸构建体在丝状真菌宿主细胞中的转录的合适启动子的实例是从以下各项的基因获得的启动子:构巢曲霉乙酰胺酶、黑曲霉中性α-淀粉酶、黑曲霉酸稳定性α-淀粉酶、黑曲霉或泡盛曲霉葡糖淀粉酶(glaA)、米曲霉TAKA淀粉酶、米曲霉碱性蛋白酶、米曲霉丙糖磷酸异构酶、尖镰孢(Fusarium oxysporum)胰蛋白酶样蛋白酶(WO 96/00787)、镶片镰孢菌(Fusarium venenatum)淀粉葡糖苷酶(WO 00/56900)、镶片镰孢菌Daria(Fusarium venenatum Daria)(WO 00/56900)、镶片镰孢Quinn(Fusarium venenatumQuinn)(WO 00/56900)、米黑根毛霉(Rhizomucor miehei)脂肪酶、米黑根毛霉天冬氨酸蛋白酶、里氏木霉β-葡糖苷酶、里氏木霉纤维二糖水解酶I、里氏木霉纤维二糖水解酶II、里氏木霉内切葡聚糖酶I、里氏木霉内切葡聚糖酶II、里氏木霉内切葡聚糖酶III、里氏木霉内切葡聚糖酶IV、里氏木霉内切葡聚糖酶V、里氏木霉木聚糖酶I、里氏木霉木聚糖酶II、里氏木霉β-木糖苷酶,以及NA2tpi启动子(修饰的启动子,其来自曲霉属中性α-淀粉酶基因,其中未翻译的前导序列由曲霉属丙糖磷酸异构酶基因的未翻译的前导序列替换;非限制性实例包括修饰的启动子,其来自黑曲霉中性α-淀粉酶的基因,其中未翻译的前导序列由构巢曲霉或米曲霉丙糖磷酸异构酶基因的未翻译的前导序列替换);以及其突变型启动子、截短型启动子、以及杂合型启动子。
控制序列也可以是被宿主细胞识别以终止转录的适合转录终止子序列。该终止子序列被可操作地连接至编码该多肽的多核苷酸的3’-末端。可以使用在所选的酵母细胞中具有功能的任何终止子。该终止子可以与所选的天然终止子相同或与其具有高水平的序列一致性(例如,至少约80%、至少约85%、至少约90%、至少约95%或至少约99%)。在某些实施例中,3-HP途径基因被连接至一种终止子,该终止子包括对于该宿主细胞而言天然的天然GAL10基因的功能部分或与天然GAL10终止子具有至少80%、至少85%、至少90%或至少95%序列一致性的序列。
细菌宿主细胞的适合终止子可以从针对以下各项的基因获得:克劳氏芽孢杆菌(Bacillus clausii)碱性蛋白酶(aprH)、地衣芽孢杆菌α-淀粉酶(amyL)和大肠杆菌(Escherichia coli)核糖体RNA(rrnB)。
酵母宿主细胞的适合的终止子可以获得自以下各项的基因:烯醇酶(例如,酿酒酵母(S.cerevisiae)或东方伊萨酵母烯醇酶)、细胞色素C(例如,酿酒酵母或东方伊萨酵母细胞色素(CYC1))、甘油醛-3磷酸脱氢酶(例如,酿酒酵母或东方伊萨酵母甘油醛-3-磷酸脱氢酶(gpd))、PDC1、XR、XDH、转醛醇酶(TAL)、转酮醇酶(TKL)、核糖5-磷酸-酮醇异构酶(RKI)、CYB2、以及半乳糖基因家族(尤其是GAL10终止子)。Romanos等人(1992,同上)描述了酵母宿主细胞的其他有用的终止子。
丝状真菌宿主细胞的适合终止子可以从以下各项的基因中获得:构巢曲霉邻氨基苯甲酸合酶、黑曲霉葡糖淀粉酶、黑曲霉α-葡糖苷酶、米曲霉TAKA淀粉酶以及尖镰孢胰蛋白酶样蛋白酶。
控制序列还可以是启动子下游和基因的编码序列上游的mRNA稳定子区,其增加该基因的表达。
适合的mRNA稳定区的实例是从以下获得的:苏云金芽孢杆菌cryIIIA基因(WO 94/25612)和枯草芽孢杆菌SP82基因(Hue等人,1995,Journal of Bacteriology[细菌学杂志]177:3465-3471)。
控制序列也可以是适合的前导子序列,其中转录时,所述前导子序列是对由宿主细胞翻译重要的mRNA的非翻译区。该前导子序列可操作地连接至编码该多肽的多核苷酸的5’-末端。可以使用在选择的酵母细胞中具有功能的任何前导子序列。
酵母宿主细胞的适合前导序列获得自以下各项的基因:烯醇酶(例如,酿酒酵母或东方伊萨酵母烯醇酶(ENO-1))、3-磷酸甘油酸激酶(例如,酿酒酵母或东方伊萨酵母3-磷酸甘油酸激酶)、α-因子(例如,酿酒酵母或东方伊萨酵母α-因子)以及醇脱氢酶/甘油醛-3-磷酸脱氢酶(例如,酿酒酵母或东方伊萨酵母醇脱氢酶/甘油醛-3磷酸脱氢酶(ADH2/GAP))。
用于丝状真菌宿主细胞的优选前导序列是从米曲霉TAKA淀粉酶和构巢曲霉丙糖磷酸异构酶的基因获得。
控制序列还可以是一种聚腺苷酸化序列,可操作地连接至该多核苷酸的3’-末端并且当转录时由宿主细胞识别为将聚腺苷酸残基添加至所转录的mRNA的信号的序列。可以使用在选择的宿主细胞中具有功能的任何聚腺苷酸化序列。对于酵母细胞有用的聚腺苷酸化序列描述于以下文献:Guo和Sherman,1995,Mol.Cellular Biol.[分子细胞生物学]15:5983-5990。丝状真菌宿主细胞的优选多腺苷酸化序列是从以下各项的基因中获得的:构巢曲霉邻氨基苯甲酸合酶、黑曲霉葡糖淀粉酶、黑曲霉α-葡萄糖苷酶、米曲霉TAKA淀粉酶以及尖镰孢胰蛋白酶样蛋白酶。
也可能令人希望的是添加调控序列,该调控序列允许相对于宿主细胞的生长而调节多肽的表达。调节系统的实例是引起将响应于化学或物理刺激(包含调节性化合物的存在)而开启或关闭的基因表达的那些系统。在原核系统中的调节系统包括lac、tac、以及trp操纵基因系统。在酵母中,可以使用ADH2系统或GAL1系统。在丝状真菌中,可以使用黑曲霉葡糖淀粉酶启动子、米曲霉TAKAα-淀粉酶启动子、以及米曲霉葡糖淀粉酶启动子。调控序列的其他例子是允许基因扩增的那些。在真核系统中,这些调控序列包括在甲氨蝶呤存在下被扩增的二氢叶酸还原酶基因以及用重金属扩增的金属硫蛋白基因。在这些情况下,编码该多肽的多核苷酸将与调控序列可操作地连接。
这些载体可以包含一个或多个(例如,两个、若干个)允许方便地选择转化细胞、转染细胞、转导细胞等细胞的选择性标记。选择性标记是这样一种基因,该基因的产物提供了杀生物剂抗性或病毒抗性、重金属抗性、营养缺陷型的原养型等。
细菌性选择性标记的实例是地衣芽孢杆菌或枯草芽孢杆菌dal基因,或赋予抗生素抗性(例如氨苄青霉素、氯霉素、卡那霉素、新霉素、大观霉素或四环素抗性)的标记。用于酵母宿主细胞的适合的标记包括但不限于ADE2、HIS3、LEU2、LYS2、MET3、TRP1以及URA3。用于在丝状真菌宿主细胞中使用的选择性标记包括但不限于amdS(乙酰胺酶)、argB(鸟氨酸氨甲酰基转移酶)、bar(草胺膦乙酰转移酶)、hph(潮霉素磷酸转移酶)、niaD(硝酸还原酶)、pyrG(乳清苷-5’磷酸脱羧酶)、sC(硫酸腺苷基转移酶)、以及trpC(邻氨基苯甲酸合酶),连同其等效物。优选在曲霉属细胞中使用的是构巢曲霉或米曲霉amdS和pyrG基因以及吸水链霉菌(Streptomyces hygroscopicus)bar基因。
这些载体可以包含一个或多个(例如,两个、若干个)允许将该载体整合进宿主细胞的基因组中或在该细胞中独立于基因组而自主复制的元件。
对于整合到该宿主细胞基因组中,该载体可以依靠编码该多肽的多核苷酸序列或者用于通过同源或非同源重组整合到该基因组中的该载体的任何其他元件。可替代地,该载体可以包含用于指导通过同源重组而整合到宿主细胞基因组中的一个或多个染色体中的一个或多个精确位置的另外的多核苷酸。为了增加在精确位置整合的可能性,这些整合的元件应包含足够数量的核酸,例如100至10,000个碱基对、400至10,000个碱基对、以及800至10,000个碱基对,这些碱基对与相应的靶序列具有高度的序列一致性以提高同源重组的可能性。这些整合元件可以是与宿主细胞基因组内的靶序列同源的任何序列。此外,这些整合元件可以是非编码多核苷酸或编码多核苷酸。另一方面,该载体可以通过非同源重组整合至宿主细胞的基因组中。潜在整合位点包括本领域所描述的那些(例如,参见US2012/0135481)。
对于自主复制,载体可以进一步包含使该载体能够在所讨论的酵母细胞中自主复制的复制起点。复制起点可以是在细胞中起作用的介导自主复制的任何质粒复制子。术语“复制起点(origin of replication)”或“质粒复制子(plasmid replicator)”意指使得质粒或载体可在体内复制的多核苷酸。
细菌的复制起点的实例是允许在大肠杆菌内进行复制的质粒pBR322、pUC19、pACYC177、以及pACYC184以及允许在芽孢杆菌内进行复制的pUB110、pE194、pTA1060、以及pAMβ1的复制起点。
用于酵母宿主细胞中的复制起点的实例是2微米复制起点、ARS1、ARS4、ARS1和CEN3的组合以及ARS4和CEN6的组合。
在丝状真菌细胞中,有用的复制起点的实例是AMA1和ANS1(Gems等人,1991,Gene[基因]98:61-67;Cullen等人,1987,Nucleic Acids Res.[核酸研究]15:9163-9175;WO00/24883)。可以根据WO 00/24883中披露的方法完成AMA1基因的分离和包含该基因的质粒或载体的构建。
可以将在此描述的多核苷酸的多于一个的拷贝插入到宿主细胞中以增加多肽的产生。通过将序列的至少一个另外的拷贝整合到酵母细胞基因组中或者通过包含一个与该多核苷酸一起的可扩增的选择性标记基因可以获得多核苷酸的增加的拷贝数目,其中通过在适当的选择性试剂的存在下培养细胞可以选择包含选择性标记基因的经扩增的拷贝的细胞、以及由此该多核苷酸的另外的拷贝。
用于连接以上所描述的元件以构建在此描述的重组表达载体的程序是本领域普通技术人员熟知的(参见例如,Sambrook等人,1989,同上)。
本领域已知的用于制备包含一个或多个3-HP途径基因的重组细胞的另外的程序和技术描述于例如WO 2012/074818中,将其内容通过引用结合在此。
主动3-HP途径
在此描述的宿主细胞,例如包含对于上述丙酮酸还原酶基因的破坏的突变细胞可进一步包含主动3-HP途径。)。3-HP途径、3-HP途径基因以及用于发酵3-HP的相应的工程转化体是本领域已知的(例如,美国公开号2012/0135481;美国专利号6,852,517;美国专利号7,309,597;美国公开号2001/0021978;美国公开号2008/0199926;WO 02/42418;以及WO10/031083;将其内容以其全文通过引用结合在此)。若干已知的3-HP途径的综述示于图1中。
在某些实施例中,在此提供的重组细胞具有通过丙二酸半醛中间体进行的主动3-HP途径。本领域已知若干种通过丙二酸半醛中间体进行的3-HP途径。例如,这些细胞具有以下主动3-HP途径,该主动3-HP途径通过PEP或丙酮酸、OAA、天冬氨酸、β-丙氨酸以及丙二酸半醛中间体而进行(参见,例如,US 7,186,541,图55)。在这些实施例中,这些重组细胞包含一套3-HP途径基因,这些基因包括丙酮酸羧化酶(PYC)、PEP羧化酶(PPC)、天冬氨酸转氨酶(AAT)、天冬氨酸1-脱羧酶(ADC)、β-丙氨酸转氨酶(BAAT)、氨基丁酸转氨酶(gabT)、3-HP脱氢酶(3-HPDH)、3-羟基异丁酸脱氢酶(HIBADH)以及4-羟基丁酸脱氢酶基因中的一种或多种。这些3-HP途径基因还可以包括PEP羧激酶(PCK)基因,该基因已被修饰为产生以下多肽,该多肽优选地催化PEP转化为OAA(天然的PCK基因通常产生以下多肽,该多肽优选地催化OAA至PEP的逆反应)。
在另一个实例中,这些重组细胞可以具有通过PEP或丙酮酸、OAA以及丙二酸半醛中间体而进行的主动3-HP途径(参见,例如,美国公开号2010/0021978,图1)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括PPC、PYC、2-酮酸脱羧酶、α-酮戊二酸(AKG)脱羧酶(KGD)、支链α-酮酸脱羧酶(BCKA)、吲哚丙酮酸脱羧酶(IPDA)、3-HPDH、HIBADH以及4-羟基丁酸脱氢酶基因中的一种或多种。这些3-HP途径基因还可以包括一种PCK基因,该基因已被修饰为产生以下多肽,该多肽优选地催化PEP转化为OAA。另外,这些3-HP途径基因包括一种PDC基因和/或苯甲酰甲酸脱羧酶基因,该基因已被修饰为编码以下多肽,该多肽能够催化OAA转化为丙二酸半醛。
在另一个实例中,这些重组细胞可以具有通过PEP或丙酮酸、OAA、丙二酰辅酶A以及丙二酸半醛中间体而进行的主动3-HP途径,其中丙二酸半醛中间体是任选的(参见,例如,美国公开号2010/0021978,图2)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括PPC、PYC、OAA甲酸裂解酶、丙二酰辅酶A还原酶、辅酶A酰化丙二酸半醛脱氢酶、3-HPDH、HIBADH以及4-羟基丁酸脱氢酶基因中的一种或多种。这些3-HP途径基因还可以包括一种PCK基因,该基因已被修饰为产生以下多肽,该多肽优选地催化PEP转化为OAA。另外,这些3-HP途径基因可以包括一种OAA脱氢酶基因,该基因是通过将2-酮酸脱氢酶基因修饰以产生一种多肽而得到,该多肽催化OAA转化为丙二酰辅酶A。
在另一个实例中,这些重组细胞可以具有通过丙酮酸、乙酰辅酶A、丙二酰辅酶A以及丙二酸半醛中间体而进行的主动3-HP途径,其中丙二酸半醛中间体是任选的(参见例如,US 7,186,541;图44)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括丙酮酸脱氢酶(PDH)、乙酰辅酶A羧化酶(ACC)、丙二酰辅酶A还原酶、辅酶A酰化丙二酸半醛脱氢酶、3-HPDH、HIBADH以及4-羟基丁酸脱氢酶基因中的一种或多种。
在另一个实例中,这些重组细胞可以具有以下主动3-HP途径,该主动3-HP途径通过丙酮酸、丙氨酸、β-丙氨酸、β-丙氨酰辅酶A、丙烯酰辅酶A、3-HP辅酶A以及丙二酸半醛中间体而进行,其中β-丙氨酰辅酶A、丙烯酰辅酶A、3-HP辅酶A以及丙二酸半醛中间体是任选的(可以经由丙二酸半醛中间体或经由β-丙氨酰辅酶A、丙烯酰辅酶A和3-HP辅酶A中间体将β-丙氨酸转化为3-HP)(参见例如,美国专利7,309,597,图1)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括丙氨酸脱氢酶、丙酮酸/丙氨酸转氨酶、丙氨酸2,3氨基变位酶、辅酶A转移酶、辅酶A合成酶、β-丙氨酰辅酶A解氨酶、3-HP辅酶A脱水酶、3-HP辅酶A水解酶、3-羟基异丁酰辅酶A水解酶、BAAT、3-HPDH、HIBADH以及4-羟基丁酸脱氢酶基因中的一种或多种。
其他不利用丙二酸半醛中间体的途径也是本领域已知的。例如,这些重组细胞可以具有以下主动3-HP途径,该主动3-HP途径通过PEP或丙酮酸、OAA和苹果酸中间体而进行(参见例如,美国公开号2010/0021978;图4)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括PPC、PYC、苹果酸脱氢酶以及苹果酸脱羧酶基因中的一种或多种。这些3-HP途径基因还可以包括一种PCK基因,该基因已被修饰为产生以下多肽,该多肽优选地催化PEP转化为OAA。
在另一个实例中,这些重组细胞可以具有以下主动3-HP途径,该主动3-HP途径通过丙酮酸、乳酸、乳酰辅酶A、丙烯酰辅酶A以及3-HP辅酶A中间体而进行(参见例如,WO 02/042418,图1)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括LDH、辅酶A转移酶、辅酶A合成酶、乳酰辅酶A脱水酶、3-HP辅酶A脱水酶、3-HP辅酶A水解酶以及3-羟基异丁酰辅酶A水解酶基因中的一种或多种。
在另一个实例中,这些重组细胞可以具有以下主动3-HP途径,该主动3-HP途径通过甘油和3-HPA中间体而进行(参见例如,美国专利6,852,517)。在这些实施例中,这些细胞包括一套3-HP途径基因,这些基因包括甘油脱水酶和醛脱氢酶基因中的一种或多种。
在另一个实例中,这些重组细胞可以具有以下主动3-HP途径,该主动3-HP途径通过PEP或丙酮酸、OAA、天冬氨酸、β-丙氨酸、β-丙氨酰辅酶A、丙烯酰辅酶A、3-HP辅酶A以及丙氨酸中间体而进行,其中OAA、天冬氨酸和丙氨酸中间体是任选的(可以经由OAA和天冬氨酸或经由丙氨酸将PEP或丙酮酸转化为β-丙氨酸)(参见WO 02/042418,图54;美国专利7,309,597,图1)。在这些实施例中,这些细胞包含一套3-HP途径基因,这些基因包括PPC、PYC、AAT、ADC、辅酶A转移酶、辅酶A合成酶、β-丙氨酰辅酶A解氨酶、3-HP-辅酶A脱水酶、3-HP-辅酶A水解酶、3-羟基异丁酰辅酶A水解酶、丙氨酸脱氢酶、丙酮酸/丙氨酸转氨酶以及AAM基因中的一种或多种。这些3-HP途径基因还可以包括一种PCK基因,该基因已被修饰为产生以下多肽,该多肽优选地催化PEP转化为OAA。
在某些实施例中,在此提供的重组细胞表达一种或多种编码选自下组的酶的3-HP途径基因,该组由以下各项组成:ACC(催化乙酰辅酶A转化为丙二酰辅酶A),丙氨酸2,3氨基变位酶(AAM,催化丙氨酸转化为β-丙氨酸),丙氨酸脱氢酶(催化丙酮酸转化为丙氨酸),醛脱氢酶(催化3-HPA转化为3-HP),KGD(催化OAA转化为丙二酸半醛),AAT(催化OAA转化为天冬氨酸),ADC(催化天冬氨酸转化为β-丙氨酸),BCKA(催化OAA转化为丙二酸半醛),BAAT(催化β-丙氨酸转化为丙二酸半醛),4-氨基丁酸转氨酶(gabT,催化β-丙氨酸转化为丙二酸半醛),β-丙氨酰辅酶A解氨酶(催化β-丙氨酰辅酶A转化为丙烯酰辅酶A),辅酶A酰化丙二酸半醛脱氢酶(催化丙二酰辅酶A转化为丙二酸半醛),辅酶A合成酶(催化β-丙氨酸转化为β-丙氨酰辅酶A或将乳酸转化为乳酰辅酶A),辅酶A转移酶(催化β-丙氨酸转化为β-丙氨酰辅酶A和/或将乳酸转化为乳酰辅酶A),甘油脱水酶(催化甘油转化为3-HPA),IPDA(催化OAA转化为丙二酸半醛),LDH(催化丙酮酸转化为乳酸),乳酰辅酶A脱水酶(催化乳酰辅酶A转化为丙烯酰辅酶A),苹果酸脱羧酶(催化苹果酸转化为3-HP),苹果酸脱氢酶(催化OAA转化为苹果酸),丙二酰辅酶A还原酶(催化丙二酰辅酶A转化为丙二酸半醛或3-HP),OAA甲酸裂解酶(亦称丙酮酸-甲酸裂解酶和酮酸甲酸裂解酶,催化OAA转化为丙二酰辅酶A),OAA脱氢酶(催化OAA转化为丙二酰辅酶A);PPC(催化PEP转化为OAA),丙酮酸/丙氨酸转氨酶(催化丙酮酸转化为丙氨酸),PYC(催化丙酮酸转化为OAA),PDH(催化丙酮酸转化为乙酰辅酶A),2-酮酸脱羧酶(催化OAA转化为丙二酸半醛),3-HP辅酶A脱水酶(亦称丙烯酰辅酶A水合酶,催化丙烯酰辅酶A转化为3-HP辅酶A),3-HPDH(催化丙二酸半醛转化为3-HP),3-HP辅酶A水解酶(催化3-HP辅酶A转化为3-HP),HIBADH(催化丙二酸半醛转化为3-HP),3-羟基异丁酰辅酶A水解酶(催化3-HP辅酶A转化为3-HP)以及4-羟基丁酸脱氢酶(催化丙二酸半醛转化为3-HP)。对于这些酶活性中的每种酶活性而言,感兴趣的反应(括号中)可以是内源或异源活性的结果。
可以使用任何适合的3-HP途径基因(内源的或异源的)并以足以产生在所选的主动3-HP途径中涉及的酶的量表达。伴随现在多于550个物种可获得的全基因组序列(其中多于一半的这些序列可在公共数据库(例如NCBI)中获得)(包括395个微生物基因组以及多种酵母、真菌、植物和哺乳动物基因组),对于所选的宿主而言,编码在此教导的所选3-HP途径的酶活性的基因的鉴定在本领域是常规且熟知的。例如,已知基因的适合的同源物、直向同源物、旁系同源物和非直向同源物基因替代,和在生物之间的遗传改变的互换可以在与选择的宿主相关的或远缘的宿主中来鉴定。
对于没有已知基因组序列的重组细胞而言,可以典型地使用本领域已知的技术获得感兴趣的基因的序列(作为过表达候选物或作为插入位点)。可以利用常规实验设计测试各种基因的表达和各种酶的活性,包括在3-HP途径中起作用的基因和酶。可以进行实验,其中单独地在细胞中和在酶区块(直至并优选包括所有途径酶,针对改进3-HP生产的需要(或希望)而建立)中表达每种酶。一种说明性实验设计测试了每种单独的酶以及每个独特的酶对的表达,并且可以进一步测试所有需要的酶或每个独特的酶组合的表达。如应该领会的,可以采取多种方法。
可以通过引入编码一种或多种参与3-HP途径的酶的异源多核苷酸而产生本发明的重组宿主细胞,如下所述。如本领域普通技术人员应该领会的,在一些情况下(例如,取决于宿主的选择),鉴于宿主细胞可以具有来自一个或多个途径基因的内源酶活性,示于3-HP途径中的每个基因的异源表达可能并不为3-HP生产所需要。例如,如果选择的宿主缺乏3-HP途径的一种或多种酶,则将一种或多种缺陷酶的异源多核苷酸引入该宿主中,用于进行随后的表达。可替代地,如果选择的宿主展现出一些途径基因的内源表达,但是缺乏其他基因的内源表达,则需要缺乏的一种或多种酶的编码多核苷酸,以实现生物合成3-HP。因此,可以通过引入异源多核苷酸而产生重组宿主细胞,以获得希望的生物合成途径的酶活性,或可以通过引入一种或多种希望的异源多核苷酸而获得希望的生物合成途径,这些希望的异源多核苷酸与一种或多种内源酶一起产生希望的产物,例如3-HP。
取决于所选重组宿主生物的3-HP途径成分,本发明的宿主细胞将包括至少一种异源多核苷酸以及任选地直到所有的3-HP途径的编码异源多核苷酸。例如,可以通过异源表达相应的多核苷酸而在缺乏3-HP途径酶的宿主中建立3-HP生物合成。在缺乏3-HP途径的所有酶的宿主中,可以包括异源表达该途径中的所有酶,尽管应该理解的是即使该宿主包括至少一种途径酶,仍可以表达途径的所有酶。
如在此使用,“丙酮酸羧化酶基因”或“PYC基因”是指编码具有丙酮酸羧化酶活性的多肽的任何基因,意味着能够催化丙酮酸、CO2和ATP转化为OAA、ADP和磷酸。在某些实施例中,PYC基因可以来自酵母来源。例如,该PYC基因可以来自东方伊萨酵母PYC基因,该基因编码阐述于SEQ ID NO:2中的氨基酸序列。在其他实施例中,该基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:2的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自东方伊萨酵母的PYC基因可以包括阐述于SEQ ID NO:1中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:1中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该PYC基因可以来自细菌来源。例如,该PYC基因可以来自仅使用PYC而不使用PPC(参见下文)用于进行回补的几种细菌物种之一(例如类球红细菌(R.sphaeroides))或来自具有PYC和PPC两者的细菌物种(例如菜豆根瘤菌(R.etli))。由类球红细菌和菜豆根瘤菌的PYC基因编码的氨基酸序列分别阐述于SEQ ID NO:3和4中。PYC基因可以来自编码SEQ IDNO:3或4的氨基酸序列的基因或来自编码以下氨基酸序列的基因,该氨基酸序列与SEQ IDNO:3或4的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。可替代地,该PYC基因可以来自编码不依赖乙酰辅酶A进行活化的酶的PYC基因,例如编码阐述于SEQ ID NO:5(羧基转移酶亚单位)或SEQID NO:6(生物素羧化酶亚单位)中的氨基酸序列的荧光假单胞菌(P.fluorescens)PYC基因、编码阐述于SEQ ID NO:7中的氨基酸序列的谷氨酸棒杆菌(C.glutamicum)PYC基因或编码以下氨基酸序列的基因,该氨基酸序列与SEQ ID NO:5、6或7的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%、或至少99%序列一致性。PYC基因还可以来自编码不被天冬氨酸抑制的酶的PYC基因,例如编码阐述于SEQ ID NO:8中的氨基酸序列的苜蓿中华根瘤菌(S.meliloti)PYC基因(萨奥尔(Sauer)FEMS微生物学评论(FEMS Microbiol Rev)29:765(2005)),或来自编码以下氨基酸序列的基因,该氨基酸序列与SEQ ID NO:8的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
如在此使用,“PEP羧化酶基因”或“PPC基因”是指编码具有PEP羧化酶活性的多肽的任何基因,意味着能够催化PEP和CO2转化为OAA和磷酸。在某些实施例中,PPC基因可以来自细菌PPC基因。例如,该PPC基因可以来自编码阐述于SEQ ID NO:10中的氨基酸序列或以下氨基酸序列的大肠杆菌PPC基因,该氨基酸序列与SEQ ID NO:10的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自大肠杆菌的PPC基因可以包括阐述于SEQ ID NO:9中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:9中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,PPC基因可以来自“A”型PPC,发现于许多古细菌和数目有限的细菌中,不被乙酰辅酶A激活且较不受天冬氨酸的抑制。例如,PPC基因可以来自编码阐述于SEQ ID NO:11中的氨基酸序列的热自养甲烷杆菌(M.thermoautotrophicum)PPC A基因、编码阐述于SEQ ID NO:12中的氨基酸序列的产气荚膜梭菌(C.perfringens)PPC A基因或编码以下氨基酸序列的基因,该氨基酸序列与SEQ ID NO:11或12的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在这些实施例的某些实施例中,该基因可以已经经历对天然基因的一个或多个突变,以便产生具有改进的特征的酶。例如,该基因可以被突变为编码以下PPC多肽,该多肽与天然多肽相比对天冬氨酸反馈具有增加的抗性。在其他实施例中,该PPC基因可以来自植物来源。
如在此使用,“天冬氨酸转氨酶基因”或“AAT基因”是指编码具有天冬氨酸转氨酶活性的多肽的任何基因,意味着能够催化OAA转化为天冬氨酸。具有天冬氨酸转氨酶活性的酶被分类为EC 2.6.1.1。在某些实施例中,AAT基因可以来自酵母来源,例如东方伊萨酵母或酿酒酵母。例如,该AAT基因可以来自编码阐述于SEQ ID NO:14中的氨基酸序列的东方伊萨酵母AAT基因或编码阐述于SEQ ID NO:15中的氨基酸序列的酿酒酵母AAT2基因。在其他实施例中,该基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:14或15的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自东方伊萨酵母的AAT基因可以包括阐述于SEQ ID NO:13中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:13中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该AAT基因可以来自细菌来源。例如,该AAT基因可以来自大肠杆菌aspC基因,该基因编码包括阐述于SEQ ID NO:16中的氨基酸序列的多肽。在其他实施例中,该基因可以编码以下氨基酸序列,该氨基酸序列与SEQ IDNO:16的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
如在此使用,“天冬氨酸脱羧酶基因”或“ADC基因”是指编码具有天冬氨酸脱羧酶活性的多肽的任何基因,意味着能够催化天冬氨酸转化为β-丙氨酸。具有天冬氨酸脱羧酶活性的酶被分类为EC 4.1.1.11。在某些实施例中,ADC基因可以来自细菌来源。
在一些实施例中,该ADC基因可以来自除虫链霉菌(S.avermitilis)panD基因,该基因编码阐述于SEQ ID NO:17中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:17的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自除虫链霉菌的ADC基因可以包括阐述于SEQ ID NO:130、145、146或147的任一项中的核苷酸序列;或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:130、145、146或147的任一项中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以来自丙酮丁醇梭菌(C.acetobutylicum)panD基因,该基因编码阐述于SEQ ID NO:18中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:18的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自丙酮丁醇梭菌的ADC基因可以包括阐述于SEQ ID NO:131中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:131中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以来自幽门螺杆菌(H.pylori)ADC基因,该基因编码阐述于SEQ ID NO:133中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:133的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自幽门螺杆菌的ADC基因可以包括阐述于SEQ ID NO:133中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:133中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以来自芽孢杆菌属物种(Bacillus sp.)TS25 ADC基因,该基因编码阐述于SEQ ID NO:135中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:135的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自芽孢杆菌属物种TS25的ADC基因可以包括阐述于SEQ ID NO:134中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:134中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以来自谷氨酸棒杆菌ADC基因,该基因编码阐述于SEQ ID NO:137中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:137的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自谷氨酸棒杆菌的ADC基因可以包括阐述于SEQ ID NO:136中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:136中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以来自地衣芽孢杆菌ADC基因,该基因编码阐述于SEQ ID NO:139中的氨基酸序列。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:139的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自地衣芽孢杆菌(B.licheniformis)的ADC基因可以包括阐述于SEQ ID NOs:138、148、149、150或151的任一项中的核苷酸序列;或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NOs:138、148、149、150或151的任一项中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该ADC基因可以衍生自昆虫纲(昆虫)ADC基因,例如,如WO 2015/017721中所述(将其内容通过引用结合在此)。例如,在一些实施例中,该ADC基因可以编码以下各项的氨基酸序列:SEQ ID NO:162的埃及伊蚊(Aedes aegypti)ADC、SEQ ID NO:163的云南致倦库蚊(Culex quinquefasciatus)ADC、SEQ ID NO:164的冈比亚按蚊(Anophelesgambiae)ADC、SEQ ID NO:165的赤拟谷盗(Tribolium castaneum)ADC、SEQ ID NO:166的红缘皮蠹(Attagenus smirnovi)ADC、SEQ ID NO:167的豌豆蚜(Acyrthosiphon pisum)ADC、SEQ ID NO:168的瑟车利亚果蝇(Drosophila sechellia)ADC、SEQ ID NO:169的黑腹果蝇(Drosophila melanogaster)ADC、SEQ ID NO:170的大斑蝶(Danaus plexippus)ADC、SEQID NO:171的亚库巴果蝇(Drosophila yakuba)ADC、SEQ ID NO:172的埃瑞克塔果蝇(Drosophila erecta)ADC、SEQ ID NO:173的柑橘凤蝶(Papilio xuthus)ADC、SEQ ID NO:174的黑翅果蝇(Drosophila persimilis)ADC、SEQ ID NO:175的家蚕(Bombyx mori)ADC、SEQ ID NO:176的嗜凤梨果蝇(Drosophila ananassae)ADC、SEQ ID NO:177的漠海威果蝇(Drosophila mojavensis)ADC、SEQ ID NO:178的格瑞姆肖果蝇(Drosophila grimshawi)ADC、SEQ ID NO:179的桦尺蠖(Biston betularia)ADC、SEQ ID NO:180的威尔斯托尼果蝇(Drosophila willistoni)ADC、SEQ ID NO:181的西方蜜蜂(Apis mellifera)ADC、SEQ IDNO:182的维尔利斯果蝇(Drosophila virilis)ADC、SEQ ID NO:183的丽蝇蛹集金小蜂(Nasonia vitripennis)ADC、SEQ ID NO:184的偏瞳蔽眼蝶(Bicyclus anynana)ADC、SEQID NO:185的印度跳蚁(Harpegnathos saltator)ADC、SEQ ID NO:186的切叶蚁(Acromyrmex echinatior)ADC、SEQ ID NO:187的佛罗里达弓背蚁(Camponotusfloridanus)ADC、SEQ ID NO:188的人虱(Pediculus humanus)ADC、SEQ ID NO:189的大头美切叶蚁(Atta cephalotes)ADC、SEQ ID NO:190的油菜花露尾甲(Meligethes aeneus)ADC、或SEQ ID NO:191的红火蚁(Solenopsis invicta)ADC。在另一个方面,是包含双壳纲、鳃足纲、腹足纲、或头索纲(Leptocardii)的天冬氨酸1-脱羧酶(ADC)的重组细胞,该天冬氨酸1-脱羧酶(ADC)如SEQ ID NO:192的蚤状溞(Daphnia pulex)ADC、SEQ ID NO:193的霸王莲花青螺(Lottia gigantea)ADC、SEQ ID NO:194的佛罗里达文昌鱼(Branchiostomafloridae)ADC、或SEQ ID NO:195的太平洋牡蛎(Crassostrea gigas)ADC。在一些实施例中,该ADC基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:162-195中任一个的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
如在此使用,“β-丙氨酸转氨酶基因”或“BAAT基因”是指编码具有β-丙氨酸转氨酶活性的多肽的任何基因,意味着能够催化β-丙氨酸转化为丙二酸半醛。具有β-丙氨酸转氨酶活性的酶被分类为EC 2.6.1.19。在某些实施例中,BAAT基因可以来自酵母来源。例如,BAAT基因可以来自pyd4基因的东方伊萨酵母同系物,其编码阐述于SEQ ID NO:20中的氨基酸序列。在一些实施例中,该BAAT基因可以编码以下氨基酸序列,该氨基酸序列与SEQ IDNO:20的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自东方伊萨酵母的BAAT基因可以包括阐述于SEQ ID NO:19中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:19中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该BAAT基因可以来自克鲁维酵母(S.kluyveri)pyd4基因,该基因编码阐述于SEQ ID NO:21中的氨基酸序列。在一些实施例中,该BAAT基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:21的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自克鲁维酵母的BAAT基因可以包括阐述于SEQ ID NO:142中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ IDNO:142中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该BAAT基因可以来自细菌来源。例如,BAAT基因可以来自除虫链霉菌BAAT基因,该基因编码阐述于SEQ ID NO:22中的氨基酸序列。在一些实施例中,该BAAT基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:22的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自除虫链霉菌的BAAT基因可以包括阐述于SEQ ID NO:140中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:140中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
BAAT基因还可以是“4-氨基丁酸转氨酶”或“gabT基因”,意指它对4-氨基丁酸以及β-丙氨酸具有天然活性。可替代地,可以通过随机或定向工程化来自细菌或酵母来源的天然gabT基因而得到BAAT基因,以编码具有BAAT活性的多肽。例如,BAAT基因可以来自除虫链霉菌gabT基因,该gabT基因编码阐述于SEQ ID NO:23中的氨基酸序列。在一些实施例中,来自除虫链霉菌的BAAT基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:23的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,BAAT基因可以来自酿酒酵母gabT基因UGA1,该基因编码阐述于SEQ ID NO:24中的氨基酸序列。在一些实施例中,来自酿酒酵母的BAAT基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:24的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自酿酒酵母的BAAT基因可以包括阐述于SEQ ID NO:141中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:141中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在某些实施例中,3-HPDH基因可以来自酵母来源。例如,3-HPDH基因可以来自YMR226C基因的东方伊萨酵母同系物,其编码阐述于SEQ ID NO:26中的氨基酸序列。在一些实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:26的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自东方伊萨酵母的3-HPDH基因可以包括阐述于SEQ ID NO:25中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ IDNO:25中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,3-HPDH基因可以来自酿酒酵母YMR226C基因,该YMR226C基因编码阐述于SEQ ID NO:129中的氨基酸序列。在一些实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:129的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自酿酒酵母的3-HPDH基因可以包括阐述于SEQID NO:144中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:144中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:197的近平滑假丝酵母(Candida parapsilosis)氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:199的汉逊德巴利酵母(Debaryomyces hansenii)氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:201的季也蒙酵母(Meyerozyma guilliermondii)氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ IDNO:203的季也蒙酵母(Meyerozyma guilliermondii)氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在其他实施例中,该3-HPDH基因可以来自细菌来源。例如,3-HPDH基因可以来自大肠杆菌ydfG基因,该基因编码SEQ ID NO:27中的氨基酸序列。在一些实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与SEQ ID NO:27的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自大肠杆菌的3-HPDH基因可以包括阐述于SEQ ID NO:143中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:143中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,3-HPDH基因可以来自勤奋生金球菌(M.sedula)丙二酸半醛还原酶基因,该基因编码阐述于SEQ ID NO:29中的氨基酸序列。在一些实施例中,该3-HPDH基因可以编码以下氨基酸序列,该氨基酸序列与阐述于SEQ ID NO:29中的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在某些实施例中,来自勤奋生金球菌的3-HPDH基因可以包括阐述于SEQ ID NO:152中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ ID NO:152中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
在某些实施例中,与NADP(H)相比,3-HPDH基因可以是具有针对NAD(H)的增加的特异性的天然的或工程化的基因。具有针对NAD(H)的增加的特异性的3-HPDA变体描述于WO2013/049073(将其内容通过引用结合在此)中。
如在此使用,“3-羟基异丁酸脱氢酶基因”或“HIBADH基因”是指编码具有3-羟基异丁酸脱氢酶活性的多肽的任何基因,意味着能够催化3-羟基异丁酸转化为甲基丙二酸半醛。具有3-羟基异丁酸脱氢酶活性的酶被分类为EC 1.1.1.31。一些3-羟基异丁酸脱氢酶还具有3-HPDH活性。在某些实施例中,HIBADH基因可以来自细菌来源。例如,HIBADH基因可以来自编码阐述于SEQ ID NO:28中的氨基酸序列的粪产碱菌(A.faecalis)M3A基因、分别编码阐述于SEQ ID NO:30或SEQ ID NO:31中的氨基酸序列的恶臭假单胞菌(P.putida)KT2440或E23440 mmsB基因或编码阐述于SEQ ID NO:32中的氨基酸序列的铜绿假单胞菌(P.aeruginosa)PAO1 mmsB基因。在一些实施例中,HIBADH基因可以编码以下氨基酸序列,该氨基酸序列与阐述于SEQ ID NO:28、30、31或32中的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
如在此使用,“4-羟基丁酸脱氢酶基因”是指编码具有4-羟基丁酸脱氢酶活性的多肽的任何基因,意味着能够催化4-羟基丁酸转化为丁二酸半醛。具有4-羟基丁酸脱氢酶活性的酶被分类为EC 1.1.1.61。一些4-羟基丁酸脱氢酶还具有3-HPDH活性。在某些实施例中,4-羟基丁酸脱氢酶基因可以来自细菌来源。例如,4-羟基丁酸脱氢酶基因可以来自编码阐述于SEQ ID NO:33中的氨基酸序列的真氧产碱杆菌(R.eutropha)H16 4hbd基因或编码阐述于SEQ ID NO:34中的氨基酸序列的克氏梭菌(C.kluyveri)DSM 555hbd基因。在其他实施例中,该基因可以编码以下氨基酸序列,该氨基酸序列与阐述于SEQ ID NO:33或34中的氨基酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。
如在此使用,“PEP羧激酶基因”或“PCK基因”是指编码具有PEP羧激酶活性的多肽的任何基因,意味着能够催化PEP、CO2、和ADP或GDP转化为OAA和ATP或GTP,或反之亦然。具有PEP羧激酶活性的酶被分类为EC 4.1.1.32(利用GTP/GDP)和EC 4.1.1.49(利用ATP/ADP)。在某些实施例中,PCK基因可以来自酵母来源。在其他实施例中,PCK基因可以来自细菌来源,并且在这些实例的某些实施例中,该基因可以来自以下细菌,在该细菌体内PCK反应有利于OAA的产生而非其中脱羧作用是优势的更常见形式的反应。例如,PCK基因可以来自编码阐述于SEQ ID NO:35中的氨基酸序列的产琥拍酸曼氏杆菌(M.succiniciproducens)PCK基因、编码阐述于SEQ ID NO:36中的氨基酸序列的产琥珀酸厌氧螺菌(A.succiniciproducens)PCK基因、编码阐述于SEQ ID NO:37中的氨基酸序列的产琥珀酸放线杆菌(A.succinogenes)PCK基因或编码阐述于SEQ ID NO:38中的氨基酸序列的真氧产碱杆菌PCK基因。在其他实施例中,PCK基因已经经历对它来自其中的天然基因的一个或多个突变,这样使得所得基因编码以下多肽,该多肽优选地催化PEP转化为OAA。例如,PCK基因可以来自编码阐述于SEQ ID NO:39中的氨基酸序列的大肠杆菌K12菌株PCK基因,其中该基因已被突变为优选地催化PEP转化为OAA。在其他实施例中,通过PEP羧基转磷酸酶催化PEP至OAA的转化,例如发现于丙酸细菌(例如,谢氏丙酸杆菌(P.shermanii)、伍氏醋酸杆菌(A.woodii))中的,这些丙酸细菌使用无机磷酸和二磷酸而非ATP/ADP或GTP/GDP。
如在此使用,“苹果酸脱氢酶基因”是指编码具有苹果酸脱氢酶活性的多肽的任何基因,意味着能够催化OAA转化为苹果酸。在某些实施例中,苹果酸脱氢酶基因可以来自细菌或酵母来源。
如在此使用,“苹果酸脱羧酶基因”是指编码具有苹果酸脱羧酶活性的多肽的任何基因,意味着能够催化苹果酸转化为3-HP。已知苹果酸脱羧酶活性不天然地发生。因此,可以通过将一个或多个突变掺入天然来源基因中而得到苹果酸脱羧酶基因,该天然来源基因编码具有乙酰乳酸脱羧酶活性的多肽。具有乙酰乳酸脱羧酶活性的多肽催化2-羟基-2-甲基-3-氧代丁酸转化为2-乙偶姻,并被分类为EC 4.1.1.5。在某些实施例中,苹果酸脱羧酶基因可以来自细菌来源。例如,苹果酸脱羧酶基因可以来自编码阐述于SEQ ID NO:40中的氨基酸序列的乳酸乳球菌(L.lactis)aldB基因、编码阐述于SEQ ID NO:41中的氨基酸序列的嗜热链球菌(S.thermophilus)aldB基因、编码阐述于SEQ ID NO:42中的氨基酸序列的短芽孢杆菌(B.brevis)aldB基因或编码阐述于SEQ ID NO:43中的氨基酸序列的产气肠杆菌(E.aerogenes)budA基因。
如在此使用,“α-酮戊二酸(AKG)脱羧酶基因”或“KGD基因”是指编码具有α-酮戊二酸脱羧酶活性的多肽的任何基因,意味着能够催化α-酮戊二酸(2-氧化戊二酸)转化为丁二酸半醛。具有AKG脱羧酶活性的酶被分类为EC 4.1.1.71。可以使用KGD基因得到以下基因,该基因编码能够催化OAA转化为丙二酸半醛的多肽。这一活性可以发现于天然KGD基因中,或可以通过将一个或多个突变掺入天然KGD基因中而得到该活性。在某些实施例中,KGD基因可以来自细菌来源。例如,KGD基因可以来自编码阐述于SEQ ID NO:44中的氨基酸序列的结核分枝杆菌(M.tuberculosis)KGD基因、编码阐述于SEQ ID NO:45中的氨基酸序列的大豆慢生根瘤菌(B.japonicum)KGD基因或编码阐述于SEQ ID NO:46中的氨基酸序列的百脉根根瘤菌(M.loti)(又称百脉根瘤菌(Rhizobium loti))KGD基因。
如在此使用,“支链α-酮酸脱羧酶基因”或“BCKA基因”是指编码具有支链α-酮酸脱羧酶活性的多肽的任何基因,该活性可用于使一系列长度为三至六个碳的α-酮酸脱羧。具有BCKA活性的酶被分类为EC 4.1.1.72。可以使用BCKA基因得到以下基因,该基因编码能够催化OAA转化为丙二酸半醛的多肽。这一活性可以发现于天然BCKA基因中,或可以通过将一个或多个突变掺入天然BCKA基因中而得到该活性。在某些实施例中,BCKA基因可以来自细菌来源。例如,BCKA基因可以来自乳酸乳球菌kdcA基因,该kdcA基因编码阐述于SEQ ID NO:47中的氨基酸序列。
如在此使用,“吲哚丙酮酸脱羧酶基因”或“IPDA基因”是指编码具有吲哚丙酮酸脱羧酶活性的多肽的任何基因,意味着能够催化吲哚丙酮酸转化为吲哚乙醛。具有IPDA活性的酶被分类为EC 4.1.1.74。可以使用IPDA基因得到以下基因,该基因编码能够催化OAA转化为丙二酸半醛的多肽。这一活性可以发现于天然IPDA基因中,或可以通过将一个或多个突变掺入天然IPDA基因中而得到该活性。在某些实施例中,吲哚丙酮酸脱羧酶基因可以来自酵母、细菌或植物来源。
如在此使用,“丙酮酸脱羧酶基因”或“PDC基因”是指编码具有丙酮酸脱羧酶活性的多肽的任何基因,意味着能够催化丙酮酸转化为乙醛。具有PDC活性的酶被分类为EC4.1.1.1。在优选实施例中,被掺入如在此提供的重组细胞中的PDC基因已经经历对它来自其中的天然基因的一个或多个突变,这样使得所得基因编码能够催化OAA转化为丙二酸半醛的多肽。在某些实施例中,PDC基因可以来自酵母来源。例如,PDC基因可以来自编码阐述于SEQ ID NO:49中的氨基酸序列的东方伊萨酵母PDC基因、编码阐述于SEQ ID NO:50中的氨基酸序列的酿酒酵母PDC1基因或编码阐述于SEQ ID NO:51中的氨基酸序列的乳酸克鲁维酵母(K.lactis)PDC基因。在某些实施例中,来自东方伊萨酵母PDC基因的PDC基因可以包括阐述于SEQ ID NO:48中的核苷酸序列或以下核苷酸序列,该核苷酸序列与阐述于SEQ IDNO:48中的核苷酸序列具有至少50%、至少60%、至少70%、至少80%、至少85%、至少90%、至少95%、至少97%或至少99%序列一致性。在其他实施例中,PDC基因可以来自细菌来源。例如,PDC基因可以来自编码阐述于SEQ ID NO:52中的氨基酸序列的运动发酵单胞菌(Z.mobilis)PDC基因或编码阐述于SEQ ID NO:53中的氨基酸序列的巴氏醋杆菌(A.pasteurianus)PDC基因。
如在此使用,“苯甲酰甲酸脱羧酶”基因是指编码具有苯甲酰甲酸脱羧酶活性的多肽的任何基因,意味着能够催化苯甲酰甲酸转化为苯甲醛。具有苯甲酰甲酸脱羧酶活性的酶被分类为EC 4.1.1.7。在优选实施例中,被掺入如在此提供的重组细胞中的苯甲酰甲酸脱羧酶基因已经经历对它来自其中的天然基因的一个或多个突变,这样使得所得基因编码能够催化OAA转化为丙二酸半醛的多肽。在某些实施例中,苯甲酰甲酸脱羧酶基因可以来自细菌来源。例如,苯甲酰甲酸脱羧酶基因可以来自编码阐述于SEQ ID NO:54中的氨基酸序列的恶臭假单胞菌mdlC基因、编码阐述于SEQ ID NO:55中的氨基酸序列的铜绿假单胞菌mdlC基因、编码阐述于SEQ ID NO:56中的氨基酸序列的施氏假单胞菌(P.stutzeri)dpgB基因或编码阐述于SEQ ID NO:57中的氨基酸序列的荧光假单胞菌(P.fluorescens)ilvB-1基因。
如在此使用,“OAA甲酸裂解酶基因”是指编码具有OAA甲酸裂解酶活性的多肽的任何基因,意味着能够催化酰化物酮酸转化为其相应的辅酶A衍生物。由OAA甲酸裂解酶基因编码的多肽对丙酮酸或对另一种酮酸可以具有活性。在某些实施例中,OAA甲酸裂解酶基因编码一种将OAA转化为丙二酰辅酶A的多肽。
如在此使用,“丙二酰辅酶A还原酶基因”是指编码具有丙二酰辅酶A还原酶活性的多肽的任何基因,意味着能够催化丙二酰辅酶A转化为丙二酸半醛(亦称辅酶A酰化丙二酸半醛脱氢酶活性)。在某些实施例中,丙二酰辅酶A还原酶基因可以来自双功能丙二酰辅酶A还原酶基因,该双功能丙二酰辅酶A还原酶基因还具有催化丙二酸半醛转化为3-HP的能力。在这些实施例的某些实施例中,丙二酰辅酶A还原酶基因可以来自细菌来源。例如,丙二酰辅酶A还原酶基因可以来自编码阐述于SEQ ID NO:58中的氨基酸序列的橙色绿屈挠菌(C.aurantiacus)丙二酰辅酶A还原酶基因、编码阐述于SEQ ID NO:59中的氨基酸序列的卡氏玫瑰弯菌(R.castenholzii)丙二酰辅酶A还原酶基因或编码阐述于SEQ ID NO:60中的氨基酸序列的赤细菌属物种(Erythrobacter sp.)NAP1丙二酰辅酶A还原酶基因。在其他实施例中,丙二酰辅酶A还原酶基因可以来自以下丙二酰辅酶A还原酶基因,该基因编码仅催化丙二酰辅酶A转化为丙二酸半醛的多肽。例如,丙二酰辅酶A还原酶基因可以来自编码阐述于SEQ ID NO:61中的氨基酸序列的勤奋生金球菌Msed_0709基因或编码阐述于SEQ ID NO:62中的氨基酸序列的东工大硫化叶菌(S.tokodaii)丙二酰辅酶A还原酶基因。
如在此使用,“丙酮酸脱氢酶基因”或“PDH基因”是指编码具有丙酮酸脱氢酶活性的多肽的任何基因,意味着能够催化丙酮酸转化为乙酰辅酶A。在某些实施例中,PDH基因可以来自酵母来源。例如,PDH基因可以来自酿酒酵母LAT1、PDA1、PDB1或LPD基因,这些基因分别编码阐述于SEQ ID NO:63-66中的氨基酸序列。在其他实施例中,PDH基因可以来自细菌来源。例如,PDH基因可以来自大肠杆菌菌株K12亚株MG1655 aceE、aceF或lpd基因,这些基因分别编码阐述于SEQ ID NO:67-69中的氨基酸序列,或者枯草芽孢杆菌(B.subtilis)pdhA、pdhB、pdhC或pdhD基因,这些基因分别编码阐述于SEQ ID NO:70-73中的氨基酸序列。
如在此使用,“乙酰辅酶A羧化酶基因”或“ACC基因”是指编码具有乙酰辅酶A羧化酶活性的多肽的任何基因,意味着能够催化乙酰辅酶A转化为丙二酰辅酶A。具有乙酰辅酶A羧化酶活性的酶被分类为EC 6.4.1.2。在某些实施例中,乙酰辅酶A羧化酶基因可以来自酵母来源。例如,乙酰辅酶A羧化酶基因可以来自酿酒酵母ACC1基因,该基因编码阐述于SEQID NO:74中的氨基酸序列。在其他实施例中,乙酰辅酶A羧化酶基因可以来自细菌来源。例如,乙酰辅酶A羧化酶基因可以来自分别编码阐述于SEQ ID NO:75-78中的氨基酸序列的大肠杆菌accA、accB、accC或accD基因或分别编码阐述于SEQ ID NO:79-82中的氨基酸序列的橙色绿屈挠菌accA、accB、accC或accD基因。
如在此使用,“丙氨酸脱氢酶基因”是指编码具有丙氨酸脱氢酶基因活性的多肽的任何基因,意味着能够催化丙酮酸NAD依赖性还原氨化为丙氨酸。具有丙氨酸脱氢酶活性的酶被分类为EC 1.4.1.1。在某些实施例中,丙氨酸脱氢酶基因可以来自细菌来源。例如,丙氨酸脱氢酶基因可以来自枯草芽孢杆菌丙氨酸脱氢酶基因,该枯草芽孢杆菌丙氨酸脱氢酶基因编码阐述于SEQ ID NO:83中的氨基酸序列。
如在此使用,“丙酮酸/丙氨酸转氨酶基因”是指编码具有丙酮酸/丙氨酸转氨酶活性的多肽的任何基因,意味着能够催化丙酮酸和L-谷氨酸转化为丙氨酸和2-氧化戊二酸。在某些实施例中,丙酮酸/丙氨酸转氨酶基因来自酵母来源。例如,丙酮酸/丙氨酸转氨酶基因可以来自编码阐述于SEQ ID NO:84中的氨基酸序列的粟酒裂殖酵母(S.pombe)丙酮酸/丙氨酸转氨酶基因或编码阐述于SEQ ID NO:85中的氨基酸序列的酿酒酵母ALT2基因。
如在此使用,“丙氨酸2,3氨基变位酶基因”或“AAM基因”是指编码具有丙氨酸2,3氨基变位酶活性的多肽的任何基因,意味着能够催化丙氨酸转化为β-丙氨酸。已知丙氨酸2,3氨基变位酶活性不天然地发生。因此,可通过将一个或多个突变掺入天然来源基因中而得到丙氨酸2,3氨基变位酶基因,该天然来源基因编码具有类似活性(例如赖氨酸2,3氨基变位酶活性)的多肽(参见例如,美国专利7,309,597)。在某些实施例中,该天然来源基因可以是编码阐述于SEQ ID NO:86中的氨基酸序列的枯草芽孢杆菌赖氨酸2,3氨基变位酶基因、编码阐述于SEQ ID NO:87中的氨基酸序列的牙龈卟啉单胞菌(P.gingivalis)赖氨酸2,3氨基变位酶基因或编码阐述于SEQ ID NO:88中的氨基酸序列的具核梭杆菌(F.nucleatum)(ATCC-10953)赖氨酸2,3氨基变位酶基因。
如在此使用,“辅酶A转移酶基因”是指编码具有辅酶A转移酶活性的多肽的任何基因,在一个实例中,该活性包括催化β-丙氨酸转化为β-丙氨酰辅酶A和/或催化乳酸转化为乳酰辅酶A的能力。在某些实施例中,辅酶A转移酶基因可以来自酵母来源。在其他实施例中,辅酶A转移酶基因可以来自细菌来源。例如,辅酶A转移酶基因可以来自埃氏巨球形菌(M.elsdenii)辅酶A转移酶基因,该埃氏巨球形菌辅酶A转移酶基因编码阐述于SEQ ID NO:89中的氨基酸序列。
如在此使用,“辅酶A合成酶基因”是指编码具有辅酶A合成酶活性的多肽的任何基因。在一个实例中,这一活性包括催化β-丙氨酸转化为β-丙氨酰辅酶A的能力。在另一个实例中,这一活性包括催化乳酸转化为乳酰辅酶A的能力。在某些实施例中,辅酶A合成酶基因可以来自酵母来源。例如,辅酶A合成酶基因可以来自酿酒酵母辅酶A合成酶基因。在其他实施例中,辅酶A合成酶基因可以来自细菌来源。例如,辅酶A合成酶基因可以来自大肠杆菌辅酶A合成酶、类球红细菌或肠道沙门菌(S.enterica)辅酶A合成酶基因。
如在此使用,“β-丙氨酰辅酶A解氨酶基因”是指编码具有β-丙氨酰辅酶A解氨酶活性的多肽的任何基因,意味着能够催化β-丙氨酰辅酶A转化为丙烯酰辅酶A。在某些实施例中,β-丙氨酰辅酶A解氨酶基因可以来自细菌来源,例如丙酸梭菌(C.propionicum)β-丙氨酰辅酶A解氨酶基因,该基因编码阐述于SEQ ID NO:90中的氨基酸序列。
如在此使用,“3-HP辅酶A脱水酶基因”或“丙烯酰辅酶A水合酶基因”是指编码具有3-HP辅酶A脱水酶活性的多肽的任何基因,意味着能够催化丙烯酰辅酶A转化为3-HP辅酶A。具有3-HP辅酶A脱水酶活性的酶被分类为EC 4.2.1.116。在某些实施例中,3-HP辅酶A脱水酶基因可以来自酵母或真菌来源,例如大豆疫霉菌(P.sojae)3-HP辅酶A脱水酶基因,该基因编码阐述于SEQ ID NO:91中的氨基酸序列。在其他实施例中,3-HP辅酶A脱水酶基因可以来自细菌来源。例如,3-HP辅酶A脱水酶基因可以来自编码阐述于SEQ ID NO:92中的氨基酸序列的橙色绿屈挠菌3-HP辅酶A脱水酶基因、编码阐述于SEQ ID NO:93中的氨基酸序列的深红红螺菌(R.rubrum)3-HP辅酶A脱水酶基因或编码阐述于SEQ ID NO:94中的氨基酸序列的有蒴红螺菌(R.capsulates)3-HP辅酶A脱水酶基因。在仍其他实施例中,3-HP辅酶A脱水酶基因可以来自哺乳动物来源。例如,3-HP辅酶A脱水酶基因可以来自智人(H.sapiens)-HP辅酶A脱水酶基因,该智人-HP辅酶A脱水酶基因编码阐述于SEQ ID NO:95中的氨基酸序列。
如在此使用,“3-HP辅酶A水解酶基因”是指编码具有3-HP辅酶A水解酶活性的多肽的任何基因,意味着能够催化3-HP辅酶A转化为3-HP。在某些实施例中,3-HP辅酶A水解酶基因可以来自酵母或真菌来源。在其他实施例中,3-HP辅酶A水解酶基因可以来自细菌来源。
如在此使用,“3-羟基异丁酰辅酶A水解酶基因”是指编码具有3-羟基异丁酰辅酶A水解酶活性的多肽的任何基因,在一个实例中,该活性包括催化3-HP辅酶A转化为3-HP的能力。在某些实施例中,3-羟基异丁酰辅酶A水解酶基因可以来自细菌来源,例如编码阐述于SEQ ID NO:96中的氨基酸序列的荧光假单胞菌3-羟基异丁酰辅酶A水解酶基因或编码阐述于SEQ ID NO:97中的氨基酸序列的蜡状芽孢杆菌(B.cereus)3-羟基异丁酰辅酶A水解酶基因。在其他实施例中,3-羟基异丁酰辅酶A水解酶基因可以来自哺乳动物来源,例如智人3-羟基异丁酰辅酶A水解酶基因,该基因编码阐述于SEQ ID NO:98中的氨基酸序列。
如在此使用,“乳酸脱氢酶基因”或“LDH基因”是指编码具有乳酸脱氢酶活性的多肽的任何基因,意味着能够催化丙酮酸转化为乳酸。在某些实施例中,LDH基因可以来自真菌、细菌或哺乳动物来源。
如在此使用,“乳酰辅酶A脱水酶基因”是指编码具有乳酰辅酶A脱水酶活性的多肽的任何基因,意味着能够催化乳酰辅酶A转化为丙烯酰辅酶A。在某些实施例中,乳酰辅酶A脱水酶基因可以来自细菌来源。例如,乳酰辅酶A脱水酶基因可以来自埃氏巨球形菌乳酰辅酶A脱水酶E1、EIIa或EIIb亚单位基因,这些基因编码阐述于SEQ ID NO:99-101中的氨基酸序列。
如在此使用,“醛脱氢酶基因”是指编码具有醛脱氢酶活性的多肽的任何基因,在一个实例中,该活性包括催化3-HPA转化为3-HP的能力(反之亦然)。在某些实施例中,醛脱氢酶基因可以来自酵母来源,例如编码阐述于SEQ ID NO:102中的氨基酸序列的酿酒酵母醛脱氢酶基因或编码阐述于SEQ ID NO:122、124或126中的氨基酸序列的东方伊萨酵母醛脱氢酶基因。在其他实施例中,醛脱氢酶基因可以来自细菌来源,例如编码阐述于SEQ IDNO:103中的氨基酸序列的大肠杆菌aldH基因或编码阐述于SEQ ID NO:104中的氨基酸序列的肺炎克雷伯氏菌(K.pneumoniae)醛脱氢酶基因。
如在此使用,“甘油脱水酶基因”是指编码具有甘油脱水酶活性的多肽的任何基因,意味着能够催化甘油转化为3-HPA。在某些实施例中,甘油脱水酶基因可以来自细菌来源,例如肺炎克雷伯氏菌或弗氏柠檬酸杆菌甘油脱水酶基因。
可以使用本领域已知的方法或如在此描述检测所选主动3-HP途径的酶及其活性。这些检测方法可包括使用特异抗体、形成酶产物、或酶底物的消失。参见,例如Sambrook等人,Molecular Cloning:A Laboratory Manual,Third Ed.,Cold Spring HarborLaboratory,New York(2001)[分子克隆实验指南,第三版,冷泉港实验室,纽约(2001)];Ausubel等人,Current Protocols in Molecular Biology,John Wiley and Sons,Baltimore,MD(1999)[当前分子生物学,约翰威利父子公司,巴尔的摩,马里兰州(1999)];以及Hanai等人,Appl.Environ.Microbiol.[应用及环境微生物学]73:7814-7818(2007))。
在此描述的重组细胞可以进一步包含脂肪酶或酯酶活性,例如归因于表达一种编码脂肪酶或酯酶(EC 3.1.1.-)的异源多核苷酸。可以使用此类细胞生产3-HP的酯,例如3-羟基丙酸甲酯、3-羟基丙酸乙酯、3-羟基丙酸丙酯、3-羟基丙酸丁酯或2-乙基己基3-羟基丙酸酯。这些细胞可以进一步包含酯酶活性,例如归因于表达一种编码酯酶的异源多核苷酸。可以使用此类细胞生产聚合的3-HP。这些细胞可以进一步包含醇脱氢酶(EC 1.1.1.1)活性、醛脱氢酶(EC 1.2.1.-)活性或两者,例如归因于表达一种编码醇脱氢酶、醛脱氢酶或两者的异源多核苷酸。可以使用此类细胞生产1,3-丙二醇。
这些重组细胞还可以包括一个或多个(例如,两个、若干个)基因破坏,例如以将糖代谢从不希望的产物转移至3-HP。在一些方面,当在相同条件下培养时,与没有这一个或多个破坏的细胞相比,这些重组宿主细胞产生更大量的3-HP。在一些方面,使这些被破坏的内源基因中的一个或多个失活。
在某些实施例中,在此提供的重组细胞包含编码在乙醇生产中涉及的酶的一个或多个内源基因的破坏,包括例如丙酮酸脱羧酶(PDC,将丙酮酸转化为乙醛)和/或醇脱氢酶(ADH,将乙醛转化为乙醇)基因。这些修饰降低了细胞生产乙醇的能力,从而使3-HP生产最大化。然而,在某些实施例中,在此提供的重组细胞可以被工程化为同时生产3-HP和乙醇。在那些实施例中,优选地不破坏编码在乙醇发酵中涉及的酶的内源基因,并且在某些实施例中,这些细胞可以包括一个或多个增加乙醇产生的异源基因。
在一些实施例中,这些重组细胞可以包括对编码PDC的内源基因的破坏,该PDC与SEQ ID NO:154具有至少75%,例如至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一些实施例中,该内源基因编码具有以下氨基酸序列的PDC,该氨基酸序列包括SEQ ID NO:154或由其组成。在一些实施例中,编码PDC的内源基因的编码序列与SEQ ID NO:153具有至少75%,例如至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一些实施例中,编码PDC的内源基因的编码序列包括SEQ ID NO:153或由其组成。在一些实施例中,使编码PDC的内源基因失活。
在某些实施例中,在此提供的重组细胞包含一个或多个内源基因的破坏,这一个或多个内源基因编码在生产替代发酵产物(例如甘油)或其他副产物(例如乙酸或二醇)中涉及的酶。例如,在此提供的细胞可以包括以下一个或多个的基因中的破坏:甘油3-磷酸脱氢酶(GPD,催化二羟丙酮磷酸反应为甘油3-磷酸),甘油3-磷酸酶(GPP,催化甘油-3磷酸转化为甘油),甘油激酶(催化甘油3-磷酸转化为甘油),二羟丙酮激酶(催化二羟丙酮磷酸转化为二羟丙酮),甘油脱氢酶(催化二羟丙酮转化为甘油),醛脱氢酶(ALD,例如,将乙醛转化为乙酸或将3-HP转化为3-HPA)、以及丁二醇脱氢酶(催化丁二醇转化为乙偶姻,反之亦然)。
在一些实施例中,这些重组细胞可以包括对编码GPD的内源基因的破坏,该PDC与SEQ ID NO:156具有至少75%,例如至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一些实施例中,该内源基因编码具有以下氨基酸序列的GPD,该氨基酸序列包括SEQ ID NO:156或由其组成。在一些实施例中,编码GPD的内源基因的编码序列与SEQ ID NO:155具有至少75%,例如至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。在一些实施例中,编码GPD的内源基因的编码序列包括SEQ ID NO:155或由其组成。在一些实施例中,使编码GPD的内源基因失活。
在某些实施例中,在此提供的重组细胞包含一个或多个内源基因的破坏,这一个或多个内源基因编码催化3-HP途径中的逆反应的酶,包括例如PEP羧激酶(PCK)、具有OAA脱羧酶活性的酶或CYB2A或CYB2B(催化乳酸转化为丙酮酸)。PCK催化PEP转化为OAA,反之亦然,但是对OAA至PEP的反应展现出偏好。为了减少OAA至PEP的转化,可以破坏天然PCK基因的一个或多个拷贝。在某些实施例中,其中的一个或多个天然PCK基因已经被破坏的细胞可以表达一个或多个异源PCK基因,这一个或多个异源PCK基因已被突变为编码以下多肽,该多肽有利于将PEP转化为OAA。OAA脱羧酶催化OAA转化为丙酮酸。已经鉴定了具有OAA脱羧酶活性的酶,例如由大肠杆菌中的恩纳-杜道夫醛缩酶(Entner-Doudoroff aldolase,eda)基因编码的酶以及酵母和真菌中的苹果酸酶(MAE)。为了减少OAA脱羧酶活性,可以破坏编码具有OAA脱羧酶活性的酶的天然基因的一个或多个拷贝。在某些实施例中,其中的一个或多个天然OAA脱羧基因已经被破坏的细胞可以表达一个或多个异源OAA脱羧基因,这一个或多个异源OAA脱羧基因已被突变为编码以下多肽,该多肽催化丙酮酸转化为OAA。
在某些实施例中,在此提供的重组细胞包含对选自dse2、scw11、eaf3、sed1和sam2的一种或多种基因的破坏(参见,例如Suzuki等人,J.Biosci.Bioeng.[生物科学与生物工程杂志]2013,115,467-474;以及Dato等人,Microbial Cell Factories[微生物细胞工厂],2014,13,147)。
在某些实施例中,在此提供的重组细胞包含一个或多个内源基因的破坏,这一个或多个内源基因编码在与3-HP途径产物或中间体进行的不希望的反应中涉及的酶。此类基因的实例包括编码将3-HP转化为3-HP的醛的酶的那些基因,已知这些醛对某些细胞而言是有毒的。
在某些实施例中,在此提供的重组细胞包含一个或多个内源基因的破坏,这一个或多个内源基因编码对3-HP途径具有中性作用的酶,包括例如GAL6(将半乳糖转化为葡萄糖的GAL系统的负调节物)。中性基因的破坏允许在不影响天然途径的情况下插入一个或多个异源基因。
还可以使用建模设计基因破坏,这些基因破坏另外地优化途径的利用(参见,例如,U.S.2002/0012939、U.S.2003/0224363、U.S.2004/0029149、U.S.2004/0072723、U.S.2003/0059792、U.S.2002/0168654、U.S.2004/0009466、以及美国专利7,127,379)。建模分析允许可靠地预测将代谢移向更有效地生产3-HP对细胞生长的影响。用于鉴定并设计有利于生物合成希望的产物的代谢改变的的一种示例性计算方法是OptKnock计算框架(OptKnock computational framework),Burgard等人,2003,Biotechnol.Bioeng.[生物技术与生物工程]84:647-657。
上文详细描述了用于破坏基因的示例性方法。
生产3-HP以及相关化合物的方法
在此描述的包含主动3-HP途径的宿主细胞可以用于生产3-HP。在一个方面,是一种产生3-HP的方法,该方法包括:(a)在适合的条件下,在可发酵的培养基中培养重组宿主细胞以产生3-HP,该重组宿主细胞包含主动3-HP途径和对在此描述的编码丙酮酸还原酶(例如,SEQ ID NO:205的丙酮酸还原酶)的内源基因的破坏;并且(b)回收3-HP。
可以使用本领域熟知的方法,在适合生产3-HP的营养培养基中培养包含主动3-HP途径的重组细胞。例如,可以通过在适合的发酵培养基中和在允许产生3-HP的条件下,进行摇瓶培养,以及在实验室或工业发酵罐中进行小规模或大规模发酵(包括连续、分批、分批补料或固态发酵)来培养这些细胞。
这些重组细胞可以在一种可发酵的培养基中产生3-HP,该可发酵培养基包括一种或多种(例如,两种、若干种)糖,这些糖是例如葡萄糖、果糖、蔗糖、纤维二糖、木糖、木酮糖、阿拉伯糖、甘露糖、半乳糖和/或可溶的低聚糖。碳源可以是十二碳糖(例如蔗糖)、己糖(例如葡萄糖或果糖)、多糖或葡萄糖的其他聚合物、葡萄糖寡聚物(例如麦芽糖、麦芽三糖和异麦芽三糖)、潘糖以及果糖寡聚物。如果该细胞被修饰为赋予发酵戊糖的能力,则该发酵培养基可以包括戊糖,例如木糖、木聚糖或木糖的其他寡聚物和/或阿拉伯糖。此类戊糖可以适合地是包含半纤维素的生物质的水解产物。在一些实施例中,该细胞不能发酵戊糖和/或该可发酵的培养基包括低于1%的戊糖。在一些情况下,该可发酵的培养基来自天然来源,例如甘蔗、淀粉、或纤维素;并且可以是通过酶水解(糖化作用)对这种来源进行预处理的结果。在一些方面,该可发酵的培养基包括甘蔗汁。适合的培养基从商业供应商处可获得或者可根据公开的组成(例如,在美国典型培养物保藏中心目录中)来制备,或可以从可商购的成分制备。
除了来自一种或多种(例如,两种、若干种)糖的适当的碳源之外,该可发酵的培养基可以包含本领域普通技术人员已知的其他营养素或刺激物,例如大量营养素(如,氮源)以及微量营养素(如,维生素、矿物盐、以及金属辅因子)。在一些方面,碳源优先地可以补充有至少一种碳源,例如酵母提取物、N2、蛋白胨(例如,BactoTM蛋白胨)、或大豆蛋白胨(例如,BactoTM大豆蛋白胨)。维生素的非限制性实例包括多种维生素、生物素、泛酸盐、烟酸、内消旋肌醇、硫胺素、吡哆醇、对氨基苯甲酸、叶酸、核黄素、以及维生素A、维生素B、维生素C、维生素D和维生素E。矿物盐和金属辅因子的实例包括但不限于Na、P、K、Mg、S、Ca、Fe、Zn、Mn、Co以及Cu。
在一些实施例中,可以在化学成分确定的培养基中培养本发明的重组细胞。在一个实例中,该培养基包含大约5g/L硫酸铵,大约3g/L磷酸二氢钾,大约0.5g/L硫酸镁、痕量元素和维生素以及大约150g/L葡萄糖。在培养过程中可以允许pH自由变化,或必要的话,可以缓冲,以防止pH跌至预定的水平之下或升至预定的水平之上。在某些实施例中,用足够的细胞接种该发酵培养基,这些细胞是产生约1.0的OD600的评估对象。除非另外明确指明,如在此使用,OD600是指使用型号DU600分光光度计(贝克曼库尔特公司(Beckman Coulter))在600nm的波长下以1cm路长测量的光密度。
用于生产3-HP的方法的具体条件可以由本领域普通技术人员根据在此的教导确定。在这些方法的一些方面,将这些细胞培养约12小时至约216小时,例如约24小时至约144小时、或约36小时至约96小时。温度典型地在约26℃至约60℃之间,例如约34℃至约50℃。
视情况而定,可以在厌氧、基本上厌氧(微好氧)、或好氧条件下进行培养。简言之,厌氧的是指一种缺少氧气的环境,基本上厌氧的(微好氧的)是指一种其中氧气的浓度比空气低的环境,并且好氧是指一种其中氧气浓度大致等于或大于空气的氧气浓度的环境。基本上缺氧条件包括例如使得在培养基中的溶氧浓度保持在小于10%的饱和度的培养、分批发酵或连续发酵。基本上缺氧条件还包括使细胞在维持有小于1%氧气的气氛的密封室内的液体培养基中或固体琼脂上生长或静止。例如可以通过向培养物鼓吹N2/CO2混合物或其他一种或多种适合的非氧气体来维持氧气的百分比。在一些实施例中,在缺氧条件或基本上缺氧条件下进行培养。
在一个实例中,在生产期过程中,发酵培养基中的细胞浓度典型地在约1至40,例如在从2至20、或从3至10克干细胞/升发酵培养基的范围内。希望的话,摄氧率(OUR)可以贯穿整个发酵变化作为过程控制(参见例如,WO 03/102200)。在一些实施例中,在微好氧条件下培养在此提供的重组细胞,这些微好氧条件由从2至45mmol/L/hr,例如2至25、2至20、2至15、2至10、10至45、15至40、20至35或25至35mmol/L/hr的摄氧率表征。在某些实施例中,当在由从2至25mmol/L/hr的摄氧率表征的微好氧条件下培养时,在此提供的重组细胞表现尤其好。可以在生产期过程中缓冲培养基,这样使得将pH维持在约3.0至约7.0或从约4.0至约6.0的范围内。适合的缓冲剂是当酸形成时中和它的碱性材料,并且包括例如氢氧化钙、碳酸钙、氢氧化钠、氢氧化钾、碳酸钾、碳酸钠、碳酸铵、氨、氢氧化铵等。通常,那些在常规的发酵过程中使用的缓冲剂在此也是适合的。
在利用缓冲发酵的那些实施例中,当酸性发酵产物形成时,可以将它们中和成相应的盐。在这些实施例中,回收酸包括再生游离酸。这可以通过以下方式完成:移除细胞并用强酸(例如硫酸)酸化发酵液。这导致形成盐副产物。例如,在将钙盐用作中和剂并且将硫酸用作酸化剂的情况下,将石膏产生为盐副产物。将这一副产物与发酵液分离,并且使用例如液-液提取、蒸馏、吸收以及其他技术回收酸(参见例如,T.B.Vickroy,Vol.3,Chapter38of Comprehensive Biotechnology,(ed.M.Moo-Young),Pergamon,Oxford,1985[T.B.Vickroy,综合生物技术,第3卷,第38章,M.Moo-Young编辑,帕加马,牛津,1985];Datta等人,1995,FEMS Microbiol.Rev.[FEMS微生物学评论]16:221-231;美国专利4,275,234、4,771,001、5,132,456、5,420,304、5,510,526、5,641,406和5,831,122以及WO 93/00440)。
在其他实施例中,在培养过程中,可以允许发酵培养基的pH从处于或高于3-HP的pKa的起始pH(典型地是4.5或更高,比如5.5、6.0或6.5)降至处于或低于酸发酵产物的pKa,例如低于4.5或4.0,如在约1.5至约4.5的范围内、在从约2.0至约4.0的范围内或在从约2.0至约3.5的范围内。
在仍其他实施例中,可以通过在发酵过程之前或开始时,将发酵液的pH调节为处于或低于产物酸的pKa而进行发酵,以产生该产物酸。之后,贯穿整个培养,可以将pH维持为处于或低于该产物酸的pKa。在某些实施例中,可以将pH维持为低于4.5或4.0,例如在约1.5至约4.5的范围内、在约2.0至约4.0的范围内或在约2.0至约3.5的范围内。
在此描述的这些方法可以利用任何适合的发酵操作模式。例如,分批模式发酵可以与一个封闭系统一起使用,其中培养基和重组宿主细胞(在发酵的开始设置的)除了试剂(某些例如用于pH控制、泡沫控制,或过程支持所需的其他试剂)之外,没有另外的投入。在此描述的过程还可以在补料分批-模式或连续模式中利用,如上所提及。
可以在若干生物反应器构型中实践在此描述的这些方法,例如搅拌槽、鼓泡塔、气升式反应器以及本领域普通技术人员已知的其他反应器。视情况而定,能以游离细胞培养或固定化细胞培养的方式执行这些方法。可以使用任何用于支持固定化细胞培养的材料,例如藻酸盐、纤维床、或菱形材料,例如温石棉、蒙脱石KSF和蒙脱石K-10。
在这些方法的一个方面,生产的3-HP的滴度大于约5g/L,例如大于约10g/L、25g/L、50g/L、75g/L、100g/L、125g/L、150g/L、160g/L、170g/L、180g/L、190g/L、200g/L、210g/L、225g/L、250g/L、275g/L、300g/L、325g/L、350g/L、400g/L或500g/L;或在约10g/L与约500g/L之间,例如约50g/L与约350g/L之间、约100g/L与约300g/L之间、约150g/L与约250g/L之间、约175g/L与约225g/L之间或约190g/L与约210g/L之间。在一个实施例中,以大于约0.01克/克碳水化合物的滴度生产3-HP,例如大于约0.02、0.05、0.75、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、或1.0克/克碳水化合物。
在于此提供的这些方法的某些实施例中,这些重组细胞产生相对较低水平的乙醇。在某些实施例中,可以按10%或更少的产量,优选按2%或更少的产量生产乙醇。在这些实施例的某些实施例中,产生的乙醇检测不出。然而,在其他实施例中,可以同时生产3-HP和乙醇。在这些实施例中,可以按大于10%、大于25%或大于50%的产量生产乙醇。
可以使用本领域已知的任何程序从发酵培养基中任选地回收3-HP,这些程序包括但不限于色谱法(例如,尺寸排阻色谱法、吸附色谱法、离子交换色谱法)、电泳程序、溶解度差异、渗透、蒸馏、提取(例如,液液提取)、渗透蒸发、萃取过滤、膜过滤、膜分离、反渗透或超滤。在一个方面,从其他发酵材料中分离3-HP并通过蒸馏的常规方法加以纯化。因此,在一个方面,该方法进一步包括通过蒸馏纯化回收的3-HP。
还可以通过上述程序(例如,色谱法、电泳程序、溶解度差异、蒸馏或提取)将杂质(污染物)化学转化为更易于从3-HP中去除的产物和/或通过将杂质直接化学转化为3-HP而纯化重组3-HP。例如,在一个方面,该方法进一步包括通过使用本领域已知的化学技术将-丙氨酸污染物转化为3-HP而纯化回收的3-HP。
在这些方法的一些方面,在可任选地纯化之前和/或之后的重组3-HP制剂是基本上纯的。关于生产3-HP的方法,“基本上纯的”意指回收的制剂包含不超过15%的杂质,其中杂质意指除3-HP以外的化合物。在一种变体中,提供了一种基本上纯的制剂,其中该制剂包含至多25%杂质、或至多20%杂质、或至多10%杂质、或至多5%杂质、或至多3%杂质、或至多1%杂质、或至多0.5%杂质。
可以将使用在此披露的方法所生产的3-HP转化为其他有机化合物(例如,如US 8,030,045和WO 03/082795中所述,将其内容通过引用结合在此)。例如,可以将3-HP氢化以形成1,3丙二醇(一种有价值的聚酯单体)。也可以使用在体外或在体内具有氧化还原酶活性的多肽从3-HP生产丙二醇(例如使用如上所述的一种或多种另外的酶活性)。可以使用任何方法氢化有机酸(例如3-HP),例如用于氢化丁二酸和/或乳酸的那些方法。例如,可以使用金属催化剂氢化3-HP。因此,在一个方面,是一种生产1,3-丙二醇的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞(例如,包含主动3-HP途径的重组宿主细胞),其中该重组细胞进一步包含氧化还原酶活性,以生产1,3-丙二醇;并且(b)回收1,3-丙二醇。在另一个方面,是一种生产1,3-丙二醇的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞(例如,包含主动3-HP途径的重组宿主细胞),以生产3-HP;(b)回收该3-HP;(c)在适合的条件下,氢化该3-HP以生产1,3-丙二醇;并且(d)回收1,3-丙二醇。
3-HP还可以被转化为3-HP的酯,例如3-羟基丙酸甲酯、3-羟基丙酸乙酯、3-羟基丙酸丙酯、3-羟基丙酸丁酯或2-乙基己基3-羟基丙酸酯。在一个方面,是一种生产3-HP酯的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞(例如,包含主动3-HP的重组宿主细胞),其中该重组细胞还包含脂肪酶或酯酶活性,以生产3-HP酯;并且(b)回收3-HP酯。在另一个方面,是一种生产3-HP酯的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞(例如,包含主动3-HP途径的重组宿主细胞),以生产3-HP;(b)回收该3-HP;(c)在适合的条件下,酯化该3-HP以产生3-HP酯;并且(d)回收3-HP酯。在此描述的重组宿主细胞也可用于使用本领域已知的技术在体外或体内产生聚合的3-HP(例如,Zhou等人,Metab.Eng.[代谢工程]2011,13(6),777-785;以及US 8,030,045;将其内容通过引用结合在此)。
可以将通过任何在此描述的方法生产的3-HP转化为丙烯酸。可以使用本领域已知的技术通过将3-HP化学脱水而产生丙烯酸,例如在催化剂(例如,固体氧化物脱水催化剂,如二氧化钛或氧化铝)的存在下加热。
在另一个方面,是一种生产丙烯酸或其盐的方法,该方法包括:(a)在适合的条件下,在培养基中培养在此描述的重组细胞(例如,包含主动3-HP途径的重组宿主细胞),以生产3-HP;(b)回收该3-HP;(c)在适合的条件下,将该3-HP脱水,以产生丙烯酸或其盐;并且(d)回收该丙烯酸或其盐。
可以使用本领域已知的方法进行适合的测定,这些测定用于测试在此描述的生产方法和细胞的3-HP、乳酸盐/乳酸酯及其衍生物的产生。例如,可以通过多种方法(例如HPLC(高效液相色谱)、GC-MS(气相色谱-质谱法)和LC-MS(液相色谱-质谱法))或使用本领域熟知的常规程序的其他适合的分析方法对终产物3-HP和中间体(例如,丙酮酸)以及其他有机化合物进行分析。还可以用培养上清液测试发酵液中的3-HP的释放。可以使用例如针对葡萄糖和醇类的折光率检测器、以及针对有机酸的UV检测器通过HPLC(Lin等人,Biotechnol.Bioeng.[生物技术与生物工程]90:775-779(2005))、或使用本领域熟知的其他适合的测定和检测方法量化在发酵培养基中的副产物和残余的糖(例如,葡萄糖)。
本发明可进一步通过下述编号段落描述:
[1]一种重组酵母细胞,包含(1)能够产生3-HP的主动3-HP途径和(2)对编码丙酮酸还原酶的内源基因的破坏,其中:
(a)该丙酮酸还原酶与SEQ ID NO:205具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性;
(b)编码该丙酮酸还原酶的内源基因的编码序列在至少低严格条件下,例如中严格条件下、中-高严格条件下、高严格条件下、或非常高严格条件下与SEQ ID NO:204的全长互补链杂交;或者
(c)编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。
[2]如段落[1]所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。
[3]如段落[1]或[2]所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205相差不超过十个氨基酸,例如不超过五个氨基酸、不超过四个氨基酸、不超过三个氨基酸,不超过两个氨基酸,或一个氨基酸。
[4]如段落[1]所述的重组细胞,其中该细胞包含对编码丙酮酸还原酶的内源基因的破坏,该丙酮酸还原酶包含SEQ ID NO:205或者由其组成。
[5]如段落[1]-[4]中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%或100%序列一致性。
[6]如段落[1]所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列包括SEQ ID NO:204或由其组成。
[7]如段落[1]-[6]中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列在至少低严格条件下,例如中严格条件下、中-高严格条件下、高严格条件下、或非常高严格条件下与SEQ ID NO:204的全长互补链杂交。
[8]如段落[1]-[7]中任一项所述的重组细胞,其中该破坏发生于编码该丙酮酸还原酶的内源基因的编码序列中。
[9]如段落[1]-[8]中任一项所述的重组细胞,其中该破坏发生于编码该丙酮酸还原酶的内源基因的控制序列中。
[10]如段落[1]-[9]中任一项所述的重组细胞,其中当在相同条件下培养时,与缺乏对编码该丙酮酸还原酶的内源基因的破坏的亲本菌株相比,细胞产生更少的D-乳酸盐/D-乳酸酯(例如,减少至少25%、减少50%、减少至少60%、减少至少70%、减少至少80%、减少至少90%、或减少100%)。
[11]如段落[1]-[10]中任一项所述的重组细胞,其中当在相同条件下培养时,与缺乏对编码该丙酮酸还原酶的内源基因的破坏的亲本菌株相比,该细胞产生更少的丙酮酸还原酶(例如,减少至少25%、减少至少50%、减少至少60%、减少至少70%、减少至少80%、减少至少90%、或减少100%)。
[12]如段落[1]-[11]中任一项所述的重组细胞,其中将编码该丙酮酸还原酶的内源基因失活。
[13]如段落[1]-[12]中任一项所述的重组细胞,其中该细胞包含选自以下各项的一种或多种(例如两种、若干种)异源多核苷酸:
编码丙酮酸脱氢酶(PDH)的异源多核苷酸;
编码乙酰辅酶A羧化酶(ACC)的异源多核苷酸;
编码丙二酰辅酶A还原酶的异源多核苷酸;和
编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。
[14]如段落[1]-[12]中任一项所述的重组细胞,其中该细胞包含选自以下各项的一种或多种(例如两种、若干种)异源多核苷酸:
编码PEP羧化酶(PPC)的异源多核苷酸;
编码丙酮酸羧化酶(PYC)的异源多核苷酸;
编码天冬氨酸转氨酶(AAT)的异源多核苷酸;
编码天冬氨酸1-脱羧酶(ADC)的异源多核苷酸;
编码β-丙氨酸转氨酶(BAAT)或氨基丁酸转氨酶(gabT)的异源多核苷酸;和
编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。
[15]如段落[1]-[14]中任一项所述的重组细胞,其中该细胞是克拉布特里阴性或克拉布特里阳性的。
[16]如段落[1]-[15]中任一项所述的重组细胞,其中该细胞属于选自以下各项的属:伊萨酵母属、假丝酵母属、克鲁维酵母属、毕赤酵母属、裂殖酵母属、有孢圆酵母属、接合酵母属和酵母属。
[17]如段落[16]所述的重组细胞,其中该细胞选自东方伊萨酵母、郎比可假丝酵母以及布拉迪酵母。
[18]段落[1]-[17]中任一项所述的重组细胞,其中该细胞是CB1酵母细胞。
[19]如段落[1]-[18]中任一项所述的重组细胞,其中该酵母细胞不能发酵戊糖。
[20]如段落[1]-[19]中任一项所述的重组细胞,其中所述细胞进一步包含对一个或多个内源性dse2、scw11、eaf3、sed1或sam2基因和/或编码PDC、ADH、GAL6、CYB2A、CYB2B、GPD、GPP、ALD或PCK的一个或多个内源基因的破坏。
[21]如段落[1]-[20]中任一项所述的重组细胞,其中所述细胞进一步包含对编码PDC的内源基因的破坏。
[22]如段落[1]-[21]中任一项所述的重组细胞,其中所述细胞进一步包含对编码GPD的内源基因的破坏。
[23]一种组合物,包含如段落[1]-[22]中任一项所述的重组细胞。
[24]如段落[23]所述的组合物,其中该组合物包括可发酵的培养基。
[25]如段落[24]所述的组合物,其中该可发酵的培养基包括蔗糖、葡萄糖和/或果糖。
[26]如段落[24]或[25]中任一项所述的组合物,其中该可发酵的培养基包括少于1%的戊糖。
[27]如段落[23]-[26]中任一项所述的组合物,进一步包括3-HP。
[28]如段落[26]所述的组合物,其中3-HP的滴度大于约1g/L,例如大于约2g/L、5g/L、10g/L、15g/L、20g/L、25g/L、30g/L、35g/L、40g/L、45g/L、50g/L、55g/L、60g/L、65g/L、70g/L、75g/L、80g/L、85g/L、90g/L、95g/L、100g/L、125g/L、150g/L、200g/L或250g/L。
[29]如段落[23]-[28]中任一项所述的组合物,其中该培养基的pH低于6.5,例如在约1.5至约4.5、约2.0至约4.0或约2.0至约3.5的范围内。
[30]一种生产3-HP的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如段落[1]-[22]中任一项所述的重组酵母细胞,以生产3-HP;并且
(b)回收该3-HP。
[31]如段落[30]所述的方法,其中该可发酵的培养基包括蔗糖、葡萄糖和/或果糖。
[32]如段落[30]或[31]所述的方法,其中该可发酵的培养基包括少于1%的戊糖。
[33]如段落[30]-[32]中任一项所述的方法,其中产生的3-HP的滴度大于约1g/L,例如大于约2g/L、5g/L、10g/L、15g/L、20g/L、25g/L、30g/L、35g/L、40g/L、45g/L、50g/L、55g/L、60g/L、65g/L、70g/L、75g/L、80g/L、85g/L、90g/L、95g/L、100g/L、125g/L、150g/L、200g/L或250g/L。
[34]如段落[30]-[33]中任一项所述的方法,其中所得3-HP是基本上纯的。
[35]如段落[30]-[34]中任一项所述的方法,其中发酵结束时的最终pH小于6.5,例如在约1.5至约4.5、约2.0至约4.0,或约2.0至约3.5的范围内。
[36]一种生产丙烯酸或其盐的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如段落[1]-[22]中任一项所述的重组细胞,以生产3-HP;
(b)回收该3-HP;
(c)在适合的条件下,将该3-HP脱水,以生产丙烯酸或其盐;并且
(d)回收该丙烯酸或其盐。
[37]一种生产3-HP酯的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如段落[1]-[22]中任一项所述的重组细胞,以生产3-HP;
(b)回收该3-HP;
(c)在适合的条件下,酯化该3-HP以产生3-HP酯;并且
(d)回收该3-HP酯
[38]一种生产1,3-丙二醇的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如段落[1]-[22]中任一项所述的重组细胞,以生产3-HP;
(b)回收该3-HP;
(c)在适合的条件下,氢化该3-HP以生产1,3-丙二醇;并且
(d)回收该1,3-丙二醇。
[39]如段落[30]-[38]中任一项所述的方法,其中重组细胞是CNB1细胞,该CNB1细胞被培养在包含少于1%戊糖的发酵培养基中。
[40]一种用于获得如段落[1]-[22]中任一项所述的重组宿主细胞的方法,该方法包括:
(a)培养亲本菌株;
(b)(i)用一个或多个3-HP途径基因转化该亲本菌株以在(a)的亲本菌株中提供主动3-HP途径;
(b)(ii)破坏在(a)的亲本菌株中编码丙酮酸还原酶的内源基因;并且
(c)分离生成自(b)(i)和(b)(ii)的突变菌株。
[41]如段落[40]所述的方法,其中步骤(b)(i)发生在步骤(b)(ii)之前。
[42]如段落[40]所述的方法,其中步骤(b)(ii)发生在步骤(b)(i)之前。
通过说明提供了以下实例,并且并不旨在使限制本发明。
实例
用作缓冲液和底物的化学品是至少试剂级的商业产品。
培养基和溶液
LiOAc/TE溶液是由8份无菌水、1份1M LiOAc、以及1份10X TE构成。
2X诺布尔琼脂板培养基是由10g的蒂福科琼脂诺布尔(贝迪公司(Becton,Dickinson and Company))和250ml的去离子水构成,并且然后通过高压灭菌进行灭菌。
PEG/LiOAc/TE溶液是由8份50%PEG3350、1份1M LiOAc、以及1份10X TE构成。
50%PEG3350是通过将100g的PEG3350添加至150mL的水中并且进行加热和搅拌直至溶解来制备的。然后将体积用水加至200mL,并且然后通过高压灭菌进行灭菌。
2X SD ura-培养基是由6.66g的无氨基酸的酵母氮源、1.54g的ura-DO补充物(克隆技术实验室公司(Clontech Laboratories,Inc.))、20g的右旋糖、以及补足至1升的去离子水构成。将所得溶液进行过滤灭菌。
通过将250mL的2X-Ura选择培养基和2X诺布尔琼脂板培养基(在高压灭菌后,冷却至65℃)混合,并且然后将熔化的培养基倒入皮氏培养皿并且允许冷却至室温来制备SDura-板。
TAE缓冲液由4.84g的Tris碱、1.14mL的冰醋酸、以及2mL的0.5M EDTA(pH 8.0)、以及加至1升的去离子水构成。
TBE缓冲液由10.8g的Tris碱、5.5g的硼酸、4mL的0.5M EDTA(pH 8.0)、以及补足至1升的去离子水构成。
10X TE(200mL)是由2.42g的Tris碱、以及4mL的0.5M EDTA(pH 8.0)构成。使用5MHCl将pH调节至7.5,并且将溶液通过高压灭菌进行灭菌。
YP+10%葡萄糖培养基是由500mL的YP肉汤和100mL的无菌50%葡萄糖构成。
YPD培养基是由500mL的YP肉汤和100mL的无菌50%葡萄糖构成。
YP肉汤是由10g的酵母提取物、20g的蛋白胨、以及补足至1升的去离子水构成。通过高压灭菌灭菌该溶液。
YPD板是由10g酵母提取物、20g细菌蛋白胨和20g细菌培养用琼脂、以及补足至1升的去离子水构成。高压灭菌后,每升添加40ml的无菌50%葡萄糖,并且倒平板。
2X Yt+amp板是由16g的胰蛋白胨、10g的酵母提取物、5g的NaCl、15g的细菌培养用琼脂、以及补足至1升的去离子水构成。在高压灭菌后,每升添加100mg的氨比西林并且倒平板。
FOA板是由250mL的2X FOA液体培养基和250mL的2X-SD FOA平板培养基构成,熔融并冷却至65℃。
2X-SD FOA液体培养基是由6.66g不含有氨基酸的酵母氮碱、1.54g ura-DO补充物(Clontech,山景城(Mountain View),加利福尼亚州(CA),美国)、20g右旋糖、50mg尿嘧啶、2mg尿苷、以及2g 5-FOA(5-氟乳清酸,一水合物;多伦多研究化学(Toronto ResearchChemicals),北约克(North York),安大略省(ON),加拿大)以及补足至1L的水构成。将所得溶液进行过滤以灭菌。
2X-SD FOA平板培养基是由10g细菌培养用琼脂和250mL水构成。将所得溶液进行高压蒸汽处理以灭菌。
表1:实例中使用的引物序列。
Figure GDA0003149663890000611
Figure GDA0003149663890000621
实例1:丙酮酸还原酶的鉴定
丙酮酸还原酶活性测定用于跟踪在48小时收获的分级发酵液的活性,该分级发酵液收获自包含主动3-HP途径的CNB1衍生酵母细胞(参见,WO 2012/074818)。
将来自750mL发酵液的细胞沉淀重新悬浮于包含1%蛋白酶抑制混合剂(罗氏诊断公司(Roche Diagnostics))的100ml PBS中。将细胞悬浮液转移至具有大约15ml体积的裂解基质Y(Lysing Matrix Y)(0.5mm氧化钇-稳定的锆球;MP生物医学(MP Biomedicals))的50ml锥形管中,并且使用
Figure GDA0003149663890000622
破坏器(MP生物医学(MP Biomedicals))进行细胞裂解。按6.5m/s2进行三个60秒打浆循环,其中循环之间在冰上冷却5分钟。将试管以8370×g和在4℃下离心15分钟,并且将每个上清液转移至新鲜的管中。然后将上清液以28,880×g、在4℃下另外离心45min。然后将可溶性蛋白质级分用100%硫酸铵沉淀1hr,并且该蛋白质以10,000×g离心15分钟。将沉淀储存于-20℃。
将沉淀解冻并重悬浮于FPLC(AKTA纯化仪,GE医疗集团(GE Healthcare))起始缓冲液:20mM Tris pH 8。需要添加100ml缓冲液来溶解蛋白质,并准备30ml用于分级。使用Econo-Pac 10Dg脱盐柱(伯乐公司(Bio Rad))将蛋白质脱盐。将脱盐蛋白加载到Q琼脂糖HP(Q Sepharose HP)(GE医疗集团(GE Healthcare))50ml柱上,并且蛋白质用超过10CV的0%至60%的缓冲液B(20mM Tris pH 8,1M NaCl)进行梯度洗脱,然后用超过2CV的100%缓冲液B进行洗脱。
对所有级分进行活性测定(如实例6所描述),并且鉴定活性级分且将其汇集以供进一步分级。蛋白质从Q琼脂糖Hp柱中以大约300mM NaCl洗脱。将蛋白质使用重力柱经缓冲液交换进入20mM Tris pH 7.5,并加载到源15Phe 4.6/100PE柱(Source 15Phe 4.6/100PEcolumn)(GE医疗集团(GE Healthcare))上。蛋白质用超过20CV的梯度洗脱至100%缓冲液B(20mM Tris pH 7.5,1M硫酸铵)中,并且感兴趣的蛋白质以大约500mM的硫酸铵浓度洗脱。如上文所描述,将级分针对活性进行测试,并且然后通过速度真空进行浓缩并脱盐(如上文所描述),以在SDS-Page标准凝胶8%-16%Tris-HCl(伯乐公司(Bio Rad))上进行跑胶。通过MADLI(布鲁克公司的Autoflex(Bruker Autoflex))鉴定并送出了四条条带用于肽质量指纹图谱ID。一些条带与已知的、与丙酮酸还原酶不相似的酶具有同源性,并且两条条带是有未知的功能。相应于g4240(SEQ ID NO:205)的蛋白质具有NADH/NADPH辅因子的识别序列,并且被选择用于如下所述的破坏和进一步分析。
实例2:DNA转化的程序
基于以下具体程序,进行以下实例中描述的将DNA转化进宿主基因组中以产生重组菌株。
将四mL的YP+10%葡萄糖培养基添加至14mL法尔肯管(Falcon tube)中,并且使用无菌环将所需菌株接种进该培养基中。使培养物在250rpm摇动下在37℃生长过夜(约16小时)。将1mL过夜培养物添加至250mL带挡板的包含50mL液体YP+10%葡萄糖培养基的摇瓶中。使摇瓶培养物在250rpm摇动下在37℃生长。以大约以小时计的间隔抽取培养物的小等分部分,并且测量OD600。使培养物生长直到OD600为0.6-1.0。
将细胞通过在室温在2279x g离心来进行收获,将沉淀重悬浮在25mL无菌水中,然后在室温在2279x g进行离心。将沉淀重悬浮在1mL无菌水中,并且将重悬浮的细胞转移至1.5mL试管,并且然后在16,100x g进行沉淀。将细胞重悬浮在1mL LiOAc/TE溶液中,并且然后在16,100x g进行沉淀。然后将细胞沉淀重悬浮在500μL LiOAc/TE溶液中。
将以下组分添加至1.5mL试管中:100μL以上细胞,10μL新鲜煮沸的然后冰冻的鲑鱼精子DNA(安捷伦科技(Agilent Technologies),圣克拉拉(Santa Clara),加利福尼亚州,美国),以及10μL所希望的、线性化的转化DNA。还制备了用水代替DNA的对照反应。向各转化反应中依次添加600μL的PEG/LiOAc/TE溶液、40μL DMSO,并且将反应倒置数次以混合。°将转化反应孵育于42℃水浴中持续5分钟,并且将细胞在5,400x g沉淀1min。将细胞重悬浮在水中,分为两部分,并且将转化反应的各一半铺于ura选择性培养基平板上。将平板放置在37℃下。生长18至24hr后,菌落是可见的。
实例3:克隆丙酮酸还原酶基因座的左侧破坏构建体
为了产生靶向同源重组(URA3选择性标记的5’半端至CNB1酵母的丙酮酸还原酶基因座)的质粒(WO 2012/074818),用引物1209140和1209141对丙酮酸还原酶的DNA区5'进行PCR扩增。
引物1209140包含与pMHCT246目标载体(WO 2015/017721)同源的5'区、HpaI位点,且与CNB1丙酮酸还原酶5'侧翼DNA具有同源性。引物1209141是一个反向互补序列引物,具有与pMHCT246目标载体同源的5'区、PacI和NotI位点,且与CNB1丙酮酸还原酶5'侧翼DNA具有同源性。
每个PCR反应(50μL)包含从菌株MBIN500(WO 2012/074818)分离的10mg基因组DNA(可制备,例如,使用来自中心生物技术公司(
Figure GDA0003149663890000641
Biotechnologies)的MasterPureTM酵母DNA纯化试剂盒),1X HF缓冲液(赛默飞世尔科技公司(ThermoScientific)),引物1209140和1209141各100pmol,dATP、dCTP、dGTP和dTTP各200μM,以及一个单位的Phusion HS II DNA聚合酶(赛默飞世尔科技公司)。建立了8个相同的反应,并在
Figure GDA0003149663890000642
(艾本德科技公司(Eppendorf Scientific))中进行梯度PCR,其被编程为:一个循环,在98℃下持续30秒;随后为35个循环,每个循环在98℃下持续10秒,梯度退火温度持续30秒,以及72℃持续1分钟,最后在72℃延伸10分钟。退火温度梯度为50℃、51.4℃、53.8℃、57.5℃、62℃、65.9℃、68.5℃和70.0℃,每个温度下有一管。热循环后,通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳分离PCR反应产物,其中每个梯度条件在单独的泳道中。凝胶的可视化示出,所有八个退火温度均成功地扩增了产物,因此,从所有泳道的凝胶中切下940bp的PCR产物,并且根据制造商的说明书,使用
Figure GDA0003149663890000645
Extract II试剂盒(马歇雷纳格尔公司(Macherey-Nagel),伯利恒(Bethlehem),宾夕法尼亚州,美国)对其进行一并纯化。
为了产生5'侧翼DNA的目标载体,质粒pMHCT246(WO 2015/017721)用HpaI和PacI消化,并且所得片段通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳来分离,其中从凝胶中切下大约4kbp的条带,并根据制造商的说明书使用
Figure GDA0003149663890000643
Extract II试剂盒进行纯化。
使用IN-FUSIONTM HD克隆试剂盒(克隆技术公司(Clontech))将丙酮酸还原酶5'侧翼DNA和pMHCT246载体片段组合,其总反应体积为10μL,由150ng的pMhCt246 HpaI和PacI载体片段、70ng 5'丙酮酸还原酶侧翼DNA的PCR产物和1X In-Fusion HD反应缓冲液(克隆技术公司)构成。将反应物在37℃下孵育15分钟,在50℃下孵育15分钟,并且然后放置在冰上。根据制造商的说明书,用2μl的反应物转化StellarTM感受态细胞。将转化反应物分散到2XYT+amp平板上并在37℃下孵育过夜。从选择平板中分离出假定的转化体菌落,并且使用
Figure GDA0003149663890000644
从每一菌落中制备质粒DNA,并用MfeI和NotI筛选所需的丙酮酸还原酶侧翼DNA的适当插入,且将所希望的插入(对于具有预期的消化模式的一个分离质粒)通过DNA测序确认并指定为pMHCT260b。
质粒pMHCT260b(图2)是基于pUC19(Yanisch-Perron,C.,Vieira,J.和Messing,J.(1985)Gene[基因],33,103-119)的载体,具有5'丙酮酸还原酶侧翼DNA靶向序列、URA3启动子和URA3 ORF的5'片段。
实例4:克隆丙酮酸还原酶基因座的右侧破坏构建体
为了产生靶向同源重组(URA3选择性标记的3’半端至CNB1酵母的丙酮酸还原酶基因座)的质粒(WO 2012/074818),用引物1209142和1209143对丙酮酸还原酶的DNA区3'进行PCR扩增。
引物1209142包含与pMHCT247目标载体同源的5'区、Notl位点,且与CNB1丙酮酸还原酶3'侧翼DNA具有同源性。引物1209143是一个反向互补序列引物,具有与pMHCT247目标载体同源的5'区、SacII位点,且与CNB1丙酮酸还原酶3'侧翼DNA具有同源性。
每个PCR反应(50μL)包含从菌株MBIN500(WO 2012074818)分离的10mg基因组DNA,1X ThermoPol反应缓冲液(新英格兰生物实验室(New England Biolabs)),引物1209142和1209143各100pmol,dATP、dCTP、dGTP和dTTP各200μM,2μL的100mM MgSO4,以及2个单位的
Figure GDA0003149663890000651
(外切-)DNA聚合酶(新英格兰生物实验室)。建立了8个相同的反应,并在
Figure GDA0003149663890000652
(艾本德科技公司)中进行梯度PCR,其被编程为:一个循环,在94℃下持续2分钟;随后为34个循环,每个循环在94℃下持续30秒,梯度退火温度持续30秒,以及72℃持续1分钟,最后在72℃延伸10分钟。退火温度梯度为50℃、51.4℃、53.8℃、57.5℃、62℃、65.9℃、68.5℃和70.0℃,每个温度下有一管。热循环后,通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳分离PCR反应产物,其中每个梯度条件在单独的泳道中。凝胶的可视化示出,具有最低退火温度的六个反应成功地扩增了产物,因此从这六个泳道的凝胶中切下大约470bp的PCR产物,并且根据制造商的说明书,使用NUCLEOSPINExtract II试剂盒(马歇雷纳格尔公司,伯利恒,宾夕法尼亚州,美国)对其进行一并纯化。
质粒pMHCT247(图3)类似于pMHCT239(WO 2015/017721),除了与苹果酸脱氢酶(mdhB)基因座(SEQ ID NO:206)的3'DNA具有同源性的DNA片段,该片段插入了PDC终止子的3’中,以允许靶向mdhB基因座。
为了产生3'侧翼DNA的目标载体,质粒pMHCT247用PmeI和SacII消化,并且所得片段通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳来分离,其中从凝胶中切下大约3.8kbp的条带,并根据制造商的说明书使用
Figure GDA0003149663890000653
Extract II试剂盒进行纯化。
使用IN-FUSIONTM HD克隆试剂盒(克隆技术公司)将丙酮酸还原酶3'侧翼DNA和pMHCT247载体片段组合,其总反应体积为10μL,由50ng的pMhCt247 PmeI和SacII载体片段、21ng 3'丙酮酸还原酶侧翼DNA的PCR产物和1X In-Fusion HD反应缓冲液(克隆技术公司)构成。将反应物在37℃下孵育15分钟,在50℃下孵育15分钟,并且然后放置在冰上。根据制造商的说明书,用2μl的反应物来转化StellarTM感受态细胞。将转化反应物分散到2X YT+amp平板上并在37℃下孵育过夜。从选择平板中分离出假定的转化体菌落,并且使用
Figure GDA0003149663890000661
从每一菌落中制备质粒DNA,并用MfeI和NotI筛选所需的丙酮酸还原酶侧翼DNA的适当插入,且将所希望的插入(对于具有预期的消化模式的一个分离质粒)通过DNA测序确认并指定为pMHCT261。
质粒pMHCT261(图4)是基于pUC19(Yanisch-Perron,C.,Vieira,J.和Messing,J.(1985)Gene[基因],33,103-119)的载体,具有URA3 ORF的3'片段、URA3终止子、URA3启动子和3'丙酮酸还原酶侧翼DNA靶向序列。实例5:酵母宿主细胞中丙酮酸还原酶基因座的破坏
本实例描述了具有丙酮酸还原酶基因的破坏的酵母菌株的构建,该丙酮酸还原酶基因包含SEQ ID NO:204的编码序列,其编码SEQ ID NO:205的丙酮酸还原酶。
在转化前,将大约25μg的pMHCT260b(同上)用HpaI、SacII和BsaI消化以从pUC19骨架载体释放所希望的转化DNA。同样地,将大约μg的pMHCT261(同上)用PfoI、SacII和BsaI消化以从pUC19骨架载体释放所希望的转化DNA。针对pMHCT260b,通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳,将大约2.3kbp的条带(包含破坏盒的所希望的左侧)从载体DNA分离,将其从凝胶上切下,并据制造商的说明书,使用
Figure GDA0003149663890000662
Extract II试剂盒进行纯化。针对pMHCT261,通过在TAE缓冲液中进行0.9%琼脂糖凝胶电泳,将大约2.0kbp的条带(包含破坏盒的所希望的右侧)从载体DNA分离,将其从凝胶上切下,并据制造商的说明书,使用
Figure GDA0003149663890000663
Extract II试剂盒进行纯化。将各DNA在20μl制造商提供的洗脱缓冲液中洗脱,预热至65℃。使用4μl纯化的pMHCT260b片段和4μl纯化的pMHCT261片段转化菌株Ckle213(包含主动3-HP途径,并且衍生自CNB1酵母菌株,如WO 2012/074818所描述)。
在30℃下,在SD ura-板上选择转化体。三天后挑出二十四个转化体,并且针对单菌落在SD ura-板上重新划线,并且在37℃下生长两天。然后从由每个初始转化体产生的条纹中的每个挑出单菌落并且在SD ura-板上重新划线。
在单菌落纯化和生长一轮后,进行PCR以检验如在此描述的发生的所希望的靶向整合。使用引物1209724和612909连同引物612908和1209725证实了丙酮酸还原酶基因座的一个等位基因的正确破坏。引物1209724结合pMHCT260b中的5'丙酮酸还原酶侧翼DNA的基因组5',而引物612909结合URA3 ORF并在反义方向扩增。用这些引物通过PCR产生大约2.0kbp的条带表明,所希望的整合事件的发生,导致丙酮酸还原酶ORF被URA3选择性标记所替换。引物612908结合URA3 ORF,而引物1209725结合pMHCT261中的3'丙酮酸还原酶侧翼DNA的基因组3'并在反义方向扩增。用这些引物通过PCR产生大约1.4kbp的条带表明,所希望的整合事件的发生,导致丙酮酸还原酶ORF被URA3选择性标记所替换。
通过将小量的酵母再悬浮于10μL的水中来制备PCR的模板DNA。然后添加四十μL的Y-裂解缓冲液(ZYMO研究公司(Zymo Research))和2μL的酶解酶(ZYMO研究公司),并且将再悬浮的酵母细胞在37℃下孵育30分钟。然后将这些管转移到4℃直到PCR。
这些PCR(25μL)是由待筛选的菌株的1μL模板DNA(如上所描述),1X
Figure GDA0003149663890000671
Taq反应缓冲液(新英格兰生物实验室有限公司(New England Biolabs,Inc.)),0.4μM的正义引物,0.4μM的反义引物,各300μM的dATP、dCTP、dGTP、以及dTTP,和2.5个单位的
Figure GDA0003149663890000672
Taq DNA聚合酶(新英格兰生物实验室有限公司)构成。用
Figure GDA0003149663890000673
Figure GDA0003149663890000674
进行PCR,其被编程为:1个循环,在94℃下持续4分钟;随后32个循环,每个循环在94℃下持续20秒、60℃下持续20秒、以及65℃下持续2分钟;以及最终延长,在65℃下持续10分钟。热循环后,在TBE缓冲液中,将该PCR产物通过0.9%琼脂糖凝胶电泳来分离,并且如以上所描述的将带的大小可视化并进行解释。具有所需条带的两个独立分离的转化体被指定为yMhCt230和yMhCt231。菌株yMhCt230和yMhCt231对于丙酮酸还原酶基因座的缺失是杂合的。
为了获得这些菌株的ura-衍生物,将yMHCT230和yMHCT231在SD ura-培养基中生长过夜。将100μl的过夜培养物添加至4ml的YPD培养基中,并在37℃下生长五小时。将125ul的培养物平铺于FOA平板上。将FOA抗性菌落进行划线以得到YPD平板的单个菌落,然后从这些条纹的每一个中挑取单个菌落并划线至YPD平板。
在FOA选择和单菌落分离之后,如在此所描述,使用PCR(使用引物1209724和1209725)来确认URA3选择性标记的所希望的环出去除(loop-out removal)。、引物1209724结合pMHCT260b中的5'丙酮酸还原酶侧翼DNA的基因组5',而引物1209725结合pMHCT261中的3'丙酮酸还原酶侧翼DNA的基因组3',并在反义方向扩增。用这些引物通过PCR产生大约2.3kbp的条带表明该URA3选择性标记的所希望的环出的发生,而用这些引物通过PCR产生大约2.6kbp的条带表明存在野生型丙酮酸还原酶基因座。大约3.7kbp条带的产生表明URA3选择性标记没有从丙酮酸还原酶基因座的破坏拷贝中去除。
通过将小量的酵母再悬浮于10μL的水中来制备PCR的模板DNA。然后添加四十μL的Y-裂解缓冲液(ZYMO研究公司(Zymo Research))和2μL的酶解酶(ZYMO研究公司),并且将再悬浮的酵母细胞在37℃下孵育30分钟。然后将这些管转移到4℃直到PCR。
这些PCR(25μL)是由待筛选的菌株的1μL模板DNA(如上所描述),1X
Figure GDA0003149663890000681
Taq反应缓冲液(新英格兰生物实验室有限公司(New England Biolabs,Inc.)),0.4μM的正义引物,0.4μM的反义引物,各300μM的dATP、dCTP、dGTP、以及dTTP,和2.5个单位的
Figure GDA0003149663890000682
Taq DNA聚合酶(新英格兰生物实验室有限公司)构成。用
Figure GDA0003149663890000683
Figure GDA0003149663890000684
进行PCR,其被编程为:1个循环,在94℃下持续4分钟;随后32个循环,每个循环在94℃下持续20秒、60℃下持续20秒、以及65℃下持续4分钟;以及最终延长,在65℃下持续10分钟。热循环后,在TBE缓冲液中,将该PCR产物通过0.9%琼脂糖凝胶电泳来分离,并且如以上所描述的将带的大小可视化并进行解释。来自具有所希望的2.3kbp和2.6kbp的双条带的yMHCT230的一个FOA抗性菌落(表明在标记环出后对于丙酮酸还原酶缺失是杂合的)被指定为yMhCt232。同样地,来自具有所希望的双条带的yMHCT231的一个FOA抗性菌落被指定为yMhCt233。
为了产生对于丙酮酸还原酶基因座的缺失是纯合的菌株,用如上所述制备的4μl的纯化的pMHCT260b片段和4μl的纯化的pMHCT261片段来各自转化yMHCT232和yMHCT233。在37℃下,在SD ura-板上选择转化体。五天后,从各转化菌株中挑出三十六个转化体,并在SDura-板上重新划线以得到单菌落,并在37℃下生长两天。然后从由每个初始转化体产生的条纹中的每个挑出单菌落并且在SD ura-板上重新划线。
在转化体的这种单菌落分离之后,使用PCR来确认丙酮酸还原酶基因的缺失和URA3选择性标记的适当整合(使用如在此描述的三组引物)。使用引物1210137(在丙酮酸还原酶开放阅读框中退火)和引物1209725(结合pMHCT261中的3'丙酮酸还原酶侧翼DNA的基因组3'并在反义方向扩增)的PCR在丙酮酸还原酶缺失的条件下不产生任何大小的条带,但是在丙酮酸还原酶基因存在的条件下产生670bp的条带。使用引物1209724(结合pMHCT260b中的5'丙酮酸还原酶侧翼DNA的基因组5')和引物1210138(在丙酮酸还原酶开放阅读框中退火并在反义方向扩增)的PCR在丙酮酸还原酶缺失的条件下不产生任何大小的条带,但是在丙酮酸还原酶基因存在的条件下产生大约1.1kbp的条带。使用引物1209724(结合pMHCT260b中5'丙酮酸还原酶侧翼DNA的基因组5')和引物1209725(结合pMHCT261中3'丙酮酸还原酶侧翼DNA的基因组3'并在反义方向扩增)的PCR在丙酮酸还原酶被URA3选择性标记破坏的条件下产生约大3.7kbp的条带,而大约2.3kbp的带表明丙酮酸还原酶基因座的存在,该丙酮酸还原酶基因座在选择性标记盒的环出后仅包含ura3启动子区。
通过将小量的酵母再悬浮于10μL的水中来制备PCR的模板DNA。然后添加四十μL的Y-裂解缓冲液(ZYMO研究公司(Zymo Research))和2μL的酶解酶(ZYMO研究公司),并且将再悬浮的酵母细胞在37℃下孵育30分钟。然后将这些管转移到4℃直到PCR。
这些PCR(25μL)是由待筛选的菌株的1μL模板DNA(如上所描述),1X
Figure GDA0003149663890000691
Taq反应缓冲液(新英格兰生物实验室有限公司(New England Biolabs,Inc.)),0.4μM的正义引物,0.4μM的反义引物,各300μM的dATP、dCTP、dGTP、以及dTTP,和2.5个单位的
Figure GDA0003149663890000692
Taq DNA聚合酶(新英格兰生物实验室有限公司)构成。在
Figure GDA0003149663890000693
Figure GDA0003149663890000694
中进行PCR,其被编程为:1个循环,在94℃下持续4分钟;随后32个循环,每个循环在94℃下持续20秒、60℃下持续20秒、以及65℃下持续1分钟(对于引物1210137+引物12097254的PCR)或4分钟(对于引物1209724+引物1210138或引物1209724+引物1209725的PCR);最后在65℃延伸10分钟。热循环后,在TBE缓冲液中,将该PCR产物通过0.9%琼脂糖凝胶电泳来分离,并且如以上所描述的将带的大小可视化并进行解释。来自yMHCT232的两个转化体被指定为yMhCt234和yMHCT235,它们具有用于破坏丙酮酸还原酶基因座的全部两个拷贝的所希望的条带。类似地,来自yMHCT233的两个转化体被指定为yMhCt236和yMHCT237,它们具有所希望的条带。
实例6:突变菌株(具有丙酮酸还原酶基因座的破坏)的丙酮酸还原酶活性制备用于酶测定的粗无细胞提取物(CFE):
来自突变菌株MhCt235和MhCt236(实例5)以及对照菌株Ckle210(ura加上Ckle213的亲本,其是来自如WO 2012/074818所述的CNB1酵母菌株的MhCt235和MhCt236的祖先)的粗无细胞提取物是通过离心、弃上清液以及用等体积的磷酸盐缓冲盐水(PBS)洗涤细胞沉淀来进行各自别收集。在2个时间点(16和40hr)测定发酵的酶活性。为了制备粗无细胞提取物(CFE),将各细胞沉淀用含1%蛋白酶抑制剂混合物(罗氏诊断公司(RocheDiagnostics))和10mM磷酸钾(pH 7)的PBS重悬浮至OD600为25。将每一细胞悬浮液转移至具有2.4g的裂解基质Y(0.5mm氧化钇-稳定的锆球,Mp生物医学公司(MP Biomedicals))的2.0mL微量离心管,并且使用
Figure GDA0003149663890000695
破坏器(MP生物医学公司)在6.5/50秒的设定下进行3轮细胞裂解。在每轮之间将样品试管在冰上冷却3分钟。在裂解后,将样品在微量离心机中以最大速度在4℃离心10分钟。将上清液转移至新管中,并且保持在冰上或储存于-20℃直到使用。使用BCA蛋白测定试剂盒(赛默飞世尔科技公司(Thermo Fisher ScientificInc.))并且以牛血清白蛋白作为标准品,根据厂商提供的说明书来确定裂解物中的总蛋白浓度。
丙酮酸还原酶活性测定:
在上文指定细胞的CFE中的依赖NADH的丙酮酸还原酶活性确定如下:制备原料反应混合溶液,当其在测定反应混合物中与CFE组合时,提供以下:100mM Tris(pH 8.0);500μM NADH;5mM丙酮酸。将180μL的该混合物添加至96-孔微量滴定板的孔中,并且添加20μL的适当稀释的CFE以开始反应。使用SpectraMax 340Pc酶标仪在340nm对NADH的氧化进行监测。通过LC/MS(具有安捷伦6410三重四极杆LC/MS检测器)的安捷伦(Agilent)1200系列HPLC证实了乳酸盐/乳酸酯的形成,并通过D和L乳酸盐/乳酸酯测定试剂盒(生物视野公司(Biovision))确认了D型立体异构体。
在16hr和24hr时所得的丙酮酸还原酶活性示于图5(体外产生D-乳酸盐/D-乳酸酯的速率)。突变菌株MhCt235和MhCt236(其包含破坏的丙酮酸还原酶)示出体外无丙酮酸的还原活性。
实例7:乳酸盐/乳酸酯产生于具有丙酮酸还原酶基因座的破坏的突变菌株
将突变菌株MhCt235和MhCt236(实例5)以及对照菌株Ckle210(ura加上Ckle213的亲本,其是来自如WO 2012/074818所述的CNB1酵母菌株的MhCt235和MhCt236的祖先)使用种子繁殖阶段进行培养,随后使用与WO 2014/085330中描述的类似方法在3L生物反应器(欧谱(Applikon),福斯特市(Foster City),加利福尼亚州,美国)中进行单级发酵。
通过80hr,从发酵中所得的D-乳酸盐/D-乳酸酯的形成示于图6。突变菌株MhCt235和MhCt236(其包含破坏的丙酮酸还原酶)示出没有可检测的D-乳酸盐/D-乳酸酯,而亲本菌株Ckle210累积了高达1.4g/L的D-乳酸盐/D-乳酸酯(基于四次独立发酵的平均值)。
虽然出于清楚理解的目的,已经通过说明以及实例的方式相当详细描述了上文,本领域普通技术人员将清楚的是,可以实施任何等效方面或修饰。因此,该说明和实例不应当解释为限制本发明的范围。
序列表
<110> 诺维信公司(Novozymes A/S)
Frias, Janice
Barbier, Guillaume
<120> 用于生产3-羟基丙酸的突变宿主细胞(Mutant Host Cells For The ProductionOf 3-Hydroxypropionic Acid)
<130> 13059-WO-PCT
<150> US 62/126,377
<151> 2015-02-27
<160> 216
<170> PatentIn版本3.5(PatentIn version 3.5)
<210> 1
<211> 3543
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 1
atgtcaactg tggaagatca ctcctcccta cataaattga gaaaggaatc tgagattctt 60
tccaatgcaa acaaaatctt agtggctaat agaggtgaaa ttccaattag aattttcagg 120
tcagcccatg aattgtcaat gcatactgtg gcgatctatt cccatgaaga tcggttgtcc 180
atgcataggt tgaaggccga cgaggcttat gcaatcggta agactggtca atattcgcca 240
gttcaagctt atctacaaat tgacgaaatt atcaaaatag caaaggaaca tgatgtttcc 300
atgatccatc caggttatgg tttcttatct gaaaactccg aattcgcaaa gaaggttgaa 360
gaatccggta tgatttgggt tgggcctcct gctgaagtta ttgattctgt tggtgacaag 420
gtttctgcaa gaaatttggc aattaaatgt gacgttcctg ttgttcctgg taccgatggt 480
ccaattgaag acattgaaca ggctaaacag tttgtggaac aatatggtta tcctgtcatt 540
ataaaggctg catttggtgg tggtggtaga ggtatgagag ttgttagaga aggtgatgat 600
atagttgatg ctttccaaag agcgtcatct gaagcaaagt ctgcctttgg taatggtact 660
tgttttattg aaagattttt ggataagcca aaacatattg aggttcaatt attggctgat 720
aattatggta acacaatcca tctctttgaa agagattgtt ctgttcaaag aagacatcaa 780
aaggttgttg aaattgcacc tgccaaaact ttacctgttg aagttagaaa tgctatatta 840
aaggatgctg taacgttagc taaaaccgct aactatagaa atgctggtac tgcagaattt 900
ttagttgatt cccaaaacag acattatttt attgaaatta atccaagaat tcaagttgaa 960
catacaatta ctgaagaaat cacgggtgtt gatattgttg ccgctcaaat tcaaattgct 1020
gcaggtgcat cattggaaca attgggtcta ttacaaaaca aaattacaac tagaggtttt 1080
gcaattcaat gtagaattac aaccgaggat cctgctaaga attttgcccc agatacaggt 1140
aaaattgagg tttatagatc tgcaggtggt aacggtgtca gattagatgg tggtaatggg 1200
tttgccggtg ctgttatatc tcctcattat gactcgatgt tggttaaatg ttcaacatct 1260
ggttctaact atgaaattgc cagaagaaag atgattagag ctttagttga atttagaatc 1320
agaggtgtca agaccaatat tcctttctta ttggcattgc taactcatcc agttttcatt 1380
tcgggtgatt gttggacaac ttttattgat gatacccctt cgttattcga aatggtttct 1440
tcaaagaata gagcccaaaa attattggca tatattggtg acttgtgtgt caatggttct 1500
tcaattaaag gtcaaattgg tttccctaaa ttgaacaagg aagcagaaat cccagatttg 1560
ttggatccaa atgatgaggt tattgatgtt tctaaacctt ctaccaatgg tctaagaccg 1620
tatctattaa agtatggacc agatgcgttt tccaaaaaag ttcgtgaatt cgatggttgt 1680
atgattatgg ataccacctg gagagatgca catcaatcat tattggctac aagagttaga 1740
actattgatt tactgagaat tgctccaacg actagtcatg ccttacaaaa tgcatttgca 1800
ttagaatgtt ggggtggcgc aacatttgat gttgcgatga ggttcctcta tgaagatcct 1860
tgggagagat taagacaact tagaaaggca gttccaaata ttcctttcca aatgttattg 1920
agaggtgcta atggtgttgc ttattcgtca ttacctgata atgcaattga tcattttgtt 1980
aagcaagcaa aggataatgg tgttgatatt ttcagagtct ttgatgcttt gaacgatttg 2040
gaacaattga aggttggtgt tgatgctgtc aagaaagccg gaggtgttgt tgaagctaca 2100
gtttgttact caggtgatat gttaattcca ggtaaaaagt ataacttgga ttattattta 2160
gagactgttg gaaagattgt ggaaatgggt acccatattt taggtattaa ggatatggct 2220
ggcacgttaa agccaaaggc tgctaagttg ttgattggct cgatcagatc aaaataccct 2280
gacttggtta tccatgtcca tacccatgac tctgctggta ccggtatttc aacttatgtt 2340
gcatgcgcat tggcaggtgc cgacattgtc gattgtgcaa tcaattcgat gtctggttta 2400
acctctcaac cttcaatgag tgcttttatt gctgctttag atggtgatat cgaaactggt 2460
gttccagaac attttgcaag acaattagat gcatactggg cagaaatgag attgttatac 2520
tcatgtttcg aagccgactt gaagggacca gacccagaag tttataaaca tgaaattcca 2580
ggtggacagt tgactaacct aatcttccaa gcccaacaag ttggtttggg tgaacaatgg 2640
gaagaaacta agaagaagta tgaagatgct aacatgttgt tgggtgatat tgtcaaggtt 2700
accccaacct ccaaggttgt tggtgattta gcccaattta tggtttctaa taaattagaa 2760
aaagaagatg ttgaaaaact tgctaatgaa ttagatttcc cagattcagt tcttgatttc 2820
tttgaaggat taatgggtac accatatggt ggattcccag agcctttgag aacaaatgtc 2880
atttccggca agagaagaaa attaaagggt agaccaggtt tagaattaga acctttcaac 2940
ctcgaggaaa tcagagaaaa tttggtttcc agatttggtc caggtattac tgaatgtgat 3000
gttgcatctt ataacatgta tccaaaggtt tacgagcaat atcgtaaggt ggttgaaaaa 3060
tatggtgatt tatctgtttt accaacaaaa gcatttttgg ctcctccaac tattggtgaa 3120
gaagttcatg tggaaattga gcaaggtaag actttgatta ttaagttatt agccatttct 3180
gacttgtcta aatctcatgg tacaagagaa gtatactttg aattgaatgg tgaaatgaga 3240
aaggttacaa ttgaagataa aacagctgca attgagactg ttacaagagc aaaggctgac 3300
ggacacaatc caaatgaagt tggtgcgcca atggctggtg tcgttgttga agttagagtg 3360
aagcatggaa cagaagttaa gaagggtgat ccattagccg ttttgagtgc aatgaaaatg 3420
gaaatggtta tttctgctcc tgttagtggt agggtcggtg aagtttttgt caacgaaggc 3480
gattccgttg atatgggtga tttgcttgtg aaaattgcca aagatgaagc gccagcagct 3540
taa 3543
<210> 2
<211> 1180
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 2
Met Ser Thr Val Glu Asp His Ser Ser Leu His Lys Leu Arg Lys Glu
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Ser Glu Ile Leu Ser Asn Ala Asn Lys Ile Leu Val Ala Asn Arg Gly
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Glu Ile Pro Ile Arg Ile Phe Arg Ser Ala His Glu Leu Ser Met His
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Thr Val Ala Ile Tyr Ser His Glu Asp Arg Leu Ser Met His Arg Leu
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Lys Ala Asp Glu Ala Tyr Ala Ile Gly Lys Thr Gly Gln Tyr Ser Pro
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Val Gln Ala Tyr Leu Gln Ile Asp Glu Ile Ile Lys Ile Ala Lys Glu
85 90 95
His Asp Val Ser Met Ile His Pro Gly Tyr Gly Phe Leu Ser Glu Asn
100 105 110
Ser Glu Phe Ala Lys Lys Val Glu Glu Ser Gly Met Ile Trp Val Gly
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Pro Pro Ala Glu Val Ile Asp Ser Val Gly Asp Lys Val Ser Ala Arg
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Asn Leu Ala Ile Lys Cys Asp Val Pro Val Val Pro Gly Thr Asp Gly
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Pro Ile Glu Asp Ile Glu Gln Ala Lys Gln Phe Val Glu Gln Tyr Gly
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Tyr Pro Val Ile Ile Lys Ala Ala Phe Gly Gly Gly Gly Arg Gly Met
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Arg Val Val Arg Glu Gly Asp Asp Ile Val Asp Ala Phe Gln Arg Ala
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Ser Ser Glu Ala Lys Ser Ala Phe Gly Asn Gly Thr Cys Phe Ile Glu
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Arg Phe Leu Asp Lys Pro Lys His Ile Glu Val Gln Leu Leu Ala Asp
225 230 235 240
Asn Tyr Gly Asn Thr Ile His Leu Phe Glu Arg Asp Cys Ser Val Gln
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Arg Arg His Gln Lys Val Val Glu Ile Ala Pro Ala Lys Thr Leu Pro
260 265 270
Val Glu Val Arg Asn Ala Ile Leu Lys Asp Ala Val Thr Leu Ala Lys
275 280 285
Thr Ala Asn Tyr Arg Asn Ala Gly Thr Ala Glu Phe Leu Val Asp Ser
290 295 300
Gln Asn Arg His Tyr Phe Ile Glu Ile Asn Pro Arg Ile Gln Val Glu
305 310 315 320
His Thr Ile Thr Glu Glu Ile Thr Gly Val Asp Ile Val Ala Ala Gln
325 330 335
Ile Gln Ile Ala Ala Gly Ala Ser Leu Glu Gln Leu Gly Leu Leu Gln
340 345 350
Asn Lys Ile Thr Thr Arg Gly Phe Ala Ile Gln Cys Arg Ile Thr Thr
355 360 365
Glu Asp Pro Ala Lys Asn Phe Ala Pro Asp Thr Gly Lys Ile Glu Val
370 375 380
Tyr Arg Ser Ala Gly Gly Asn Gly Val Arg Leu Asp Gly Gly Asn Gly
385 390 395 400
Phe Ala Gly Ala Val Ile Ser Pro His Tyr Asp Ser Met Leu Val Lys
405 410 415
Cys Ser Thr Ser Gly Ser Asn Tyr Glu Ile Ala Arg Arg Lys Met Ile
420 425 430
Arg Ala Leu Val Glu Phe Arg Ile Arg Gly Val Lys Thr Asn Ile Pro
435 440 445
Phe Leu Leu Ala Leu Leu Thr His Pro Val Phe Ile Ser Gly Asp Cys
450 455 460
Trp Thr Thr Phe Ile Asp Asp Thr Pro Ser Leu Phe Glu Met Val Ser
465 470 475 480
Ser Lys Asn Arg Ala Gln Lys Leu Leu Ala Tyr Ile Gly Asp Leu Cys
485 490 495
Val Asn Gly Ser Ser Ile Lys Gly Gln Ile Gly Phe Pro Lys Leu Asn
500 505 510
Lys Glu Ala Glu Ile Pro Asp Leu Leu Asp Pro Asn Asp Glu Val Ile
515 520 525
Asp Val Ser Lys Pro Ser Thr Asn Gly Leu Arg Pro Tyr Leu Leu Lys
530 535 540
Tyr Gly Pro Asp Ala Phe Ser Lys Lys Val Arg Glu Phe Asp Gly Cys
545 550 555 560
Met Ile Met Asp Thr Thr Trp Arg Asp Ala His Gln Ser Leu Leu Ala
565 570 575
Thr Arg Val Arg Thr Ile Asp Leu Leu Arg Ile Ala Pro Thr Thr Ser
580 585 590
His Ala Leu Gln Asn Ala Phe Ala Leu Glu Cys Trp Gly Gly Ala Thr
595 600 605
Phe Asp Val Ala Met Arg Phe Leu Tyr Glu Asp Pro Trp Glu Arg Leu
610 615 620
Arg Gln Leu Arg Lys Ala Val Pro Asn Ile Pro Phe Gln Met Leu Leu
625 630 635 640
Arg Gly Ala Asn Gly Val Ala Tyr Ser Ser Leu Pro Asp Asn Ala Ile
645 650 655
Asp His Phe Val Lys Gln Ala Lys Asp Asn Gly Val Asp Ile Phe Arg
660 665 670
Val Phe Asp Ala Leu Asn Asp Leu Glu Gln Leu Lys Val Gly Val Asp
675 680 685
Ala Val Lys Lys Ala Gly Gly Val Val Glu Ala Thr Val Cys Tyr Ser
690 695 700
Gly Asp Met Leu Ile Pro Gly Lys Lys Tyr Asn Leu Asp Tyr Tyr Leu
705 710 715 720
Glu Thr Val Gly Lys Ile Val Glu Met Gly Thr His Ile Leu Gly Ile
725 730 735
Lys Asp Met Ala Gly Thr Leu Lys Pro Lys Ala Ala Lys Leu Leu Ile
740 745 750
Gly Ser Ile Arg Ser Lys Tyr Pro Asp Leu Val Ile His Val His Thr
755 760 765
His Asp Ser Ala Gly Thr Gly Ile Ser Thr Tyr Val Ala Cys Ala Leu
770 775 780
Ala Gly Ala Asp Ile Val Asp Cys Ala Ile Asn Ser Met Ser Gly Leu
785 790 795 800
Thr Ser Gln Pro Ser Met Ser Ala Phe Ile Ala Ala Leu Asp Gly Asp
805 810 815
Ile Glu Thr Gly Val Pro Glu His Phe Ala Arg Gln Leu Asp Ala Tyr
820 825 830
Trp Ala Glu Met Arg Leu Leu Tyr Ser Cys Phe Glu Ala Asp Leu Lys
835 840 845
Gly Pro Asp Pro Glu Val Tyr Lys His Glu Ile Pro Gly Gly Gln Leu
850 855 860
Thr Asn Leu Ile Phe Gln Ala Gln Gln Val Gly Leu Gly Glu Gln Trp
865 870 875 880
Glu Glu Thr Lys Lys Lys Tyr Glu Asp Ala Asn Met Leu Leu Gly Asp
885 890 895
Ile Val Lys Val Thr Pro Thr Ser Lys Val Val Gly Asp Leu Ala Gln
900 905 910
Phe Met Val Ser Asn Lys Leu Glu Lys Glu Asp Val Glu Lys Leu Ala
915 920 925
Asn Glu Leu Asp Phe Pro Asp Ser Val Leu Asp Phe Phe Glu Gly Leu
930 935 940
Met Gly Thr Pro Tyr Gly Gly Phe Pro Glu Pro Leu Arg Thr Asn Val
945 950 955 960
Ile Ser Gly Lys Arg Arg Lys Leu Lys Gly Arg Pro Gly Leu Glu Leu
965 970 975
Glu Pro Phe Asn Leu Glu Glu Ile Arg Glu Asn Leu Val Ser Arg Phe
980 985 990
Gly Pro Gly Ile Thr Glu Cys Asp Val Ala Ser Tyr Asn Met Tyr Pro
995 1000 1005
Lys Val Tyr Glu Gln Tyr Arg Lys Val Val Glu Lys Tyr Gly Asp
1010 1015 1020
Leu Ser Val Leu Pro Thr Lys Ala Phe Leu Ala Pro Pro Thr Ile
1025 1030 1035
Gly Glu Glu Val His Val Glu Ile Glu Gln Gly Lys Thr Leu Ile
1040 1045 1050
Ile Lys Leu Leu Ala Ile Ser Asp Leu Ser Lys Ser His Gly Thr
1055 1060 1065
Arg Glu Val Tyr Phe Glu Leu Asn Gly Glu Met Arg Lys Val Thr
1070 1075 1080
Ile Glu Asp Lys Thr Ala Ala Ile Glu Thr Val Thr Arg Ala Lys
1085 1090 1095
Ala Asp Gly His Asn Pro Asn Glu Val Gly Ala Pro Met Ala Gly
1100 1105 1110
Val Val Val Glu Val Arg Val Lys His Gly Thr Glu Val Lys Lys
1115 1120 1125
Gly Asp Pro Leu Ala Val Leu Ser Ala Met Lys Met Glu Met Val
1130 1135 1140
Ile Ser Ala Pro Val Ser Gly Arg Val Gly Glu Val Phe Val Asn
1145 1150 1155
Glu Gly Asp Ser Val Asp Met Gly Asp Leu Leu Val Lys Ile Ala
1160 1165 1170
Lys Asp Glu Ala Pro Ala Ala
1175 1180
<210> 3
<211> 1154
<212> PRT
<213> 类球红细菌(Rhodobacter sphaeroides)
<400> 3
Met Ala Glu Phe Arg Lys Ile Leu Ile Ala Asn Arg Gly Glu Ile Ala
1 5 10 15
Ile Arg Val Met Arg Ala Ala Asn Glu Met Gly Lys Lys Thr Val Ala
20 25 30
Val Tyr Ala Glu Glu Asp Lys Leu Ser Leu His Arg Phe Lys Ala Asp
35 40 45
Glu Ala Tyr Arg Ile Gly Glu Gly Leu Ser Pro Val Gly Ala Tyr Leu
50 55 60
Ser Ile Pro Glu Ile Ile Arg Val Ala Gln Met Ser Gly Ala Asp Ala
65 70 75 80
Ile His Pro Gly Tyr Gly Leu Leu Ser Glu Asn Pro Asp Phe Val Glu
85 90 95
Ala Cys Asp Ala Ala Gly Ile Ala Phe Ile Gly Pro Lys Ala Glu Thr
100 105 110
Met Arg Ala Leu Gly Asp Lys Ala Ser Ala Arg Arg Val Ala Met Ala
115 120 125
Ala Gly Val Pro Val Ile Pro Ala Thr Glu Val Leu Gly Asp Asp Met
130 135 140
Glu Glu Ile Lys Arg Gln Ala Ala Glu Ile Gly Tyr Pro Leu Met Leu
145 150 155 160
Lys Ala Ser Trp Gly Gly Gly Gly Arg Gly Met Arg Pro Ile Thr Ser
165 170 175
Glu Ala Glu Leu Ala Asp Lys Val Arg Glu Gly Arg Arg Glu Ala Glu
180 185 190
Ala Ala Phe Gly Asn Gly Glu Gly Tyr Leu Glu Lys Met Ile Gln Arg
195 200 205
Ala Arg His Val Glu Val Gln Ile Leu Gly Asp Lys Tyr Gly Ala Ile
210 215 220
Tyr His Leu Tyr Glu Arg Asp Cys Thr Val Gln Arg Arg Asn Gln Lys
225 230 235 240
Val Val Glu Arg Ala Pro Ala Pro Tyr Leu Thr Glu Glu Gln Arg Thr
245 250 255
Glu Ile Cys Glu Leu Gly Arg Arg Ile Cys Ala His Val Asn Tyr Glu
260 265 270
Cys Ala Gly Thr Val Glu Phe Leu Met Asp Met Asp Ser Glu Lys Phe
275 280 285
Tyr Phe Ile Glu Val Asn Pro Arg Val Gln Val Glu His Thr Val Thr
290 295 300
Glu Glu Val Thr Gly Ile Asp Ile Val Gln Ser Gln Ile Arg Ile Ala
305 310 315 320
Glu Gly Ala Thr Leu Ala Glu Ala Thr Gly Cys Pro Ser Gln Asp Asp
325 330 335
Ile Lys Leu Ser Gly His Ala Leu Gln Cys Arg Val Thr Thr Glu Asp
340 345 350
Pro Gln Asn Asn Phe Ile Pro Asp Tyr Gly Arg Leu Thr Ala Tyr Arg
355 360 365
Ser Ala Thr Gly Met Gly Ile Arg Leu Asp Gly Gly Thr Ala Tyr Ala
370 375 380
Gly Gly Val Ile Thr Arg Tyr Tyr Asp Ser Leu Leu Val Lys Val Thr
385 390 395 400
Ala Trp Ala Pro Thr Pro Glu Lys Ala Ile Ala Arg Met Asp Arg Ala
405 410 415
Leu Arg Glu Phe Arg Ile Arg Gly Val Ala Thr Asn Ile Ala Phe Val
420 425 430
Glu Asn Leu Leu Lys His Pro Ser Phe Leu Asp Tyr Ser Tyr Thr Thr
435 440 445
Lys Phe Ile Asp Thr Thr Pro Asp Leu Phe Asn Phe Lys Pro Arg Arg
450 455 460
Asp Arg Ala Thr Lys Ile Leu Thr Tyr Ile Ala Asp Ile Thr Val Asn
465 470 475 480
Gly His Pro Glu Thr Ala Gly Arg Val Arg Pro Ser Ala Glu Leu Lys
485 490 495
Asp Pro Lys Ala Pro Glu Pro Lys Gly Ala Pro Gln Pro Gly Thr Arg
500 505 510
Thr Leu Leu Glu Glu Lys Gly Pro Gln Ala Val Ala Asp Trp Met Ala
515 520 525
Ala Gln Thr Arg Val Leu Met Thr Asp Thr Thr Met Arg Asp Gly His
530 535 540
Gln Ser Leu Leu Ala Thr Arg Met Arg Ser Ile Asp Met Ile Lys Val
545 550 555 560
Thr Pro Ala Tyr Ala Ala Asn Leu Gly Gly Leu Phe Ser Val Glu Cys
565 570 575
Trp Gly Gly Ala Thr Phe Asp Val Ala Tyr Arg Phe Leu Gln Glu Cys
580 585 590
Pro Trp Gln Arg Leu Arg Asp Ile Arg Ala Arg Leu Pro Asn Val Met
595 600 605
Thr Gln Met Leu Leu Arg Ala Ser Asn Gly Val Gly Tyr Thr Asn Tyr
610 615 620
Pro Asp Asn Val Val Gln Glu Phe Val Arg Gln Ala Ala Glu Thr Gly
625 630 635 640
Val Asp Val Phe Arg Val Phe Asp Ser Leu Asn Trp Val Glu Asn Met
645 650 655
Arg Val Ala Met Asp Ala Val Ile Glu Ala Asn Lys Val Cys Glu Gly
660 665 670
Thr Ile Cys Tyr Thr Gly Asp Leu Leu Asp Pro Asp Arg Ser Lys Tyr
675 680 685
Asp Leu Asn Tyr Tyr Val Gly Met Gly Arg Ala Leu Arg Asp Ala Gly
690 695 700
Ala His Val Leu Gly Leu Lys Asp Met Ala Gly Leu Leu Lys Pro Ala
705 710 715 720
Ala Ala Arg Val Leu Val Lys Ala Leu Lys Glu Glu Val Gly Leu Pro
725 730 735
Ile His Phe His Thr His Asp Thr Ser Gly Ile Ala Gly Ala Thr Val
740 745 750
Leu Ala Ala Cys Asp Ala Gly Val Asp Ala Val Asp Ala Ala Met Asp
755 760 765
Ala Phe Ser Gly Gly Thr Ser Gln Pro Cys Leu Gly Ser Ile Val Glu
770 775 780
Ala Leu Lys His Thr Asp Arg Asp Thr Gly Leu Asp Ile Ala Ala Ile
785 790 795 800
Arg Glu Ile Ser Asp Tyr Trp Gly His Val Arg Gln Gln Tyr Ser Ala
805 810 815
Phe Glu Ser Gly Leu Pro Ser Pro Ala Ser Glu Val Tyr Leu His Glu
820 825 830
Met Pro Gly Gly Gln Phe Thr Asn Leu Lys Ala Gln Ala Arg Ser Met
835 840 845
Gly Leu Glu Glu Arg Trp Ser Glu Val Ala Gln Ala Tyr Ala Asp Ala
850 855 860
Asn Arg Met Phe Gly Asp Ile Val Lys Val Thr Pro Ser Ser Lys Val
865 870 875 880
Val Gly Asp Met Ala Leu Met Met Val Ala Gln Gly Leu Thr Arg Glu
885 890 895
Glu Val Glu Asp Pro Glu Val Glu Val Ser Phe Pro Asp Ser Val Val
900 905 910
Asp Met Leu Lys Gly Asn Leu Gly Gln Pro His Gly Gly Trp Pro Glu
915 920 925
Pro Ile Leu Lys Lys Val Leu Lys Gly Glu Ala Pro Ser Thr Glu Arg
930 935 940
Pro Gly Ala His Leu Pro Pro Val Asp Ile Ala Ala Ala Arg Glu Lys
945 950 955 960
Leu Leu Ser Glu Ile Lys Gln Gly Asp Asp Asp Pro Leu Asp Thr Ala
965 970 975
Val Asp Ala Glu Asp Leu Asn Gly Tyr Leu Met Tyr Pro Lys Val Phe
980 985 990
Thr Asp Tyr Arg Ala Arg His Arg Ile Tyr Gly Pro Val Arg Thr Leu
995 1000 1005
Pro Thr Arg Thr Phe Phe Tyr Gly Met Glu Pro Gly Glu Glu Ile
1010 1015 1020
Ser Ala Glu Ile Asp Pro Gly Lys Thr Leu Glu Ile Arg Leu Ser
1025 1030 1035
Ala Val Gly Glu Thr Ser Asp Asp Gly Asp Ala Lys Val Phe Phe
1040 1045 1050
Glu Leu Asn Gly Gln Pro Arg Val Ile Arg Val Ala Asn Arg Ala
1055 1060 1065
Val Lys Ala Lys Thr Ala Thr Arg Pro Lys Ala Gln Asp Gly Asn
1070 1075 1080
Pro Ala His Val Gly Ala Pro Met Pro Gly Ser Val Ala Ser Val
1085 1090 1095
Ala Val Ser Ala Gly Gln Lys Val Lys Pro Gly Asp Leu Leu Val
1100 1105 1110
Thr Ile Glu Ala Met Lys Met Glu Thr Gly Leu His Ala Asp Arg
1115 1120 1125
Ala Ala Thr Val Lys Ala Val His Val Gly Pro Gly Ala Gln Ile
1130 1135 1140
Glu Ala Lys Asp Leu Leu Val Glu Leu Glu Asp
1145 1150
<210> 4
<211> 1154
<212> PRT
<213> 菜豆根瘤菌(Rhizobium etli)
<400> 4
Met Pro Ile Ser Lys Ile Leu Val Ala Asn Arg Ser Glu Ile Ala Ile
1 5 10 15
Arg Val Phe Arg Ala Ala Asn Glu Leu Gly Ile Lys Thr Val Ala Ile
20 25 30
Trp Ala Glu Glu Asp Lys Leu Ala Leu His Arg Phe Lys Ala Asp Glu
35 40 45
Ser Tyr Gln Val Gly Arg Gly Pro His Leu Ala Arg Asp Leu Gly Pro
50 55 60
Ile Glu Ser Tyr Leu Ser Ile Asp Glu Val Ile Arg Val Ala Lys Leu
65 70 75 80
Ser Gly Ala Asp Ala Ile His Pro Gly Tyr Gly Leu Leu Ser Glu Ser
85 90 95
Pro Glu Phe Val Asp Ala Cys Asn Lys Ala Gly Ile Ile Phe Ile Gly
100 105 110
Pro Lys Ala Asp Thr Met Arg Gln Leu Gly Asn Lys Val Ala Ala Arg
115 120 125
Asn Leu Ala Ile Ser Val Gly Val Pro Val Val Pro Ala Thr Glu Pro
130 135 140
Leu Pro Asp Asp Met Ala Glu Val Ala Lys Met Ala Ala Ala Ile Gly
145 150 155 160
Tyr Pro Val Met Leu Lys Ala Ser Trp Gly Gly Gly Gly Arg Gly Met
165 170 175
Arg Val Ile Arg Ser Glu Ala Asp Leu Ala Lys Glu Val Thr Glu Ala
180 185 190
Lys Arg Glu Ala Met Ala Ala Phe Gly Lys Asp Glu Val Tyr Leu Glu
195 200 205
Lys Leu Val Glu Arg Ala Arg His Val Glu Ser Gln Ile Leu Gly Asp
210 215 220
Thr His Gly Asn Val Val His Leu Phe Glu Arg Asp Cys Ser Val Gln
225 230 235 240
Arg Arg Asn Gln Lys Val Val Glu Arg Ala Pro Ala Pro Tyr Leu Ser
245 250 255
Glu Ala Gln Arg Gln Glu Leu Ala Ala Tyr Ser Leu Lys Ile Ala Gly
260 265 270
Ala Thr Asn Tyr Ile Gly Ala Gly Thr Val Glu Tyr Leu Met Asp Ala
275 280 285
Asp Thr Gly Lys Phe Tyr Phe Ile Glu Val Asn Pro Arg Ile Gln Val
290 295 300
Glu His Thr Val Thr Glu Val Val Thr Gly Ile Asp Ile Val Lys Ala
305 310 315 320
Gln Ile His Ile Leu Asp Gly Ala Ala Ile Gly Thr Pro Gln Ser Gly
325 330 335
Val Pro Asn Gln Glu Asp Ile Arg Leu Asn Gly His Ala Leu Gln Cys
340 345 350
Arg Val Thr Thr Glu Asp Pro Glu His Asn Phe Ile Pro Asp Tyr Gly
355 360 365
Arg Ile Thr Ala Tyr Arg Ser Ala Ser Gly Phe Gly Ile Arg Leu Asp
370 375 380
Gly Gly Thr Ser Tyr Ser Gly Ala Ile Ile Thr Arg Tyr Tyr Asp Pro
385 390 395 400
Leu Leu Val Lys Val Thr Ala Trp Ala Pro Asn Pro Leu Glu Ala Ile
405 410 415
Ser Arg Met Asp Arg Ala Leu Arg Glu Phe Arg Ile Arg Gly Val Ala
420 425 430
Thr Asn Leu Thr Phe Leu Glu Ala Ile Ile Gly His Pro Lys Phe Arg
435 440 445
Asp Asn Ser Tyr Thr Thr Arg Phe Ile Asp Thr Thr Pro Glu Leu Phe
450 455 460
Gln Gln Val Lys Arg Gln Asp Arg Ala Thr Lys Leu Leu Thr Tyr Leu
465 470 475 480
Ala Asp Val Thr Val Asn Gly His Pro Glu Ala Lys Asp Arg Pro Lys
485 490 495
Pro Leu Glu Asn Ala Ala Arg Pro Val Val Pro Tyr Ala Asn Gly Asn
500 505 510
Gly Val Lys Asp Gly Thr Lys Gln Leu Leu Asp Thr Leu Gly Pro Lys
515 520 525
Lys Phe Gly Glu Trp Met Arg Asn Glu Lys Arg Val Leu Leu Thr Asp
530 535 540
Thr Thr Met Arg Asp Gly His Gln Ser Leu Leu Ala Thr Arg Met Arg
545 550 555 560
Thr Tyr Asp Ile Ala Arg Ile Ala Gly Thr Tyr Ser His Ala Leu Pro
565 570 575
Asn Leu Leu Ser Leu Glu Cys Trp Gly Gly Ala Thr Phe Asp Val Ser
580 585 590
Met Arg Phe Leu Thr Glu Asp Pro Trp Glu Arg Leu Ala Leu Ile Arg
595 600 605
Glu Gly Ala Pro Asn Leu Leu Leu Gln Met Leu Leu Arg Gly Ala Asn
610 615 620
Gly Val Gly Tyr Thr Asn Tyr Pro Asp Asn Val Val Lys Tyr Phe Val
625 630 635 640
Arg Gln Ala Ala Lys Gly Gly Ile Asp Leu Phe Arg Val Phe Asp Cys
645 650 655
Leu Asn Trp Val Glu Asn Met Arg Val Ser Met Asp Ala Ile Ala Glu
660 665 670
Glu Asn Lys Leu Cys Glu Ala Ala Ile Cys Tyr Thr Gly Asp Ile Leu
675 680 685
Asn Ser Ala Arg Pro Lys Tyr Asp Leu Lys Tyr Tyr Thr Asn Leu Ala
690 695 700
Val Glu Leu Glu Lys Ala Gly Ala His Ile Ile Ala Val Lys Asp Met
705 710 715 720
Ala Gly Leu Leu Lys Pro Ala Ala Ala Lys Val Leu Phe Lys Ala Leu
725 730 735
Arg Glu Ala Thr Gly Leu Pro Ile His Phe His Thr His Asp Thr Ser
740 745 750
Gly Ile Ala Ala Ala Thr Val Leu Ala Ala Val Glu Ala Gly Val Asp
755 760 765
Ala Val Asp Ala Ala Met Asp Ala Leu Ser Gly Asn Thr Ser Gln Pro
770 775 780
Cys Leu Gly Ser Ile Val Glu Ala Leu Ser Gly Ser Glu Arg Asp Pro
785 790 795 800
Gly Leu Asp Pro Ala Trp Ile Arg Arg Ile Ser Phe Tyr Trp Glu Ala
805 810 815
Val Arg Asn Gln Tyr Ala Ala Phe Glu Ser Asp Leu Lys Gly Pro Ala
820 825 830
Ser Glu Val Tyr Leu His Glu Met Pro Gly Gly Gln Phe Thr Asn Leu
835 840 845
Lys Glu Gln Ala Arg Ser Leu Gly Leu Glu Thr Arg Trp His Gln Val
850 855 860
Ala Gln Ala Tyr Ala Asp Ala Asn Gln Met Phe Gly Asp Ile Val Lys
865 870 875 880
Val Thr Pro Ser Ser Lys Val Val Gly Asp Met Ala Leu Met Met Val
885 890 895
Ser Gln Asp Leu Thr Val Ala Asp Val Val Ser Pro Asp Arg Glu Val
900 905 910
Ser Phe Pro Glu Ser Val Val Ser Met Leu Lys Gly Asp Leu Gly Gln
915 920 925
Pro Pro Ser Gly Trp Pro Glu Ala Leu Gln Lys Lys Ala Leu Lys Gly
930 935 940
Glu Lys Pro Tyr Thr Val Arg Pro Gly Ser Leu Leu Lys Glu Ala Asp
945 950 955 960
Leu Asp Ala Glu Arg Lys Val Ile Glu Lys Lys Leu Glu Arg Glu Val
965 970 975
Ser Asp Phe Glu Phe Ala Ser Tyr Leu Met Tyr Pro Lys Val Phe Thr
980 985 990
Asp Phe Ala Leu Ala Ser Asp Thr Tyr Gly Pro Val Ser Val Leu Pro
995 1000 1005
Thr Pro Ala Tyr Phe Tyr Gly Leu Ala Asp Gly Glu Glu Leu Phe
1010 1015 1020
Ala Asp Ile Glu Lys Gly Lys Thr Leu Val Ile Val Asn Gln Ala
1025 1030 1035
Val Ser Ala Thr Asp Ser Gln Gly Met Val Thr Val Phe Phe Glu
1040 1045 1050
Leu Asn Gly Gln Pro Arg Arg Ile Lys Val Pro Asp Arg Ala His
1055 1060 1065
Gly Ala Thr Gly Ala Ala Val Arg Arg Lys Ala Glu Pro Gly Asn
1070 1075 1080
Ala Ala His Val Gly Ala Pro Met Pro Gly Val Ile Ser Arg Val
1085 1090 1095
Phe Val Ser Ser Gly Gln Ala Val Asn Ala Gly Asp Val Leu Val
1100 1105 1110
Ser Ile Glu Ala Met Lys Met Glu Thr Ala Ile His Ala Glu Lys
1115 1120 1125
Asp Gly Thr Ile Ala Glu Val Leu Val Lys Ala Gly Asp Gln Ile
1130 1135 1140
Asp Ala Lys Asp Leu Leu Ala Val Tyr Gly Gly
1145 1150
<210> 5
<211> 602
<212> PRT
<213> 荧光假单胞菌(Pseudomonas fluorescens)
<400> 5
Met Thr Lys Lys Ile Phe Val Thr Asp Thr Ile Leu Arg Asp Ala His
1 5 10 15
Gln Ser Leu Leu Ala Thr Arg Met Arg Thr Glu Asp Met Leu Pro Ile
20 25 30
Cys Asp Lys Leu Asp Lys Val Gly Tyr Trp Ser Leu Glu Cys Trp Gly
35 40 45
Gly Ala Thr Phe Asp Ala Cys Val Arg Phe Leu Lys Glu Asp Pro Trp
50 55 60
Glu Arg Leu Arg Gln Leu Arg Ala Ala Leu Pro Asn Thr Arg Leu Gln
65 70 75 80
Met Leu Leu Arg Gly Gln Asn Leu Leu Gly Tyr Arg His Tyr Ser Asp
85 90 95
Asp Val Val Lys Ala Phe Val Ala Lys Ala Ala Val Asn Gly Ile Asp
100 105 110
Val Phe Arg Ile Phe Asp Ala Met Asn Asp Val Arg Asn Leu Arg Val
115 120 125
Ala Ile Glu Ala Val Lys Ala Ala Gly Lys His Ala Gln Gly Thr Ile
130 135 140
Ala Tyr Thr Thr Ser Pro Val His Thr Ile Asp Ala Phe Val Ala Gln
145 150 155 160
Ala Lys Gln Met Glu Ala Met Gly Cys Asp Ser Val Ala Ile Lys Asp
165 170 175
Met Ala Gly Leu Leu Thr Pro Tyr Ala Thr Gly Glu Leu Val Arg Ala
180 185 190
Leu Lys Ala Glu Gln Ser Leu Pro Val Phe Ile His Ser His Asp Thr
195 200 205
Ala Gly Leu Ala Ala Met Cys Gln Leu Lys Ala Ile Glu Asn Gly Ala
210 215 220
Asp His Ile Asp Thr Ala Ile Ser Ser Phe Ala Ser Gly Thr Ser His
225 230 235 240
Pro Gly Thr Glu Ser Met Val Ala Ala Leu Lys Gly Thr Glu Phe Asp
245 250 255
Thr Gly Leu Asn Leu Glu Leu Leu Gln Glu Ile Gly Leu Tyr Phe Tyr
260 265 270
Ala Val Arg Lys Lys Tyr His Gln Phe Glu Ser Glu Phe Thr Ala Val
275 280 285
Asp Thr Arg Val Gln Val Asn Gln Val Pro Gly Gly Met Ile Ser Asn
290 295 300
Leu Ala Asn Gln Leu Lys Glu Gln Gly Ala Leu Asn Arg Met Gly Glu
305 310 315 320
Val Leu Ala Glu Ile Pro Arg Val Arg Glu Asp Leu Gly Phe Pro Pro
325 330 335
Leu Val Thr Pro Thr Ser Gln Ile Val Gly Thr Gln Ala Phe Phe Asn
340 345 350
Val Leu Ala Gly Glu Arg Tyr Lys Thr Ile Thr Asn Glu Val Lys Leu
355 360 365
Tyr Leu Gln Gly Gly Tyr Gly Lys Ala Pro Gly Thr Val Asn Glu Lys
370 375 380
Leu Arg Arg Gln Ala Ile Gly Ser Glu Glu Val Ile Asp Val Arg Pro
385 390 395 400
Ala Asp Leu Leu Lys Pro Glu Met Thr Lys Leu Arg Ala Asp Ile Gly
405 410 415
Ala Leu Ala Lys Ser Glu Glu Asp Val Leu Thr Phe Ala Met Phe Pro
420 425 430
Asp Ile Gly Arg Lys Phe Leu Glu Glu Arg Ala Ala Gly Thr Leu Thr
435 440 445
Pro Glu Val Leu Leu Pro Ile Pro Glu Ala Gly Lys Val Ala Ser Ala
450 455 460
Gly Gly Glu Gly Val Pro Thr Glu Phe Val Ile Asp Val His Gly Glu
465 470 475 480
Thr Tyr Arg Val Asp Ile Thr Gly Val Gly Val Lys Ala Glu Gly Lys
485 490 495
Arg His Phe Tyr Leu Ser Ile Asp Gly Met Pro Glu Glu Val Val Phe
500 505 510
Glu Pro Leu Asn Glu Phe Val Gly Gly Gly Ser Ser Lys Arg Lys Gln
515 520 525
Ala Ser Ala Pro Gly His Val Ser Thr Thr Met Pro Gly Asn Ile Val
530 535 540
Asp Val Leu Val Lys Glu Gly Asp Thr Val Lys Ala Gly Gln Ala Val
545 550 555 560
Leu Ile Thr Glu Ala Met Lys Met Glu Thr Glu Val Gln Ala Ala Ile
565 570 575
Ala Gly Lys Val Thr Ala Ile His Val Ala Lys Gly Asp Arg Val Asn
580 585 590
Pro Gly Glu Ile Leu Ile Glu Ile Glu Gly
595 600
<210> 6
<211> 481
<212> PRT
<213> 荧光假单胞菌(Pseudomonas fluorescens)
<400> 6
Met Val Pro Pro Ala Gln Gly Asn Leu Gln Val Ile Thr Lys Ile Leu
1 5 10 15
Ile Ala Asn Arg Gly Glu Ile Ala Val Arg Ile Val Arg Ala Cys Ala
20 25 30
Glu Met Gly Ile Arg Ser Val Ala Ile Tyr Ser Asp Ala Asp Arg His
35 40 45
Ala Leu His Val Lys Arg Ala Asp Glu Ala His Ser Ile Gly Ala Glu
50 55 60
Pro Leu Ala Gly Tyr Leu Asn Pro Arg Lys Leu Val Asn Leu Ala Val
65 70 75 80
Glu Thr Gly Cys Asp Ala Leu His Pro Gly Tyr Gly Phe Leu Ser Glu
85 90 95
Asn Ala Glu Leu Ala Asp Ile Cys Ala Glu Arg Gly Ile Lys Phe Ile
100 105 110
Gly Pro Ser Ala Glu Val Ile Arg Arg Met Gly Asp Lys Thr Glu Ala
115 120 125
Arg Arg Ser Met Ile Lys Ala Gly Val Pro Val Thr Pro Gly Thr Glu
130 135 140
Gly Asn Val Ser Gly Ile Glu Glu Ala Leu Ser Glu Gly Asp Arg Ile
145 150 155 160
Gly Tyr Pro Val Met Leu Lys Ala Thr Ser Gly Gly Gly Gly Arg Gly
165 170 175
Ile Arg Arg Cys Asn Ser Arg Glu Glu Leu Glu Gln Asn Phe Pro Arg
180 185 190
Val Ile Ser Glu Ala Thr Lys Ala Phe Gly Ser Ala Glu Val Phe Leu
195 200 205
Glu Lys Cys Ile Val Asn Pro Lys His Ile Glu Ala Gln Ile Leu Gly
210 215 220
Asp Ser Phe Gly Asn Val Val His Leu Phe Glu Arg Asp Cys Ser Ile
225 230 235 240
Gln Arg Arg Asn Gln Lys Leu Ile Glu Ile Ala Pro Ser Pro Gln Leu
245 250 255
Thr Pro Glu Gln Arg Ala Tyr Ile Gly Asp Leu Ser Val Arg Ala Ala
260 265 270
Lys Ala Val Gly Tyr Glu Asn Ala Gly Thr Val Glu Phe Leu Leu Ala
275 280 285
Glu Gly Glu Val Tyr Phe Met Glu Met Asn Thr Arg Val Gln Val Glu
290 295 300
His Thr Ile Thr Glu Glu Ile Thr Gly Ile Asp Ile Val Arg Glu Gln
305 310 315 320
Ile Arg Ile Ala Ser Gly Leu Pro Leu Ser Val Lys Gln Glu Asp Ile
325 330 335
Gln His Arg Gly Phe Ala Leu Gln Phe Arg Ile Asn Ala Glu Asp Pro
340 345 350
Lys Asn Asn Phe Leu Pro Ser Phe Gly Lys Ile Thr Arg Tyr Tyr Ala
355 360 365
Pro Gly Gly Pro Gly Val Arg Thr Asp Thr Ala Ile Tyr Thr Gly Tyr
370 375 380
Thr Ile Pro Pro Phe Tyr Asp Ser Met Cys Leu Lys Leu Val Val Trp
385 390 395 400
Ala Leu Thr Trp Glu Glu Ala Met Asp Arg Gly Leu Arg Ala Leu Asp
405 410 415
Asp Met Arg Leu Gln Gly Val Lys Thr Thr Ala Ala Tyr Tyr Gln Glu
420 425 430
Ile Leu Arg Asn Pro Glu Phe Arg Ser Gly Gln Phe Asn Thr Ser Phe
435 440 445
Val Glu Ser His Pro Glu Leu Thr Asn Tyr Ser Ile Lys Arg Lys Pro
450 455 460
Glu Glu Leu Ala Leu Ala Ile Ala Ala Ala Ile Ala Ala His Ala Gly
465 470 475 480
Leu
<210> 7
<211> 1140
<212> PRT
<213> 谷氨酸棒杆菌(Corynebacterium glutamicum)
<400> 7
Met Ser Thr His Thr Ser Ser Thr Leu Pro Ala Phe Lys Lys Ile Leu
1 5 10 15
Val Ala Asn Arg Gly Glu Ile Ala Val Arg Ala Phe Arg Ala Ala Leu
20 25 30
Glu Thr Gly Ala Ala Thr Val Ala Ile Tyr Pro Arg Glu Asp Arg Gly
35 40 45
Ser Phe His Arg Ser Phe Ala Ser Glu Ala Val Arg Ile Gly Thr Glu
50 55 60
Gly Ser Pro Val Lys Ala Tyr Leu Asp Ile Asp Glu Ile Ile Gly Ala
65 70 75 80
Ala Lys Lys Val Lys Ala Asp Ala Ile Tyr Pro Gly Tyr Gly Phe Leu
85 90 95
Ser Glu Asn Ala Gln Leu Ala Arg Glu Cys Ala Glu Asn Gly Ile Thr
100 105 110
Phe Ile Gly Pro Thr Pro Glu Val Leu Asp Leu Thr Gly Asp Lys Ser
115 120 125
Arg Ala Val Thr Ala Ala Lys Lys Ala Gly Leu Pro Val Leu Ala Glu
130 135 140
Ser Thr Pro Ser Lys Asn Ile Asp Glu Ile Val Lys Ser Ala Glu Gly
145 150 155 160
Gln Thr Tyr Pro Ile Phe Val Lys Ala Val Ala Gly Gly Gly Gly Arg
165 170 175
Gly Met Arg Phe Val Ala Ser Pro Asp Glu Leu Arg Lys Leu Ala Thr
180 185 190
Glu Ala Ser Arg Glu Ala Glu Ala Ala Phe Gly Asp Gly Ala Val Tyr
195 200 205
Val Glu Arg Ala Val Ile Asn Pro Gln His Ile Glu Val Gln Ile Leu
210 215 220
Gly Asp His Thr Gly Glu Val Val His Leu Tyr Glu Arg Asp Cys Ser
225 230 235 240
Leu Gln Arg Arg His Gln Lys Val Val Glu Ile Ala Pro Ala Gln His
245 250 255
Leu Asp Pro Glu Leu Arg Asp Arg Ile Cys Ala Asp Ala Val Lys Phe
260 265 270
Cys Arg Ser Ile Gly Tyr Gln Gly Ala Gly Thr Val Glu Phe Leu Val
275 280 285
Asp Glu Lys Gly Asn His Val Phe Ile Glu Met Asn Pro Arg Ile Gln
290 295 300
Val Glu His Thr Val Thr Glu Glu Val Thr Glu Val Asp Leu Val Lys
305 310 315 320
Ala Gln Met Arg Leu Ala Ala Gly Ala Thr Leu Lys Glu Leu Gly Leu
325 330 335
Thr Gln Asp Lys Ile Lys Thr His Gly Ala Ala Leu Gln Cys Arg Ile
340 345 350
Thr Thr Glu Asp Pro Asn Asn Gly Phe Arg Pro Asp Thr Gly Thr Ile
355 360 365
Thr Ala Tyr Arg Ser Pro Gly Gly Ala Gly Val Arg Leu Asp Gly Ala
370 375 380
Ala Gln Leu Gly Gly Glu Ile Thr Ala His Phe Asp Ser Met Leu Val
385 390 395 400
Lys Met Thr Cys Arg Gly Ser Asp Phe Glu Thr Ala Val Ala Arg Ala
405 410 415
Gln Arg Ala Leu Ala Glu Phe Thr Val Ser Gly Val Ala Thr Asn Ile
420 425 430
Gly Phe Leu Arg Ala Leu Leu Arg Glu Glu Asp Phe Thr Ser Lys Arg
435 440 445
Ile Ala Thr Gly Phe Ile Ala Asp His Pro His Leu Leu Gln Ala Pro
450 455 460
Pro Ala Asp Asp Glu Gln Gly Arg Ile Leu Asp Tyr Leu Ala Asp Val
465 470 475 480
Thr Val Asn Lys Pro His Gly Val Arg Pro Lys Asp Val Ala Ala Pro
485 490 495
Ile Asp Lys Leu Pro Asn Ile Lys Asp Leu Pro Leu Pro Arg Gly Ser
500 505 510
Arg Asp Arg Leu Lys Gln Leu Gly Pro Ala Ala Phe Ala Arg Asp Leu
515 520 525
Arg Glu Gln Asp Ala Leu Ala Val Thr Asp Thr Thr Phe Arg Asp Ala
530 535 540
His Gln Ser Leu Leu Ala Thr Arg Val Arg Ser Phe Ala Leu Lys Pro
545 550 555 560
Ala Ala Glu Ala Val Ala Lys Leu Thr Pro Glu Leu Leu Ser Val Glu
565 570 575
Ala Trp Gly Gly Ala Thr Tyr Asp Val Ala Met Arg Phe Leu Phe Glu
580 585 590
Asp Pro Trp Asp Arg Leu Asp Glu Leu Arg Glu Ala Met Pro Asn Val
595 600 605
Asn Ile Gln Met Leu Leu Arg Gly Arg Asn Thr Val Gly Tyr Thr Pro
610 615 620
Tyr Pro Asp Ser Val Cys Arg Ala Phe Val Lys Glu Ala Ala Ser Ser
625 630 635 640
Gly Val Asp Ile Phe Arg Ile Phe Asp Ala Leu Asn Asp Val Ser Gln
645 650 655
Met Arg Pro Ala Ile Asp Ala Val Leu Glu Thr Asn Thr Ala Val Ala
660 665 670
Glu Val Ala Met Ala Tyr Ser Gly Asp Leu Ser Asp Pro Asn Glu Lys
675 680 685
Leu Tyr Thr Leu Asp Tyr Tyr Leu Lys Met Ala Glu Glu Ile Val Lys
690 695 700
Ser Gly Ala His Ile Leu Ala Ile Lys Asp Met Ala Gly Leu Leu Arg
705 710 715 720
Pro Ala Ala Val Thr Lys Leu Val Thr Ala Leu Arg Arg Glu Phe Asp
725 730 735
Leu Pro Val His Val His Thr His Asp Thr Ala Gly Gly Gln Leu Ala
740 745 750
Thr Tyr Phe Ala Ala Ala Gln Ala Gly Ala Asp Ala Val Asp Gly Ala
755 760 765
Ser Ala Pro Leu Ser Gly Thr Thr Ser Gln Pro Ser Leu Ser Ala Ile
770 775 780
Val Ala Ala Phe Ala His Thr Arg Arg Asp Thr Gly Leu Ser Leu Glu
785 790 795 800
Ala Val Ser Asp Leu Glu Pro Tyr Trp Glu Ala Val Arg Gly Leu Tyr
805 810 815
Leu Pro Phe Glu Ser Gly Thr Pro Gly Pro Thr Gly Arg Val Tyr Arg
820 825 830
His Glu Ile Pro Gly Gly Gln Leu Ser Asn Leu Arg Ala Gln Ala Thr
835 840 845
Ala Leu Gly Leu Ala Asp Arg Phe Glu Leu Ile Glu Asp Asn Tyr Ala
850 855 860
Ala Val Asn Glu Met Leu Gly Arg Pro Thr Lys Val Thr Pro Ser Ser
865 870 875 880
Lys Val Val Gly Asp Leu Ala Leu His Leu Val Gly Ala Gly Val Asp
885 890 895
Pro Ala Asp Phe Ala Ala Asp Pro Gln Lys Tyr Asp Ile Pro Asp Ser
900 905 910
Val Ile Ala Phe Leu Arg Gly Glu Leu Gly Asn Pro Pro Gly Gly Trp
915 920 925
Pro Glu Pro Leu Arg Thr Arg Ala Leu Glu Gly Arg Ser Glu Gly Lys
930 935 940
Ala Pro Leu Thr Glu Val Pro Glu Glu Glu Gln Ala His Leu Asp Ala
945 950 955 960
Asp Asp Ser Lys Glu Arg Arg Asn Ser Leu Asn Arg Leu Leu Phe Pro
965 970 975
Lys Pro Thr Glu Glu Phe Leu Glu His Arg Arg Arg Phe Gly Asn Thr
980 985 990
Ser Ala Leu Asp Asp Arg Glu Phe Phe Tyr Gly Leu Val Glu Gly Arg
995 1000 1005
Glu Thr Leu Ile Arg Leu Pro Asp Val Arg Thr Pro Leu Leu Val
1010 1015 1020
Arg Leu Asp Ala Ile Ser Glu Pro Asp Asp Lys Gly Met Arg Asn
1025 1030 1035
Val Val Ala Asn Val Asn Gly Gln Ile Arg Pro Met Arg Val Arg
1040 1045 1050
Asp Arg Ser Val Glu Ser Val Thr Ala Thr Ala Glu Lys Ala Asp
1055 1060 1065
Ser Ser Asn Lys Gly His Val Ala Ala Pro Phe Ala Gly Val Val
1070 1075 1080
Thr Val Thr Val Ala Glu Gly Asp Glu Val Lys Ala Gly Asp Ala
1085 1090 1095
Val Ala Ile Ile Glu Ala Met Lys Met Glu Ala Thr Ile Thr Ala
1100 1105 1110
Ser Val Asp Gly Lys Ile Asp Arg Val Val Val Pro Ala Ala Thr
1115 1120 1125
Lys Val Glu Gly Gly Asp Leu Ile Val Val Val Ser
1130 1135 1140
<210> 8
<211> 1152
<212> PRT
<213> 苜蓿中华根瘤菌(Sinorhizobium meliloti)
<400> 8
Met Ser Ile Ser Lys Ile Leu Val Ala Asn Arg Ser Glu Ile Ala Ile
1 5 10 15
Arg Val Phe Arg Ala Ala Asn Glu Leu Gly Leu Lys Thr Val Ala Ile
20 25 30
Trp Ala Glu Glu Asp Lys Leu Ala Leu His Arg Phe Lys Ala Asp Glu
35 40 45
Ser Tyr Gln Val Gly Arg Gly Pro His Leu Pro Arg Asp Leu Gly Pro
50 55 60
Ile Met Ser Tyr Leu Ser Ile Asp Glu Val Ile Arg Val Ala Lys Leu
65 70 75 80
Ser Gly Ala Asp Ala Ile His Pro Gly Tyr Gly Leu Leu Ser Glu Ser
85 90 95
Pro Glu Phe Ala Glu Ala Cys Ala Ala Asn Gly Ile Thr Phe Ile Gly
100 105 110
Pro Lys Pro Glu Thr Met Arg Gln Leu Gly Asn Lys Val Ala Ala Arg
115 120 125
Asn Leu Ala Ile Ser Ile Gly Val Pro Val Val Pro Ala Thr Glu Pro
130 135 140
Leu Pro Asp Asp Pro Glu Glu Ile Lys Arg Leu Ala Glu Glu Ile Gly
145 150 155 160
Tyr Pro Val Met Leu Lys Ala Ser Trp Gly Gly Gly Gly Arg Gly Met
165 170 175
Arg Ala Ile Arg Asp Pro Lys Asp Leu Ile Arg Glu Val Thr Glu Ala
180 185 190
Lys Arg Glu Ala Lys Ala Ala Phe Gly Lys Asp Glu Val Tyr Leu Glu
195 200 205
Lys Leu Val Glu Arg Ala Arg His Val Glu Ser Gln Ile Leu Gly Asp
210 215 220
Thr His Gly Asn Val Val His Leu Phe Glu Arg Asp Cys Ser Ile Gln
225 230 235 240
Arg Arg Asn Gln Lys Val Val Glu Arg Ala Pro Ala Pro Tyr Leu Asn
245 250 255
Asp Ala Gln Arg Gln Glu Leu Ala Asp Tyr Ser Leu Lys Ile Ala Arg
260 265 270
Ala Thr Asn Tyr Ile Gly Ala Gly Thr Val Glu Tyr Leu Met Asp Ser
275 280 285
Asp Thr Gly Lys Phe Tyr Phe Ile Glu Val Asn Pro Arg Ile Gln Val
290 295 300
Glu His Thr Val Thr Glu Val Val Thr Gly Ile Asp Ile Val Lys Ala
305 310 315 320
Gln Ile His Ile Leu Asp Gly Phe Ala Ile Gly Ala Pro Glu Ser Gly
325 330 335
Val Pro Arg Gln Glu Asp Ile Arg Leu Asn Gly His Ala Leu Gln Cys
340 345 350
Arg Ile Thr Thr Glu Asp Pro Glu Gln Asn Phe Ile Pro Asp Tyr Gly
355 360 365
Arg Ile Thr Ala Tyr Arg Gly Ala Thr Gly Phe Gly Ile Arg Leu Asp
370 375 380
Gly Gly Thr Ala Tyr Ser Gly Ala Val Ile Thr Arg Tyr Tyr Asp Pro
385 390 395 400
Leu Leu Glu Lys Val Thr Ala Trp Ala Pro Asn Pro Gly Glu Ala Ile
405 410 415
Gln Arg Met Ile Arg Ala Leu Arg Glu Phe Arg Ile Arg Gly Val Ala
420 425 430
Thr Asn Leu Thr Phe Leu Glu Ala Ile Ile Ser His Pro Lys Phe His
435 440 445
Asp Asn Ser Tyr Thr Thr Arg Phe Ile Asp Thr Thr Pro Glu Leu Phe
450 455 460
Gln Gln Val Lys Arg Gln Asp Arg Ala Thr Lys Leu Leu Thr Tyr Leu
465 470 475 480
Ala Asp Val Thr Val Asn Gly His Pro Glu Val Lys Gly Arg Pro Lys
485 490 495
Pro Ser Asp Asp Ile Ala Ala Pro Val Val Pro Phe Thr Gly Gly Asp
500 505 510
Val Lys Pro Gly Thr Lys Gln Arg Leu Asp Gln Leu Gly Pro Lys Lys
515 520 525
Phe Ala Glu Trp Val Lys Ala Gln Pro Glu Val Leu Ile Thr Asp Thr
530 535 540
Thr Met Arg Asp Gly His Gln Ser Leu Leu Ala Thr Arg Met Arg Thr
545 550 555 560
Tyr Asp Ile Ala Arg Ile Ala Gly Thr Tyr Ala Arg Ala Leu Pro Asn
565 570 575
Leu Phe Ser Leu Glu Cys Trp Gly Gly Ala Thr Phe Asp Val Ser Met
580 585 590
Arg Phe Leu Thr Glu Asp Pro Trp Glu Arg Leu Ala Met Val Arg Glu
595 600 605
Gly Ala Pro Asn Leu Leu Leu Gln Met Leu Leu Arg Gly Ala Asn Gly
610 615 620
Val Gly Tyr Lys Asn Tyr Pro Asp Asn Val Val Lys Tyr Phe Val Arg
625 630 635 640
Gln Ala Ala Lys Gly Gly Ile Asp Val Phe Arg Val Phe Asp Cys Leu
645 650 655
Asn Trp Val Glu Asn Met Arg Val Ala Met Asp Ala Val Ala Glu Glu
660 665 670
Asp Arg Ile Cys Glu Ala Ala Ile Cys Tyr Thr Gly Asp Ile Leu Asn
675 680 685
Ser Ala Arg Pro Lys Tyr Asp Leu Lys Tyr Tyr Thr Ala Leu Ala Ala
690 695 700
Glu Leu Glu Lys Ala Gly Ala His Met Ile Ala Val Lys Asp Met Ala
705 710 715 720
Gly Leu Leu Lys Pro Ala Ala Ala Arg Val Leu Phe Lys Ala Leu Lys
725 730 735
Glu Ala Thr Gly Leu Pro Ile His Phe His Thr His Asp Thr Ser Gly
740 745 750
Ile Ala Ala Ala Thr Val Leu Ala Ala Val Glu Ser Gly Val Asp Val
755 760 765
Val Asp Ala Ala Met Asp Ala Leu Ser Gly Asn Thr Ser Gln Pro Cys
770 775 780
Leu Gly Ser Ile Val Glu Ala Leu Ser Gly Ser Glu Arg Asp Pro Gly
785 790 795 800
Leu Asp Pro Glu Trp Ile Arg Arg Ile Ser Phe Tyr Trp Glu Ala Val
805 810 815
Arg His Gln Tyr Ala Ala Phe Glu Ser Asp Leu Lys Gly Pro Ala Ser
820 825 830
Glu Val Tyr Leu His Glu Met Pro Gly Gly Gln Phe Thr Asn Leu Lys
835 840 845
Glu Gln Ala Arg Ser Leu Gly Leu Glu Thr Arg Trp His Glu Val Ala
850 855 860
Gln Ala Tyr Ala Asp Ala Asn Arg Met Phe Gly Asp Ile Val Lys Val
865 870 875 880
Thr Pro Ser Ser Lys Val Val Gly Asp Met Ala Leu Met Met Val Ser
885 890 895
Gln Asp Leu Thr Val Ala Asp Val Glu Asn Pro Gly Lys Asp Ile Ala
900 905 910
Phe Pro Glu Ser Val Val Ser Met Leu Lys Gly Asp Leu Gly Gln Pro
915 920 925
Pro Gly Gly Trp Pro Glu Ala Leu Gln Lys Lys Ala Leu Lys Gly Glu
930 935 940
Glu Pro Tyr Asp Ala Arg Pro Gly Ser Leu Leu Glu Asp Ala Asp Leu
945 950 955 960
Asp Ala Glu Arg Lys Gly Ile Glu Glu Lys Leu Gly Arg Glu Val Thr
965 970 975
Asp Phe Glu Phe Ala Ser Tyr Leu Met Tyr Pro Lys Val Phe Thr Asp
980 985 990
Tyr Ala Val Ala Cys Glu Thr Tyr Gly Pro Val Ser Val Leu Pro Thr
995 1000 1005
Pro Ala Tyr Phe Tyr Gly Met Ala Pro Gly Glu Glu Leu Phe Ala
1010 1015 1020
Asp Ile Glu Lys Gly Lys Thr Leu Val Ile Leu Asn Gln Ala Gln
1025 1030 1035
Gly Glu Ile Asp Glu Lys Gly Met Val Lys Met Phe Phe Glu Met
1040 1045 1050
Asn Gly Gln Pro Arg Ser Ile Lys Val Pro Asp Arg Asn Arg Gly
1055 1060 1065
Ala Ser Ala Ala Val Arg Arg Lys Ala Glu Ala Gly Asn Ala Ala
1070 1075 1080
His Leu Gly Ala Pro Met Pro Gly Val Ile Ser Thr Val Ala Val
1085 1090 1095
Ala Ser Gly Gln Ser Val Lys Ala Gly Asp Val Leu Leu Ser Ile
1100 1105 1110
Glu Ala Met Lys Met Glu Thr Ala Leu His Ala Glu Lys Asp Gly
1115 1120 1125
Val Ile Ser Glu Val Leu Val Arg Ala Gly Asp Gln Ile Asp Ala
1130 1135 1140
Lys Asp Leu Leu Val Val Phe Gly Gly
1145 1150
<210> 9
<211> 2652
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 9
atgaacgaac aatattccgc attgcgtagt aatgtcagta tgctcggcaa agtgctggga 60
gaaaccatca aggatgcgtt gggagaacac attcttgaac gcgtagaaac tatccgtaag 120
ttgtcgaaat cttcacgcgc tggcaatgat gctaaccgcc aggagttgct caccacctta 180
caaaatttgt cgaacgacga gctgctgccc gttgcgcgtg cgtttagtca gttcctgaac 240
ctggccaaca ccgccgagca ataccacagc atttcgccga aaggcgaagc tgccagcaac 300
ccggaagtga tcgcccgcac cctgcgtaaa ctgaaaaacc agccggaact gagcgaagac 360
accatcaaaa aagcagtgga atcgctgtcg ctggaactgg tcctcacggc tcacccaacc 420
gaaattaccc gtcgtacact gatccacaaa atggtggaag tgaacgcctg tttaaaacag 480
ctcgataaca aagatatcgc tgactacgaa cacaaccagc tgatgcgtcg cctgcgccag 540
ttgatcgccc agtcatggca taccgatgaa atccgtaagc tgcgtccaag cccggtagat 600
gaagccaaat ggggctttgc cgtagtggaa aacagcctgt ggcaaggcgt accaaattac 660
ctgcgcgaac tgaacgaaca actggaagag aacctcggct acaaactgcc cgtcgaattt 720
gttccggtcc gttttacttc gtggatgggc ggcgaccgcg acggcaaccc gaacgtcact 780
gccgatatca cccgccacgt cctgctactc agccgctgga aagccaccga tttgttcctg 840
aaagatattc aggtgctggt ttctgaactg tcgatggttg aagcgacccc tgaactgctg 900
gcgctggttg gcgaagaagg tgccgcagaa ccgtatcgct atctgatgaa aaacctgcgt 960
tctcgcctga tggcgacaca ggcatggctg gaagcgcgcc tgaaaggcga agaactgcca 1020
aaaccagaag gcctgctgac acaaaacgaa gaactgtggg aaccgctcta cgcttgctac 1080
cagtcacttc aggcgtgtgg catgggtatt atcgccaacg gcgatctgct cgacaccctg 1140
cgccgcgtga aatgtttcgg cgtaccgctg gtccgtattg atatccgtca ggagagcacg 1200
cgtcataccg aagcgctggg cgagctgacc cgctacctcg gtatcggcga ctacgaaagc 1260
tggtcagagg ccgacaaaca ggcgttcctg atccgcgaac tgaactccaa acgtccgctt 1320
ctgccgcgca actggcaacc aagcgccgaa acgcgcgaag tgctcgatac ctgccaggtg 1380
attgccgaag caccgcaagg ctccattgcc gcctacgtga tctcgatggc gaaaacgccg 1440
tccgacgtac tggctgtcca cctgctgctg aaagaagcgg gtatcgggtt tgcgatgccg 1500
gttgctccgc tgtttgaaac cctcgatgat ctgaacaacg ccaacgatgt catgacccag 1560
ctgctcaata ttgactggta tcgtggcctg attcagggca aacagatggt gatgattggc 1620
tattccgact cagcaaaaga tgcgggagtg atggcagctt cctgggcgca atatcaggca 1680
caggatgcat taatcaaaac ctgcgaaaaa gcgggtattg agctgacgtt gttccacggt 1740
cgcggcggtt ccattggtcg cggcggcgca cctgctcatg cggcgctgct gtcacaaccg 1800
ccaggaagcc tgaaaggcgg cctgcgcgta accgaacagg gcgagatgat ccgctttaaa 1860
tatggtctgc cagaaatcac cgtcagcagc ctgtcgcttt ataccggggc gattctggaa 1920
gccaacctgc tgccaccgcc ggagccgaaa gagagctggc gtcgcattat ggatgaactg 1980
tcagtcatct cctgcgatgt ctaccgcggc tacgtacgtg aaaacaaaga ttttgtgcct 2040
tacttccgct ccgctacgcc ggaacaagaa ctgggcaaac tgccgttggg ttcacgtccg 2100
gcgaaacgtc gcccaaccgg cggcgtcgag tcactacgcg ccattccgtg gatcttcgcc 2160
tggacgcaaa accgtctgat gctccccgcc tggctgggtg caggtacggc gctgcaaaaa 2220
gtggtcgaag acggcaaaca gagcgagctg gaggctatgt gccgcgattg gccattcttc 2280
tcgacgcgtc tcggcatgct ggagatggtc ttcgccaaag cagacctgtg gctggcggaa 2340
tactatgacc aacgcctggt agacaaagca ctgtggccgt taggtaaaga gttacgcaac 2400
ctgcaagaag aagacatcaa agtggtgctg gcgattgcca acgattccca tctgatggcc 2460
gatctgccgt ggattgcaga gtctattcag ctacggaata tttacaccga cccgctgaac 2520
gtattgcagg ccgagttgct gcaccgctcc cgccaggcag aaaaagaagg ccaggaaccg 2580
gatcctcgcg tcgaacaagc gttaatggtc actattgccg ggattgcggc aggtatgcgt 2640
aataccggct aa 2652
<210> 10
<211> 883
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 10
Met Asn Glu Gln Tyr Ser Ala Leu Arg Ser Asn Val Ser Met Leu Gly
1 5 10 15
Lys Val Leu Gly Glu Thr Ile Lys Asp Ala Leu Gly Glu His Ile Leu
20 25 30
Glu Arg Val Glu Thr Ile Arg Lys Leu Ser Lys Ser Ser Arg Ala Gly
35 40 45
Asn Asp Ala Asn Arg Gln Glu Leu Leu Thr Thr Leu Gln Asn Leu Ser
50 55 60
Asn Asp Glu Leu Leu Pro Val Ala Arg Ala Phe Ser Gln Phe Leu Asn
65 70 75 80
Leu Ala Asn Thr Ala Glu Gln Tyr His Ser Ile Ser Pro Lys Gly Glu
85 90 95
Ala Ala Ser Asn Pro Glu Val Ile Ala Arg Thr Leu Arg Lys Leu Lys
100 105 110
Asn Gln Pro Glu Leu Ser Glu Asp Thr Ile Lys Lys Ala Val Glu Ser
115 120 125
Leu Ser Leu Glu Leu Val Leu Thr Ala His Pro Thr Glu Ile Thr Arg
130 135 140
Arg Thr Leu Ile His Lys Met Val Glu Val Asn Ala Cys Leu Lys Gln
145 150 155 160
Leu Asp Asn Lys Asp Ile Ala Asp Tyr Glu His Asn Gln Leu Met Arg
165 170 175
Arg Leu Arg Gln Leu Ile Ala Gln Ser Trp His Thr Asp Glu Ile Arg
180 185 190
Lys Leu Arg Pro Ser Pro Val Asp Glu Ala Lys Trp Gly Phe Ala Val
195 200 205
Val Glu Asn Ser Leu Trp Gln Gly Val Pro Asn Tyr Leu Arg Glu Leu
210 215 220
Asn Glu Gln Leu Glu Glu Asn Leu Gly Tyr Lys Leu Pro Val Glu Phe
225 230 235 240
Val Pro Val Arg Phe Thr Ser Trp Met Gly Gly Asp Arg Asp Gly Asn
245 250 255
Pro Asn Val Thr Ala Asp Ile Thr Arg His Val Leu Leu Leu Ser Arg
260 265 270
Trp Lys Ala Thr Asp Leu Phe Leu Lys Asp Ile Gln Val Leu Val Ser
275 280 285
Glu Leu Ser Met Val Glu Ala Thr Pro Glu Leu Leu Ala Leu Val Gly
290 295 300
Glu Glu Gly Ala Ala Glu Pro Tyr Arg Tyr Leu Met Lys Asn Leu Arg
305 310 315 320
Ser Arg Leu Met Ala Thr Gln Ala Trp Leu Glu Ala Arg Leu Lys Gly
325 330 335
Glu Glu Leu Pro Lys Pro Glu Gly Leu Leu Thr Gln Asn Glu Glu Leu
340 345 350
Trp Glu Pro Leu Tyr Ala Cys Tyr Gln Ser Leu Gln Ala Cys Gly Met
355 360 365
Gly Ile Ile Ala Asn Gly Asp Leu Leu Asp Thr Leu Arg Arg Val Lys
370 375 380
Cys Phe Gly Val Pro Leu Val Arg Ile Asp Ile Arg Gln Glu Ser Thr
385 390 395 400
Arg His Thr Glu Ala Leu Gly Glu Leu Thr Arg Tyr Leu Gly Ile Gly
405 410 415
Asp Tyr Glu Ser Trp Ser Glu Ala Asp Lys Gln Ala Phe Leu Ile Arg
420 425 430
Glu Leu Asn Ser Lys Arg Pro Leu Leu Pro Arg Asn Trp Gln Pro Ser
435 440 445
Ala Glu Thr Arg Glu Val Leu Asp Thr Cys Gln Val Ile Ala Glu Ala
450 455 460
Pro Gln Gly Ser Ile Ala Ala Tyr Val Ile Ser Met Ala Lys Thr Pro
465 470 475 480
Ser Asp Val Leu Ala Val His Leu Leu Leu Lys Glu Ala Gly Ile Gly
485 490 495
Phe Ala Met Pro Val Ala Pro Leu Phe Glu Thr Leu Asp Asp Leu Asn
500 505 510
Asn Ala Asn Asp Val Met Thr Gln Leu Leu Asn Ile Asp Trp Tyr Arg
515 520 525
Gly Leu Ile Gln Gly Lys Gln Met Val Met Ile Gly Tyr Ser Asp Ser
530 535 540
Ala Lys Asp Ala Gly Val Met Ala Ala Ser Trp Ala Gln Tyr Gln Ala
545 550 555 560
Gln Asp Ala Leu Ile Lys Thr Cys Glu Lys Ala Gly Ile Glu Leu Thr
565 570 575
Leu Phe His Gly Arg Gly Gly Ser Ile Gly Arg Gly Gly Ala Pro Ala
580 585 590
His Ala Ala Leu Leu Ser Gln Pro Pro Gly Ser Leu Lys Gly Gly Leu
595 600 605
Arg Val Thr Glu Gln Gly Glu Met Ile Arg Phe Lys Tyr Gly Leu Pro
610 615 620
Glu Ile Thr Val Ser Ser Leu Ser Leu Tyr Thr Gly Ala Ile Leu Glu
625 630 635 640
Ala Asn Leu Leu Pro Pro Pro Glu Pro Lys Glu Ser Trp Arg Arg Ile
645 650 655
Met Asp Glu Leu Ser Val Ile Ser Cys Asp Val Tyr Arg Gly Tyr Val
660 665 670
Arg Glu Asn Lys Asp Phe Val Pro Tyr Phe Arg Ser Ala Thr Pro Glu
675 680 685
Gln Glu Leu Gly Lys Leu Pro Leu Gly Ser Arg Pro Ala Lys Arg Arg
690 695 700
Pro Thr Gly Gly Val Glu Ser Leu Arg Ala Ile Pro Trp Ile Phe Ala
705 710 715 720
Trp Thr Gln Asn Arg Leu Met Leu Pro Ala Trp Leu Gly Ala Gly Thr
725 730 735
Ala Leu Gln Lys Val Val Glu Asp Gly Lys Gln Ser Glu Leu Glu Ala
740 745 750
Met Cys Arg Asp Trp Pro Phe Phe Ser Thr Arg Leu Gly Met Leu Glu
755 760 765
Met Val Phe Ala Lys Ala Asp Leu Trp Leu Ala Glu Tyr Tyr Asp Gln
770 775 780
Arg Leu Val Asp Lys Ala Leu Trp Pro Leu Gly Lys Glu Leu Arg Asn
785 790 795 800
Leu Gln Glu Glu Asp Ile Lys Val Val Leu Ala Ile Ala Asn Asp Ser
805 810 815
His Leu Met Ala Asp Leu Pro Trp Ile Ala Glu Ser Ile Gln Leu Arg
820 825 830
Asn Ile Tyr Thr Asp Pro Leu Asn Val Leu Gln Ala Glu Leu Leu His
835 840 845
Arg Ser Arg Gln Ala Glu Lys Glu Gly Gln Glu Pro Asp Pro Arg Val
850 855 860
Glu Gln Ala Leu Met Val Thr Ile Ala Gly Ile Ala Ala Gly Met Arg
865 870 875 880
Asn Thr Gly
<210> 11
<211> 483
<212> PRT
<213> 热自养甲烷杆菌(Methanobacterium thermoautotrophicum)
<400> 11
Met Lys Val Pro Arg Cys Met Ser Thr Gln His Pro Asp Asn Val Asn
1 5 10 15
Pro Pro Phe Phe Ala Glu Glu Pro Glu Leu Gly Gly Glu Asp Glu Ile
20 25 30
Arg Glu Ala Tyr Tyr Val Phe Ser His Leu Gly Cys Asp Glu Gln Met
35 40 45
Trp Asp Cys Glu Gly Lys Glu Val Asp Asn Tyr Val Val Lys Lys Leu
50 55 60
Leu Thr Lys Tyr Gln Ala Phe Phe Arg Asp His Val Leu Gly Glu Asp
65 70 75 80
Leu Arg Leu Thr Leu Arg Val Pro Asn Pro Thr Val Glu Arg Ala Glu
85 90 95
Ala Lys Ile Leu Leu Glu Thr Leu Glu Ser Ile Pro Arg Ser Tyr Asp
100 105 110
Thr Ala Ser Leu Phe Tyr Gly Met Asp Ala Ala Pro Val Phe Glu Val
115 120 125
Ile Leu Pro Met Thr Ser Ser Ser Ser Cys Leu Asn Arg Ile His Ser
130 135 140
Tyr Tyr Leu Asp Phe Val Lys Gly Lys Glu Arg Leu Gln Leu Ala Asp
145 150 155 160
Gly Val Thr Val Lys Glu Trp Ile Gly Glu Phe Arg Pro Asp Glu Ile
165 170 175
Asn Val Ile Pro Leu Phe Glu Asp His Glu Gly Met Leu Asn Ala Ala
180 185 190
Lys Ile Thr Gly Glu Tyr Leu Asp Gly Lys Asp Ile Gln Glu Gln Arg
195 200 205
Val Phe Leu Ala Arg Ser Asp Pro Ala Met Asn Tyr Gly Met Ile Ser
210 215 220
Ala Thr Leu Leu Asn Arg Ile Ala Leu Ser Asp Phe Arg Asp Leu Glu
225 230 235 240
Glu Glu Ser Gly Val Lys Leu Tyr Pro Ile Ile Gly Met Gly Ser Ala
245 250 255
Pro Phe Arg Gly Asn Leu Arg Pro Asp Asn Val Glu Asp Val Thr Trp
260 265 270
Glu Tyr Arg Gly Ala Tyr Thr Phe Thr Val Gln Ser Ser Phe Lys Tyr
275 280 285
Asp His Glu Pro Ser Asp Val Ile Arg Gly Ile Lys Lys Leu Arg Ser
290 295 300
Val Lys Pro Gly Arg Ala Ala Glu Ile Glu Arg Glu Ser Val Leu Glu
305 310 315 320
Ile Ile Ser Ala Tyr Cys Arg Glu Tyr Arg Arg Gln Val Met Asp Leu
325 330 335
Val Asp Ile Ile Asn Arg Val Ala Arg Tyr Val Pro Gly Arg Arg Lys
340 345 350
Arg Lys Leu His Ile Gly Leu Phe Gly Tyr Ser Arg Ser Met Gly Asn
355 360 365
Val Ser Leu Pro Arg Ala Ile Thr Phe Thr Ala Ala Leu Tyr Ser Leu
370 375 380
Gly Val Pro Pro Glu Leu Leu Gly Phe Asn Ala Leu Ser Ser Gly Asp
385 390 395 400
Leu Glu Phe Ile Glu Glu Val Tyr Pro Gly Leu Gly Arg Asp Leu His
405 410 415
Asp Ala Ala Arg Tyr Ala Asn Pro Glu Ser Pro Phe Leu Ser Pro Glu
420 425 430
Val Lys Ser Ser Phe Glu Glu Tyr Leu Glu Pro Glu Tyr Asp Glu Gly
435 440 445
His Met Lys Thr Thr Glu Glu Ile Ile Arg Ala Leu Arg Ile Asn Arg
450 455 460
Thr Ala Asn Leu Gln Glu Leu Ile Leu Glu Ala Ala Ser Gln Arg Lys
465 470 475 480
Phe Leu Gly
<210> 12
<211> 537
<212> PRT
<213> 产气荚膜梭菌(Clostridium perfringens)
<400> 12
Met Lys Ile Pro Cys Ser Met Met Thr Gln His Pro Asp Asn Val Glu
1 5 10 15
Thr Tyr Ile Ser Ile Gln Gln Glu Pro Ala Glu Ala Ile Lys Gly Leu
20 25 30
Thr Pro Gln Asp Lys Gly Gly Leu Gly Ile Glu Glu Val Met Ile Asp
35 40 45
Phe Glu Gly Lys Leu Thr Pro Tyr His Gln Thr Ser Gln Ile Ala Leu
50 55 60
Gly Leu Ile Ser Asn Gly Ile Ile Pro Gly Lys Asp Val Arg Val Thr
65 70 75 80
Pro Arg Ile Pro Asn Ala Asn Lys Glu Ser Val Phe Arg Gln Leu Met
85 90 95
Ser Ile Met Ser Ile Ile Glu Thr Asn Val Gln Ser Lys Glu Leu Thr
100 105 110
Gly Thr Pro Ala Ile Ser Glu Val Val Val Pro Met Ile Glu Thr Gly
115 120 125
Lys Glu Ile Ser Glu Phe Gln Asp Arg Val Asn Ser Val Val Asp Met
130 135 140
Gly Asn Lys Asn Tyr Lys Thr Lys Leu Asp Leu Asn Ser Val Arg Ile
145 150 155 160
Ile Pro Leu Val Glu Asp Val Pro Ala Leu Ala Asn Ile Asp Arg Ile
165 170 175
Leu Asp Glu His Tyr Glu Ile Glu Lys Ser Lys Gly His Ile Leu Lys
180 185 190
Asp Leu Arg Ile Met Ile Ala Arg Ser Asp Thr Ala Met Ser Tyr Gly
195 200 205
Leu Ile Ser Gly Val Leu Ser Val Leu Met Ala Val Asp Gly Ala Tyr
210 215 220
Lys Trp Gly Glu Lys His Gly Val Thr Ile Ser Pro Ile Leu Gly Cys
225 230 235 240
Gly Ser Leu Pro Phe Arg Gly His Phe Ser Glu Glu Asn Ile Asp Glu
245 250 255
Ile Leu Ala Thr Tyr Ser Gly Ile Lys Thr Phe Thr Phe Gln Ser Ala
260 265 270
Leu Arg Tyr Asp His Gly Glu Glu Ala Thr Lys His Ala Val Arg Glu
275 280 285
Leu Lys Glu Lys Ile Ala Gln Ser Lys Pro Arg Asn Phe Ser Glu Glu
290 295 300
Asp Lys Asp Leu Met Lys Glu Phe Ile Gly Ile Cys Ser Lys His Tyr
305 310 315 320
Leu Gln Thr Phe Leu Lys Val Ile Asp Thr Val Ser Phe Val Ser Asp
325 330 335
Phe Ile Pro Lys Asn Arg Asp Arg Leu Thr Lys Ala Lys Thr Gly Leu
340 345 350
Glu Tyr Asn Arg Glu Val Ala Asn Leu Asp Asn Val Ala Asp Leu Val
355 360 365
Lys Asp Glu Val Leu Lys Gln Glu Ile Leu Ser Ile Asp Asn Ser Lys
370 375 380
Glu Tyr Ala Val Pro Arg Ala Ile Ser Phe Thr Gly Ala Met Tyr Thr
385 390 395 400
Leu Gly Met Pro Pro Glu Leu Met Gly Met Gly Arg Ala Leu Asn Glu
405 410 415
Ile Lys Thr Lys Tyr Gly Gln Glu Gly Ile Asp Lys Leu Leu Glu Ile
420 425 430
Tyr Pro Ile Leu Arg Lys Asp Leu Ala Phe Ala Ala Arg Phe Ala Asn
435 440 445
Gly Gly Val Ser Lys Lys Ile Ile Asp Glu Glu Ala Arg Gln Glu Tyr
450 455 460
Lys Glu Asp Met Lys Tyr Val Asn Glu Ile Leu Asn Leu Gly Leu Asp
465 470 475 480
Tyr Asp Phe Leu Asn Glu Asn Glu Phe Tyr His Thr Leu Leu Lys Thr
485 490 495
Thr Lys Pro Ile Ile Met His Leu Met Gly Leu Glu Glu Asn Val Met
500 505 510
Arg Asn Ser Thr Glu Glu Leu Lys Ile Leu Asn Glu Trp Ile Val Arg
515 520 525
Met Gly Lys Val Arg Gly Ser Ile Gly
530 535
<210> 13
<211> 1260
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 13
atgtccagag gcttctttac tgagaacatt acgcaattgc caccagaccc tttgtttggt 60
cttaaggcca ggttcagcaa tgactcacgt gaaaacaagg tcgatttagg tattggggca 120
tatagggacg acaacggtaa gccatggatc ttaccatctg tcaggttggc cgaaaacttg 180
attcagaact ccccagacta caaccatgag tacctaccaa tcggtggact tgctgatttc 240
acttctgctg cggcaagagt tgtatttgga ggcgattcta aagccatttc gcaaaaccgt 300
cttgtctcca tccagagttt gtcaggtaca ggtgccttac atgttgctgg tctatttatc 360
aagcgccaat acaagtctct tgatggcact tccgaagacc ctctaatata tctatcggaa 420
cctacatggg ccaaccacgt tcaaatcttt gaagttattg gtctcaagcc tgtattctat 480
ccatattggc atgccgcaag caagaccttg gatctgaagg gctacttaaa ggcaataaac 540
gatgctccag aagggtcggt ttttgtattg catgcaacgg ctcataaccc tactggtttg 600
gatccaacac aagaacaatg gatggagatt ttggccgcta taagtgccaa aaagcatctg 660
ccattatttg attgtgcata tcagggtttc acctccgggt ctctagatag agatgcttgg 720
gctgttcgag aagctgtcaa caatgacaag tacgaattcc cgggaattat tgtctgtcaa 780
tcgtttgcga aaaatgttgg catgtatggt gaacggattg gtgcagttca tattgttcta 840
cctgaatcag acgcttccct aaacagcgcc atcttctccc aattgcaaaa gacaatcaga 900
tcggagattt ccaatccacc aggatacggt gcaaagattg tgtctaaagt tttgaacact 960
ccggaacttt acaaacagtg ggagcaagat ttgatcacca tgtcttcgag aatcactgca 1020
atgagaaagg agctagtaaa tgagctcgag cgtcttggaa cccctggcac ttggagacac 1080
atcaccgagc aacagggtat gttttccttt actggtttga acccggagca ggttgccaag 1140
ctagagaagg agcatggtgt ttatcttgtt cgtagtggac gtgcaagtat tgcaggcctc 1200
aacatgggaa acgtcaagta tgttgccaag gccattgact ctgtcgtgag agacctttag 1260
<210> 14
<211> 419
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 14
Met Ser Arg Gly Phe Phe Thr Glu Asn Ile Thr Gln Leu Pro Pro Asp
1 5 10 15
Pro Leu Phe Gly Leu Lys Ala Arg Phe Ser Asn Asp Ser Arg Glu Asn
20 25 30
Lys Val Asp Leu Gly Ile Gly Ala Tyr Arg Asp Asp Asn Gly Lys Pro
35 40 45
Trp Ile Leu Pro Ser Val Arg Leu Ala Glu Asn Leu Ile Gln Asn Ser
50 55 60
Pro Asp Tyr Asn His Glu Tyr Leu Pro Ile Gly Gly Leu Ala Asp Phe
65 70 75 80
Thr Ser Ala Ala Ala Arg Val Val Phe Gly Gly Asp Ser Lys Ala Ile
85 90 95
Ser Gln Asn Arg Leu Val Ser Ile Gln Ser Leu Ser Gly Thr Gly Ala
100 105 110
Leu His Val Ala Gly Leu Phe Ile Lys Arg Gln Tyr Lys Ser Leu Asp
115 120 125
Gly Thr Ser Glu Asp Pro Leu Ile Tyr Leu Ser Glu Pro Thr Trp Ala
130 135 140
Asn His Val Gln Ile Phe Glu Val Ile Gly Leu Lys Pro Val Phe Tyr
145 150 155 160
Pro Tyr Trp His Ala Ala Ser Lys Thr Leu Asp Leu Lys Gly Tyr Leu
165 170 175
Lys Ala Ile Asn Asp Ala Pro Glu Gly Ser Val Phe Val Leu His Ala
180 185 190
Thr Ala His Asn Pro Thr Gly Leu Asp Pro Thr Gln Glu Gln Trp Met
195 200 205
Glu Ile Leu Ala Ala Ile Ser Ala Lys Lys His Leu Pro Leu Phe Asp
210 215 220
Cys Ala Tyr Gln Gly Phe Thr Ser Gly Ser Leu Asp Arg Asp Ala Trp
225 230 235 240
Ala Val Arg Glu Ala Val Asn Asn Asp Lys Tyr Glu Phe Pro Gly Ile
245 250 255
Ile Val Cys Gln Ser Phe Ala Lys Asn Val Gly Met Tyr Gly Glu Arg
260 265 270
Ile Gly Ala Val His Ile Val Leu Pro Glu Ser Asp Ala Ser Leu Asn
275 280 285
Ser Ala Ile Phe Ser Gln Leu Gln Lys Thr Ile Arg Ser Glu Ile Ser
290 295 300
Asn Pro Pro Gly Tyr Gly Ala Lys Ile Val Ser Lys Val Leu Asn Thr
305 310 315 320
Pro Glu Leu Tyr Lys Gln Trp Glu Gln Asp Leu Ile Thr Met Ser Ser
325 330 335
Arg Ile Thr Ala Met Arg Lys Glu Leu Val Asn Glu Leu Glu Arg Leu
340 345 350
Gly Thr Pro Gly Thr Trp Arg His Ile Thr Glu Gln Gln Gly Met Phe
355 360 365
Ser Phe Thr Gly Leu Asn Pro Glu Gln Val Ala Lys Leu Glu Lys Glu
370 375 380
His Gly Val Tyr Leu Val Arg Ser Gly Arg Ala Ser Ile Ala Gly Leu
385 390 395 400
Asn Met Gly Asn Val Lys Tyr Val Ala Lys Ala Ile Asp Ser Val Val
405 410 415
Arg Asp Leu
<210> 15
<211> 418
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 15
Met Ser Ala Thr Leu Phe Asn Asn Ile Glu Leu Leu Pro Pro Asp Ala
1 5 10 15
Leu Phe Gly Ile Lys Gln Arg Tyr Gly Gln Asp Gln Arg Ala Thr Lys
20 25 30
Val Asp Leu Gly Ile Gly Ala Tyr Arg Asp Asp Asn Gly Lys Pro Trp
35 40 45
Val Leu Pro Ser Val Lys Ala Ala Glu Lys Leu Ile His Asn Asp Ser
50 55 60
Ser Tyr Asn His Glu Tyr Leu Gly Ile Thr Gly Leu Pro Ser Leu Thr
65 70 75 80
Ser Asn Ala Ala Lys Ile Ile Phe Gly Thr Gln Ser Asp Ala Phe Gln
85 90 95
Glu Asp Arg Val Ile Ser Val Gln Ser Leu Ser Gly Thr Gly Ala Leu
100 105 110
His Ile Ser Ala Lys Phe Phe Ser Lys Phe Phe Pro Asp Lys Leu Val
115 120 125
Tyr Leu Ser Lys Pro Thr Trp Ala Asn His Met Ala Ile Phe Glu Asn
130 135 140
Gln Gly Leu Lys Thr Ala Thr Tyr Pro Tyr Trp Ala Asn Glu Thr Lys
145 150 155 160
Ser Leu Asp Leu Asn Gly Phe Leu Asn Ala Ile Gln Lys Ala Pro Glu
165 170 175
Gly Ser Ile Phe Val Leu His Ser Cys Ala His Asn Pro Thr Gly Leu
180 185 190
Asp Pro Thr Ser Glu Gln Trp Val Gln Ile Val Asp Ala Ile Ala Ser
195 200 205
Lys Asn His Ile Ala Leu Phe Asp Thr Ala Tyr Gln Gly Phe Ala Thr
210 215 220
Gly Asp Leu Asp Lys Asp Ala Tyr Ala Val Arg Leu Gly Val Glu Lys
225 230 235 240
Leu Ser Thr Val Ser Pro Val Phe Val Cys Gln Ser Phe Ala Lys Asn
245 250 255
Ala Gly Met Tyr Gly Glu Arg Val Gly Cys Phe His Leu Ala Leu Thr
260 265 270
Lys Gln Ala Gln Asn Lys Thr Ile Lys Pro Ala Val Thr Ser Gln Leu
275 280 285
Ala Lys Ile Ile Arg Ser Glu Val Ser Asn Pro Pro Ala Tyr Gly Ala
290 295 300
Lys Ile Val Ala Lys Leu Leu Glu Thr Pro Glu Leu Thr Glu Gln Trp
305 310 315 320
His Lys Asp Met Val Thr Met Ser Ser Arg Ile Thr Lys Met Arg His
325 330 335
Ala Leu Arg Asp His Leu Val Lys Leu Gly Thr Pro Gly Asn Trp Asp
340 345 350
His Ile Val Asn Gln Cys Gly Met Phe Ser Phe Thr Gly Leu Thr Pro
355 360 365
Gln Met Val Lys Arg Leu Glu Glu Thr His Ala Val Tyr Leu Val Ala
370 375 380
Ser Gly Arg Ala Ser Ile Ala Gly Leu Asn Gln Gly Asn Val Glu Tyr
385 390 395 400
Val Ala Lys Ala Ile Asp Glu Val Val Arg Phe Tyr Thr Ile Glu Ala
405 410 415
Lys Leu
<210> 16
<211> 396
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 16
Met Phe Glu Asn Ile Thr Ala Ala Pro Ala Asp Pro Ile Leu Gly Leu
1 5 10 15
Ala Asp Leu Phe Arg Ala Asp Glu Arg Pro Gly Lys Ile Asn Leu Gly
20 25 30
Ile Gly Val Tyr Lys Asp Glu Thr Gly Lys Thr Pro Val Leu Thr Ser
35 40 45
Val Lys Lys Ala Glu Gln Tyr Leu Leu Glu Asn Glu Thr Thr Lys Asn
50 55 60
Tyr Leu Gly Ile Asp Gly Ile Pro Glu Phe Gly Arg Cys Thr Gln Glu
65 70 75 80
Leu Leu Phe Gly Lys Gly Ser Ala Leu Ile Asn Asp Lys Arg Ala Arg
85 90 95
Thr Ala Gln Thr Pro Gly Gly Thr Gly Ala Leu Arg Val Ala Ala Asp
100 105 110
Phe Leu Ala Lys Asn Thr Ser Val Lys Arg Val Trp Val Ser Asn Pro
115 120 125
Ser Trp Pro Asn His Lys Ser Val Phe Asn Ser Ala Gly Leu Glu Val
130 135 140
Arg Glu Tyr Ala Tyr Tyr Asp Ala Glu Asn His Thr Leu Asp Phe Asp
145 150 155 160
Ala Leu Ile Asn Ser Leu Asn Glu Ala Gln Ala Gly Asp Val Val Leu
165 170 175
Phe His Gly Cys Cys His Asn Pro Thr Gly Ile Asp Pro Thr Leu Glu
180 185 190
Gln Trp Gln Thr Leu Ala Gln Leu Ser Val Glu Lys Gly Trp Leu Pro
195 200 205
Leu Phe Asp Phe Ala Tyr Gln Gly Phe Ala Arg Gly Leu Glu Glu Asp
210 215 220
Ala Glu Gly Leu Arg Ala Phe Ala Ala Met His Lys Glu Leu Ile Val
225 230 235 240
Ala Ser Ser Tyr Ser Lys Asn Phe Gly Leu Tyr Asn Glu Arg Val Gly
245 250 255
Ala Cys Thr Leu Val Ala Ala Asp Ser Glu Thr Val Asp Arg Ala Phe
260 265 270
Ser Gln Met Lys Ala Ala Ile Arg Ala Asn Tyr Ser Asn Pro Pro Ala
275 280 285
His Gly Ala Ser Val Val Ala Thr Ile Leu Ser Asn Asp Ala Leu Arg
290 295 300
Ala Ile Trp Glu Gln Glu Leu Thr Asp Met Arg Gln Arg Ile Gln Arg
305 310 315 320
Met Arg Gln Leu Phe Val Asn Thr Leu Gln Glu Lys Gly Ala Asn Arg
325 330 335
Asp Phe Ser Phe Ile Ile Lys Gln Asn Gly Met Phe Ser Phe Ser Gly
340 345 350
Leu Thr Lys Glu Gln Val Leu Arg Leu Arg Glu Glu Phe Gly Val Tyr
355 360 365
Ala Val Ala Ser Gly Arg Val Asn Val Ala Gly Met Thr Pro Asp Asn
370 375 380
Met Ala Pro Leu Cys Glu Ala Ile Val Ala Val Leu
385 390 395
<210> 17
<211> 139
<212> PRT
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 17
Met Leu Arg Thr Met Phe Lys Ser Lys Ile His Arg Ala Thr Val Thr
1 5 10 15
Gln Ala Asp Leu His Tyr Val Gly Ser Val Thr Ile Asp Ala Asp Leu
20 25 30
Leu Asp Ala Ala Asp Leu Leu Pro Gly Glu Leu Val His Ile Val Asp
35 40 45
Ile Thr Asn Gly Ala Arg Leu Glu Thr Tyr Val Ile Glu Gly Glu Arg
50 55 60
Gly Ser Gly Val Val Gly Ile Asn Gly Ala Ala Ala His Leu Val His
65 70 75 80
Pro Gly Asp Leu Val Ile Ile Ile Ser Tyr Ala Gln Val Ser Asp Ala
85 90 95
Glu Ala Arg Ala Leu Arg Pro Arg Val Val His Val Asp Arg Asp Asn
100 105 110
Arg Val Val Ala Leu Gly Ala Asp Pro Ala Glu Pro Val Pro Gly Ser
115 120 125
Asp Gln Ala Arg Ser Pro Gln Ala Val Thr Ala
130 135
<210> 18
<211> 127
<212> PRT
<213> 丙酮丁醇梭菌(Clostridium acetobutylicum)
<400> 18
Met His Leu Asn Met Leu Lys Ser Lys Ile His Arg Ala Thr Val Val
1 5 10 15
Gln Ala Asp Leu Asn Tyr Val Gly Ser Ile Thr Ile Asp Arg Asn Leu
20 25 30
Met Asp Lys Ala Asn Ile Leu Glu Tyr Glu Lys Val Glu Ile Ala Asn
35 40 45
Ile Asn Asn Gly Ala Arg Phe Glu Thr Tyr Val Ile Ala Gly Glu Ala
50 55 60
Gly Ser Gly Ile Ile Cys Leu Asn Gly Ala Ala Ala Arg Cys Ala Gln
65 70 75 80
Ala Gly Asp Lys Val Ile Ile Met Cys Tyr Cys Ser Leu Thr Pro Glu
85 90 95
Glu Ala Ser Glu His Arg Pro Lys Val Val Phe Val Asn Asp Asp Asn
100 105 110
Ser Ile Ser Asn Val Thr Glu Tyr Glu Lys His Gly Thr Ile Gly
115 120 125
<210> 19
<211> 1404
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 19
atgtctatta gtgaaaaata ttttcctcaa gaacctcaat ctcctgaatt gaaaactgca 60
attccaggtc ctcaatcaaa ggcaaagctc gaggaattat ctgctgtcta tgatacaaag 120
gctgcatatt ttgttaccga ctactacaaa tctcttggta actatattgt ggatgcagat 180
ggcaacaagc tactagattc ttattgccaa atctcttcta tcgcattggg ttacaataat 240
ccagcattat taaaagtagc acattctgat gaaatgacag ttgctttatg taacagacct 300
gctttggcat gttttccatc cactgattac tatgaaatac taaagaaggg attgttgtcc 360
gttgctccaa agggattaga taaggtttgt actgcacaca cgggatctga tgccaatgaa 420
atggcattta aggctgcatt tttgtttcaa gcaagtaaga agagaggtga caaaccattt 480
accagcgaag agctggaatc tgtcatggag aacaagttgc caggcacctc tgacatggtt 540
atcctgtcat ttgaaaaagg gttccatggt agattgtttg gatctttatc taccactaga 600
tctaaagcta ttcacaaact ggatattcct gcgtttgaat ggccaaaggc tccattccct 660
cagttaaagt atcctctgga tcaattccaa gctgaaaaca aagcagaaga agaaagatgt 720
ttgaaggctt tagaggaaat tattgtcaac tctcctgcca aaattgcagc tgcaatcatt 780
gaaccggtcc aatctgaagg tggtgataat catgcttcac cagaattctt ccaaggtatt 840
agagaaatca ccaaaaagca cggtgtcatt cttattgttg atgaagttca aacaggaggt 900
ggtgcttctg gtaagatgtg gttacatgaa cactatggca ttgtcccaga catcatgact 960
ttttctaaaa aaatgcaaaa tgcaggtttc tttttcagtg aagcaggtct tgctggggac 1020
caaccattca gacaattcaa tacctggtgc ggtgatccat caaaagctct aattgcaaga 1080
accataattg aagaaattaa agataagaac ctattgacta gtgttaccga aacaggtgac 1140
tacctatatt caaagctcga agcaatttca gcaaagtatg acaaaatgat caacttgaga 1200
ggtaagggaa gaggtttctt tattgcattt gatgccccaa caccggagtt aagaaacaaa 1260
tttattgctg aatgtaagaa attaggttta aacattggtg gatgcggtga acaaggtgtt 1320
agattgagac ctgcattagt ttttgaaaag aagcatgctg atatcttagc ctccattatt 1380
gatcaagctt tttccaaaat ttaa 1404
<210> 20
<211> 467
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 20
Met Ser Ile Ser Glu Lys Tyr Phe Pro Gln Glu Pro Gln Ser Pro Glu
1 5 10 15
Leu Lys Thr Ala Ile Pro Gly Pro Gln Ser Lys Ala Lys Leu Glu Glu
20 25 30
Leu Ser Ala Val Tyr Asp Thr Lys Ala Ala Tyr Phe Val Thr Asp Tyr
35 40 45
Tyr Lys Ser Leu Gly Asn Tyr Ile Val Asp Ala Asp Gly Asn Lys Leu
50 55 60
Leu Asp Ser Tyr Cys Gln Ile Ser Ser Ile Ala Leu Gly Tyr Asn Asn
65 70 75 80
Pro Ala Leu Leu Lys Val Ala His Ser Asp Glu Met Thr Val Ala Leu
85 90 95
Cys Asn Arg Pro Ala Leu Ala Cys Phe Pro Ser Thr Asp Tyr Tyr Glu
100 105 110
Ile Leu Lys Lys Gly Leu Leu Ser Val Ala Pro Lys Gly Leu Asp Lys
115 120 125
Val Cys Thr Ala His Thr Gly Ser Asp Ala Asn Glu Met Ala Phe Lys
130 135 140
Ala Ala Phe Leu Phe Gln Ala Ser Lys Lys Arg Gly Asp Lys Pro Phe
145 150 155 160
Thr Ser Glu Glu Leu Glu Ser Val Met Glu Asn Lys Leu Pro Gly Thr
165 170 175
Ser Asp Met Val Ile Leu Ser Phe Glu Lys Gly Phe His Gly Arg Leu
180 185 190
Phe Gly Ser Leu Ser Thr Thr Arg Ser Lys Ala Ile His Lys Leu Asp
195 200 205
Ile Pro Ala Phe Glu Trp Pro Lys Ala Pro Phe Pro Gln Leu Lys Tyr
210 215 220
Pro Leu Asp Gln Phe Gln Ala Glu Asn Lys Ala Glu Glu Glu Arg Cys
225 230 235 240
Leu Lys Ala Leu Glu Glu Ile Ile Val Asn Ser Pro Ala Lys Ile Ala
245 250 255
Ala Ala Ile Ile Glu Pro Val Gln Ser Glu Gly Gly Asp Asn His Ala
260 265 270
Ser Pro Glu Phe Phe Gln Gly Ile Arg Glu Ile Thr Lys Lys His Gly
275 280 285
Val Ile Leu Ile Val Asp Glu Val Gln Thr Gly Gly Gly Ala Ser Gly
290 295 300
Lys Met Trp Leu His Glu His Tyr Gly Ile Val Pro Asp Ile Met Thr
305 310 315 320
Phe Ser Lys Lys Met Gln Asn Ala Gly Phe Phe Phe Ser Glu Ala Gly
325 330 335
Leu Ala Gly Asp Gln Pro Phe Arg Gln Phe Asn Thr Trp Cys Gly Asp
340 345 350
Pro Ser Lys Ala Leu Ile Ala Arg Thr Ile Ile Glu Glu Ile Lys Asp
355 360 365
Lys Asn Leu Leu Thr Ser Val Thr Glu Thr Gly Asp Tyr Leu Tyr Ser
370 375 380
Lys Leu Glu Ala Ile Ser Ala Lys Tyr Asp Lys Met Ile Asn Leu Arg
385 390 395 400
Gly Lys Gly Arg Gly Phe Phe Ile Ala Phe Asp Ala Pro Thr Pro Glu
405 410 415
Leu Arg Asn Lys Phe Ile Ala Glu Cys Lys Lys Leu Gly Leu Asn Ile
420 425 430
Gly Gly Cys Gly Glu Gln Gly Val Arg Leu Arg Pro Ala Leu Val Phe
435 440 445
Glu Lys Lys His Ala Asp Ile Leu Ala Ser Ile Ile Asp Gln Ala Phe
450 455 460
Ser Lys Ile
465
<210> 21
<211> 475
<212> PRT
<213> 克鲁维酵母(Saccharomyces kluyveri)
<400> 21
Met Pro Ser Tyr Ser Val Ala Glu Leu Tyr Tyr Pro Asp Glu Pro Thr
1 5 10 15
Glu Pro Lys Ile Ser Thr Ser Ser Tyr Pro Gly Pro Lys Ala Lys Gln
20 25 30
Glu Leu Glu Lys Leu Ser Asn Val Phe Asp Thr Arg Ala Ala Tyr Leu
35 40 45
Leu Ala Asp Tyr Tyr Lys Ser Arg Gly Asn Tyr Ile Val Asp Gln Asp
50 55 60
Gly Asn Val Leu Leu Asp Val Tyr Ala Gln Ile Ser Ser Ile Ala Leu
65 70 75 80
Gly Tyr Asn Asn Pro Glu Ile Leu Lys Val Ala Lys Ser Asp Ala Met
85 90 95
Ser Val Ala Leu Ala Asn Arg Pro Ala Leu Ala Cys Phe Pro Ser Asn
100 105 110
Asp Tyr Gly Gln Leu Leu Glu Asp Gly Leu Leu Lys Ala Ala Pro Gln
115 120 125
Gly Gln Asp Lys Ile Trp Thr Ala Leu Ser Gly Ser Asp Ala Asn Glu
130 135 140
Thr Ala Phe Lys Ala Cys Phe Met Tyr Gln Ala Ala Lys Lys Arg Asn
145 150 155 160
Gly Arg Ser Phe Ser Thr Glu Glu Leu Glu Ser Val Met Asp Asn Gln
165 170 175
Leu Pro Gly Thr Ser Glu Met Val Ile Cys Ser Phe Glu Lys Gly Phe
180 185 190
His Gly Arg Leu Phe Gly Ser Leu Ser Thr Thr Arg Ser Lys Pro Ile
195 200 205
His Lys Leu Asp Ile Pro Ala Phe Asn Trp Pro Lys Ala Pro Phe Pro
210 215 220
Asp Leu Lys Tyr Pro Leu Glu Glu Asn Lys Glu Ala Asn Lys Ala Glu
225 230 235 240
Glu Ser Ser Cys Ile Glu Lys Phe Ser Gln Ile Val Gln Glu Trp Gln
245 250 255
Gly Lys Ile Ala Ala Val Ile Ile Glu Pro Ile Gln Ser Glu Gly Gly
260 265 270
Asp Asn His Ala Ser Ser Asp Phe Phe Gln Lys Leu Arg Glu Ile Thr
275 280 285
Ile Glu Asn Gly Ile Leu Met Ile Val Asp Glu Val Gln Thr Gly Val
290 295 300
Gly Ala Thr Gly Lys Met Trp Ala His Glu His Trp Asn Leu Ser Asn
305 310 315 320
Pro Pro Asp Leu Val Thr Phe Ser Lys Lys Phe Gln Ala Ala Gly Phe
325 330 335
Tyr Tyr His Asp Pro Lys Leu Gln Pro Asp Gln Pro Phe Arg Gln Phe
340 345 350
Asn Thr Trp Cys Gly Asp Pro Ser Lys Ala Leu Ile Ala Lys Val Ile
355 360 365
Tyr Glu Glu Ile Val Lys His Asp Leu Val Thr Arg Thr Ala Glu Val
370 375 380
Gly Asn Tyr Leu Phe Asn Arg Leu Glu Lys Leu Phe Glu Gly Lys Asn
385 390 395 400
Tyr Ile Gln Asn Leu Arg Gly Lys Gly Gln Gly Thr Tyr Ile Ala Phe
405 410 415
Asp Phe Gly Thr Ser Ser Glu Arg Asp Ser Phe Leu Ser Arg Leu Arg
420 425 430
Cys Asn Gly Ala Asn Val Ala Gly Cys Gly Asp Ser Ala Val Arg Leu
435 440 445
Arg Pro Ser Leu Thr Phe Glu Glu Lys His Ala Asp Val Leu Val Ser
450 455 460
Ile Phe Asp Lys Thr Leu Arg Gln Leu Tyr Gly
465 470 475
<210> 22
<211> 451
<212> PRT
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 22
Met Thr Pro Gln Pro Asn Pro Gln Val Gly Ala Ala Val Lys Ala Ala
1 5 10 15
Asp Arg Ala His Val Phe His Ser Trp Ser Ala Gln Glu Leu Ile Asp
20 25 30
Pro Leu Ala Val Ala Gly Ala Glu Gly Ser Tyr Phe Trp Asp Tyr Asp
35 40 45
Gly Arg Arg Tyr Leu Asp Phe Thr Ser Gly Leu Val Phe Thr Asn Ile
50 55 60
Gly Tyr Gln His Pro Lys Val Val Ala Ala Ile Gln Glu Gln Ala Ala
65 70 75 80
Ser Leu Thr Thr Phe Ala Pro Ala Phe Ala Val Glu Ala Arg Ser Glu
85 90 95
Ala Ala Arg Leu Ile Ala Glu Arg Thr Pro Gly Asp Leu Asp Lys Ile
100 105 110
Phe Phe Thr Asn Gly Gly Ala Asp Ala Ile Glu His Ala Val Arg Met
115 120 125
Ala Arg Ile His Thr Gly Arg Pro Lys Val Leu Ser Ala Tyr Arg Ser
130 135 140
Tyr His Gly Gly Thr Gln Gln Ala Val Asn Ile Thr Gly Asp Pro Arg
145 150 155 160
Arg Trp Ala Ser Asp Ser Ala Ser Ala Gly Val Val His Phe Trp Ala
165 170 175
Pro Tyr Leu Tyr Arg Ser Arg Phe Tyr Ala Glu Thr Glu Gln Gln Glu
180 185 190
Cys Glu Arg Ala Leu Glu His Leu Glu Thr Thr Ile Ala Phe Glu Gly
195 200 205
Pro Gly Thr Ile Ala Ala Ile Val Leu Glu Thr Val Pro Gly Thr Ala
210 215 220
Gly Ile Met Val Pro Pro Pro Gly Tyr Leu Ala Gly Val Arg Glu Leu
225 230 235 240
Cys Asp Lys Tyr Gly Ile Val Phe Val Leu Asp Glu Val Met Ala Gly
245 250 255
Phe Gly Arg Thr Gly Glu Trp Phe Ala Ala Asp Leu Phe Asp Val Thr
260 265 270
Pro Asp Leu Met Thr Phe Ala Lys Gly Val Asn Ser Gly Tyr Val Pro
275 280 285
Leu Gly Gly Val Ala Ile Ser Gly Lys Ile Ala Glu Thr Phe Gly Lys
290 295 300
Arg Ala Tyr Pro Gly Gly Leu Thr Tyr Ser Gly His Pro Leu Ala Cys
305 310 315 320
Ala Ala Ala Val Ala Thr Ile Asn Val Met Ala Glu Glu Gly Val Val
325 330 335
Glu Asn Ala Ala Asn Leu Gly Ala Arg Val Ile Glu Pro Gly Leu Arg
340 345 350
Glu Leu Ala Glu Arg His Pro Ser Val Gly Glu Val Arg Gly Val Gly
355 360 365
Met Phe Trp Ala Leu Glu Leu Val Lys Asp Arg Glu Thr Arg Glu Pro
370 375 380
Leu Val Pro Tyr Asn Ala Ala Gly Glu Ala Asn Ala Pro Met Ala Ala
385 390 395 400
Phe Gly Ala Ala Ala Lys Ala Asn Gly Leu Trp Pro Phe Ile Asn Met
405 410 415
Asn Arg Thr His Val Val Pro Pro Cys Asn Val Thr Glu Ala Glu Ala
420 425 430
Lys Glu Gly Leu Ala Ala Leu Asp Ala Ala Leu Ser Val Ala Asp Glu
435 440 445
Tyr Thr Val
450
<210> 23
<211> 419
<212> PRT
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 23
Met Ser Ala Leu Ser Pro His Leu Arg Gln Ala Thr Pro Val Val Ala
1 5 10 15
Val Arg Gly Glu Gly Val His Leu Tyr Gly Glu Asp Gly Arg Arg Tyr
20 25 30
Leu Asp Phe Thr Ala Gly Ile Gly Val Thr Ser Thr Gly His Cys His
35 40 45
Pro Arg Val Val Ala Ala Ala Gln Glu Gln Ala Gly Thr Leu Val His
50 55 60
Gly Gln Tyr Thr Thr Val Leu His Pro Pro Leu Arg Arg Leu Val Asp
65 70 75 80
Arg Leu Gly Glu Val Leu Pro Ala Gly Leu Asp Ser Leu Phe Phe Thr
85 90 95
Asn Ser Gly Ser Glu Ala Val Glu Ala Ala Leu Arg Leu Ala Arg Gln
100 105 110
Ala Thr Gly Arg Pro Asn Val Leu Val Cys His Gly Gly Phe His Gly
115 120 125
Arg Thr Val Ala Ala Ala Ala Met Thr Thr Ser Gly Thr Arg Phe Arg
130 135 140
Ser Gly Phe Ser Pro Leu Met Ser Gly Val Val Val Thr Pro Phe Pro
145 150 155 160
Thr Ala Phe Arg Tyr Gly Trp Asp Glu Glu Thr Ala Thr Arg Phe Ala
165 170 175
Leu Gln Glu Leu Asp Tyr Thr Leu Arg Thr Ile Ser Ser Pro Asp Asp
180 185 190
Thr Ala Ala Ile Ile Val Glu Pro Val Leu Gly Glu Gly Gly Tyr Val
195 200 205
Pro Ala Thr Arg Ala Phe Leu Glu Gly Leu Arg Glu Arg Ala Asp Arg
210 215 220
His Gly Phe Val Leu Ile Leu Asp Glu Val Gln Thr Gly Val Gly Arg
225 230 235 240
Thr Gly Arg Phe Trp Gly His Asp His Phe Gly Val Thr Pro Asp Ile
245 250 255
Leu Ile Thr Ala Lys Gly Leu Ala Ser Gly Phe Pro Leu Ser Gly Ile
260 265 270
Ala Ala Ser Ala Glu Leu Met Gly Lys Ala Trp Pro Gly Ser Gln Gly
275 280 285
Gly Thr Tyr Gly Ala Asn Ala Val Ala Cys Ala Ala Ala Cys Ala Thr
290 295 300
Leu Asp Val Val Arg Asp Glu Lys Leu Val Asp Asn Ala Glu Ala Met
305 310 315 320
Gly Ala Arg Leu Arg Ala Gly Leu Ala Ala Val Ala Ala Thr Thr Pro
325 330 335
Ala Ile Gly Asp Val Arg Gly Leu Gly Leu Met Leu Ala Ser Glu Phe
340 345 350
Val Thr Glu Asp Gly Gly Pro Asp Pro Glu Thr Ala Ala Arg Val Gln
355 360 365
Arg Ala Ala Val Asp Glu Gly Leu Leu Leu Leu Leu Cys Gly Ala Trp
370 375 380
Asn Gln Val Val Arg Met Ile Pro Ala Leu Val Ile Asp Glu Ala Glu
385 390 395 400
Val Asp Glu Gly Leu Arg Ala Trp Ser Ala Ala Val Glu Val Gly Val
405 410 415
Pro Ala Arg
<210> 24
<211> 471
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 24
Met Ser Ile Cys Glu Gln Tyr Tyr Pro Glu Glu Pro Thr Lys Pro Thr
1 5 10 15
Val Lys Thr Glu Ser Ile Pro Gly Pro Glu Ser Gln Lys Gln Leu Lys
20 25 30
Glu Leu Gly Glu Val Phe Asp Thr Arg Pro Ala Tyr Phe Leu Ala Asp
35 40 45
Tyr Glu Lys Ser Leu Gly Asn Tyr Ile Thr Asp Val Asp Gly Asn Thr
50 55 60
Tyr Leu Asp Leu Tyr Ala Gln Ile Ser Ser Ile Ala Leu Gly Tyr Asn
65 70 75 80
Asn Pro Ala Leu Ile Lys Ala Ala Gln Ser Pro Glu Met Ile Arg Ala
85 90 95
Leu Val Asp Arg Pro Ala Leu Gly Asn Phe Pro Ser Lys Asp Leu Asp
100 105 110
Lys Ile Leu Lys Gln Ile Leu Lys Ser Ala Pro Lys Gly Gln Asp His
115 120 125
Val Trp Ser Gly Leu Ser Gly Ala Asp Ala Asn Glu Leu Ala Phe Lys
130 135 140
Ala Ala Phe Ile Tyr Tyr Arg Ala Lys Gln Arg Gly Tyr Asp Ala Asp
145 150 155 160
Phe Ser Glu Lys Glu Asn Leu Ser Val Met Asp Asn Asp Ala Pro Gly
165 170 175
Ala Pro His Leu Ala Val Leu Ser Phe Lys Arg Ala Phe His Gly Arg
180 185 190
Leu Phe Ala Ser Gly Ser Thr Thr Cys Ser Lys Pro Ile His Lys Leu
195 200 205
Asp Phe Pro Ala Phe His Trp Pro His Ala Glu Tyr Pro Ser Tyr Gln
210 215 220
Tyr Pro Leu Asp Glu Asn Ser Asp Ala Asn Arg Lys Glu Asp Asp His
225 230 235 240
Cys Leu Ala Ile Val Glu Glu Leu Ile Lys Thr Trp Ser Ile Pro Val
245 250 255
Ala Ala Leu Ile Ile Glu Pro Ile Gln Ser Glu Gly Gly Asp Asn His
260 265 270
Ala Ser Lys Tyr Phe Leu Gln Lys Leu Arg Asp Ile Thr Leu Lys Tyr
275 280 285
Asn Val Val Tyr Ile Ile Asp Glu Val Gln Thr Gly Val Gly Ala Thr
290 295 300
Gly Lys Leu Trp Cys His Glu Tyr Ala Asp Ile Gln Pro Pro Val Asp
305 310 315 320
Leu Val Thr Phe Ser Lys Lys Phe Gln Ser Ala Gly Tyr Phe Phe His
325 330 335
Asp Pro Lys Phe Ile Pro Asn Lys Pro Tyr Arg Gln Phe Asn Thr Trp
340 345 350
Cys Gly Glu Pro Ala Arg Met Ile Ile Ala Gly Ala Ile Gly Gln Glu
355 360 365
Ile Ser Asp Lys Lys Leu Thr Glu Gln Cys Ser Arg Val Gly Asp Tyr
370 375 380
Leu Phe Lys Lys Leu Glu Gly Leu Gln Lys Lys Tyr Pro Glu Asn Phe
385 390 395 400
Gln Asn Leu Arg Gly Lys Gly Arg Gly Thr Phe Ile Ala Trp Asp Leu
405 410 415
Pro Thr Gly Glu Lys Arg Asp Leu Leu Leu Lys Lys Leu Lys Leu Asn
420 425 430
Gly Cys Asn Val Gly Gly Cys Ala Val His Ala Val Arg Leu Arg Pro
435 440 445
Ser Leu Thr Phe Glu Glu Lys His Ala Asp Ile Phe Ile Glu Ala Leu
450 455 460
Ala Lys Ser Val Asn Glu Leu
465 470
<210> 25
<211> 813
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 25
atgtttggta atatttccca aagacttgca ggcaagaaca tcctaattac aggtgcgtcc 60
actggtatcg gataccatac agcaaagtat tttgcagaag ctgcaaatgg agacttgaag 120
ttggttttgg ctgcaagaag aaaggagaag ctggaggcac taaaggcaga cttgcttgcc 180
aagtatccat ccatcaaagt ccatattgag agtttggatg tctccaaaac ggaaaccatt 240
gcacctttct taaaaggttt acctgaggaa ttttcaattg tcgacgtgtt ggtcaacaat 300
gcaggtaagg cgcttggttt ggatccaatt ggctctgtcg atccaaagga cgtggatgaa 360
atgttccaga ccaatgtttt gggtatgatt caattgaccc agttggttgt acagcaaatg 420
aaggagagaa actccgggga cattgtccaa ctaggttcag tggctggtag aaacccatac 480
ccaggtggtg gtatctactg tgcctccaag gccgcattga gatcttttac acatgtattg 540
agagaggaat tgattaatac caagattaga gtgattgaaa tcgagcctgg aaatgttgca 600
actgaggaat tttctttgac cagattcaaa ggtgataagt ccaaggccga aaaggtctat 660
gagggaaccg agccattgta tggtaccgat attgcagaat tgattctatt tgcagtttct 720
agacctcaaa acactgttat tgcagaaaca cttgtttttg ctagtaacca agcttctgct 780
taccatattt tcagaggatc attagataaa tag 813
<210> 26
<211> 270
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 26
Met Phe Gly Asn Ile Ser Gln Arg Leu Ala Gly Lys Asn Ile Leu Ile
1 5 10 15
Thr Gly Ala Ser Thr Gly Ile Gly Tyr His Thr Ala Lys Tyr Phe Ala
20 25 30
Glu Ala Ala Asn Gly Asp Leu Lys Leu Val Leu Ala Ala Arg Arg Lys
35 40 45
Glu Lys Leu Glu Ala Leu Lys Ala Asp Leu Leu Ala Lys Tyr Pro Ser
50 55 60
Ile Lys Val His Ile Glu Ser Leu Asp Val Ser Lys Thr Glu Thr Ile
65 70 75 80
Ala Pro Phe Leu Lys Gly Leu Pro Glu Glu Phe Ser Ile Val Asp Val
85 90 95
Leu Val Asn Asn Ala Gly Lys Ala Leu Gly Leu Asp Pro Ile Gly Ser
100 105 110
Val Asp Pro Lys Asp Val Asp Glu Met Phe Gln Thr Asn Val Leu Gly
115 120 125
Met Ile Gln Leu Thr Gln Leu Val Val Gln Gln Met Lys Glu Arg Asn
130 135 140
Ser Gly Asp Ile Val Gln Leu Gly Ser Val Ala Gly Arg Asn Pro Tyr
145 150 155 160
Pro Gly Gly Gly Ile Tyr Cys Ala Ser Lys Ala Ala Leu Arg Ser Phe
165 170 175
Thr His Val Leu Arg Glu Glu Leu Ile Asn Thr Lys Ile Arg Val Ile
180 185 190
Glu Ile Glu Pro Gly Asn Val Ala Thr Glu Glu Phe Ser Leu Thr Arg
195 200 205
Phe Lys Gly Asp Lys Ser Lys Ala Glu Lys Val Tyr Glu Gly Thr Glu
210 215 220
Pro Leu Tyr Gly Thr Asp Ile Ala Glu Leu Ile Leu Phe Ala Val Ser
225 230 235 240
Arg Pro Gln Asn Thr Val Ile Ala Glu Thr Leu Val Phe Ala Ser Asn
245 250 255
Gln Ala Ser Ala Tyr His Ile Phe Arg Gly Ser Leu Asp Lys
260 265 270
<210> 27
<211> 248
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 27
Met Ile Val Leu Val Thr Gly Ala Thr Ala Gly Phe Gly Glu Cys Ile
1 5 10 15
Thr Arg Arg Phe Ile Gln Gln Gly His Lys Val Ile Ala Thr Gly Arg
20 25 30
Arg Gln Glu Arg Leu Gln Glu Leu Lys Asp Glu Leu Gly Asp Asn Leu
35 40 45
Tyr Ile Ala Gln Leu Asp Val Arg Asn Arg Ala Ala Ile Glu Glu Met
50 55 60
Leu Ala Ser Leu Pro Ala Glu Trp Cys Asn Ile Asp Ile Leu Val Asn
65 70 75 80
Asn Ala Gly Leu Ala Leu Gly Met Glu Pro Ala His Lys Ala Ser Val
85 90 95
Glu Asp Trp Glu Thr Met Ile Asp Thr Asn Asn Lys Gly Leu Val Tyr
100 105 110
Met Thr Arg Ala Val Leu Pro Gly Met Val Glu Arg Asn His Gly His
115 120 125
Ile Ile Asn Ile Gly Ser Thr Ala Gly Ser Trp Pro Tyr Ala Gly Gly
130 135 140
Asn Val Tyr Gly Ala Thr Lys Ala Phe Val Arg Gln Phe Ser Leu Asn
145 150 155 160
Leu Arg Thr Asp Leu His Gly Thr Ala Val Arg Val Thr Asp Ile Glu
165 170 175
Pro Gly Leu Val Gly Gly Thr Glu Phe Ser Asn Val Arg Phe Lys Gly
180 185 190
Asp Asp Gly Lys Ala Glu Lys Thr Tyr Gln Asn Thr Val Ala Leu Thr
195 200 205
Pro Glu Asp Val Ser Glu Ala Val Trp Trp Val Ser Thr Leu Pro Ala
210 215 220
His Val Asn Ile Asn Thr Leu Glu Met Met Pro Val Thr Gln Ser Tyr
225 230 235 240
Ala Gly Leu Asn Val His Arg Gln
245
<210> 28
<211> 298
<212> PRT
<213> 粪产碱菌(Alcaligenes faecalis)
<400> 28
Met Ser Asn Thr Ile Ala Phe Ile Gly Leu Gly His Met Gly Lys Pro
1 5 10 15
Met Ala Leu Asn Leu Leu Lys Ala Gly His Ser Leu Asn Val Phe Asp
20 25 30
Leu Asn Ala Gln Ala Met Gln Glu Leu Gln Ala Ala Gly Ala Gln Val
35 40 45
Gly Glu Ser Ala Val Gln Ile Ala Gln Asp Ala Gln Met Val Phe Thr
50 55 60
Met Leu Pro Ala Gly Arg His Val Arg Gln Val Tyr Glu Gly Glu Asn
65 70 75 80
Gly Leu Leu Gln Thr Val Ala Pro Gly Thr Val Leu Val Asp Cys Ser
85 90 95
Thr Ile Asp Ala Gln Thr Ser Gln Asp Leu Ala Ala Lys Ala Ser Lys
100 105 110
Leu Gly Leu Phe Met Leu Asp Ala Pro Val Ser Gly Gly Thr Gly Gly
115 120 125
Ala Ile Ala Gly Thr Leu Thr Phe Met Val Gly Gly Glu Asp Gln Ala
130 135 140
Leu Glu Lys Ala Arg Pro Tyr Leu Asp Ala Met Gly Lys Asn Ile Phe
145 150 155 160
His Ala Gly Lys Ala Gly Ala Gly Gln Val Ala Lys Ile Cys Asn Asn
165 170 175
Met Leu Leu Gly Ile Leu Met Ala Gly Thr Ala Glu Ala Leu Ala Leu
180 185 190
Gly Val Ala His Gly Leu Asp Pro Ala Val Leu Ser Thr Ile Met Ala
195 200 205
Arg Ser Ser Gly Arg Asn Trp Ala Thr Glu Leu Tyr Asn Pro Trp Pro
210 215 220
Gly Val Met Pro Asp Val Pro Ala Ser Arg Asp Tyr Gln Gly Gly Phe
225 230 235 240
Ala Thr Gly Leu Met Leu Lys Asp Leu Gly Leu Ala Ala Asp Ala Ala
245 250 255
Val Ser Gln Asn Ser Ala Thr Pro Leu Gly Glu Leu Ala Arg Asn Leu
260 265 270
Phe Ala Leu His Ala Ala Gln Gly Gln Asn Ala Gly Leu Asp Phe Ser
275 280 285
Ser Ile Leu Asn Leu Tyr Arg Gln Lys His
290 295
<210> 29
<211> 314
<212> PRT
<213> 勤奋生金球菌(Metallosphaera sedula)
<400> 29
Met Thr Glu Lys Val Ser Val Val Gly Ala Gly Val Ile Gly Val Gly
1 5 10 15
Trp Ala Thr Leu Phe Ala Ser Lys Gly Tyr Ser Val Ser Leu Tyr Thr
20 25 30
Glu Lys Lys Glu Thr Leu Asp Lys Gly Ile Glu Lys Leu Arg Asn Tyr
35 40 45
Val Gln Val Met Lys Asn Asn Ser Gln Ile Thr Glu Asp Val Asn Thr
50 55 60
Val Ile Ser Arg Val Ser Pro Thr Thr Asn Leu Asp Glu Ala Val Arg
65 70 75 80
Gly Ala Asn Phe Val Ile Glu Ala Val Ile Glu Asp Tyr Asp Ala Lys
85 90 95
Lys Lys Ile Phe Gly Tyr Leu Asp Ser Val Leu Asp Lys Glu Val Ile
100 105 110
Leu Ala Ser Ser Thr Ser Gly Leu Leu Ile Thr Glu Val Gln Lys Ala
115 120 125
Met Ser Lys His Pro Glu Arg Ala Val Ile Ala His Pro Trp Asn Pro
130 135 140
Pro His Leu Leu Pro Leu Val Glu Ile Val Pro Gly Glu Lys Thr Ser
145 150 155 160
Met Glu Val Val Glu Arg Thr Lys Ser Leu Met Glu Lys Leu Asp Arg
165 170 175
Ile Val Val Val Leu Lys Lys Glu Ile Pro Gly Phe Ile Gly Asn Arg
180 185 190
Leu Ala Phe Ala Leu Phe Arg Glu Ala Val Tyr Leu Val Asp Glu Gly
195 200 205
Val Ala Thr Val Glu Asp Ile Asp Lys Val Met Thr Ala Ala Ile Gly
210 215 220
Leu Arg Trp Ala Phe Met Gly Pro Phe Leu Thr Tyr His Leu Gly Gly
225 230 235 240
Gly Glu Gly Gly Leu Glu Tyr Phe Phe Asn Arg Gly Phe Gly Tyr Gly
245 250 255
Ala Asn Glu Trp Met His Thr Leu Ala Lys Tyr Asp Lys Phe Pro Tyr
260 265 270
Thr Gly Val Thr Lys Ala Ile Gln Gln Met Lys Glu Tyr Ser Phe Ile
275 280 285
Lys Gly Lys Thr Phe Gln Glu Ile Ser Lys Trp Arg Asp Glu Lys Leu
290 295 300
Leu Lys Val Tyr Lys Leu Val Trp Glu Lys
305 310
<210> 30
<211> 295
<212> PRT
<213> 恶臭假单胞菌(Pseudomonas putida)
<400> 30
Met Arg Ile Ala Phe Ile Gly Leu Gly Asn Met Gly Ala Pro Met Ala
1 5 10 15
Arg Asn Leu Ile Lys Ala Gly His Gln Leu Asn Leu Phe Asp Leu Asn
20 25 30
Lys Ala Val Leu Ala Glu Leu Ala Glu Leu Gly Gly Gln Ile Ser Pro
35 40 45
Ser Pro Lys Asp Ala Ala Ala Asn Ser Glu Leu Val Ile Thr Met Leu
50 55 60
Pro Ala Ala Ala His Val Arg Ser Val Tyr Leu Asn Glu Asp Gly Val
65 70 75 80
Leu Ala Gly Ile Arg Pro Gly Thr Pro Thr Val Asp Cys Ser Thr Ile
85 90 95
Asp Pro Gln Thr Ala Arg Asp Val Ser Lys Ala Ala Ala Ala Lys Gly
100 105 110
Val Asp Met Gly Asp Ala Pro Val Ser Gly Gly Thr Gly Gly Ala Ala
115 120 125
Ala Gly Thr Leu Thr Phe Met Val Gly Ala Ser Thr Glu Leu Phe Ala
130 135 140
Ser Leu Lys Pro Val Leu Glu Gln Met Gly Arg Asn Ile Val His Cys
145 150 155 160
Gly Glu Val Gly Thr Gly Gln Ile Ala Lys Ile Cys Asn Asn Leu Leu
165 170 175
Leu Gly Ile Ser Met Ile Gly Val Ser Glu Ala Met Ala Leu Gly Asn
180 185 190
Ala Leu Gly Ile Asp Thr Lys Val Leu Ala Gly Ile Ile Asn Ser Ser
195 200 205
Thr Gly Arg Cys Trp Ser Ser Asp Thr Tyr Asn Pro Trp Pro Gly Ile
210 215 220
Ile Glu Thr Ala Pro Ala Ser Arg Gly Tyr Thr Gly Gly Phe Gly Ala
225 230 235 240
Glu Leu Met Leu Lys Asp Leu Gly Leu Ala Thr Glu Ala Ala Arg Gln
245 250 255
Ala His Gln Pro Val Ile Leu Gly Ala Val Ala Gln Gln Leu Tyr Gln
260 265 270
Ala Met Ser Leu Arg Gly Glu Gly Gly Lys Asp Phe Ser Ala Ile Val
275 280 285
Glu Gly Tyr Arg Lys Lys Asp
290 295
<210> 31
<211> 295
<212> PRT
<213> 恶臭假单胞菌(Pseudomonas putida)
<400> 31
Met Arg Ile Ala Phe Ile Gly Leu Gly Asn Met Gly Ala Pro Met Ala
1 5 10 15
Arg Asn Leu Ile Lys Ala Gly His Gln Leu Asn Leu Phe Asp Leu Asn
20 25 30
Lys Thr Val Leu Ala Glu Leu Ala Glu Leu Gly Gly Gln Ile Ser Pro
35 40 45
Ser Pro Lys Asp Ala Ala Ala Ser Ser Glu Leu Val Ile Thr Met Leu
50 55 60
Pro Ala Ala Ala His Val Arg Ser Val Tyr Leu Asn Asp Asp Gly Val
65 70 75 80
Leu Ala Gly Ile Arg Pro Gly Thr Pro Thr Val Asp Cys Ser Thr Ile
85 90 95
Asp Pro Gln Thr Ala Arg Asp Val Ser Lys Ala Ala Ala Ala Lys Gly
100 105 110
Val Asp Met Gly Asp Ala Pro Val Ser Gly Gly Thr Gly Gly Ala Ala
115 120 125
Ala Gly Thr Leu Thr Phe Met Val Gly Ala Ser Ala Glu Leu Phe Ala
130 135 140
Ser Leu Lys Pro Val Leu Glu Gln Met Gly Arg Asn Ile Val His Cys
145 150 155 160
Gly Glu Val Gly Thr Gly Gln Ile Ala Lys Ile Cys Asn Asn Leu Leu
165 170 175
Leu Gly Ile Ser Met Ile Gly Val Ser Glu Ala Met Ala Leu Gly Asn
180 185 190
Ala Leu Gly Ile Asp Thr Lys Val Leu Ala Gly Ile Ile Asn Ser Ser
195 200 205
Thr Gly Arg Cys Trp Ser Ser Asp Thr Tyr Asn Pro Trp Pro Gly Ile
210 215 220
Ile Glu Thr Ala Pro Ala Ser Arg Gly Tyr Thr Gly Gly Phe Gly Ala
225 230 235 240
Glu Leu Met Leu Lys Asp Leu Gly Leu Ala Thr Glu Ala Ala Arg Gln
245 250 255
Ala His Gln Pro Val Ile Leu Gly Ala Val Ala Gln Gln Leu Tyr Gln
260 265 270
Ala Met Ser Leu Arg Gly Glu Gly Gly Lys Asp Phe Ser Ala Ile Val
275 280 285
Glu Gly Tyr Arg Lys Lys Asp
290 295
<210> 32
<211> 298
<212> PRT
<213> 铜绿假单胞菌(Pseudomonas aeruginosa)
<400> 32
Met Thr Asp Ile Ala Phe Leu Gly Leu Gly Asn Met Gly Gly Pro Met
1 5 10 15
Ala Ala Asn Leu Leu Lys Ala Gly His Arg Val Asn Val Phe Asp Leu
20 25 30
Gln Pro Lys Ala Val Leu Gly Leu Val Glu Gln Gly Ala Gln Gly Ala
35 40 45
Asp Ser Ala Leu Gln Cys Cys Glu Gly Ala Glu Val Val Ile Ser Met
50 55 60
Leu Pro Ala Gly Gln His Val Glu Ser Leu Tyr Leu Gly Asp Asp Gly
65 70 75 80
Leu Leu Ala Arg Val Ala Gly Lys Pro Leu Leu Ile Asp Cys Ser Thr
85 90 95
Ile Ala Pro Glu Thr Ala Arg Lys Val Ala Glu Ala Ala Ala Ala Lys
100 105 110
Gly Leu Thr Leu Leu Asp Ala Pro Val Ser Gly Gly Val Gly Gly Ala
115 120 125
Arg Ala Gly Thr Leu Ser Phe Ile Val Gly Gly Pro Ala Glu Gly Phe
130 135 140
Ala Arg Ala Arg Pro Val Leu Glu Asn Met Gly Arg Asn Ile Phe His
145 150 155 160
Ala Gly Asp His Gly Ala Gly Gln Val Ala Lys Ile Cys Asn Asn Met
165 170 175
Leu Leu Gly Ile Leu Met Ala Gly Thr Ala Glu Ala Leu Ala Leu Gly
180 185 190
Val Lys Asn Gly Leu Asp Pro Ala Val Leu Ser Glu Val Met Lys Gln
195 200 205
Ser Ser Gly Gly Asn Trp Ala Leu Asn Leu Tyr Asn Pro Trp Pro Gly
210 215 220
Val Met Pro Gln Ala Pro Ala Ser Asn Gly Tyr Ala Gly Gly Phe Gln
225 230 235 240
Val Arg Leu Met Asn Lys Asp Leu Gly Leu Ala Leu Ala Asn Ala Gln
245 250 255
Ala Val Gln Ala Ser Thr Pro Leu Gly Ala Leu Ala Arg Asn Leu Phe
260 265 270
Ser Leu His Ala Gln Ala Asp Ala Glu His Glu Gly Leu Asp Phe Ser
275 280 285
Ser Ile Gln Lys Leu Tyr Arg Gly Lys Asp
290 295
<210> 33
<211> 382
<212> PRT
<213> 真氧产碱杆菌(Ralstonia eutropha)
<400> 33
Met Ala Phe Ile Tyr Tyr Leu Thr His Ile His Leu Asp Phe Gly Ala
1 5 10 15
Val Ser Leu Leu Lys Ser Glu Cys Glu Arg Ile Gly Ile Arg Arg Pro
20 25 30
Leu Leu Val Thr Asp Lys Gly Val Val Ala Ala Gly Val Ala Gln Arg
35 40 45
Ala Ile Asp Ala Met Gln Gly Leu Gln Val Ala Val Phe Asp Glu Thr
50 55 60
Pro Ser Asn Pro Thr Glu Ala Met Val Arg Lys Ala Ala Ala Gln Tyr
65 70 75 80
Arg Glu Ala Gly Cys Asp Gly Leu Val Ala Val Gly Gly Gly Ser Ser
85 90 95
Ile Asp Leu Ala Lys Gly Ile Ala Ile Leu Ala Thr His Glu Gly Glu
100 105 110
Leu Thr Thr Tyr Ala Thr Ile Glu Gly Gly Ser Ala Arg Ile Thr Asp
115 120 125
Lys Ala Ala Pro Leu Ile Ala Val Pro Thr Thr Ser Gly Thr Gly Ser
130 135 140
Glu Val Ala Arg Gly Ala Ile Ile Ile Leu Asp Asp Gly Arg Lys Leu
145 150 155 160
Gly Phe His Ser Trp His Leu Leu Pro Lys Ser Ala Val Cys Asp Pro
165 170 175
Glu Leu Thr Leu Gly Leu Pro Ala Gly Leu Thr Ala Ala Thr Gly Met
180 185 190
Asp Ala Ile Ala His Cys Ile Glu Thr Phe Leu Ala Pro Ala Phe Asn
195 200 205
Pro Pro Ala Asp Gly Ile Ala Leu Asp Gly Leu Glu Arg Gly Trp Gly
210 215 220
His Ile Glu Arg Ala Thr Arg Asp Gly Gln Asp Arg Asp Ala Arg Leu
225 230 235 240
Asn Met Met Ser Ala Ser Met Gln Gly Ala Met Ala Phe Gln Lys Gly
245 250 255
Leu Gly Cys Val His Ser Leu Ser His Pro Leu Gly Gly Leu Lys Ile
260 265 270
Asp Gly Arg Thr Gly Leu His His Gly Thr Leu Asn Ala Val Val Met
275 280 285
Pro Ala Val Leu Arg Phe Asn Ala Asp Ala Pro Thr Val Val Arg Asp
290 295 300
Asp Arg Tyr Ala Arg Leu Arg Arg Ala Met His Leu Pro Asp Gly Ala
305 310 315 320
Asp Ile Ala Gln Ala Val His Asp Met Thr Val Arg Leu Gly Leu Pro
325 330 335
Thr Gly Leu Arg Gln Met Gly Val Thr Glu Asp Met Phe Asp Lys Val
340 345 350
Ile Ala Gly Ala Leu Val Asp His Cys His Lys Thr Asn Pro Lys Glu
355 360 365
Ala Ser Ala Ala Asp Tyr Arg Arg Met Leu Glu Gln Ser Met
370 375 380
<210> 34
<211> 371
<212> PRT
<213> 克氏梭菌(Clostridium kluyveri)
<400> 34
Met Lys Leu Leu Lys Leu Ala Pro Asp Val Tyr Lys Phe Asp Thr Ala
1 5 10 15
Glu Glu Phe Met Lys Tyr Phe Lys Val Gly Lys Gly Asp Phe Ile Leu
20 25 30
Thr Asn Glu Phe Leu Tyr Lys Pro Phe Leu Glu Lys Phe Asn Asp Gly
35 40 45
Ala Asp Ala Val Phe Gln Glu Lys Tyr Gly Leu Gly Glu Pro Ser Asp
50 55 60
Glu Met Ile Asn Asn Ile Ile Lys Asp Ile Gly Asp Lys Gln Tyr Asn
65 70 75 80
Arg Ile Ile Ala Val Gly Gly Gly Ser Val Ile Asp Ile Ala Lys Ile
85 90 95
Leu Ser Leu Lys Tyr Thr Asp Asp Ser Leu Asp Leu Phe Glu Gly Lys
100 105 110
Val Pro Leu Val Lys Asn Lys Glu Leu Ile Ile Val Pro Thr Thr Cys
115 120 125
Gly Thr Gly Ser Glu Val Thr Asn Val Ser Val Ala Glu Leu Lys Arg
130 135 140
Arg His Thr Lys Lys Gly Ile Ala Ser Asp Glu Leu Tyr Ala Thr Tyr
145 150 155 160
Ala Val Leu Val Pro Glu Phe Ile Lys Gly Leu Pro Tyr Lys Phe Phe
165 170 175
Val Thr Ser Ser Val Asp Ala Leu Ile His Ala Thr Glu Ala Tyr Val
180 185 190
Ser Pro Asn Ala Asn Pro Tyr Thr Asp Met Phe Ser Val Lys Ala Met
195 200 205
Glu Leu Ile Leu Asn Gly Tyr Met Gln Met Val Glu Lys Gly Asn Asp
210 215 220
Tyr Arg Val Glu Ile Ile Glu Asp Phe Val Ile Gly Ser Asn Tyr Ala
225 230 235 240
Gly Ile Ala Phe Gly Asn Ala Gly Val Gly Ala Val His Ala Leu Ser
245 250 255
Tyr Pro Ile Gly Gly Asn Tyr His Val Pro His Gly Glu Ala Asn Tyr
260 265 270
Leu Phe Phe Thr Glu Ile Phe Lys Thr Tyr Tyr Glu Lys Asn Pro Asn
275 280 285
Gly Lys Ile Lys Asp Val Asn Lys Leu Leu Ala Gly Ile Leu Lys Cys
290 295 300
Asp Glu Ser Glu Ala Tyr Asp Ser Leu Ser Gln Leu Leu Asp Lys Leu
305 310 315 320
Leu Ser Arg Lys Pro Leu Arg Glu Tyr Gly Met Lys Glu Glu Glu Ile
325 330 335
Glu Thr Phe Ala Asp Ser Val Ile Glu Gly Gln Gln Arg Leu Leu Val
340 345 350
Asn Asn Tyr Glu Pro Phe Ser Arg Glu Asp Ile Val Asn Thr Tyr Lys
355 360 365
Lys Leu Tyr
370
<210> 35
<211> 538
<212> PRT
<213> 产琥拍酸曼氏杆菌(Mannheimia succiniciproducens)
<400> 35
Met Thr Asp Leu Asn Gln Leu Thr Gln Glu Leu Gly Ala Leu Gly Ile
1 5 10 15
His Asp Val Gln Glu Val Val Tyr Asn Pro Ser Tyr Glu Leu Leu Phe
20 25 30
Ala Glu Glu Thr Lys Pro Gly Leu Glu Gly Tyr Glu Lys Gly Thr Val
35 40 45
Thr Asn Gln Gly Ala Val Ala Val Asn Thr Gly Ile Phe Thr Gly Arg
50 55 60
Ser Pro Lys Asp Lys Tyr Ile Val Leu Asp Asp Lys Thr Lys Asp Thr
65 70 75 80
Val Trp Trp Thr Ser Glu Lys Val Lys Asn Asp Asn Lys Pro Met Ser
85 90 95
Gln Asp Thr Trp Asn Ser Leu Lys Gly Leu Val Ala Asp Gln Leu Ser
100 105 110
Gly Lys Arg Leu Phe Val Val Asp Ala Phe Cys Gly Ala Asn Lys Asp
115 120 125
Thr Arg Leu Ala Val Arg Val Val Thr Glu Val Ala Trp Gln Ala His
130 135 140
Phe Val Thr Asn Met Phe Ile Arg Pro Ser Ala Glu Glu Leu Lys Gly
145 150 155 160
Phe Lys Pro Asp Phe Val Val Met Asn Gly Ala Lys Cys Thr Asn Pro
165 170 175
Asn Trp Lys Glu Gln Gly Leu Asn Ser Glu Asn Phe Val Ala Phe Asn
180 185 190
Ile Thr Glu Gly Val Gln Leu Ile Gly Gly Thr Trp Tyr Gly Gly Glu
195 200 205
Met Lys Lys Gly Met Phe Ser Met Met Asn Tyr Phe Leu Pro Leu Arg
210 215 220
Gly Ile Ala Ser Met His Cys Ser Ala Asn Val Gly Lys Asp Gly Asp
225 230 235 240
Thr Ala Ile Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr Thr Leu Ser
245 250 255
Thr Asp Pro Lys Arg Gln Leu Ile Gly Asp Asp Glu His Gly Trp Asp
260 265 270
Asp Glu Gly Val Phe Asn Phe Glu Gly Gly Cys Tyr Ala Lys Thr Ile
275 280 285
Asn Leu Ser Ala Glu Asn Glu Pro Asp Ile Tyr Gly Ala Ile Lys Arg
290 295 300
Asp Ala Leu Leu Glu Asn Val Val Val Leu Asp Asn Gly Asp Val Asp
305 310 315 320
Tyr Ala Asp Gly Ser Lys Thr Glu Asn Thr Arg Val Ser Tyr Pro Ile
325 330 335
Tyr His Ile Gln Asn Ile Val Lys Pro Val Ser Lys Ala Gly Pro Ala
340 345 350
Thr Lys Val Ile Phe Leu Ser Ala Asp Ala Phe Gly Val Leu Pro Pro
355 360 365
Val Ser Lys Leu Thr Pro Glu Gln Thr Lys Tyr Tyr Phe Leu Ser Gly
370 375 380
Phe Thr Ala Lys Leu Ala Gly Thr Glu Arg Gly Ile Thr Glu Pro Thr
385 390 395 400
Pro Thr Phe Ser Ala Cys Phe Gly Ala Ala Phe Leu Ser Leu His Pro
405 410 415
Thr Gln Tyr Ala Glu Val Leu Val Lys Arg Met Gln Glu Ser Gly Ala
420 425 430
Glu Ala Tyr Leu Val Asn Thr Gly Trp Asn Gly Thr Gly Lys Arg Ile
435 440 445
Ser Ile Lys Asp Thr Arg Gly Ile Ile Asp Ala Ile Leu Asp Gly Ser
450 455 460
Ile Asp Lys Ala Glu Met Gly Ser Leu Pro Ile Phe Asp Phe Ser Ile
465 470 475 480
Pro Lys Ala Leu Pro Gly Val Asn Pro Ala Ile Leu Asp Pro Arg Asp
485 490 495
Thr Tyr Ala Asp Lys Ala Gln Trp Glu Glu Lys Ala Gln Asp Leu Ala
500 505 510
Gly Arg Phe Val Lys Asn Phe Glu Lys Tyr Thr Gly Thr Ala Glu Gly
515 520 525
Gln Ala Leu Val Ala Ala Gly Pro Lys Ala
530 535
<210> 36
<211> 532
<212> PRT
<213> 产琥珀酸厌氧螺菌(Anaerobiospirillum succiniciproducens)
<400> 36
Met Ser Leu Ser Glu Ser Leu Ala Lys Tyr Gly Ile Thr Gly Ala Thr
1 5 10 15
Asn Ile Val His Asn Pro Ser His Glu Glu Leu Phe Ala Ala Glu Thr
20 25 30
Gln Ala Ser Leu Glu Gly Phe Glu Lys Gly Thr Val Thr Glu Met Gly
35 40 45
Ala Val Asn Val Met Thr Gly Val Tyr Thr Gly Arg Ser Pro Lys Asp
50 55 60
Lys Phe Ile Val Lys Asn Glu Ala Ser Lys Glu Ile Trp Trp Thr Ser
65 70 75 80
Asp Glu Phe Lys Asn Asp Asn Lys Pro Val Thr Glu Glu Ala Trp Ala
85 90 95
Gln Leu Lys Ala Leu Ala Gly Lys Glu Leu Ser Asn Lys Pro Leu Tyr
100 105 110
Val Val Asp Leu Phe Cys Gly Ala Asn Glu Asn Thr Arg Leu Lys Ile
115 120 125
Arg Phe Val Met Glu Val Ala Trp Gln Ala His Phe Val Thr Asn Met
130 135 140
Phe Ile Arg Pro Thr Glu Glu Glu Leu Lys Gly Phe Glu Pro Asp Phe
145 150 155 160
Val Val Leu Asn Ala Ser Lys Ala Lys Val Glu Asn Phe Lys Glu Leu
165 170 175
Gly Leu Asn Ser Glu Thr Ala Val Val Phe Asn Leu Ala Glu Lys Met
180 185 190
Gln Ile Ile Leu Asn Thr Trp Tyr Gly Gly Glu Met Lys Lys Gly Met
195 200 205
Phe Ser Met Met Asn Phe Tyr Leu Pro Leu Gln Gly Ile Ala Ala Met
210 215 220
His Cys Ser Ala Asn Thr Asp Leu Glu Gly Lys Asn Thr Ala Ile Phe
225 230 235 240
Phe Gly Leu Ser Gly Thr Gly Lys Thr Thr Leu Ser Thr Asp Pro Lys
245 250 255
Arg Leu Leu Ile Gly Asp Asp Glu His Gly Trp Asp Asp Asp Gly Val
260 265 270
Phe Asn Phe Glu Gly Gly Cys Tyr Ala Lys Val Ile Asn Leu Ser Lys
275 280 285
Glu Asn Glu Pro Asp Ile Trp Gly Ala Ile Lys Arg Asn Ala Leu Leu
290 295 300
Glu Asn Val Thr Val Asp Ala Asn Gly Lys Val Asp Phe Ala Asp Lys
305 310 315 320
Ser Val Thr Glu Asn Thr Arg Val Ser Tyr Pro Ile Phe His Ile Lys
325 330 335
Asn Ile Val Lys Pro Val Ser Lys Ala Pro Ala Ala Lys Arg Val Ile
340 345 350
Phe Leu Ser Ala Asp Ala Phe Gly Val Leu Pro Pro Val Ser Ile Leu
355 360 365
Ser Lys Glu Gln Thr Lys Tyr Tyr Phe Leu Ser Gly Phe Thr Ala Lys
370 375 380
Leu Ala Gly Thr Glu Arg Gly Ile Thr Glu Pro Thr Pro Thr Phe Ser
385 390 395 400
Ser Cys Phe Gly Ala Ala Phe Leu Thr Leu Pro Pro Thr Lys Tyr Ala
405 410 415
Glu Val Leu Val Lys Arg Met Glu Ala Ser Gly Ala Lys Ala Tyr Leu
420 425 430
Val Asn Thr Gly Trp Asn Gly Thr Gly Lys Arg Ile Ser Ile Lys Asp
435 440 445
Thr Arg Gly Ile Ile Asp Ala Ile Leu Asp Gly Ser Ile Asp Thr Ala
450 455 460
Asn Thr Ala Thr Ile Pro Tyr Phe Asn Phe Thr Val Pro Thr Glu Leu
465 470 475 480
Lys Gly Val Asp Thr Lys Ile Leu Asp Pro Arg Asn Thr Tyr Ala Asp
485 490 495
Ala Ser Glu Trp Glu Val Lys Ala Lys Asp Leu Ala Glu Arg Phe Gln
500 505 510
Lys Asn Phe Lys Lys Phe Glu Ser Leu Gly Gly Asp Leu Val Lys Ala
515 520 525
Gly Pro Gln Leu
530
<210> 37
<211> 538
<212> PRT
<213> 产琥珀酸放线杆菌(Actinobacillus succinogenes)
<400> 37
Met Thr Asp Leu Asn Lys Leu Val Lys Glu Leu Asn Asp Leu Gly Leu
1 5 10 15
Thr Asp Val Lys Glu Ile Val Tyr Asn Pro Ser Tyr Glu Gln Leu Phe
20 25 30
Glu Glu Glu Thr Lys Pro Gly Leu Glu Gly Phe Asp Lys Gly Thr Leu
35 40 45
Thr Thr Leu Gly Ala Val Ala Val Asp Thr Gly Ile Phe Thr Gly Arg
50 55 60
Ser Pro Lys Asp Lys Tyr Ile Val Cys Asp Glu Thr Thr Lys Asp Thr
65 70 75 80
Val Trp Trp Asn Ser Glu Ala Ala Lys Asn Asp Asn Lys Pro Met Thr
85 90 95
Gln Glu Thr Trp Lys Ser Leu Arg Glu Leu Val Ala Lys Gln Leu Ser
100 105 110
Gly Lys Arg Leu Phe Val Val Glu Gly Tyr Cys Gly Ala Ser Glu Lys
115 120 125
His Arg Ile Gly Val Arg Met Val Thr Glu Val Ala Trp Gln Ala His
130 135 140
Phe Val Lys Asn Met Phe Ile Arg Pro Thr Asp Glu Glu Leu Lys Asn
145 150 155 160
Phe Lys Ala Asp Phe Thr Val Leu Asn Gly Ala Lys Cys Thr Asn Pro
165 170 175
Asn Trp Lys Glu Gln Gly Leu Asn Ser Glu Asn Phe Val Ala Phe Asn
180 185 190
Ile Thr Glu Gly Ile Gln Leu Ile Gly Gly Thr Trp Tyr Gly Gly Glu
195 200 205
Met Lys Lys Gly Met Phe Ser Met Met Asn Tyr Phe Leu Pro Leu Lys
210 215 220
Gly Val Ala Ser Met His Cys Ser Ala Asn Val Gly Lys Asp Gly Asp
225 230 235 240
Val Ala Ile Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr Thr Leu Ser
245 250 255
Thr Asp Pro Lys Arg Gln Leu Ile Gly Asp Asp Glu His Gly Trp Asp
260 265 270
Glu Ser Gly Val Phe Asn Phe Glu Gly Gly Cys Tyr Ala Lys Thr Ile
275 280 285
Asn Leu Ser Gln Glu Asn Glu Pro Asp Ile Tyr Gly Ala Ile Arg Arg
290 295 300
Asp Ala Leu Leu Glu Asn Val Val Val Arg Ala Asp Gly Ser Val Asp
305 310 315 320
Phe Asp Asp Gly Ser Lys Thr Glu Asn Thr Arg Val Ser Tyr Pro Ile
325 330 335
Tyr His Ile Asp Asn Ile Val Arg Pro Val Ser Lys Ala Gly His Ala
340 345 350
Thr Lys Val Ile Phe Leu Thr Ala Asp Ala Phe Gly Val Leu Pro Pro
355 360 365
Val Ser Lys Leu Thr Pro Glu Gln Thr Glu Tyr Tyr Phe Leu Ser Gly
370 375 380
Phe Thr Ala Lys Leu Ala Gly Thr Glu Arg Gly Val Thr Glu Pro Thr
385 390 395 400
Pro Thr Phe Ser Ala Cys Phe Gly Ala Ala Phe Leu Ser Leu His Pro
405 410 415
Ile Gln Tyr Ala Asp Val Leu Val Glu Arg Met Lys Ala Ser Gly Ala
420 425 430
Glu Ala Tyr Leu Val Asn Thr Gly Trp Asn Gly Thr Gly Lys Arg Ile
435 440 445
Ser Ile Lys Asp Thr Arg Gly Ile Ile Asp Ala Ile Leu Asp Gly Ser
450 455 460
Ile Glu Lys Ala Glu Met Gly Glu Leu Pro Ile Phe Asn Leu Ala Ile
465 470 475 480
Pro Lys Ala Leu Pro Gly Val Asp Pro Ala Ile Leu Asp Pro Arg Asp
485 490 495
Thr Tyr Ala Asp Lys Ala Gln Trp Gln Val Lys Ala Glu Asp Leu Ala
500 505 510
Asn Arg Phe Val Lys Asn Phe Val Lys Tyr Thr Ala Asn Pro Glu Ala
515 520 525
Ala Lys Leu Val Gly Ala Gly Pro Lys Ala
530 535
<210> 38
<211> 618
<212> PRT
<213> 真氧产碱杆菌(Ralstonia eutropha)
<400> 38
Met Asn His Pro Ser Met Gln Gly Thr Thr Ala Leu Asn Val Pro Ala
1 5 10 15
Trp Val Arg Asn Gln Lys Leu Val Ala Trp Val Ala Glu Ile Ala Ala
20 25 30
Leu Thr Lys Pro Glu Arg Ile His Trp Cys Asp Gly Ser Gln Glu Glu
35 40 45
Tyr Asp Arg Leu Cys Glu Gln Met Val Ala Ala Gly Thr Leu Lys Arg
50 55 60
Leu Asn Pro Ala Lys Arg Lys Asn Ser Tyr Leu Ala Leu Ser Asp Pro
65 70 75 80
Ser Asp Val Ala Arg Val Glu Asp Arg Thr Phe Ile Cys Ser Gln Lys
85 90 95
Lys Glu Asp Ala Gly Pro Thr Asn Asn Trp Val Ala Pro Ala Glu Met
100 105 110
Arg Thr Thr Leu Asn Gly Leu Phe Asp Gly Cys Met Arg Gly Arg Thr
115 120 125
Leu Tyr Val Val Pro Phe Ser Met Gly Pro Leu Gly Ser Pro Ile Ala
130 135 140
His Ile Gly Val Glu Leu Ser Asp Ser Pro Tyr Val Ala Val Asn Met
145 150 155 160
Arg Ile Met Thr Arg Met Gly Lys Ala Val Tyr Asp Val Leu Gly Thr
165 170 175
Asp Gly Asp Phe Val Pro Cys Val His Thr Val Gly Lys Pro Leu Ala
180 185 190
Ala Gly Glu Lys Asp Val Pro Trp Pro Cys Asn Pro Thr Lys Tyr Ile
195 200 205
Val His Phe Pro Glu Ser Arg Glu Ile Trp Ser Phe Gly Ser Gly Tyr
210 215 220
Gly Gly Asn Ala Leu Leu Gly Lys Lys Cys Phe Ala Leu Arg Ile Ala
225 230 235 240
Ser Thr Met Gly Arg Asp Glu Gly Trp Leu Ala Glu His Met Leu Ile
245 250 255
Leu Gly Val Thr Ser Pro Glu Gly Lys Lys Phe His Val Ala Ala Ala
260 265 270
Phe Pro Ser Ala Cys Gly Lys Thr Asn Phe Ala Met Leu Ile Pro Pro
275 280 285
Lys Gly Phe Glu Gly Trp Lys Val Thr Thr Ile Gly Asp Asp Ile Ala
290 295 300
Trp Ile Lys Pro Gly Lys Asp Gly Arg Leu Tyr Ala Ile Asn Pro Glu
305 310 315 320
Ala Gly Tyr Phe Gly Val Ala Pro Gly Thr Ser Glu Lys Thr Asn Phe
325 330 335
Asn Ala Met Ala Thr Leu Lys Glu Asn Val Ile Phe Thr Asn Val Ala
340 345 350
Leu Thr Asp Asp Gly Asp Val Trp Trp Glu Gly Met Thr Lys Glu Ala
355 360 365
Pro Ala His Leu Thr Asp Trp Gln Gly Lys Asp Trp Thr Pro Glu Ile
370 375 380
Ala Lys Ala Thr Gly Ala Lys Ala Ala His Pro Asn Ala Arg Phe Thr
385 390 395 400
Ala Pro Ala Ser Gln Cys Pro Ser Ile Asp Glu Asn Trp Asp Asn Pro
405 410 415
Ala Gly Val Pro Ile Asp Ala Phe Ile Phe Gly Gly Arg Arg Ser Thr
420 425 430
Thr Val Pro Leu Val Thr Glu Ala Arg Asn Trp Thr Glu Gly Val Tyr
435 440 445
Met Ala Ala Thr Met Gly Ser Glu Thr Thr Ala Ala Ala Ala Gly Gln
450 455 460
Gln Gly Val Val Arg Arg Asp Pro Phe Ala Met Leu Pro Phe Cys Gly
465 470 475 480
Tyr Asn Met Ser Asp Tyr Phe Gly His Trp Leu Ala Leu Gly Gln Lys
485 490 495
Leu Glu Ala Ala Gly Ala Lys Leu Pro Lys Ile Tyr Cys Val Asn Trp
500 505 510
Phe Arg Lys Asp Ala Asp Gly Asn Phe Val Trp Pro Gly Phe Gly Glu
515 520 525
Asn Met Arg Val Leu Ser Trp Met Ile Asp Arg Val Glu Gly Lys Gly
530 535 540
Glu Gly Ala Glu His Val Phe Gly Thr Ser Pro Arg Tyr Glu Asp Leu
545 550 555 560
Asn Trp Ser Gly Val Glu Phe Ser Val Ala Gln Phe Thr Gln Val Thr
565 570 575
Ser Ile Asp Ala Asp Ala Trp Lys Gln Glu Leu Ala Leu His Asp Glu
580 585 590
Leu Phe Thr Gln Leu Lys His Asn Leu Pro Gln Ala Leu Ala Glu Ala
595 600 605
Arg Ala Ala Leu Gly Lys Arg Leu Glu Gly
610 615
<210> 39
<211> 540
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 39
Met Arg Val Asn Asn Gly Leu Thr Pro Gln Glu Leu Glu Ala Tyr Gly
1 5 10 15
Ile Ser Asp Val His Asp Ile Val Tyr Asn Pro Ser Tyr Asp Leu Leu
20 25 30
Tyr Gln Glu Glu Leu Asp Pro Ser Leu Thr Gly Tyr Glu Arg Gly Val
35 40 45
Leu Thr Asn Leu Gly Ala Val Ala Val Asp Thr Gly Ile Phe Thr Gly
50 55 60
Arg Ser Pro Lys Asp Lys Tyr Ile Val Arg Asp Asp Thr Thr Arg Asp
65 70 75 80
Thr Phe Trp Trp Ala Asp Lys Gly Lys Gly Lys Asn Asp Asn Lys Pro
85 90 95
Leu Ser Pro Glu Thr Trp Gln His Leu Lys Gly Leu Val Thr Arg Gln
100 105 110
Leu Ser Gly Lys Arg Leu Phe Val Val Asp Ala Phe Cys Gly Ala Asn
115 120 125
Pro Asp Thr Arg Leu Ser Val Arg Phe Ile Thr Glu Val Ala Trp Gln
130 135 140
Ala His Phe Val Lys Asn Met Phe Ile Arg Pro Ser Asp Glu Glu Leu
145 150 155 160
Ala Gly Phe Lys Pro Asp Phe Ile Val Met Asn Gly Ala Lys Cys Thr
165 170 175
Asn Pro Gln Trp Lys Glu Gln Gly Leu Asn Ser Glu Asn Phe Val Ala
180 185 190
Phe Asn Leu Thr Glu Arg Met Gln Leu Ile Gly Gly Thr Trp Tyr Gly
195 200 205
Gly Glu Met Lys Lys Gly Met Phe Ser Met Met Asn Tyr Leu Leu Pro
210 215 220
Leu Lys Gly Ile Ala Ser Met His Cys Ser Ala Asn Val Gly Glu Lys
225 230 235 240
Gly Asp Val Ala Val Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr Thr
245 250 255
Leu Ser Thr Asp Pro Lys Arg Arg Leu Ile Gly Asp Asp Glu His Gly
260 265 270
Trp Asp Asp Asp Gly Val Phe Asn Phe Glu Gly Gly Cys Tyr Ala Lys
275 280 285
Thr Ile Lys Leu Ser Lys Glu Ala Glu Pro Glu Ile Tyr Asn Ala Ile
290 295 300
Arg Arg Asp Ala Leu Leu Glu Asn Val Thr Val Arg Glu Asp Gly Thr
305 310 315 320
Ile Asp Phe Asp Asp Gly Ser Lys Thr Glu Asn Thr Arg Val Ser Tyr
325 330 335
Pro Ile Tyr His Ile Asp Asn Ile Val Lys Pro Val Ser Lys Ala Gly
340 345 350
His Ala Thr Lys Val Ile Phe Leu Thr Ala Asp Ala Phe Gly Val Leu
355 360 365
Pro Pro Val Ser Arg Leu Thr Ala Asp Gln Thr Gln Tyr His Phe Leu
370 375 380
Ser Gly Phe Thr Ala Lys Leu Ala Gly Thr Glu Arg Gly Ile Thr Glu
385 390 395 400
Pro Thr Pro Thr Phe Ser Ala Cys Phe Gly Ala Ala Phe Leu Ser Leu
405 410 415
His Pro Thr Gln Tyr Ala Glu Val Leu Val Lys Arg Met Gln Ala Ala
420 425 430
Gly Ala Gln Ala Tyr Leu Val Asn Thr Gly Trp Asn Gly Thr Gly Lys
435 440 445
Arg Ile Ser Ile Lys Asp Thr Arg Ala Ile Ile Asp Ala Ile Leu Asn
450 455 460
Gly Ser Leu Asp Asn Ala Glu Thr Phe Thr Leu Pro Met Phe Asn Leu
465 470 475 480
Ala Ile Pro Thr Glu Leu Pro Gly Val Asp Thr Lys Ile Leu Asp Pro
485 490 495
Arg Asn Thr Tyr Ala Ser Pro Glu Gln Trp Gln Glu Lys Ala Glu Thr
500 505 510
Leu Ala Lys Leu Phe Ile Asp Asn Phe Asp Lys Tyr Thr Asp Thr Pro
515 520 525
Ala Gly Ala Ala Leu Val Ala Ala Gly Pro Lys Leu
530 535 540
<210> 40
<211> 236
<212> PRT
<213> 乳酸乳球菌(Lactococcus lactis)
<400> 40
Met Ser Glu Ile Thr Gln Leu Phe Gln Tyr Asn Thr Leu Gly Ala Leu
1 5 10 15
Met Ala Gly Leu Tyr Glu Gly Thr Met Thr Ile Gly Glu Leu Leu Lys
20 25 30
His Gly Asp Leu Gly Ile Gly Thr Leu Asp Ser Ile Asp Gly Glu Leu
35 40 45
Ile Val Leu Asp Gly Lys Ala Tyr Gln Ala Lys Gly Asp Lys Thr Ile
50 55 60
Val Glu Leu Thr Asp Asp Ile Lys Val Pro Tyr Ala Ala Val Val Pro
65 70 75 80
His Gln Ala Glu Val Val Phe Lys Gln Lys Phe Thr Val Ser Asp Lys
85 90 95
Glu Leu Glu Asp Arg Ile Glu Ser Tyr Phe Asp Gly Gln Asn Leu Phe
100 105 110
Arg Ser Ile Lys Ile Thr Gly Lys Phe Pro Lys Met His Val Arg Met
115 120 125
Ile Pro Arg Ala Lys Ser Gly Thr Lys Phe Val Glu Val Ser Gln Asn
130 135 140
Gln Pro Glu Tyr Thr Glu Glu Asn Ile Lys Gly Thr Ile Val Gly Ile
145 150 155 160
Trp Thr Pro Glu Met Phe His Gly Val Ser Val Ala Gly Tyr His Leu
165 170 175
His Phe Ile Ser Glu Asp Phe Thr Phe Gly Gly His Val Leu Asp Phe
180 185 190
Ile Ile Asp Asn Gly Thr Val Glu Ile Gly Ala Ile Asp Gln Leu Asn
195 200 205
Gln Ser Phe Pro Val Gln Asp Arg Lys Phe Leu Phe Ala Asp Leu Asp
210 215 220
Ile Glu Ala Leu Lys Lys Asp Ile Asp Val Ala Glu
225 230 235
<210> 41
<211> 239
<212> PRT
<213> 嗜热链球菌(Streptococcus thermophilus)
<400> 41
Met Ser Glu Ala Ile Lys Leu Phe Gln Tyr Asn Thr Leu Gly Ala Leu
1 5 10 15
Met Ala Gly Leu Tyr Gly Gly Thr Leu Thr Val Gly Glu Leu Leu Glu
20 25 30
His Gly Asp Leu Gly Leu Gly Thr Leu Asp Ser Ile Asp Gly Glu Leu
35 40 45
Ile Val Leu Asp Gly Lys Ala Tyr Gln Ala Lys Gly Ser Glu Gly Lys
50 55 60
Val Glu Val Val Glu Val Ser Pro Asp Glu Lys Val Pro Tyr Ala Ala
65 70 75 80
Val Val Pro His Gln Ala Glu Val Ile Phe Arg Gln Arg Tyr Glu Met
85 90 95
Thr Asp Lys Glu Leu Glu Asp Arg Ile Glu Ser Tyr Tyr Asp Gly Val
100 105 110
Asn Leu Phe Arg Ser Ile Lys Ile Lys Gly His Phe Lys His Met His
115 120 125
Val Arg Met Ile Pro Lys Ser Asn Ala Asp Ile Lys Phe Ala Asp Val
130 135 140
Ala Thr Arg Gln Pro Glu Tyr Glu Val Asp Asp Ile Ser Gly Thr Ile
145 150 155 160
Val Gly Ile Trp Thr Pro Glu Met Phe His Gly Val Ser Val Ala Gly
165 170 175
Tyr His Leu His Phe Ile Ser Asp Asp Leu Thr Phe Gly Gly His Val
180 185 190
Met Asp Phe Val Ile Glu Asn Gly Ile Ile Glu Val Gly Pro Val Asp
195 200 205
Gln Leu Asp Gln Arg Phe Pro Val Gln Asp Arg Gln Tyr Leu Phe Ala
210 215 220
Lys Phe Asn Val Asp Glu Met Arg Lys Asp Ile Thr Lys Ala Glu
225 230 235
<210> 42
<211> 285
<212> PRT
<213> 短芽孢杆菌(Brevibacillus brevis)
<400> 42
Met Lys Lys Asn Ile Ile Thr Ser Ile Thr Ser Leu Ala Leu Val Ala
1 5 10 15
Gly Leu Ser Leu Thr Ala Phe Ala Ala Thr Thr Ala Thr Val Pro Ala
20 25 30
Pro Pro Ala Lys Gln Glu Ser Lys Pro Ala Val Ala Ala Asn Pro Ala
35 40 45
Pro Lys Asn Val Leu Phe Gln Tyr Ser Thr Ile Asn Ala Leu Met Leu
50 55 60
Gly Gln Phe Glu Gly Asp Leu Thr Leu Lys Asp Leu Lys Leu Arg Gly
65 70 75 80
Asp Met Gly Leu Gly Thr Ile Asn Asp Leu Asp Gly Glu Met Ile Gln
85 90 95
Met Gly Thr Lys Phe Tyr Gln Ile Asp Ser Thr Gly Lys Leu Ser Glu
100 105 110
Leu Pro Glu Ser Val Lys Thr Pro Phe Ala Val Thr Thr His Phe Glu
115 120 125
Pro Lys Glu Lys Thr Thr Leu Thr Asn Val Gln Asp Tyr Asn Gln Leu
130 135 140
Thr Lys Met Leu Glu Glu Lys Phe Glu Asn Lys Asn Val Phe Tyr Ala
145 150 155 160
Val Lys Leu Thr Gly Thr Phe Lys Met Val Lys Ala Arg Thr Val Pro
165 170 175
Lys Gln Thr Arg Pro Tyr Pro Gln Leu Thr Glu Val Thr Lys Lys Gln
180 185 190
Ser Glu Phe Glu Phe Lys Asn Val Lys Gly Thr Leu Ile Gly Phe Tyr
195 200 205
Thr Pro Asn Tyr Ala Ala Ala Leu Asn Val Pro Gly Phe His Leu His
210 215 220
Phe Ile Thr Glu Asp Lys Thr Ser Gly Gly His Val Leu Asn Leu Gln
225 230 235 240
Phe Asp Asn Ala Asn Leu Glu Ile Ser Pro Ile His Glu Phe Asp Val
245 250 255
Gln Leu Pro His Thr Asp Asp Phe Ala His Ser Asp Leu Thr Gln Val
260 265 270
Thr Thr Ser Gln Val His Gln Ala Glu Ser Glu Arg Lys
275 280 285
<210> 43
<211> 259
<212> PRT
<213> 产气肠杆菌(Enterobacter aerogenes)
<400> 43
Met Met Met His Ser Ser Ala Cys Asp Cys Glu Ala Ser Leu Cys Glu
1 5 10 15
Thr Leu Arg Gly Phe Ser Ala Gln His Pro Asp Ser Val Ile Tyr Gln
20 25 30
Thr Ser Leu Met Ser Ala Leu Leu Ser Gly Val Tyr Val Gly Glu Thr
35 40 45
Thr Ile Ala Asp Leu Leu Ala His Gly Asp Phe Gly Leu Gly Thr Phe
50 55 60
Asn Glu Leu Asp Gly Glu Met Ile Ala Phe Ser Ser Gln Val Tyr Gln
65 70 75 80
Leu Arg Ala Asp Gly Ser Ala Arg Ala Ala Lys Pro Glu Gln Lys Thr
85 90 95
Pro Phe Ala Val Met Thr Trp Phe Gln Pro Gln Tyr Arg Lys Thr Phe
100 105 110
Asn Gly Pro Val Ser Arg Gln Gln Ile His Asp Val Ile Asp Gln Gln
115 120 125
Ile Pro Ser Asp Asn Leu Phe Cys Val Arg Ile Asp Gly Asn Phe Arg
130 135 140
His Ala His Thr Arg Thr Val Pro Arg Gln Thr Pro Pro Tyr Arg Ala
145 150 155 160
Met Thr Asp Val Leu Asp Asp Gln Pro Val Phe Arg Phe Asn Gln Arg
165 170 175
Glu Gly Val Leu Val Gly Phe Arg Thr Pro Gln His Met Gln Gly Ile
180 185 190
Asn Val Ala Gly Tyr His Glu His Phe Ile Thr Asp Asp Arg Gln Gly
195 200 205
Gly Gly His Leu Leu Asp Tyr Gln Leu Glu Ser Gly Val Leu Thr Phe
210 215 220
Gly Glu Ile His Lys Leu Met Ile Asp Leu Pro Ala Asp Ser Ala Phe
225 230 235 240
Leu Gln Ala Asn Leu His Pro Ser Asn Leu Asp Ala Ala Ile Arg Ala
245 250 255
Val Glu Asn
<210> 44
<211> 1231
<212> PRT
<213> 结核分枝杆菌(Mycobacterium tuberculosis)
<400> 44
Met Ala Asn Ile Ser Ser Pro Phe Gly Gln Asn Glu Trp Leu Val Glu
1 5 10 15
Glu Met Tyr Arg Lys Phe Arg Asp Asp Pro Ser Ser Val Asp Pro Ser
20 25 30
Trp His Glu Phe Leu Val Asp Tyr Ser Pro Glu Pro Thr Ser Gln Pro
35 40 45
Ala Ala Glu Pro Thr Arg Val Thr Ser Pro Leu Val Ala Glu Arg Ala
50 55 60
Ala Ala Ala Ala Pro Gln Ala Pro Pro Lys Pro Ala Asp Thr Ala Ala
65 70 75 80
Ala Gly Asn Gly Val Val Ala Ala Leu Ala Ala Lys Thr Ala Val Pro
85 90 95
Pro Pro Ala Glu Gly Asp Glu Val Ala Val Leu Arg Gly Ala Ala Ala
100 105 110
Ala Val Val Lys Asn Met Ser Ala Ser Leu Glu Val Pro Thr Ala Thr
115 120 125
Ser Val Arg Ala Val Pro Ala Lys Leu Leu Ile Asp Asn Arg Ile Val
130 135 140
Ile Asn Asn Gln Leu Lys Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr
145 150 155 160
His Leu Leu Gly Tyr Ala Leu Val Gln Ala Val Lys Lys Phe Pro Asn
165 170 175
Met Asn Arg His Tyr Thr Glu Val Asp Gly Lys Pro Thr Ala Val Thr
180 185 190
Pro Ala His Thr Asn Leu Gly Leu Ala Ile Asp Leu Gln Gly Lys Asp
195 200 205
Gly Lys Arg Ser Leu Val Val Ala Gly Ile Lys Arg Cys Glu Thr Met
210 215 220
Arg Phe Ala Gln Phe Val Thr Ala Tyr Glu Asp Ile Val Arg Arg Ala
225 230 235 240
Arg Asp Gly Lys Leu Thr Thr Glu Asp Phe Ala Gly Val Thr Ile Ser
245 250 255
Leu Thr Asn Pro Gly Thr Ile Gly Thr Val His Ser Val Pro Arg Leu
260 265 270
Met Pro Gly Gln Gly Ala Ile Ile Gly Val Gly Ala Met Glu Tyr Pro
275 280 285
Ala Glu Phe Gln Gly Ala Ser Glu Glu Arg Ile Ala Glu Leu Gly Ile
290 295 300
Gly Lys Leu Ile Thr Leu Thr Ser Thr Tyr Asp His Arg Ile Ile Gln
305 310 315 320
Gly Ala Glu Ser Gly Asp Phe Leu Arg Thr Ile His Glu Leu Leu Leu
325 330 335
Ser Asp Gly Phe Trp Asp Glu Val Phe Arg Glu Leu Ser Ile Pro Tyr
340 345 350
Leu Pro Val Arg Trp Ser Thr Asp Asn Pro Asp Ser Ile Val Asp Lys
355 360 365
Asn Ala Arg Val Met Asn Leu Ile Ala Ala Tyr Arg Asn Arg Gly His
370 375 380
Leu Met Ala Asp Thr Asp Pro Leu Arg Leu Asp Lys Ala Arg Phe Arg
385 390 395 400
Ser His Pro Asp Leu Glu Val Leu Thr His Gly Leu Thr Leu Trp Asp
405 410 415
Leu Asp Arg Val Phe Lys Val Asp Gly Phe Ala Gly Ala Gln Tyr Lys
420 425 430
Lys Leu Arg Asp Val Leu Gly Leu Leu Arg Asp Ala Tyr Cys Arg His
435 440 445
Ile Gly Val Glu Tyr Ala His Ile Leu Asp Pro Glu Gln Lys Glu Trp
450 455 460
Leu Glu Gln Arg Val Glu Thr Lys His Val Lys Pro Thr Val Ala Gln
465 470 475 480
Gln Lys Tyr Ile Leu Ser Lys Leu Asn Ala Ala Glu Ala Phe Glu Thr
485 490 495
Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly
500 505 510
Ala Glu Ser Val Ile Pro Met Met Asp Ala Ala Ile Asp Gln Cys Ala
515 520 525
Glu His Gly Leu Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg
530 535 540
Leu Asn Val Leu Ala Asn Ile Val Gly Lys Pro Tyr Ser Gln Ile Phe
545 550 555 560
Thr Glu Phe Glu Gly Asn Leu Asn Pro Ser Gln Ala His Gly Ser Gly
565 570 575
Asp Val Lys Tyr His Leu Gly Ala Thr Gly Leu Tyr Leu Gln Met Phe
580 585 590
Gly Asp Asn Asp Ile Gln Val Ser Leu Thr Ala Asn Pro Ser His Leu
595 600 605
Glu Ala Val Asp Pro Val Leu Glu Gly Leu Val Arg Ala Lys Gln Asp
610 615 620
Leu Leu Asp His Gly Ser Ile Asp Ser Asp Gly Gln Arg Ala Phe Ser
625 630 635 640
Val Val Pro Leu Met Leu His Gly Asp Ala Ala Phe Ala Gly Gln Gly
645 650 655
Val Val Ala Glu Thr Leu Asn Leu Ala Asn Leu Pro Gly Tyr Arg Val
660 665 670
Gly Gly Thr Ile His Ile Ile Val Asn Asn Gln Ile Gly Phe Thr Thr
675 680 685
Ala Pro Glu Tyr Ser Arg Ser Ser Glu Tyr Cys Thr Asp Val Ala Lys
690 695 700
Met Ile Gly Ala Pro Ile Phe His Val Asn Gly Asp Asp Pro Glu Ala
705 710 715 720
Cys Val Trp Val Ala Arg Leu Ala Val Asp Phe Arg Gln Arg Phe Lys
725 730 735
Lys Asp Val Val Ile Asp Met Leu Cys Tyr Arg Arg Arg Gly His Asn
740 745 750
Glu Gly Asp Asp Pro Ser Met Thr Asn Pro Tyr Val Tyr Asp Val Val
755 760 765
Asp Thr Lys Arg Gly Ala Arg Lys Ser Tyr Thr Glu Ala Leu Ile Gly
770 775 780
Arg Gly Asp Ile Ser Met Lys Glu Ala Glu Asp Ala Leu Arg Asp Tyr
785 790 795 800
Gln Gly Gln Leu Glu Arg Val Phe Asn Glu Val Arg Glu Leu Glu Lys
805 810 815
His Gly Val Gln Pro Ser Glu Ser Val Glu Ser Asp Gln Met Ile Pro
820 825 830
Ala Gly Leu Ala Thr Ala Val Asp Lys Ser Leu Leu Ala Arg Ile Gly
835 840 845
Asp Ala Phe Leu Ala Leu Pro Asn Gly Phe Thr Ala His Pro Arg Val
850 855 860
Gln Pro Val Leu Glu Lys Arg Arg Glu Met Ala Tyr Glu Gly Lys Ile
865 870 875 880
Asp Trp Ala Phe Gly Glu Leu Leu Ala Leu Gly Ser Leu Val Ala Glu
885 890 895
Gly Lys Leu Val Arg Leu Ser Gly Gln Asp Ser Arg Arg Gly Thr Phe
900 905 910
Ser Gln Arg His Ser Val Leu Ile Asp Arg His Thr Gly Glu Glu Phe
915 920 925
Thr Pro Leu Gln Leu Leu Ala Thr Asn Ser Asp Gly Ser Pro Thr Gly
930 935 940
Gly Lys Phe Leu Val Tyr Asp Ser Pro Leu Ser Glu Tyr Ala Ala Val
945 950 955 960
Gly Phe Glu Tyr Gly Tyr Thr Val Gly Asn Pro Asp Ala Val Val Leu
965 970 975
Trp Glu Ala Gln Phe Gly Asp Phe Val Asn Gly Ala Gln Ser Ile Ile
980 985 990
Asp Glu Phe Ile Ser Ser Gly Glu Ala Lys Trp Gly Gln Leu Ser Asn
995 1000 1005
Val Val Leu Leu Leu Pro His Gly His Glu Gly Gln Gly Pro Asp
1010 1015 1020
His Thr Ser Ala Arg Ile Glu Arg Phe Leu Gln Leu Trp Ala Glu
1025 1030 1035
Gly Ser Met Thr Ile Ala Met Pro Ser Thr Pro Ser Asn Tyr Phe
1040 1045 1050
His Leu Leu Arg Arg His Ala Leu Asp Gly Ile Gln Arg Pro Leu
1055 1060 1065
Ile Val Phe Thr Pro Lys Ser Met Leu Arg His Lys Ala Ala Val
1070 1075 1080
Ser Glu Ile Lys Asp Phe Thr Glu Ile Lys Phe Arg Ser Val Leu
1085 1090 1095
Glu Glu Pro Thr Tyr Glu Asp Gly Ile Gly Asp Arg Asn Lys Val
1100 1105 1110
Ser Arg Ile Leu Leu Thr Ser Gly Lys Leu Tyr Tyr Glu Leu Ala
1115 1120 1125
Ala Arg Lys Ala Lys Asp Asn Arg Asn Asp Leu Ala Ile Val Arg
1130 1135 1140
Leu Glu Gln Leu Ala Pro Leu Pro Arg Arg Arg Leu Arg Glu Thr
1145 1150 1155
Leu Asp Arg Tyr Glu Asn Val Lys Glu Phe Phe Trp Val Gln Glu
1160 1165 1170
Glu Pro Ala Asn Gln Gly Ala Trp Pro Arg Phe Gly Leu Glu Leu
1175 1180 1185
Pro Glu Leu Leu Pro Asp Lys Leu Ala Gly Ile Lys Arg Ile Ser
1190 1195 1200
Arg Arg Ala Met Ser Ala Pro Ser Ser Gly Ser Ser Lys Val His
1205 1210 1215
Ala Val Glu Gln Gln Glu Ile Leu Asp Glu Ala Phe Gly
1220 1225 1230
<210> 45
<211> 985
<212> PRT
<213> 大豆慢生根瘤菌(Bradyrhizobium japonicum)
<400> 45
Met Ser Arg Gln Asp Ala Asn Ala Ala Phe Ala Leu Ser Ser Phe Leu
1 5 10 15
Gln Gly Thr Asn Ala Thr Tyr Ile Asp Glu Ile Tyr Ala Arg Tyr Glu
20 25 30
Lys Asp Pro Ser Ser Val Asp Ala Glu Trp Gln Glu Phe Phe Lys Ser
35 40 45
Leu Lys Asp Gln Pro Asp Asp Val Arg Arg Asn Ala Glu Gly Pro Ser
50 55 60
Trp Glu Arg Ala Asn Trp Pro Leu Thr Pro Gln Asp Asp Leu Thr Ser
65 70 75 80
Ala Leu Asp Gly Asn Trp Ala Glu Val Glu Lys Ala Val Gly Gly Lys
85 90 95
Ile Ala Ala Lys Ala Gln Ala Lys Gly Ala Asp Ile Ser Ser Ala Asp
100 105 110
Leu Leu Gln Ala Thr Arg Asp Ser Val Arg Ala Leu Met Leu Ile Arg
115 120 125
Ser Tyr Arg Met Arg Gly His Phe His Ala Lys Leu Asp Pro Leu Gly
130 135 140
Ile Glu Ala Pro Arg Asn Arg Glu Glu Leu Asp Pro Arg Thr Tyr Gly
145 150 155 160
Phe Ser Glu Ala Asp Phe Asp Arg Lys Ile Phe Leu Asp His Val Leu
165 170 175
Gly Leu Glu Tyr Gly Thr Leu Arg Glu Ile Thr Ala Ile Cys Glu Arg
180 185 190
Thr Tyr Cys Gln Thr Leu Gly Val Glu Phe Met His Ile Ser Asn Ala
195 200 205
Ala Gln Lys Ala Trp Ile Gln Glu Arg Ile Glu Gly Pro Asp Lys Glu
210 215 220
Ile Ser Phe Thr Arg Glu Gly Arg Arg Ala Ile Leu Thr Lys Leu Val
225 230 235 240
Glu Ala Glu Gly Phe Glu Lys Phe Cys Asp Thr Lys Phe Thr Gly Thr
245 250 255
Lys Arg Phe Gly Leu Asp Gly Ala Glu Ser Leu Ile Pro Ala Leu Glu
260 265 270
Gln Ile Ile Lys Arg Gly Gly Asn Leu Gly Val Lys Glu Ile Val Leu
275 280 285
Gly Met Pro His Arg Gly Arg Leu Asn Val Leu Thr Gln Val Met Gly
290 295 300
Lys Ala His Arg Ala Leu Phe His Glu Phe Lys Gly Gly Ser Ala Asn
305 310 315 320
Pro Asp Ala Val Glu Gly Ser Gly Asp Val Lys Tyr His Leu Gly Ala
325 330 335
Ser Ser Asp Arg Glu Phe Asp Gly Asn Arg Ile His Leu Ser Leu Thr
340 345 350
Ala Asn Pro Ser His Leu Glu Ile Val Asp Pro Val Val Leu Gly Lys
355 360 365
Val Arg Ala Lys Gln Asp Gln His Gly Asp Pro Pro Asp Met Arg Ile
370 375 380
Ser Val Met Pro Leu Leu Met His Gly Asp Ala Ala Phe Ala Gly Gln
385 390 395 400
Gly Val Val Ala Glu Cys Phe Gly Leu Ser Asp Leu Lys Gly Tyr Arg
405 410 415
Thr Gly Gly Ser Val His Phe Ile Val Asn Asn Gln Ile Gly Phe Thr
420 425 430
Thr Tyr Pro Arg Tyr Ser Arg Ser Ser Pro Tyr Pro Ser Asp Val Ala
435 440 445
Lys Met Ile Asp Ala Pro Ile Phe His Val Asn Gly Asp Asp Pro Glu
450 455 460
Ala Val Val Phe Ala Ala Lys Val Ala Thr Glu Phe Arg Gln Lys Phe
465 470 475 480
His Lys Pro Val Val Ile Asp Met Phe Cys Tyr Arg Arg His Gly His
485 490 495
Asn Glu Gly Asp Glu Pro Ala Phe Thr Gln Pro Val Met Tyr Lys Lys
500 505 510
Ile Ala Ala His Pro Ser Thr Leu Glu Leu Tyr Ala Arg Arg Leu Ile
515 520 525
Ser Glu Gly Val Met Thr Glu Gly Glu Val Asp Lys Ala Lys Ala Asp
530 535 540
Trp Arg Ala Arg Leu Asp Ala Glu Phe Glu Ala Gly Thr Ser Tyr Lys
545 550 555 560
Pro Asn Lys Ala Asp Trp Leu Asp Gly Lys Trp Ala Gly Phe Lys Ile
565 570 575
Ala Asp Gln Glu Glu Asp Ala Arg Arg Gly Val Thr Gly Val Asp Ile
580 585 590
Thr Ala Leu Lys Asp Ile Gly Arg Lys Ile Thr Lys Val Pro Asp Gly
595 600 605
Phe Arg Val His Arg Thr Ile Gln Arg Phe Leu Glu Asn Arg Ser Lys
610 615 620
Ala Ile Asp Ser Gly Ala Gly Ile Asp Trp Ala Thr Gly Glu Ala Leu
625 630 635 640
Ala Phe Cys Ser Leu Leu Asn Glu Asn His His Val Arg Leu Ser Gly
645 650 655
Gln Asp Ser Glu Arg Gly Thr Phe Ser Gln Arg His Ser Val Leu Ile
660 665 670
Asp Gln Glu Asp Glu Ser Arg Tyr Thr Pro Phe Asn His Leu Gly His
675 680 685
Glu Gln Gly His Tyr Glu Val Ile Asn Ser Leu Leu Ser Glu Glu Ala
690 695 700
Val Leu Gly Phe Glu Tyr Gly Tyr Ser Leu Ala Glu Pro Asn Thr Leu
705 710 715 720
Thr Leu Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn Gly Ala Gln Val
725 730 735
Val Phe Asp Gln Phe Ile Ser Ser Gly Glu Arg Lys Trp Leu Arg Met
740 745 750
Ser Gly Leu Val Cys Leu Leu Pro His Gly Tyr Glu Gly Gln Gly Pro
755 760 765
Glu His Ser Ser Ala Arg Leu Glu Arg Tyr Leu Gln Met Cys Ala Glu
770 775 780
Asp Asn Met Gln Val Val Tyr Pro Thr Thr Pro Ala Asn Tyr Phe His
785 790 795 800
Val Leu Arg Arg Gln Leu His Arg Glu Ile Arg Lys Pro Leu Ile Leu
805 810 815
Met Thr Pro Lys Ser Leu Leu Arg His Lys Arg Ala Val Ser Arg Leu
820 825 830
Glu Glu Leu Ala Lys Gly Thr Thr Phe His Arg Ile Leu Tyr Asp Asp
835 840 845
Ala Gln Met Leu Pro Thr Asp Ala Ile Lys Leu Val Pro Asp Glu Lys
850 855 860
Ile Arg Arg Ile Val Leu Cys Ser Gly Lys Val Tyr Tyr Asp Leu Tyr
865 870 875 880
Glu Glu Arg Glu Lys Arg Gly Ile Asp Asp Ile Tyr Leu Met Arg Val
885 890 895
Glu Gln Leu Tyr Pro Val Pro Leu Lys Ala Leu Val Ala Glu Leu Ser
900 905 910
Arg Phe Lys Lys Ala Glu Val Val Trp Cys Gln Glu Glu Pro Arg Asn
915 920 925
Met Gly Ala Trp His Phe Ile Glu Pro Tyr Leu Glu Trp Val Leu Asn
930 935 940
Gln Val Asn Gly Val Ser Arg Arg Pro Arg Tyr Val Gly Arg Ala Ala
945 950 955 960
Ser Ala Ala Thr Ala Thr Gly Leu Met Ser Lys His Gln Ala Gln Leu
965 970 975
Lys Ala Phe Leu Asp Glu Ala Leu Ser
980 985
<210> 46
<211> 995
<212> PRT
<213> 百脉根瘤菌(Rhizobium loti)
<400> 46
Met Ala Arg Gln Asp Gln Thr Asn Asp Gln Phe Ser Leu Thr Ser Phe
1 5 10 15
Leu Tyr Gly Gly Asn Ala Asp Tyr Ile Asp Ala Leu Tyr Ala Ala Tyr
20 25 30
Glu Asp Asp Pro Ala Ser Val Asn Pro Glu Trp Gln Glu Phe Phe Ala
35 40 45
Gly Leu Lys Asp Asp Ala Gly Asp Val Arg Arg Asn Ala Lys Gly Ala
50 55 60
Ser Trp Ala Lys Pro Ser Trp Pro Leu Gln Ala Asn Gly Glu Leu Val
65 70 75 80
Ser Ala Leu Asp Gly Asn Trp Gly Ile Val Glu Lys His Leu Glu Lys
85 90 95
Lys Val Lys Asp Lys Ala Val Thr Asn Gly Val Val Leu Ser Asp Ala
100 105 110
Asp Val His Gln Ala Thr Arg Asp Ser Val Arg Ala Ile Met Met Ile
115 120 125
Arg Ala Tyr Arg Met Arg Gly His Leu His Ala Asn Leu Asp Pro Leu
130 135 140
Gly Ile Ala Lys Pro Leu Glu Asp Tyr Asn Glu Leu Ser Pro Glu Asn
145 150 155 160
Tyr Gly Phe Thr Ala Ala Asp Tyr Asp Arg Pro Ile Phe Leu Asp Asn
165 170 175
Val Leu Gly Leu Glu Phe Gly Thr Ile Arg Gln Met Leu Glu Ile Leu
180 185 190
Thr Arg Thr Tyr Cys Ser Thr Leu Gly Val Glu Phe Met His Ile Ser
195 200 205
Asp Pro Glu Glu Lys Ala Trp Ile Gln Ala Arg Ile Glu Gly Ala Asp
210 215 220
Lys Glu Ile Ser Phe Thr Asn Thr Gly Lys Lys Ala Ile Leu Gln Lys
225 230 235 240
Leu Val Glu Ala Glu Gly Phe Glu Gln Phe Ile Asp Val Lys Tyr Lys
245 250 255
Gly Thr Lys Arg Phe Gly Leu Asp Gly Gly Glu Ala Leu Ile Pro Ala
260 265 270
Leu Glu Gln Ile Val Lys Arg Gly Gly Gln Leu Gly Met Lys Glu Ile
275 280 285
Val Leu Gly Met Ala His Arg Gly Arg Leu Asn Val Leu Ser Gln Val
290 295 300
Met Ala Lys Pro His Arg Ala Ile Phe His Glu Phe Lys Gly Gly Ser
305 310 315 320
Ala Ala Pro Asp Glu Val Glu Gly Ser Gly Asp Val Lys Tyr His Leu
325 330 335
Gly Ala Ser Ser Asp Arg Glu Phe Asp Gly Asn Lys Val His Leu Ser
340 345 350
Leu Thr Ala Asn Pro Ser His Leu Glu Ile Val Asp Pro Val Val Met
355 360 365
Gly Lys Ala Arg Ala Lys Gln Asp Tyr Leu Phe Gly Arg Gly Arg Glu
370 375 380
Glu Ile Val Pro Leu Glu Glu Arg Ala Lys Val Leu Pro Leu Leu Leu
385 390 395 400
His Gly Asp Ala Ala Phe Ala Gly Gln Gly Val Ile Ala Glu Ile Leu
405 410 415
Gly Leu Ser Gly Leu Arg Gly His Arg Val Ala Gly Thr Leu His Phe
420 425 430
Ile Ile Asn Asn Gln Ile Gly Phe Thr Thr Asn Pro Arg Phe Ser Arg
435 440 445
Ser Ser Pro Tyr Pro Ser Asp Val Ala Lys Met Ile Glu Ala Pro Ile
450 455 460
Phe His Val Asn Gly Asp Asp Pro Glu Ala Val Val His Ala Thr Lys
465 470 475 480
Val Ala Ile Glu Phe Arg Met Lys Phe His Lys Pro Val Val Val Asp
485 490 495
Met Phe Cys Tyr Arg Arg Phe Gly His Asn Glu Gly Asp Glu Pro Ala
500 505 510
Phe Thr Gln Pro Ile Met Tyr Arg Asn Ile Arg Thr His Lys Thr Thr
515 520 525
Val Gln Ile Tyr Ala Asp Arg Leu Ile Ala Glu Gly His Ile Thr Gln
530 535 540
Ala Glu Leu Asp Gln Met Lys Ala Asp Trp Arg Ala His Leu Glu Ser
545 550 555 560
Glu Trp Glu Val Gly Gln His Tyr Lys Pro Asn Lys Ala Asp Trp Leu
565 570 575
Asp Gly Ala Trp Ser Gly Leu Arg Thr Ala Asp Asn Gln Asp Glu Gln
580 585 590
Arg Arg Gly Lys Thr Ala Val Pro Val Lys Thr Leu Lys Glu Ile Gly
595 600 605
Lys Lys Leu Thr Glu Val Pro Lys Gly Phe Glu Ala His Lys Thr Ile
610 615 620
Ile Arg Phe Leu Glu Asn Arg Arg Glu Ala Ile Glu Ser Gly Glu Gly
625 630 635 640
Ile Asp Trp Ser Thr Ala Glu Ala Leu Ala Phe Gly Ala Ile Leu Leu
645 650 655
Asp Gly Asn Pro Ile Arg Leu Ser Gly Gln Asp Ser Glu Arg Gly Thr
660 665 670
Phe Ser Gln Arg His Ser Val Leu Tyr Asp Gln Arg Asp Glu Thr Arg
675 680 685
Tyr Ile Pro Leu Asn Asn Leu Ser Ala Ala Gln Ala Gly Tyr Glu Val
690 695 700
Ile Asn Ser Met Leu Ser Glu Glu Ala Val Leu Gly Phe Glu Tyr Gly
705 710 715 720
Tyr Ser Leu Ala Glu Pro Lys Ala Leu Thr Leu Trp Glu Ala Gln Phe
725 730 735
Gly Asp Phe Ala Asn Gly Ala Gln Val Val Phe Asp Gln Phe Ile Ser
740 745 750
Ser Gly Glu Arg Lys Trp Leu Arg Met Ser Gly Leu Val Cys Leu Leu
755 760 765
Pro His Gly Tyr Glu Gly Gln Gly Pro Glu His Ser Ser Ala Arg Leu
770 775 780
Glu Arg Phe Leu Gln Leu Cys Ala Glu Asp Asn Met Gln Val Ala Asn
785 790 795 800
Cys Thr Thr Pro Ala Asn Tyr Phe His Ile Leu Arg Arg Gln Leu Lys
805 810 815
Arg Asp Phe Arg Lys Pro Leu Ile Leu Met Thr Pro Lys Ser Leu Leu
820 825 830
Arg His Lys Arg Ala Val Ser Thr Leu Pro Glu Ile Ser Gly Glu Ser
835 840 845
Ser Phe His Arg Leu Leu Trp Asp Asp Ala Gln Leu Leu Pro Asn Gln
850 855 860
Pro Ile Lys Leu Thr Lys Asp Ser Lys Ile Arg Arg Val Val Leu Cys
865 870 875 880
Ser Gly Lys Val Tyr Tyr Asp Leu Tyr Glu Glu Arg Glu Lys Arg Gly
885 890 895
Ile Asn Asp Ile Tyr Leu Leu Arg Val Glu Gln Leu Tyr Pro Phe Pro
900 905 910
Ala Lys Ala Leu Ile Thr Glu Leu Ser Arg Phe Arg Asn Ala Glu Met
915 920 925
Val Trp Cys Gln Glu Glu Pro Lys Asn Met Gly Ala Trp Ser Phe Ile
930 935 940
Asp Pro Tyr Leu Glu Trp Val Leu Ala His Ile Asp Ala Lys His Gln
945 950 955 960
Arg Val Arg Tyr Thr Gly Arg Pro Ala Ala Ala Ser Pro Ala Thr Gly
965 970 975
Leu Met Ser Lys His Leu Ala Gln Leu Ala Ala Leu Leu Glu Asp Ala
980 985 990
Leu Gly Glu
995
<210> 47
<211> 547
<212> PRT
<213> 乳酸乳球菌(Lactococcus lactis)
<400> 47
Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly
1 5 10 15
Ile Glu Glu Ile Phe Gly Val Pro Gly Asp Tyr Asn Leu Gln Phe Leu
20 25 30
Asp Gln Ile Ile Ser Arg Glu Asp Met Lys Trp Ile Gly Asn Ala Asn
35 40 45
Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys
50 55 60
Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala Ile
65 70 75 80
Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu Ile
85 90 95
Val Gly Ser Pro Thr Ser Lys Val Gln Asn Asp Gly Lys Phe Val His
100 105 110
His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu
115 120 125
Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Tyr
130 135 140
Glu Ile Asp Arg Val Leu Ser Gln Leu Leu Lys Glu Arg Lys Pro Val
145 150 155 160
Tyr Ile Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro
165 170 175
Ala Leu Ser Leu Glu Lys Glu Ser Ser Thr Thr Asn Thr Thr Glu Gln
180 185 190
Val Ile Leu Ser Lys Ile Glu Glu Ser Leu Lys Asn Ala Gln Lys Pro
195 200 205
Val Val Ile Ala Gly His Glu Val Ile Ser Phe Gly Leu Glu Lys Thr
210 215 220
Val Thr Gln Phe Val Ser Glu Thr Lys Leu Pro Ile Thr Thr Leu Asn
225 230 235 240
Phe Gly Lys Ser Ala Val Asp Glu Ser Leu Pro Ser Phe Leu Gly Ile
245 250 255
Tyr Asn Gly Lys Leu Ser Glu Ile Ser Leu Lys Asn Phe Val Glu Ser
260 265 270
Ala Asp Phe Ile Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr
275 280 285
Gly Ala Phe Thr His His Leu Asp Glu Asn Lys Met Ile Ser Leu Asn
290 295 300
Ile Asp Glu Gly Ile Ile Phe Asn Lys Val Val Glu Asp Phe Asp Phe
305 310 315 320
Arg Ala Val Val Ser Ser Leu Ser Glu Leu Lys Gly Ile Glu Tyr Glu
325 330 335
Gly Gln Tyr Ile Asp Lys Gln Tyr Glu Glu Phe Ile Pro Ser Ser Ala
340 345 350
Pro Leu Ser Gln Asp Arg Leu Trp Gln Ala Val Glu Ser Leu Thr Gln
355 360 365
Ser Asn Glu Thr Ile Val Ala Glu Gln Gly Thr Ser Phe Phe Gly Ala
370 375 380
Ser Thr Ile Phe Leu Lys Ser Asn Ser Arg Phe Ile Gly Gln Pro Leu
385 390 395 400
Trp Gly Ser Ile Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gln Ile
405 410 415
Ala Asp Lys Glu Ser Arg His Leu Leu Phe Ile Gly Asp Gly Ser Leu
420 425 430
Gln Leu Thr Val Gln Glu Leu Gly Leu Ser Ile Arg Glu Lys Leu Asn
435 440 445
Pro Ile Cys Phe Ile Ile Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu
450 455 460
Ile His Gly Pro Thr Gln Ser Tyr Asn Asp Ile Pro Met Trp Asn Tyr
465 470 475 480
Ser Lys Leu Pro Glu Thr Phe Gly Ala Thr Glu Asp Arg Val Val Ser
485 490 495
Lys Ile Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala
500 505 510
Gln Ala Asp Val Asn Arg Met Tyr Trp Ile Glu Leu Val Leu Glu Lys
515 520 525
Glu Asp Ala Pro Lys Leu Leu Lys Lys Met Gly Lys Leu Phe Ala Glu
530 535 540
Gln Asn Lys
545
<210> 48
<211> 1728
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 48
atgactgaca aaatctccct aggtacttat ctgtttgaaa agttaaagga agcaggctct 60
tattccatct ttggtgttcc tggtgatttc aatttggcat tgttggacca cgtcaaggaa 120
gttgaaggca ttagatgggt cggtaacgct aacgagttga atgccggcta cgaagctgat 180
ggttatgcaa gaatcaatgg atttgcatcc ctaatcacca cctttggtgt cggtgaattg 240
tctgccgtca atgccattgc aggttcttat gctgaacacg tcccattgat ccatattgtt 300
ggtatgcctt ccttgtctgc tatgaagaac aacttgttgt tacaccatac cttgggtgac 360
acaagattcg acaacttcac cgaaatgtca aagaaaatca gtgcaaaggt tgaaattgtt 420
tacgatttgg aatcagctcc aaaattaatt aataacttga ttgaaaccgc ttatcacaca 480
aagagaccag tctacttggg acttccttcc aactttgctg atgaattggt tccagcggca 540
ttagttaagg aaaacaagtt acatttagaa gaacctctaa acaaccccgt tgctgaagaa 600
gaattcattc ataacgttgt tgaaatggtc aagaaggcag aaaaaccaat cattctcgtt 660
gacgcttgtg ctgcaagaca taacatttct aaggaagtga gagagttggc taaattgact 720
aaattccctg tcttcaccac cccaatgggt aaatctactg ttgatgaaga tgatgaagaa 780
ttctttggct tatacttggg ttctctatct gctccagatg ttaaggacat tgttggccca 840
accgattgta tcttatcctt aggtggttta ccttctgatt tcaacaccgg ttccttctca 900
tatggttaca ccactaagaa tgtcgttgaa ttccattcca actactgtaa attcaaatct 960
gcaacttatg aaaacttgat gatgaagggc gcagtccaaa gattgatcag cgaattgaag 1020
aatattaagt attccaatgt ctcaacttta tctccaccaa aatctaaatt tgcttacgaa 1080
tctgcaaagg ttgctccaga aggtatcatc actcaagatt acctgtggaa gagattatct 1140
tacttcttaa agccaagaga tatcattgtc actgaaactg gtacttcctc ctttggtgtc 1200
ttggctaccc acttaccaag agattcaaag tctatctccc aagtcttatg gggttccatt 1260
ggtttctcct taccagctgc agttggtgct gcatttgctg ctgaagatgc acacaaacaa 1320
actggcgaac aagaaagaag aactgttttg tttattggtg atggttcttt acaattgact 1380
gtccaatcaa tctcagatgc tgcaagatgg aacatcaagc catacatctt catcttaaac 1440
aacagaggtt acactatcga aaagttgatc cacggtcgtc atgaggacta caaccaaatt 1500
caaccatggg atcaccaatt gttattgaag ctctttgctg acaagaccca atatgaaaac 1560
catgttgtta aatccgctaa ggacttggac gctttgatga aggatgaagc attcaacaag 1620
gaagataaga ttagagtcat tgaattattc ttggatgaat tcgatgctcc agaaatcttg 1680
gttgctcaag ctaaattatc tgatgaaatc aactctaaag ccgcttaa 1728
<210> 49
<211> 575
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 49
Met Thr Asp Lys Ile Ser Leu Gly Thr Tyr Leu Phe Glu Lys Leu Lys
1 5 10 15
Glu Ala Gly Ser Tyr Ser Ile Phe Gly Val Pro Gly Asp Phe Asn Leu
20 25 30
Ala Leu Leu Asp His Val Lys Glu Val Glu Gly Ile Arg Trp Val Gly
35 40 45
Asn Ala Asn Glu Leu Asn Ala Gly Tyr Glu Ala Asp Gly Tyr Ala Arg
50 55 60
Ile Asn Gly Phe Ala Ser Leu Ile Thr Thr Phe Gly Val Gly Glu Leu
65 70 75 80
Ser Ala Val Asn Ala Ile Ala Gly Ser Tyr Ala Glu His Val Pro Leu
85 90 95
Ile His Ile Val Gly Met Pro Ser Leu Ser Ala Met Lys Asn Asn Leu
100 105 110
Leu Leu His His Thr Leu Gly Asp Thr Arg Phe Asp Asn Phe Thr Glu
115 120 125
Met Ser Lys Lys Ile Ser Ala Lys Val Glu Ile Val Tyr Asp Leu Glu
130 135 140
Ser Ala Pro Lys Leu Ile Asn Asn Leu Ile Glu Thr Ala Tyr His Thr
145 150 155 160
Lys Arg Pro Val Tyr Leu Gly Leu Pro Ser Asn Phe Ala Asp Glu Leu
165 170 175
Val Pro Ala Ala Leu Val Lys Glu Asn Lys Leu His Leu Glu Glu Pro
180 185 190
Leu Asn Asn Pro Val Ala Glu Glu Glu Phe Ile His Asn Val Val Glu
195 200 205
Met Val Lys Lys Ala Glu Lys Pro Ile Ile Leu Val Asp Ala Cys Ala
210 215 220
Ala Arg His Asn Ile Ser Lys Glu Val Arg Glu Leu Ala Lys Leu Thr
225 230 235 240
Lys Phe Pro Val Phe Thr Thr Pro Met Gly Lys Ser Thr Val Asp Glu
245 250 255
Asp Asp Glu Glu Phe Phe Gly Leu Tyr Leu Gly Ser Leu Ser Ala Pro
260 265 270
Asp Val Lys Asp Ile Val Gly Pro Thr Asp Cys Ile Leu Ser Leu Gly
275 280 285
Gly Leu Pro Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Gly Tyr Thr
290 295 300
Thr Lys Asn Val Val Glu Phe His Ser Asn Tyr Cys Lys Phe Lys Ser
305 310 315 320
Ala Thr Tyr Glu Asn Leu Met Met Lys Gly Ala Val Gln Arg Leu Ile
325 330 335
Ser Glu Leu Lys Asn Ile Lys Tyr Ser Asn Val Ser Thr Leu Ser Pro
340 345 350
Pro Lys Ser Lys Phe Ala Tyr Glu Ser Ala Lys Val Ala Pro Glu Gly
355 360 365
Ile Ile Thr Gln Asp Tyr Leu Trp Lys Arg Leu Ser Tyr Phe Leu Lys
370 375 380
Pro Arg Asp Ile Ile Val Thr Glu Thr Gly Thr Ser Ser Phe Gly Val
385 390 395 400
Leu Ala Thr His Leu Pro Arg Asp Ser Lys Ser Ile Ser Gln Val Leu
405 410 415
Trp Gly Ser Ile Gly Phe Ser Leu Pro Ala Ala Val Gly Ala Ala Phe
420 425 430
Ala Ala Glu Asp Ala His Lys Gln Thr Gly Glu Gln Glu Arg Arg Thr
435 440 445
Val Leu Phe Ile Gly Asp Gly Ser Leu Gln Leu Thr Val Gln Ser Ile
450 455 460
Ser Asp Ala Ala Arg Trp Asn Ile Lys Pro Tyr Ile Phe Ile Leu Asn
465 470 475 480
Asn Arg Gly Tyr Thr Ile Glu Lys Leu Ile His Gly Arg His Glu Asp
485 490 495
Tyr Asn Gln Ile Gln Pro Trp Asp His Gln Leu Leu Leu Lys Leu Phe
500 505 510
Ala Asp Lys Thr Gln Tyr Glu Asn His Val Val Lys Ser Ala Lys Asp
515 520 525
Leu Asp Ala Leu Met Lys Asp Glu Ala Phe Asn Lys Glu Asp Lys Ile
530 535 540
Arg Val Ile Glu Leu Phe Leu Asp Glu Phe Asp Ala Pro Glu Ile Leu
545 550 555 560
Val Ala Gln Ala Lys Leu Ser Asp Glu Ile Asn Ser Lys Ala Ala
565 570 575
<210> 50
<211> 563
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 50
Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln
1 5 10 15
Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser
20 25 30
Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met Arg Trp Ala Gly Asn
35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile
50 55 60
Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser
65 70 75 80
Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu
85 90 95
His Val Val Gly Val Pro Ser Ile Ser Ala Gln Ala Lys Gln Leu Leu
100 105 110
Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met
115 120 125
Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Thr
130 135 140
Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Val Thr Gln
145 150 155 160
Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val
165 170 175
Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met Ser Leu Lys Pro Asn
180 185 190
Asp Ala Glu Ser Glu Lys Glu Val Ile Asp Thr Ile Leu Ala Leu Val
195 200 205
Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Cys Ser Arg
210 215 220
His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln Phe
225 230 235 240
Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln His
245 250 255
Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu Val
260 265 270
Lys Glu Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala Leu
275 280 285
Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys
290 295 300
Asn Ile Val Glu Phe His Ser Asp His Met Lys Ile Arg Asn Ala Thr
305 310 315 320
Phe Pro Gly Val Gln Met Lys Phe Val Leu Gln Lys Leu Leu Thr Thr
325 330 335
Ile Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala Arg
340 345 350
Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gln Glu
355 360 365
Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln Glu Gly Asp Val Val
370 375 380
Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe
385 390 395 400
Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile Gly
405 410 415
Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile
420 425 430
Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln
435 440 445
Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro
450 455 460
Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Lys Leu Ile
465 470 475 480
His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln Gly Trp Asp His Leu
485 490 495
Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val
500 505 510
Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gln Asp Lys Ser Phe Asn
515 520 525
Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met Leu Pro Val Phe Asp
530 535 540
Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu Thr Ala Ala Thr Asn
545 550 555 560
Ala Lys Gln
<210> 51
<211> 563
<212> PRT
<213> 乳酸克鲁维酵母(Saccharomyces kluyveri)
<400> 51
Met Ser Glu Ile Thr Leu Gly Arg Tyr Leu Phe Glu Arg Leu Lys Gln
1 5 10 15
Val Glu Val Gln Thr Ile Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser
20 25 30
Leu Leu Asp Asn Ile Tyr Glu Val Pro Gly Met Arg Trp Ala Gly Asn
35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Leu
50 55 60
Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser
65 70 75 80
Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu
85 90 95
His Val Val Gly Val Pro Ser Val Ser Ser Gln Ala Lys Gln Leu Leu
100 105 110
Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met
115 120 125
Ser Ser Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Asn Thr
130 135 140
Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Val Ser Gln
145 150 155 160
Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Thr Val
165 170 175
Pro Ala Ser Leu Leu Asp Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn
180 185 190
Asp Pro Glu Ala Glu Glu Glu Val Ile Glu Asn Val Leu Gln Leu Ile
195 200 205
Lys Glu Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Cys Ser Arg
210 215 220
His Asp Ala Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln Phe
225 230 235 240
Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Lys His
245 250 255
Pro Arg Phe Gly Gly Val Tyr Val Gly Thr Leu Ser Ser Pro Ala Val
260 265 270
Lys Glu Ala Val Glu Ser Ala Asp Leu Val Leu Ser Val Gly Ala Leu
275 280 285
Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys
290 295 300
Asn Ile Val Glu Phe His Ser Asp Tyr Thr Lys Ile Arg Ser Ala Thr
305 310 315 320
Phe Pro Gly Val Gln Met Lys Phe Ala Leu Gln Lys Leu Leu Thr Lys
325 330 335
Val Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Pro Val Pro Ser Glu
340 345 350
Pro Glu His Asn Glu Ala Val Ala Asp Ser Thr Pro Leu Lys Gln Glu
355 360 365
Trp Val Trp Thr Gln Val Gly Glu Phe Leu Arg Glu Gly Asp Val Val
370 375 380
Ile Thr Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr His Phe
385 390 395 400
Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile Gly
405 410 415
Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile
420 425 430
Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln
435 440 445
Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro
450 455 460
Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Arg Leu Ile
465 470 475 480
His Gly Glu Thr Ala Gln Tyr Asn Cys Ile Gln Asn Trp Gln His Leu
485 490 495
Glu Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Ala Val Arg Val
500 505 510
Ser Thr Thr Gly Glu Trp Asn Lys Leu Thr Thr Asp Glu Lys Phe Gln
515 520 525
Asp Asn Thr Arg Ile Arg Leu Ile Glu Val Met Leu Pro Thr Met Asp
530 535 540
Ala Pro Ser Asn Leu Val Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn
545 550 555 560
Ala Lys Asn
<210> 52
<211> 568
<212> PRT
<213> 运动发酵单胞菌(Zymomonas mobilis)
<400> 52
Met Ser Tyr Thr Val Gly Thr Tyr Leu Ala Glu Arg Leu Val Gln Ile
1 5 10 15
Gly Leu Lys His His Phe Ala Val Ala Gly Asp Tyr Asn Leu Val Leu
20 25 30
Leu Asp Asn Leu Leu Leu Asn Lys Asn Met Glu Gln Val Tyr Cys Cys
35 40 45
Asn Glu Leu Asn Cys Gly Phe Ser Ala Glu Gly Tyr Ala Arg Ala Lys
50 55 60
Gly Ala Ala Ala Ala Val Val Thr Tyr Ser Val Gly Ala Leu Ser Ala
65 70 75 80
Phe Asp Ala Ile Gly Gly Ala Tyr Ala Glu Asn Leu Pro Val Ile Leu
85 90 95
Ile Ser Gly Ala Pro Asn Asn Asn Asp His Ala Ala Gly His Val Leu
100 105 110
His His Ala Leu Gly Lys Thr Asp Tyr His Tyr Gln Leu Glu Met Ala
115 120 125
Lys Asn Ile Thr Ala Ala Ala Glu Ala Ile Tyr Thr Pro Glu Glu Ala
130 135 140
Pro Ala Lys Ile Asp His Val Ile Lys Thr Ala Leu Arg Glu Lys Lys
145 150 155 160
Pro Val Tyr Leu Glu Ile Ala Cys Asn Ile Ala Ser Met Pro Cys Ala
165 170 175
Ala Pro Gly Pro Ala Ser Ala Leu Phe Asn Asp Glu Ala Ser Asp Glu
180 185 190
Ala Ser Leu Asn Ala Ala Val Glu Glu Thr Leu Lys Phe Ile Ala Asn
195 200 205
Arg Asp Lys Val Ala Val Leu Val Gly Ser Lys Leu Arg Ala Ala Gly
210 215 220
Ala Glu Glu Ala Ala Val Lys Phe Ala Asp Ala Leu Gly Gly Ala Val
225 230 235 240
Ala Thr Met Ala Ala Ala Lys Ser Phe Phe Pro Glu Glu Asn Pro His
245 250 255
Tyr Ile Gly Thr Ser Trp Gly Glu Val Ser Tyr Pro Gly Val Glu Lys
260 265 270
Thr Met Lys Glu Ala Asp Ala Val Ile Ala Leu Ala Pro Val Phe Asn
275 280 285
Asp Tyr Ser Thr Thr Gly Trp Thr Asp Ile Pro Asp Pro Lys Lys Leu
290 295 300
Val Leu Ala Glu Pro Arg Ser Val Val Val Asn Gly Ile Arg Phe Pro
305 310 315 320
Ser Val His Leu Lys Asp Tyr Leu Thr Arg Leu Ala Gln Lys Val Ser
325 330 335
Lys Lys Thr Gly Ala Leu Asp Phe Phe Lys Ser Leu Asn Ala Gly Glu
340 345 350
Leu Lys Lys Ala Ala Pro Ala Asp Pro Ser Ala Pro Leu Val Asn Ala
355 360 365
Glu Ile Ala Arg Gln Val Glu Ala Leu Leu Thr Pro Asn Thr Thr Val
370 375 380
Ile Ala Glu Thr Gly Asp Ser Trp Phe Asn Ala Gln Arg Met Lys Leu
385 390 395 400
Pro Asn Gly Ala Arg Val Glu Tyr Glu Met Gln Trp Gly His Ile Gly
405 410 415
Trp Ser Val Pro Ala Ala Phe Gly Tyr Ala Val Gly Ala Pro Glu Arg
420 425 430
Arg Asn Ile Leu Met Val Gly Asp Gly Ser Phe Gln Leu Thr Ala Gln
435 440 445
Glu Val Ala Gln Met Val Arg Leu Lys Leu Pro Val Ile Ile Phe Leu
450 455 460
Ile Asn Asn Tyr Gly Tyr Thr Ile Glu Val Met Ile His Asp Gly Pro
465 470 475 480
Tyr Asn Asn Ile Lys Asn Trp Asp Tyr Ala Gly Leu Met Glu Val Phe
485 490 495
Asn Gly Asn Gly Gly Tyr Asp Ser Gly Ala Gly Lys Gly Leu Lys Ala
500 505 510
Lys Thr Gly Gly Glu Leu Ala Glu Ala Ile Lys Val Ala Leu Ala Asn
515 520 525
Thr Asp Gly Pro Thr Leu Ile Glu Cys Phe Ile Gly Arg Glu Asp Cys
530 535 540
Thr Glu Glu Leu Val Lys Trp Gly Lys Arg Val Ala Ala Ala Asn Ser
545 550 555 560
Arg Lys Pro Val Asn Lys Leu Leu
565
<210> 53
<211> 557
<212> PRT
<213> 巴氏醋杆菌(Acetobacter pasteurianus)
<400> 53
Met Thr Tyr Thr Val Gly Met Tyr Leu Ala Glu Arg Leu Val Gln Ile
1 5 10 15
Gly Leu Lys His His Phe Ala Val Gly Gly Asp Tyr Asn Leu Val Leu
20 25 30
Leu Asp Gln Leu Leu Leu Asn Lys Asp Met Lys Gln Ile Tyr Cys Cys
35 40 45
Asn Glu Leu Asn Cys Gly Phe Ser Ala Glu Gly Tyr Ala Arg Ser Asn
50 55 60
Gly Ala Ala Ala Ala Val Val Thr Phe Ser Val Gly Ala Ile Ser Ala
65 70 75 80
Met Asn Ala Leu Gly Gly Ala Tyr Ala Glu Asn Leu Pro Val Ile Leu
85 90 95
Ile Ser Gly Ala Pro Asn Ser Asn Asp Gln Gly Thr Gly His Ile Leu
100 105 110
His His Thr Ile Gly Lys Thr Asp Tyr Ser Tyr Gln Leu Glu Met Ala
115 120 125
Arg Gln Val Thr Cys Ala Ala Glu Ser Ile Thr Asp Ala His Ser Ala
130 135 140
Pro Ala Lys Ile Asp His Val Ile Arg Thr Ala Leu Arg Glu Arg Lys
145 150 155 160
Pro Ala Tyr Leu Asp Ile Ala Cys Asn Ile Ala Ser Glu Pro Cys Val
165 170 175
Arg Pro Gly Pro Val Ser Ser Leu Leu Ser Glu Pro Glu Ile Asp His
180 185 190
Thr Ser Leu Lys Ala Ala Val Asp Ala Thr Val Ala Leu Leu Lys Asn
195 200 205
Arg Pro Ala Pro Val Met Leu Leu Gly Ser Lys Leu Arg Ala Ala Asn
210 215 220
Ala Leu Ala Ala Thr Glu Thr Leu Ala Asp Lys Leu Gln Cys Ala Val
225 230 235 240
Thr Ile Met Ala Ala Ala Lys Gly Phe Phe Pro Glu Asp His Ala Gly
245 250 255
Phe Arg Gly Leu Tyr Trp Gly Glu Val Ser Asn Pro Gly Val Gln Glu
260 265 270
Leu Val Glu Thr Ser Asp Ala Leu Leu Cys Ile Ala Pro Val Phe Asn
275 280 285
Asp Tyr Ser Thr Val Gly Trp Ser Gly Met Pro Lys Gly Pro Asn Val
290 295 300
Ile Leu Ala Glu Pro Asp Arg Val Thr Val Asp Gly Arg Ala Tyr Asp
305 310 315 320
Gly Phe Thr Leu Arg Ala Phe Leu Gln Ala Leu Ala Glu Lys Ala Pro
325 330 335
Ala Arg Pro Ala Ser Ala Gln Lys Ser Ser Val Pro Thr Cys Ser Leu
340 345 350
Thr Ala Thr Ser Asp Glu Ala Gly Leu Thr Asn Asp Glu Ile Val Arg
355 360 365
His Ile Asn Ala Leu Leu Thr Ser Asn Thr Thr Leu Val Ala Glu Thr
370 375 380
Gly Asp Ser Trp Phe Asn Ala Met Arg Met Thr Leu Ala Gly Ala Arg
385 390 395 400
Val Glu Leu Glu Met Gln Trp Gly His Ile Gly Trp Ser Val Pro Ser
405 410 415
Ala Phe Gly Asn Ala Met Gly Ser Gln Asp Arg Gln His Val Val Met
420 425 430
Val Gly Asp Gly Ser Phe Gln Leu Thr Ala Gln Glu Val Ala Gln Met
435 440 445
Val Arg Tyr Glu Leu Pro Val Ile Ile Phe Leu Ile Asn Asn Arg Gly
450 455 460
Tyr Val Ile Glu Ile Ala Ile His Asp Gly Pro Tyr Asn Tyr Ile Lys
465 470 475 480
Asn Trp Asp Tyr Ala Gly Leu Met Glu Val Phe Asn Ala Gly Glu Gly
485 490 495
His Gly Leu Gly Leu Lys Ala Thr Thr Pro Lys Glu Leu Thr Glu Ala
500 505 510
Ile Ala Arg Ala Lys Ala Asn Thr Arg Gly Pro Thr Leu Ile Glu Cys
515 520 525
Gln Ile Asp Arg Thr Asp Cys Thr Asp Met Leu Val Gln Trp Gly Arg
530 535 540
Lys Val Ala Ser Thr Asn Ala Arg Lys Thr Thr Leu Ala
545 550 555
<210> 54
<211> 528
<212> PRT
<213> 恶臭假单胞菌(Pseudomonas putida)
<400> 54
Met Ala Ser Val His Gly Thr Thr Tyr Glu Leu Leu Arg Arg Gln Gly
1 5 10 15
Ile Asp Thr Val Phe Gly Asn Pro Gly Ser Asn Glu Leu Pro Phe Leu
20 25 30
Lys Asp Phe Pro Glu Asp Phe Arg Tyr Ile Leu Ala Leu Gln Glu Ala
35 40 45
Cys Val Val Gly Ile Ala Asp Gly Tyr Ala Gln Ala Ser Arg Lys Pro
50 55 60
Ala Phe Ile Asn Leu His Ser Ala Ala Gly Thr Gly Asn Ala Met Gly
65 70 75 80
Ala Leu Ser Asn Ala Trp Asn Ser His Ser Pro Leu Ile Val Thr Ala
85 90 95
Gly Gln Gln Thr Arg Ala Met Ile Gly Val Glu Ala Leu Leu Thr Asn
100 105 110
Val Asp Ala Ala Asn Leu Pro Arg Pro Leu Val Lys Trp Ser Tyr Glu
115 120 125
Pro Ala Ser Ala Ala Glu Val Pro His Ala Met Ser Arg Ala Ile His
130 135 140
Met Ala Ser Met Ala Pro Gln Gly Pro Val Tyr Leu Ser Val Pro Tyr
145 150 155 160
Asp Asp Trp Asp Lys Asp Ala Asp Pro Gln Ser His His Leu Phe Asp
165 170 175
Arg His Val Ser Ser Ser Val Arg Leu Asn Asp Gln Asp Leu Asp Ile
180 185 190
Leu Val Lys Ala Leu Asn Ser Ala Ser Asn Pro Ala Ile Val Leu Gly
195 200 205
Pro Asp Val Asp Ala Ala Asn Ala Asn Ala Asp Cys Val Met Leu Ala
210 215 220
Glu Arg Leu Lys Ala Pro Val Trp Val Ala Pro Ser Ala Pro Arg Cys
225 230 235 240
Pro Phe Pro Thr Arg His Pro Cys Phe Arg Gly Leu Met Pro Ala Gly
245 250 255
Ile Ala Ala Ile Ser Gln Leu Leu Glu Gly His Asp Val Val Leu Val
260 265 270
Ile Gly Ala Pro Val Phe Arg Tyr His Gln Tyr Asp Pro Gly Gln Tyr
275 280 285
Leu Lys Pro Gly Thr Arg Leu Ile Ser Val Thr Cys Asp Pro Leu Glu
290 295 300
Ala Ala Arg Ala Pro Met Gly Asp Ala Ile Val Ala Asp Ile Gly Ala
305 310 315 320
Met Ala Ser Ala Leu Ala Asn Leu Val Glu Glu Ser Ser Arg Gln Leu
325 330 335
Pro Thr Ala Ala Pro Glu Pro Ala Lys Val Asp Gln Asp Ala Gly Arg
340 345 350
Leu His Pro Glu Thr Val Phe Asp Thr Leu Asn Asp Met Ala Pro Glu
355 360 365
Asn Ala Ile Tyr Leu Asn Glu Ser Thr Ser Thr Thr Ala Gln Met Trp
370 375 380
Gln Arg Leu Asn Met Arg Asn Pro Gly Ser Tyr Tyr Phe Cys Ala Ala
385 390 395 400
Gly Gly Leu Gly Phe Ala Leu Pro Ala Ala Ile Gly Val Gln Leu Ala
405 410 415
Glu Pro Glu Arg Gln Val Ile Ala Val Ile Gly Asp Gly Ser Ala Asn
420 425 430
Tyr Ser Ile Ser Ala Leu Trp Thr Ala Ala Gln Tyr Asn Ile Pro Thr
435 440 445
Ile Phe Val Ile Met Asn Asn Gly Thr Tyr Gly Ala Leu Arg Trp Phe
450 455 460
Ala Gly Val Leu Glu Ala Glu Asn Val Pro Gly Leu Asp Val Pro Gly
465 470 475 480
Ile Asp Phe Arg Ala Leu Ala Lys Gly Tyr Gly Val Gln Ala Leu Lys
485 490 495
Ala Asp Asn Leu Glu Gln Leu Lys Gly Ser Leu Gln Glu Ala Leu Ser
500 505 510
Ala Lys Gly Pro Val Leu Ile Glu Val Ser Thr Val Ser Pro Val Lys
515 520 525
<210> 55
<211> 528
<212> PRT
<213> 铜绿假单胞菌(Pseudomonas aeruginosa)
<400> 55
Met Lys Thr Val His Ser Ala Ser Tyr Glu Ile Leu Arg Arg His Gly
1 5 10 15
Leu Thr Thr Val Phe Gly Asn Pro Gly Ser Asn Glu Leu Pro Phe Leu
20 25 30
Lys Asp Phe Pro Glu Asp Phe Arg Tyr Ile Leu Gly Leu His Glu Gly
35 40 45
Ala Val Val Gly Met Ala Asp Gly Phe Ala Leu Ala Ser Gly Arg Pro
50 55 60
Ala Phe Val Asn Leu His Ala Ala Ala Gly Thr Gly Asn Gly Met Gly
65 70 75 80
Ala Leu Thr Asn Ala Trp Tyr Ser His Ser Pro Leu Val Ile Thr Ala
85 90 95
Gly Gln Gln Val Arg Ser Met Ile Gly Val Glu Ala Met Leu Ala Asn
100 105 110
Val Asp Ala Gly Gln Leu Pro Lys Pro Leu Val Lys Trp Ser His Glu
115 120 125
Pro Ala Cys Ala Gln Asp Val Pro Arg Ala Leu Ser Gln Ala Ile Gln
130 135 140
Thr Ala Ser Leu Pro Pro Arg Ala Pro Val Tyr Leu Ser Ile Pro Tyr
145 150 155 160
Asp Asp Trp Ala Gln Pro Ala Pro Ala Gly Val Glu His Leu Ala Ala
165 170 175
Arg Gln Val Ser Gly Ala Ala Leu Pro Ala Pro Ala Leu Leu Ala Glu
180 185 190
Leu Gly Glu Arg Leu Ser Arg Ser Arg Asn Pro Val Leu Val Leu Gly
195 200 205
Pro Asp Val Asp Gly Ala Asn Ala Asn Gly Leu Ala Val Glu Leu Ala
210 215 220
Glu Lys Leu Arg Met Pro Ala Trp Val Ala Pro Ser Ala Ser Arg Cys
225 230 235 240
Pro Phe Pro Thr Arg His Ala Cys Phe Arg Gly Val Leu Pro Ala Ala
245 250 255
Ile Ala Gly Ile Ser Arg Leu Leu Asp Gly His Asp Leu Ile Leu Val
260 265 270
Val Gly Ala Pro Val Phe Arg Tyr His Gln Phe Ala Pro Gly Asp Tyr
275 280 285
Leu Pro Ala Gly Ala Glu Leu Val Gln Val Thr Cys Asp Pro Gly Glu
290 295 300
Ala Ala Arg Ala Pro Met Gly Asp Ala Leu Val Gly Asp Ile Ala Leu
305 310 315 320
Thr Leu Glu Ala Leu Leu Glu Gln Val Arg Pro Ser Ala Arg Pro Leu
325 330 335
Pro Glu Ala Leu Pro Arg Pro Pro Ala Leu Ala Glu Glu Gly Gly Pro
340 345 350
Leu Arg Pro Glu Thr Val Phe Asp Val Ile Asp Ala Leu Ala Pro Arg
355 360 365
Asp Ala Ile Phe Val Lys Glu Ser Thr Ser Thr Val Thr Ala Phe Trp
370 375 380
Gln Arg Val Glu Met Arg Glu Pro Gly Ser Tyr Phe Phe Pro Ala Ala
385 390 395 400
Gly Gly Leu Gly Phe Gly Leu Pro Ala Ala Val Gly Ala Gln Leu Ala
405 410 415
Gln Pro Arg Arg Gln Val Ile Gly Ile Ile Gly Asp Gly Ser Ala Asn
420 425 430
Tyr Gly Ile Thr Ala Leu Trp Ser Ala Ala Gln Tyr Arg Val Pro Ala
435 440 445
Val Phe Ile Ile Leu Lys Asn Gly Thr Tyr Gly Ala Leu Arg Trp Phe
450 455 460
Ala Gly Val Leu Glu Val Pro Asp Ala Pro Gly Leu Asp Val Pro Gly
465 470 475 480
Leu Asp Phe Cys Ala Ile Ala Arg Gly Tyr Gly Val Glu Ala Leu His
485 490 495
Ala Ala Thr Arg Glu Glu Leu Glu Gly Ala Leu Lys His Ala Leu Ala
500 505 510
Ala Asp Arg Pro Val Leu Ile Glu Val Pro Thr Gln Thr Ile Glu Pro
515 520 525
<210> 56
<211> 526
<212> PRT
<213> 施氏假单胞菌(Pseudomonas stutzeri)
<400> 56
Met Ala Ser Val His Ser Ile Thr Tyr Glu Leu Leu Arg Arg Gln Gly
1 5 10 15
Ile Asp Thr Val Phe Gly Asn Pro Gly Ser Asn Glu Leu Pro Phe Leu
20 25 30
Lys Asp Phe Pro Glu Asp Phe Arg Tyr Ile Leu Ala Leu Gln Glu Ala
35 40 45
Cys Val Val Gly Ile Ala Asp Gly Tyr Ala Gln Ala Ser Arg Lys Pro
50 55 60
Ala Phe Ile Asn Leu His Ser Ala Ala Gly Thr Gly Asn Ala Met Gly
65 70 75 80
Ala Met Ser Asn Ala Trp Asn Cys His Ser Pro Leu Ile Val Thr Ala
85 90 95
Gly Gln Gln Asn Arg Ala Met Ile Gly Val Glu Ala Leu Leu Thr Asn
100 105 110
Val Asp Ala Ala Ser Leu Pro Arg Pro Leu Val Lys Trp Ser Tyr Glu
115 120 125
Pro Ala Ser Ala Ala Glu Val Pro His Ala Met Ser Arg Ala Ile His
130 135 140
Met Ala Ser Met Ala Pro Arg Gly Pro Val Tyr Leu Ser Val Pro Tyr
145 150 155 160
Asp Asp Trp Asp Lys Glu Ala Asp Pro Gln Ser His His Leu Tyr Asp
165 170 175
Arg Ser Val Asn Ser Ala Val Arg Leu Asn Asp Gln Asp Leu Glu Val
180 185 190
Leu Val Glu Ala Leu Asn Ser Ala Ser Asn Pro Ala Ile Val Leu Gly
195 200 205
Pro Asp Val Asp Ser Ala Asn Ala Asn Ala Asp Cys Val Thr Leu Ala
210 215 220
Glu Arg Leu Lys Ala Pro Val Trp Val Ala Pro Ser Ala Pro Arg Cys
225 230 235 240
Pro Phe Pro Thr Arg His Pro Cys Phe Arg Gly Leu Met Pro Ala Gly
245 250 255
Ile Ala Ala Ile Ser Gln Leu Leu Glu Gly His Asp Val Val Leu Val
260 265 270
Ile Gly Ala Pro Val Phe Arg Tyr His Gln Tyr Asp Pro Gly Gln Tyr
275 280 285
Leu Lys Pro Gly Thr Arg Leu Ile Ser Ile Thr Cys Asp Pro Leu Glu
290 295 300
Ala Ala Arg Ala Pro Met Gly Asp Ala Ile Val Ala Asp Ile Gly Thr
305 310 315 320
Met Thr Ala Ala Leu Ala Ser Arg Ile Gly Glu Ser Glu Arg Gln Leu
325 330 335
Pro Ala Val Leu Pro Ser Pro Glu Arg Val Asn Gln Asp Ala Gly Arg
340 345 350
Leu Arg Pro Glu Thr Val Phe Asp Thr Leu Asn Glu Met Ala Pro Glu
355 360 365
Asp Ala Ile Tyr Leu Asn Glu Ser Thr Ser Thr Thr Ala Gln Met Trp
370 375 380
Gln Arg Leu Asn Met Arg Asn Pro Gly Ser Tyr Tyr Phe Cys Ala Ala
385 390 395 400
Gly Gly Leu Gly Phe Ala Leu Pro Ala Ala Ile Gly Val Gln Leu Ala
405 410 415
Glu Pro Asp Arg Gln Val Ile Ala Val Ile Gly Asp Gly Ser Ala Asn
420 425 430
Tyr Ser Ile Ser Ala Leu Trp Thr Ala Ala His Tyr Asn Ile Pro Ala
435 440 445
Ile Phe Leu Ile Met Asn Asn Gly Thr Tyr Gly Ala Leu Arg Trp Phe
450 455 460
Ala Gly Val Leu Glu Ala Glu Asn Val Pro Gly Leu Asp Val Pro Gly
465 470 475 480
Ile Asp Phe Cys Ala Ile Ala Lys Gly Tyr Gly Ile Pro Ala Leu Lys
485 490 495
Ala Asp Asn Leu Glu Gln Leu Lys Gly Ser Ile His Glu Ala Leu Ser
500 505 510
Ala Lys Gly Pro Val Leu Ile Glu Val Ser Thr Val Ser Leu
515 520 525
<210> 57
<211> 528
<212> PRT
<213> 荧光假单胞菌(Pseudomonas fluorescens)
<400> 57
Met Lys Thr Val His Ser Ala Ser Tyr Asp Ile Leu Arg Gln Gln Gly
1 5 10 15
Leu Thr Thr Val Phe Gly Asn Pro Gly Ser Asn Glu Leu Pro Phe Leu
20 25 30
Lys Gly Phe Pro Glu Asp Phe Arg Tyr Ile Leu Gly Leu His Glu Gly
35 40 45
Ala Val Val Gly Met Ala Asp Gly Phe Ala Leu Ala Ser Gly Gln Pro
50 55 60
Ala Phe Val Asn Leu His Ala Ala Ala Gly Thr Gly Asn Gly Met Gly
65 70 75 80
Ala Leu Thr Asn Ala Trp Tyr Ser His Ser Pro Leu Val Ile Thr Ala
85 90 95
Gly Gln Gln Val Arg Ser Met Ile Gly Val Glu Ala Met Leu Ala Asn
100 105 110
Val Asp Ala Pro Gln Leu Pro Lys Pro Leu Val Lys Trp Ser Ala Glu
115 120 125
Pro Ala Cys Ala Glu Asp Val Pro Arg Ala Leu Ser Gln Ala Ile His
130 135 140
Met Ala Asn Gln Ala Pro Lys Gly Pro Val Tyr Leu Ser Ile Pro Tyr
145 150 155 160
Asp Asp Trp Ala Arg Pro Ala Pro Ala Gly Val Glu His Leu Ala Arg
165 170 175
Arg Gln Val Ala Thr Ala Gly Leu Pro Ser Ala Ala Gln Leu Arg Ser
180 185 190
Leu Val Gln Arg Leu Ala Ala Ala Arg Asn Pro Val Leu Val Leu Gly
195 200 205
Pro Asp Val Asp Gly Ser Arg Ser Asn His Leu Ala Val Gln Leu Ala
210 215 220
Glu Lys Leu Arg Met Pro Ala Trp Val Ala Pro Ser Ala Ser Arg Cys
225 230 235 240
Pro Phe Pro Thr Arg His Pro Ser Phe Arg Gly Val Leu Pro Ala Ala
245 250 255
Ile Ala Gly Ile Ser Arg Cys Leu Ala Asp His Asp Leu Ile Leu Val
260 265 270
Val Gly Ala Pro Val Phe Arg Tyr His Gln Phe Ala Pro Gly Asp Tyr
275 280 285
Leu Pro Ala Gly Thr Glu Leu Leu His Ile Thr Cys Asp Pro Gly Glu
290 295 300
Ala Ala Arg Ala Pro Met Gly Asp Ala Leu Val Gly Asp Ile Val Glu
305 310 315 320
Thr Leu Gln Ala Leu Val Trp Ala Leu Pro Asp Cys Asp Arg Pro Gln
325 330 335
Pro Gln Ala Leu Pro Pro Ala Ala Pro Val Glu Glu Leu Gly Gly Leu
340 345 350
Leu Arg Pro Glu Thr Val Phe Asp Val Ile Asp Glu Leu Ala Pro Lys
355 360 365
Asp Ala Ile Tyr Val Lys Glu Ser Thr Ser Thr Val Gly Ala Phe Trp
370 375 380
Gln Arg Val Glu Met Arg Glu Pro Gly Ser Tyr Tyr Phe Pro Ala Ala
385 390 395 400
Gly Gly Leu Gly Phe Gly Leu Pro Ala Ala Val Gly Val Gln Leu Ala
405 410 415
Arg Pro Glu Arg Arg Val Ile Gly Val Ile Gly Asp Gly Ser Ala Asn
420 425 430
Tyr Gly Ile Thr Ala Leu Trp Thr Ala Ala Gln Tyr Gln Ile Pro Val
435 440 445
Val Phe Ile Ile Leu Lys Asn Gly Thr Tyr Gly Ala Leu Arg Trp Phe
450 455 460
Ala Gly Val Leu Gln Val Ser Asp Ala Pro Gly Leu Asp Val Pro Gly
465 470 475 480
Leu Asp Phe Cys Ala Ile Gly Arg Gly Tyr Gly Val His Ser Val Gln
485 490 495
Ala Asn Thr Arg Glu Ala Phe Ala Gln Ala Leu Ser Glu Ala Leu Ala
500 505 510
Gly Asp Arg Pro Val Leu Ile Glu Val Pro Thr Leu Thr Ile Glu Pro
515 520 525
<210> 58
<211> 1220
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 58
Met Ala Thr Gly Glu Ser Met Ser Gly Thr Gly Arg Leu Ala Gly Lys
1 5 10 15
Ile Ala Leu Ile Thr Gly Gly Ala Gly Asn Ile Gly Ser Glu Leu Thr
20 25 30
Arg Arg Phe Leu Ala Glu Gly Ala Thr Val Ile Ile Ser Gly Arg Asn
35 40 45
Arg Ala Lys Leu Thr Ala Leu Ala Glu Met Gln Ala Glu Ala Gly Val
50 55 60
Pro Ala Lys Arg Ile Asp Leu Glu Val Met Asp Gly Ser Asp Pro Val
65 70 75 80
Ala Val Arg Ala Gly Ile Glu Ala Ile Val Ala Arg His Gly Gln Ile
85 90 95
Asp Ile Leu Val Asn Asn Ala Gly Ser Ala Gly Ala Gln Arg Arg Leu
100 105 110
Ala Glu Ile Pro Leu Thr Glu Ala Glu Leu Gly Pro Gly Ala Glu Glu
115 120 125
Thr Leu His Ala Ser Ile Ala Asn Leu Leu Gly Met Gly Trp His Leu
130 135 140
Met Arg Ile Ala Ala Pro His Met Pro Val Gly Ser Ala Val Ile Asn
145 150 155 160
Val Ser Thr Ile Phe Ser Arg Ala Glu Tyr Tyr Gly Arg Ile Pro Tyr
165 170 175
Val Thr Pro Lys Ala Ala Leu Asn Ala Leu Ser Gln Leu Ala Ala Arg
180 185 190
Glu Leu Gly Ala Arg Gly Ile Arg Val Asn Thr Ile Phe Pro Gly Pro
195 200 205
Ile Glu Ser Asp Arg Ile Arg Thr Val Phe Gln Arg Met Asp Gln Leu
210 215 220
Lys Gly Arg Pro Glu Gly Asp Thr Ala His His Phe Leu Asn Thr Met
225 230 235 240
Arg Leu Cys Arg Ala Asn Asp Gln Gly Ala Leu Glu Arg Arg Phe Pro
245 250 255
Ser Val Gly Asp Val Ala Asp Ala Ala Val Phe Leu Ala Ser Ala Glu
260 265 270
Ser Ala Ala Leu Ser Gly Glu Thr Ile Glu Val Thr His Gly Met Glu
275 280 285
Leu Pro Ala Cys Ser Glu Thr Ser Leu Leu Ala Arg Thr Asp Leu Arg
290 295 300
Thr Ile Asp Ala Ser Gly Arg Thr Thr Leu Ile Cys Ala Gly Asp Gln
305 310 315 320
Ile Glu Glu Val Met Ala Leu Thr Gly Met Leu Arg Thr Cys Gly Ser
325 330 335
Glu Val Ile Ile Gly Phe Arg Ser Ala Ala Ala Leu Ala Gln Phe Glu
340 345 350
Gln Ala Val Asn Glu Ser Arg Arg Leu Ala Gly Ala Asp Phe Thr Pro
355 360 365
Pro Ile Ala Leu Pro Leu Asp Pro Arg Asp Pro Ala Thr Ile Asp Ala
370 375 380
Val Phe Asp Trp Ala Gly Glu Asn Thr Gly Gly Ile His Trp Ile Leu
385 390 395 400
Pro Ala Thr Ser His Glu Pro Ala Pro Cys Val Ile Glu Val Asp Asp
405 410 415
Glu Arg Val Leu Asn Phe Leu Ala Asp Glu Ile Thr Gly Thr Ile Val
420 425 430
Ile Ala Ser Arg Leu Ala Arg Tyr Trp Gln Ser Gln Arg Leu Thr Pro
435 440 445
Gly Ala Arg Ala Arg Gly Pro Arg Val Ile Phe Leu Ser Asn Gly Ala
450 455 460
Asp Gln Asn Gly Asn Val Tyr Gly Arg Ile Gln Ser Ala Ala Ile Gly
465 470 475 480
Gln Leu Ile Arg Val Trp Arg His Glu Ala Glu Leu Asp Tyr Gln Arg
485 490 495
Ala Ser Ala Ala Gly Asp His Val Leu Pro Pro Val Trp Ala Asn Gln
500 505 510
Ile Val Arg Phe Ala Asn Arg Ser Leu Glu Gly Leu Glu Phe Ala Cys
515 520 525
Ala Trp Thr Ala Gln Leu Leu His Ser Gln Arg His Ile Asn Glu Ile
530 535 540
Thr Leu Asn Ile Pro Ala Asn Ile Ser Ala Thr Thr Gly Ala Arg Ser
545 550 555 560
Ala Ser Val Gly Trp Ala Glu Ser Leu Ile Gly Leu His Leu Gly Lys
565 570 575
Val Ala Leu Ile Thr Gly Gly Ser Ala Gly Ile Gly Gly Gln Ile Gly
580 585 590
Arg Leu Leu Ala Leu Ser Gly Ala Arg Val Met Leu Ala Ala Arg Asp
595 600 605
Arg His Lys Leu Glu Gln Met Gln Ala Met Ile Gln Ser Glu Leu Ala
610 615 620
Glu Val Gly Tyr Thr Asp Val Glu Asp Arg Val His Ile Ala Pro Gly
625 630 635 640
Cys Asp Val Ser Ser Glu Ala Gln Leu Ala Asp Leu Val Glu Arg Thr
645 650 655
Leu Ser Ala Phe Gly Thr Val Asp Tyr Leu Ile Asn Asn Ala Gly Ile
660 665 670
Ala Gly Val Glu Glu Met Val Ile Asp Met Pro Val Glu Gly Trp Arg
675 680 685
His Thr Leu Phe Ala Asn Leu Ile Ser Asn Tyr Ser Leu Met Arg Lys
690 695 700
Leu Ala Pro Leu Met Lys Lys Gln Gly Ser Gly Tyr Ile Leu Asn Val
705 710 715 720
Ser Ser Tyr Phe Gly Gly Glu Lys Asp Ala Ala Ile Pro Tyr Pro Asn
725 730 735
Arg Ala Asp Tyr Ala Val Ser Lys Ala Gly Gln Arg Ala Met Ala Glu
740 745 750
Val Phe Ala Arg Phe Leu Gly Pro Glu Ile Gln Ile Asn Ala Ile Ala
755 760 765
Pro Gly Pro Val Glu Gly Asp Arg Leu Arg Gly Thr Gly Glu Arg Pro
770 775 780
Gly Leu Phe Ala Arg Arg Ala Arg Leu Ile Leu Glu Asn Lys Arg Leu
785 790 795 800
Asn Glu Leu His Ala Ala Leu Ile Ala Ala Ala Arg Thr Asp Glu Arg
805 810 815
Ser Met His Glu Leu Val Glu Leu Leu Leu Pro Asn Asp Val Ala Ala
820 825 830
Leu Glu Gln Asn Pro Ala Ala Pro Thr Ala Leu Arg Glu Leu Ala Arg
835 840 845
Arg Phe Arg Ser Glu Gly Asp Pro Ala Ala Ser Ser Ser Ser Ala Leu
850 855 860
Leu Asn Arg Ser Ile Ala Ala Lys Leu Leu Ala Arg Leu His Asn Gly
865 870 875 880
Gly Tyr Val Leu Pro Ala Asp Ile Phe Ala Asn Leu Pro Asn Pro Pro
885 890 895
Asp Pro Phe Phe Thr Arg Ala Gln Ile Asp Arg Glu Ala Arg Lys Val
900 905 910
Arg Asp Gly Ile Met Gly Met Leu Tyr Leu Gln Arg Met Pro Thr Glu
915 920 925
Phe Asp Val Ala Met Ala Thr Val Tyr Tyr Leu Ala Asp Arg Asn Val
930 935 940
Ser Gly Glu Thr Phe His Pro Ser Gly Gly Leu Arg Tyr Glu Arg Thr
945 950 955 960
Pro Thr Gly Gly Glu Leu Phe Gly Leu Pro Ser Pro Glu Arg Leu Ala
965 970 975
Glu Leu Val Gly Ser Thr Val Tyr Leu Ile Gly Glu His Leu Thr Glu
980 985 990
His Leu Asn Leu Leu Ala Met Tyr Leu Glu Arg Tyr Gly Ala Arg Gln
995 1000 1005
Val Trp Ile Val Glu Thr Glu Thr Gly Ala Glu Thr Met Arg Arg
1010 1015 1020
Leu Leu His Asp His Val Glu Ala Gly Arg Leu Met Thr Ile Val
1025 1030 1035
Ala Gly Asp Gln Ile Glu Ala Ala Ile Asp Gln Ala Ile Thr Arg
1040 1045 1050
Tyr Gly Arg Pro Gly Pro Val Val Cys Thr Pro Phe Arg Pro Leu
1055 1060 1065
Pro Thr Val Pro Leu Val Gly Arg Lys Asp Ser Asp Trp Ser Thr
1070 1075 1080
Val Leu Ser Glu Ala Glu Phe Ala Glu Leu Cys Glu His Gln Leu
1085 1090 1095
Thr His His Phe Arg Val Ala Arg Lys Ile Ala Leu Ser Asp Gly
1100 1105 1110
Ala Ser Leu Ala Leu Val Thr Pro Glu Thr Thr Ala Thr Ser Thr
1115 1120 1125
Thr Glu Gln Phe Ala Leu Ala Asn Phe Ile Lys Thr Thr Leu His
1130 1135 1140
Ala Phe Thr Ala Thr Ile Gly Val Glu Ser Glu Arg Thr Ala Gln
1145 1150 1155
Arg Ile Leu Ile Asn Gln Val Asp Leu Thr Arg Arg Ala Arg Ala
1160 1165 1170
Glu Glu Pro Arg Asp Pro His Glu Arg Gln Gln Glu Leu Glu Arg
1175 1180 1185
Phe Ile Glu Ala Val Leu Leu Val Thr Ala Pro Leu Pro Pro Glu
1190 1195 1200
Ala Asp Thr Arg Tyr Ala Gly Arg Ile His Arg Gly Arg Ala Ile
1205 1210 1215
Thr Val
1220
<210> 59
<211> 1229
<212> PRT
<213> 卡氏玫瑰弯菌(Roseiflexus castenholzii)
<400> 59
Met Ser Thr Val Arg Arg Leu Glu Gly Lys Val Ala Leu Ile Thr Gly
1 5 10 15
Gly Ala Gly Asn Ile Gly Glu Val Ile Thr Arg Arg Phe Leu Ala Glu
20 25 30
Gly Ala Thr Val Val Ile Thr Gly Arg Asn Ala Glu Lys Leu Ala Val
35 40 45
Tyr Arg Arg Arg Leu Ile Asp Glu Glu Arg Val Ala Pro Glu Arg Val
50 55 60
Val Ala Leu Arg Met Asp Gly Ser Asp Ile Ala Gln Val Arg Ala Gly
65 70 75 80
Val Ala Gln Ile Val His Gly Gly Thr Asp Val Pro Ile Pro Leu His
85 90 95
Arg Ile Asp Ile Leu Val Asn Asn Ala Gly Ser Ala Gly Pro Arg Arg
100 105 110
Arg Leu Val Asp Ile Pro Leu Glu Pro Ser Glu Val Gln Pro Pro Asp
115 120 125
Ser Glu Thr Leu Ala Gln Ala Val Gly Asn Leu Val Gly Ile Thr Trp
130 135 140
Asn Leu Thr Arg Ala Ala Ala Pro His Met Pro Ser Gly Ser Ser Val
145 150 155 160
Ile Asn Ile Ser Thr Ile Phe Ser Arg Thr Asp Tyr Tyr Gly Arg Ile
165 170 175
Ala Tyr Val Ala Pro Lys Ala Ala Leu Asn Ala Leu Ser Asp Gly Leu
180 185 190
Ala Arg Glu Leu Gly Val Arg Gly Ile Arg Val Asn Thr Ile Tyr Pro
195 200 205
Gly Pro Ile Glu Ser Glu Arg Ile Tyr Thr Met Phe Gln Ala Met Asp
210 215 220
Ala Leu Lys Gly Gln Pro Glu Gly Asp Thr Ala Ser Gly Phe Leu Arg
225 230 235 240
Met Met Arg Leu Ser Arg Ile Asp Gln Asn Gly Glu Val Val Lys Arg
245 250 255
Phe Pro Ser Pro Val Asp Val Ala Asn Thr Ala Val Phe Leu Ala Ser
260 265 270
Asp Glu Ser Ala Ala Phe Thr Gly His Ala Phe Glu Val Thr His Gly
275 280 285
Met Glu Val Pro Thr Glu Ser Arg Thr Thr Phe Val Ser Arg Pro Gly
290 295 300
Leu Arg Ser Val Asp Ala Thr Gly Lys Val Ile Leu Ile Cys Ala Gly
305 310 315 320
Asp Gln Val Asp Asp Ala Val Ala Leu Ala Asp Thr Leu Arg Ser Cys
325 330 335
Arg Ala Thr Val Val Ile Gly Phe Arg Asp Pro Arg Ala Leu Glu Lys
340 345 350
Ala Ser Val Leu Leu Arg Glu Pro Arg His Ala Leu Ala Ala Asp Met
355 360 365
Tyr Gly Arg Pro Thr Met Thr Ala Glu Ala Arg Leu Val Arg Leu Asp
370 375 380
Pro Leu Asp Pro Arg Ala Ala Ala Gln Thr Leu Glu Gln Ile His Ala
385 390 395 400
Glu Leu Gly Ala Ile His His Ala Val Val Leu Pro Gly Gln Ser Arg
405 410 415
His Ala Pro Ser Ala Ser Leu Ile Glu Val Asp Asp Gln Val Val Glu
420 425 430
Arg Phe Leu His Gln Glu Leu Val Gly Thr Ile Ala Leu Ala Arg Glu
435 440 445
Leu Ala Arg Phe Trp Glu Glu Tyr Pro Ser Gly Ser Ser Met His Arg
450 455 460
Val Leu Phe Val Ser Asn Pro Asp Asp Gln Gln Gly Asn Gln Tyr Ser
465 470 475 480
His Ile Leu Arg Ala Ala Val Glu Gln Leu Val Arg Val Trp Arg His
485 490 495
Glu Ser Glu Tyr Asp Ser Val Asn Pro Ala His Gln Gln Glu Gly Gln
500 505 510
Ser Ser Ala Ala Val Trp Ala Asn Gln Leu Ile Arg Tyr Val Asn Asn
515 520 525
Glu Met Ala Asn Leu Asp Phe Thr Cys Ala Trp Val Ala Lys Leu Leu
530 535 540
Gly Ser Asp Arg Arg Ile Ala Glu Ile Asn Leu Tyr Leu Pro Glu Glu
545 550 555 560
Ile Val Gly Thr Ile Gly Val His Asn Pro Gly Phe Gly Trp Ala Glu
565 570 575
Ser Leu Phe Gly Leu His Met Gly Lys Val Ala Leu Ile Thr Gly Gly
580 585 590
Ser Ala Gly Ile Gly Gly Gln Ile Gly Arg Leu Leu Ala Leu Ser Gly
595 600 605
Ala His Val Met Leu Ala Ala Arg Asn Ala Asp Gln Leu Glu Gln Met
610 615 620
Arg Ala Ser Ile Val Arg Glu Val Arg Asp Ala Ser Tyr Pro Asp Ala
625 630 635 640
Glu Ser Arg Val Ala Ile Phe Pro Gly Ser Asp Val Ser Asp Ile Asp
645 650 655
Gly Leu Glu Arg Leu Val Asn His Thr Val Arg Val Phe Gly Lys Val
660 665 670
Asp Tyr Leu Ile Asn Asn Ala Gly Ile Ala Gly Ala Glu Glu Met Val
675 680 685
Ile Asp Met Pro Val Asp Ala Trp Arg His Thr Leu Arg Ala Asn Leu
690 695 700
Ile Ser Asn Tyr Ala Leu Leu Arg Arg Leu Ala Pro Gln Met Lys Ala
705 710 715 720
Ala Gly Gly Ala Tyr Val Leu Asn Val Ser Ser Tyr Phe Gly Gly Glu
725 730 735
Lys Tyr Val Ala Ile Pro Tyr Pro Asn Arg Ser Asp Tyr Ala Val Ser
740 745 750
Lys Ala Gly Gln Arg Ala Met Val Glu Ser Leu Ala Arg Phe Leu Gly
755 760 765
Pro Glu Ile Gln Ile Asn Ala Ile Ala Pro Gly Pro Val Glu Gly Glu
770 775 780
Arg Leu Lys Gly Ala Gly Ser Arg Pro Gly Leu Phe Met Arg Arg Ala
785 790 795 800
Arg Leu Ile Leu Glu Asn Lys Arg Leu Asn Glu Val Phe Ala Ala Leu
805 810 815
Leu Ala Ala Arg His Glu Gly Ala Thr Ile Ala Asp Leu Leu Pro Asp
820 825 830
Leu Phe Ala Asn Asp Ile Gln Ser Ile Ala Asn Ser Ala Ala Met Pro
835 840 845
Ala Pro Leu Arg Arg Leu Ala Thr Met Leu Arg Glu Thr Ser Asp Ala
850 855 860
Gly Gly Ser Ala Gln Ser Tyr Leu Met Asn Ala Thr Ile Ala Arg Lys
865 870 875 880
Leu Leu Asn Arg Leu Glu Asn Gly Gly Tyr Ile Thr Leu His Asp Arg
885 890 895
Arg Ala Leu Thr Val Glu Pro Pro Glu Pro Phe Phe Thr Glu Ala Gln
900 905 910
Ile Glu Arg Glu Ala Ile Lys Val Arg Asp Gly Ile Leu Gly Met Leu
915 920 925
His Leu Gln Arg Met Pro Thr Glu Phe Asp Val Ala Leu Ala Thr Val
930 935 940
Phe Tyr Leu Ala Asp Arg Asn Val Thr Gly Glu Thr Phe His Pro Ser
945 950 955 960
Gly Gly Leu Arg Phe Glu Arg Thr Val Thr Glu Gly Glu Leu Phe Gly
965 970 975
Lys Pro Gly Gln Gln Arg Leu Glu Arg Leu Lys Gly Ser Val Val Tyr
980 985 990
Leu Ile Gly Glu His Leu Arg Gln His Leu Val Leu Leu Ala Arg Thr
995 1000 1005
Phe Leu Asp Glu Ile His Val Ala Arg Val Val Leu Leu Thr Glu
1010 1015 1020
Thr Thr Gln Ala Ala Thr Asp Leu Ala Ala Glu Leu Ser Asp Tyr
1025 1030 1035
Glu Ala Ala Gly Arg Phe Val Val Ile Pro Thr Cys Gly Asp Ile
1040 1045 1050
Glu Gly Gly Ile Asp Arg Ala Met Ala Glu Tyr Gly Arg Pro Gly
1055 1060 1065
Pro Val Ile Ser Thr Pro Phe Arg Pro Leu Pro Asp Arg Ala Leu
1070 1075 1080
Ser Ala Arg Asn Gly Asp Trp Ser Ser Val Leu Thr Thr Ala Glu
1085 1090 1095
Phe Glu Glu Leu Val Glu Gln Gln Ile Thr His His Phe Arg Val
1100 1105 1110
Ala Arg Lys Ala Gly Leu Ile Glu Gly Ala Asn Val Thr Leu Val
1115 1120 1125
Thr Pro Pro Thr Ser Ala Arg Ser Thr Ser Glu Glu Phe Ala Leu
1130 1135 1140
Ala Asn Phe Val Lys Thr Thr Leu His Ala Leu Thr Ala Thr Ala
1145 1150 1155
Gly Ala Glu Ser Glu Arg Thr Val Pro His Val Pro Val Asn Gln
1160 1165 1170
Val Asp Leu Thr Arg Arg Ala Arg Ser Glu Glu Pro Arg Thr Pro
1175 1180 1185
Ser Glu Glu Glu Glu Glu Leu Gln Arg Phe Val Asn Ala Val Leu
1190 1195 1200
Leu Thr Ser Ala Pro Leu Pro Thr Pro Leu Glu Ser Arg Tyr Arg
1205 1210 1215
Ala Arg Ile Tyr Arg Gly Asn Ala Ile Thr Val
1220 1225
<210> 60
<211> 1217
<212> PRT
<213> 赤细菌属物种(Erythrobacter sp.)
<400> 60
Met Ser Lys Glu Gly Asn Ala Ala Lys Gly Arg Leu Glu Gly Lys Val
1 5 10 15
Ala Leu Ile Thr Gly Ala Ala Gly Asn Leu Gly Asn Glu Ile Ser Arg
20 25 30
Ala Phe Ala Arg Glu Gly Ala Phe Val Val Met Thr Gly Arg Thr Glu
35 40 45
Glu Arg Ile Ser Ala Ala Arg Glu Gln Leu Ile Ala Asp Thr Gly Val
50 55 60
Ala Pro Glu Arg Ile Asp Thr Ala Val Leu Asp Gly Gly Asn Pro Asp
65 70 75 80
Ser Ile Arg Ala Ala Met Ala Lys Leu Arg Lys Glu Tyr Gly Arg Ile
85 90 95
Asp Ile Leu Ile Asn Asn Ala Gly Ser Ala Gly Pro Lys Gln Pro Leu
100 105 110
His Asn Val Pro Leu Ser Pro Gln Glu Met Glu Ala Cys Gly Asp Thr
115 120 125
Glu Thr Val Arg Asp Ala Met Leu Asn Ile Leu Gly Val Thr Trp Asn
130 135 140
Met Ala Arg Ile Val Ala Pro Met Met Pro Val Gly Gly Ala Met Val
145 150 155 160
Asn Ile Ser Thr Ile Phe Ser His Thr Arg Tyr Tyr Gly Arg Thr Ala
165 170 175
Tyr Val Val Pro Lys Ala Ala Leu Asn Ala Leu Ser Asn Gln Leu Ala
180 185 190
Ser Glu Leu Gly Pro Arg Gly Ile Arg Val Asn Thr Val Phe Pro Gly
195 200 205
Pro Ile Glu Ser Asp Arg Ile Arg Thr Val Phe Ala Ala Met Asp Glu
210 215 220
Val Gln Ser Gln Pro Lys Asp Thr Thr Ala Asn Tyr Phe Thr Gly Arg
225 230 235 240
Met Ala Leu Thr Arg Ser Val Asn Gly Lys Val Asp Gly Lys Pro Leu
245 250 255
Pro Asn Pro Lys Asp Ile Ala Gly Thr Cys Leu Phe Leu Ala Ser Glu
260 265 270
Glu Ala Ala Gly Ile Ala Gly Glu Glu Val Asp Val Thr His Gly Leu
275 280 285
Ser Ala Asn Arg Thr Ser Ala Ser Thr Tyr Met Thr Arg Pro Ser Met
290 295 300
Arg Ser Leu Asp Gly Ala Gly Leu Asn Ile Phe Ile Val Ser Gly Glu
305 310 315 320
Asn Trp Asp Asp Ala Leu Val Ala Ala His Thr Leu Ile Gly Ser Gly
325 330 335
Ala Lys Val Arg Leu Gly Leu Ala Arg Asn Ala Asp Val Ala Gln Ala
340 345 350
Asn Ala Arg Leu Lys Ala Gln Gly Ile Gly Glu Glu Leu Thr Val Thr
355 360 365
Arg Phe Asn Arg Ala Glu Pro Asp Ala Met Glu Asp Ala Leu Ala Ala
370 375 380
Phe Ser Gly Asp Val Asp Gly Ala Ile Thr Gly Ala Ile Ile Leu Pro
385 390 395 400
Val Lys Pro Ser Gly His Phe Thr Gly Ser Leu Leu Ala Ala Asp Asp
405 410 415
Asp Thr Val Thr Lys Phe Met Asp Thr Glu Leu Val Gly Ala Ile Ala
420 425 430
Val Ser Arg Ser Leu Ala Arg Tyr Trp His Gly Arg Glu Asp Leu Gln
435 440 445
Ser Pro Pro Arg Cys Val Phe Met Thr Asn Pro Gly Asp Pro Leu Gly
450 455 460
Asn Ser Phe Ala Ser Val Leu Ser Ala Gly Ile Thr Gln Leu Ile Arg
465 470 475 480
Ile Trp Arg Asp Glu Glu Arg Val Gln Ala Gly Asn Gly Ser Thr Glu
485 490 495
His Ala Val Trp Ser Asn Gln Ile Val Arg His Thr Asn Thr Glu Asp
500 505 510
Glu Asn Thr Arg Phe Ala Ser Gly His Ala Thr Arg Val Leu Phe Arg
515 520 525
Glu Gln His Ile Ala Glu Ile Asp Leu Lys Leu Pro Ala Asn Ile Ser
530 535 540
Glu Glu Thr Gly Ser Arg Lys Ala Met Val Gly Phe Ala Glu Asn Ile
545 550 555 560
Thr Gly Leu His Leu Gly Lys Val Ala Phe Ile Thr Gly Gly Ser Ala
565 570 575
Gly Ile Gly Gly Gln Val Ala Arg Leu Leu Ala Leu Ala Gly Ala Lys
580 585 590
Val Met Met Val Ala Arg Arg Glu Ser Glu Leu Val Ala Ala Arg Asp
595 600 605
Arg Ile Val Gly Glu Leu Gln Asp Ile Gly Phe Ala Gly Val Glu Arg
610 615 620
Arg Val Lys Tyr Met Ala Asp Ile Asp Val Ser Asp Phe Ala Ser Leu
625 630 635 640
Asp Lys Ala Val Asp Ala Thr Leu Glu Glu Phe Gly Arg Ile Asp Tyr
645 650 655
Leu Ile Asn Asn Ala Gly Val Ala Gly Ala Glu Asp Met Val Ile Asp
660 665 670
Met Glu Pro Glu Ala Trp Arg Phe Thr Leu Asp Ala Asn Leu Ile Ser
675 680 685
Asn Tyr His Leu Met Gln Arg Val Val Pro Leu Met Lys Glu Gln Gly
690 695 700
Ser Gly Tyr Val Leu Asn Val Ser Ser Tyr Phe Gly Gly Glu Lys Phe
705 710 715 720
Leu Ala Val Ala Tyr Pro Asn Arg Ala Asp Tyr Gly Leu Ser Lys Ala
725 730 735
Gly Gln Arg Ala Met Val Glu Ala Phe Ser Pro Phe Leu Gly Pro Glu
740 745 750
Val Gln Cys Asn Ala Ile Ala Pro Gly Pro Val Asp Gly Asp Arg Leu
755 760 765
Ser Gly Thr Gly Gly Lys Pro Gly Leu Phe Gln Arg Arg Ala Lys Leu
770 775 780
Ile Leu Glu Asn Lys Arg Leu Asn Ala Val Tyr Ser Ala Val Ile His
785 790 795 800
Ala Ile Arg Glu Gly Gly Asp Ala Ala Lys Ile Leu Thr Arg Leu Ser
805 810 815
Arg Asn Ser Thr Ser Thr Leu Ser His Asp Ala Glu Ala Pro Glu Glu
820 825 830
Leu Arg Lys Leu Ala Leu Asp Phe Ala Ser Gln Gly Asp Gly Leu Cys
835 840 845
Thr Trp Asp Gln Tyr Leu Leu Thr Asp Ala Met Ala Gln Arg Leu Leu
850 855 860
Val Arg Leu Gln Leu Gly Gly Phe Leu Leu Gly Ser Asn Glu Trp Ala
865 870 875 880
Ser Leu Ser Ser Ser Glu Gln Thr Trp Leu Lys Leu Ser Pro Pro Asp
885 890 895
Asp Lys Pro Phe Leu Pro Ala Ala Gln Val Asp Lys Val Ala Asn Gly
900 905 910
Val Gly Lys Gly Val Ile Ser Gln Leu His Leu Gly Ala Met Pro Thr
915 920 925
Glu Ala Glu Val Ala Gln Ala Thr Val Phe Phe Leu Ala Asp Arg Ala
930 935 940
Val Ser Gly Glu Thr Phe Met Pro Ser Gly Gly Leu Arg Val Glu Arg
945 950 955 960
Ser Asn Thr Glu Arg Glu Met Phe Gly Ser Pro Lys Gln Glu Arg Ile
965 970 975
Asp Lys Met Lys Gly Lys Thr Val Trp Ile Ile Gly Glu His Leu Ser
980 985 990
Asp Tyr Val Ala Ala Thr Ile Glu Glu Leu Val Ser Gly Cys Gly Val
995 1000 1005
Ala Lys Val Val Leu Ile Ala Lys Asp Lys Ser Gly Glu Lys Ala
1010 1015 1020
Val Arg Asp Gln Leu Pro Asn Asp Leu Ser Lys Asp Ala Leu Glu
1025 1030 1035
Val Leu Ile Ala Gly Asp Gly Leu Glu Glu Ala Met Asp Glu Ala
1040 1045 1050
Leu Gly His Trp Gly Lys Pro Thr Thr Val Leu Ser Met Pro Gly
1055 1060 1065
Glu Pro Leu Pro Asp His Leu Phe Glu Gly Gly Asn Pro Leu Ser
1070 1075 1080
Thr Lys Asp Phe Ala His Met Val Glu Ala Asn Ile Thr Arg His
1085 1090 1095
Tyr Arg Val Thr Arg Lys Ala Ser Leu Tyr Asp Gly Cys Gln Val
1100 1105 1110
Val Leu Val Ser Pro Asp Val Pro Tyr Gly Ser Asp Gly Pro Gly
1115 1120 1125
Val Ala Leu Ala Asn Phe Val Lys Thr Ser Leu His Ala Phe Thr
1130 1135 1140
Ala Thr Val Ala Val Glu Asn Glu Arg Leu Val His Asp Val Pro
1145 1150 1155
Val Asn Gln Ile Asn Leu Thr Arg Arg Val Ser Ser Glu Glu Pro
1160 1165 1170
Arg Asp Ala Asp Glu His Ala Glu Glu Leu Arg Arg Phe Thr Arg
1175 1180 1185
Ala Val Leu Leu Val Gly Ala Pro Leu Pro Asp Ala Gln Asp Ser
1190 1195 1200
Arg Tyr Arg Ser Lys Ile Tyr Arg Gly Thr Ser Met Thr Val
1205 1210 1215
<210> 61
<211> 357
<212> PRT
<213> 勤奋生金球菌(Metallosphaera sedula)
<400> 61
Met Arg Arg Thr Leu Lys Ala Ala Ile Leu Gly Ala Thr Gly Leu Val
1 5 10 15
Gly Ile Glu Tyr Val Arg Met Leu Ala Asp His Pro Tyr Ile Lys Pro
20 25 30
Thr Tyr Leu Ala Gly Lys Gly Ser Val Gly Lys Pro Tyr Gly Glu Ile
35 40 45
Val Arg Trp Gln Thr Val Gly Asn Val Pro Lys Glu Val Ala Asn Gln
50 55 60
Glu Val Lys Pro Thr Asp Pro Lys Leu Met Asp Asp Val Asp Ile Ile
65 70 75 80
Phe Ser Pro Leu Pro Gln Gly Ala Ala Gly Pro Val Glu Glu Gln Phe
85 90 95
Ala Lys Leu Gly Phe Asn Val Ile Ser Asn Ser Pro Asp His Arg Phe
100 105 110
Asp Met Asp Val Pro Met Ile Ile Pro Glu Val Asn Pro His Thr Val
115 120 125
Thr Leu Ile Asp Glu Gln Arg Lys Arg Arg Asp Trp Lys Gly Phe Ile
130 135 140
Val Thr Thr Pro Leu Cys Thr Ala Gln Gly Ala Ala Ile Pro Leu Thr
145 150 155 160
Pro Ile Tyr Gln Asn Phe Lys Met Ser Gly Val Met Ile Thr Thr Met
165 170 175
Gln Ser Leu Ser Gly Ala Gly Tyr Pro Gly Ile Ala Ser Leu Asp Ile
180 185 190
Val Asp Asn Ala Leu Pro Leu Gly Asp Gly Tyr Asp Ala Lys Thr Val
195 200 205
Lys Glu Ile Thr Arg Ile Leu Ser Glu Val Lys Arg Asn Val Gln Glu
210 215 220
Pro Gly Val Asn Glu Ile Thr Leu Asp Ala Thr Thr His Arg Ile Ala
225 230 235 240
Thr Ile His Gly His Tyr Glu Val Ala Tyr Val Thr Phe Lys Glu Asp
245 250 255
Thr Asp Val Arg Lys Val Met Glu Ser Met Glu Ser Phe Lys Gly Glu
260 265 270
Pro Gln Asp Leu Lys Leu Pro Thr Ala Pro Glu Lys Pro Ile Ile Val
275 280 285
Thr Thr Gln Asp Ala Arg Pro Gln Val Phe Phe Asp Arg Trp Ala Gly
290 295 300
Asn Pro Pro Gly Met Ser Val Val Val Gly Arg Leu Lys Gln Val Asn
305 310 315 320
Pro Arg Thr Ile Arg Phe Val Ser Leu Ile His Asn Thr Val Arg Gly
325 330 335
Ala Ala Gly Gly Gly Val Leu Thr Ala Glu Leu Leu Val Glu Lys Gly
340 345 350
Tyr Ile Asp Lys Arg
355
<210> 62
<211> 356
<212> PRT
<213> 东工大硫化叶菌(Sulfolobus tokodaii)
<400> 62
Met Arg Arg Thr Leu Lys Ala Ala Ile Leu Gly Ala Thr Gly Leu Val
1 5 10 15
Gly Ile Glu Tyr Val Arg Met Leu Ser Asn His Pro Tyr Ile Lys Pro
20 25 30
Ala Tyr Leu Ala Gly Lys Gly Ser Val Gly Lys Pro Tyr Gly Glu Val
35 40 45
Val Arg Trp Gln Thr Val Gly Gln Val Pro Lys Glu Ile Ala Asp Met
50 55 60
Glu Ile Lys Pro Thr Asp Pro Lys Leu Met Asp Asp Val Asp Ile Ile
65 70 75 80
Phe Ser Pro Leu Pro Gln Gly Ala Ala Gly Pro Val Glu Glu Gln Phe
85 90 95
Ala Lys Glu Gly Phe Pro Val Ile Ser Asn Ser Pro Asp His Arg Phe
100 105 110
Asp Pro Asp Val Pro Leu Leu Val Pro Glu Leu Asn Pro His Thr Ile
115 120 125
Ser Leu Ile Asp Glu Gln Arg Lys Arg Arg Glu Trp Lys Gly Phe Ile
130 135 140
Val Thr Thr Pro Leu Cys Thr Ala Gln Gly Ala Ala Ile Pro Leu Gly
145 150 155 160
Ala Ile Phe Lys Asp Tyr Lys Met Asp Gly Ala Phe Ile Thr Thr Ile
165 170 175
Gln Ser Leu Ser Gly Ala Gly Tyr Pro Gly Ile Pro Ser Leu Asp Val
180 185 190
Val Asp Asn Ile Leu Pro Leu Gly Asp Gly Tyr Asp Ala Lys Thr Ile
195 200 205
Lys Glu Ile Phe Arg Ile Leu Ser Glu Val Lys Arg Asn Val Asp Glu
210 215 220
Pro Lys Leu Glu Asp Val Ser Leu Ala Ala Thr Thr His Arg Ile Ala
225 230 235 240
Thr Ile His Gly His Tyr Glu Val Leu Tyr Val Ser Phe Lys Glu Glu
245 250 255
Thr Ala Ala Glu Lys Val Lys Glu Thr Leu Glu Asn Phe Arg Gly Glu
260 265 270
Pro Gln Asp Leu Lys Leu Pro Thr Ala Pro Ser Lys Pro Ile Ile Val
275 280 285
Met Asn Glu Asp Thr Arg Pro Gln Val Tyr Phe Asp Arg Trp Ala Gly
290 295 300
Asp Ile Pro Gly Met Ser Val Val Val Gly Arg Leu Lys Gln Val Asn
305 310 315 320
Lys Arg Met Ile Arg Leu Val Ser Leu Ile His Asn Thr Val Arg Gly
325 330 335
Ala Ala Gly Gly Gly Ile Leu Ala Ala Glu Leu Leu Val Glu Lys Gly
340 345 350
Tyr Ile Glu Lys
355
<210> 63
<211> 482
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 63
Met Ser Ala Phe Val Arg Val Val Pro Arg Ile Ser Arg Ser Ser Val
1 5 10 15
Leu Thr Arg Ser Leu Arg Leu Gln Leu Arg Cys Tyr Ala Ser Tyr Pro
20 25 30
Glu His Thr Ile Ile Gly Met Pro Ala Leu Ser Pro Thr Met Thr Gln
35 40 45
Gly Asn Leu Ala Ala Trp Thr Lys Lys Glu Gly Asp Gln Leu Ser Pro
50 55 60
Gly Glu Val Ile Ala Glu Ile Glu Thr Asp Lys Ala Gln Met Asp Phe
65 70 75 80
Glu Phe Gln Glu Asp Gly Tyr Leu Ala Lys Ile Leu Val Pro Glu Gly
85 90 95
Thr Lys Asp Ile Pro Val Asn Lys Pro Ile Ala Val Tyr Val Glu Asp
100 105 110
Lys Ala Asp Val Pro Ala Phe Lys Asp Phe Lys Leu Glu Asp Ser Gly
115 120 125
Ser Asp Ser Lys Thr Ser Thr Lys Ala Gln Pro Ala Glu Pro Gln Ala
130 135 140
Glu Lys Lys Gln Glu Ala Pro Ala Glu Glu Thr Lys Thr Ser Ala Pro
145 150 155 160
Glu Ala Lys Lys Ser Asp Val Ala Ala Pro Gln Gly Arg Ile Phe Ala
165 170 175
Ser Pro Leu Ala Lys Thr Ile Ala Leu Glu Lys Gly Ile Ser Leu Lys
180 185 190
Asp Val His Gly Thr Gly Pro Arg Gly Arg Ile Thr Lys Ala Asp Ile
195 200 205
Glu Ser Tyr Leu Glu Lys Ser Ser Lys Gln Ser Ser Gln Thr Ser Gly
210 215 220
Ala Ala Ala Ala Thr Pro Ala Ala Ala Thr Ser Ser Thr Thr Ala Gly
225 230 235 240
Ser Ala Pro Ser Pro Ser Ser Thr Ala Ser Tyr Glu Asp Val Pro Ile
245 250 255
Ser Thr Met Arg Ser Ile Ile Gly Glu Arg Leu Leu Gln Ser Thr Gln
260 265 270
Gly Ile Pro Ser Tyr Ile Val Ser Ser Lys Ile Ser Ile Ser Lys Leu
275 280 285
Leu Lys Leu Arg Gln Ser Leu Asn Ala Thr Ala Asn Asp Lys Tyr Lys
290 295 300
Leu Ser Ile Asn Asp Leu Leu Val Lys Ala Ile Thr Val Ala Ala Lys
305 310 315 320
Arg Val Pro Asp Ala Asn Ala Tyr Trp Leu Pro Asn Glu Asn Val Ile
325 330 335
Arg Lys Phe Lys Asn Val Asp Val Ser Val Ala Val Ala Thr Pro Thr
340 345 350
Gly Leu Leu Thr Pro Ile Val Lys Asn Cys Glu Ala Lys Gly Leu Ser
355 360 365
Gln Ile Ser Asn Glu Ile Lys Glu Leu Val Lys Arg Ala Arg Ile Asn
370 375 380
Lys Leu Ala Pro Glu Glu Phe Gln Gly Gly Thr Ile Cys Ile Ser Asn
385 390 395 400
Met Gly Met Asn Asn Ala Val Asn Met Phe Thr Ser Ile Ile Asn Pro
405 410 415
Pro Gln Ser Thr Ile Leu Ala Ile Ala Thr Val Glu Arg Val Ala Val
420 425 430
Glu Asp Ala Ala Ala Glu Asn Gly Phe Ser Phe Asp Asn Gln Val Thr
435 440 445
Ile Thr Gly Thr Phe Asp His Arg Thr Ile Asp Gly Ala Lys Gly Ala
450 455 460
Glu Phe Met Lys Glu Leu Lys Thr Val Ile Glu Asn Pro Leu Glu Met
465 470 475 480
Leu Leu
<210> 64
<211> 420
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 64
Met Leu Ala Ala Ser Phe Lys Arg Gln Pro Ser Gln Leu Val Arg Gly
1 5 10 15
Leu Gly Ala Val Leu Arg Thr Pro Thr Arg Ile Gly His Val Arg Thr
20 25 30
Met Ala Thr Leu Lys Thr Thr Asp Lys Lys Ala Pro Glu Asp Ile Glu
35 40 45
Gly Ser Asp Thr Val Gln Ile Glu Leu Pro Glu Ser Ser Phe Glu Ser
50 55 60
Tyr Met Leu Glu Pro Pro Asp Leu Ser Tyr Glu Thr Ser Lys Ala Thr
65 70 75 80
Leu Leu Gln Met Tyr Lys Asp Met Val Ile Ile Arg Arg Met Glu Met
85 90 95
Ala Cys Asp Ala Leu Tyr Lys Ala Lys Lys Ile Arg Gly Phe Cys His
100 105 110
Leu Ser Val Gly Gln Glu Ala Ile Ala Val Gly Ile Glu Asn Ala Ile
115 120 125
Thr Lys Leu Asp Ser Ile Ile Thr Ser Tyr Arg Cys His Gly Phe Thr
130 135 140
Phe Met Arg Gly Ala Ser Val Lys Ala Val Leu Ala Glu Leu Met Gly
145 150 155 160
Arg Arg Ala Gly Val Ser Tyr Gly Lys Gly Gly Ser Met His Leu Tyr
165 170 175
Ala Pro Gly Phe Tyr Gly Gly Asn Gly Ile Val Gly Ala Gln Val Pro
180 185 190
Leu Gly Ala Gly Leu Ala Phe Ala His Gln Tyr Lys Asn Glu Asp Ala
195 200 205
Cys Ser Phe Thr Leu Tyr Gly Asp Gly Ala Ser Asn Gln Gly Gln Val
210 215 220
Phe Glu Ser Phe Asn Met Ala Lys Leu Trp Asn Leu Pro Val Val Phe
225 230 235 240
Cys Cys Glu Asn Asn Lys Tyr Gly Met Gly Thr Ala Ala Ser Arg Ser
245 250 255
Ser Ala Met Thr Glu Tyr Phe Lys Arg Gly Gln Tyr Ile Pro Gly Leu
260 265 270
Lys Val Asn Gly Met Asp Ile Leu Ala Val Tyr Gln Ala Ser Lys Phe
275 280 285
Ala Lys Asp Trp Cys Leu Ser Gly Lys Gly Pro Leu Val Leu Glu Tyr
290 295 300
Glu Thr Tyr Arg Tyr Gly Gly His Ser Met Ser Asp Pro Gly Thr Thr
305 310 315 320
Tyr Arg Thr Arg Asp Glu Ile Gln His Met Arg Ser Lys Asn Asp Pro
325 330 335
Ile Ala Gly Leu Lys Met His Leu Ile Asp Leu Gly Ile Ala Thr Glu
340 345 350
Ala Glu Val Lys Ala Tyr Asp Lys Ser Ala Arg Lys Tyr Val Asp Glu
355 360 365
Gln Val Glu Leu Ala Asp Ala Ala Pro Pro Pro Glu Ala Lys Leu Ser
370 375 380
Ile Leu Phe Glu Asp Val Tyr Val Lys Gly Thr Glu Thr Pro Thr Leu
385 390 395 400
Arg Gly Arg Ile Pro Glu Asp Thr Trp Asp Phe Lys Lys Gln Gly Phe
405 410 415
Ala Ser Arg Asp
420
<210> 65
<211> 366
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 65
Met Phe Ser Arg Leu Pro Thr Ser Leu Ala Arg Asn Val Ala Arg Arg
1 5 10 15
Ala Pro Thr Ser Phe Val Arg Pro Ser Ala Ala Ala Ala Ala Leu Arg
20 25 30
Phe Ser Ser Thr Lys Thr Met Thr Val Arg Glu Ala Leu Asn Ser Ala
35 40 45
Met Ala Glu Glu Leu Asp Arg Asp Asp Asp Val Phe Leu Ile Gly Glu
50 55 60
Glu Val Ala Gln Tyr Asn Gly Ala Tyr Lys Val Ser Lys Gly Leu Leu
65 70 75 80
Asp Arg Phe Gly Glu Arg Arg Val Val Asp Thr Pro Ile Thr Glu Tyr
85 90 95
Gly Phe Thr Gly Leu Ala Val Gly Ala Ala Leu Lys Gly Leu Lys Pro
100 105 110
Ile Val Glu Phe Met Ser Phe Asn Phe Ser Met Gln Ala Ile Asp His
115 120 125
Val Val Asn Ser Ala Ala Lys Thr His Tyr Met Ser Gly Gly Thr Gln
130 135 140
Lys Cys Gln Met Val Phe Arg Gly Pro Asn Gly Ala Ala Val Gly Val
145 150 155 160
Gly Ala Gln His Ser Gln Asp Phe Ser Pro Trp Tyr Gly Ser Ile Pro
165 170 175
Gly Leu Lys Val Leu Val Pro Tyr Ser Ala Glu Asp Ala Arg Gly Leu
180 185 190
Leu Lys Ala Ala Ile Arg Asp Pro Asn Pro Val Val Phe Leu Glu Asn
195 200 205
Glu Leu Leu Tyr Gly Glu Ser Phe Glu Ile Ser Glu Glu Ala Leu Ser
210 215 220
Pro Glu Phe Thr Leu Pro Tyr Lys Ala Lys Ile Glu Arg Glu Gly Thr
225 230 235 240
Asp Ile Ser Ile Val Thr Tyr Thr Arg Asn Val Gln Phe Ser Leu Glu
245 250 255
Ala Ala Glu Ile Leu Gln Lys Lys Tyr Gly Val Ser Ala Glu Val Ile
260 265 270
Asn Leu Arg Ser Ile Arg Pro Leu Asp Thr Glu Ala Ile Ile Lys Thr
275 280 285
Val Lys Lys Thr Asn His Leu Ile Thr Val Glu Ser Thr Phe Pro Ser
290 295 300
Phe Gly Val Gly Ala Glu Ile Val Ala Gln Val Met Glu Ser Glu Ala
305 310 315 320
Phe Asp Tyr Leu Asp Ala Pro Ile Gln Arg Val Thr Gly Ala Asp Val
325 330 335
Pro Thr Pro Tyr Ala Lys Glu Leu Glu Asp Phe Ala Phe Pro Asp Thr
340 345 350
Pro Thr Ile Val Lys Ala Val Lys Glu Val Leu Ser Ile Glu
355 360 365
<210> 66
<211> 499
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 66
Met Leu Arg Ile Arg Ser Leu Leu Asn Asn Lys Arg Ala Phe Ser Ser
1 5 10 15
Thr Val Arg Thr Leu Thr Ile Asn Lys Ser His Asp Val Val Ile Ile
20 25 30
Gly Gly Gly Pro Ala Gly Tyr Val Ala Ala Ile Lys Ala Ala Gln Leu
35 40 45
Gly Phe Asn Thr Ala Cys Val Glu Lys Arg Gly Lys Leu Gly Gly Thr
50 55 60
Cys Leu Asn Val Gly Cys Ile Pro Ser Lys Ala Leu Leu Asn Asn Ser
65 70 75 80
His Leu Phe His Gln Met His Thr Glu Ala Gln Lys Arg Gly Ile Asp
85 90 95
Val Asn Gly Asp Ile Lys Ile Asn Val Ala Asn Phe Gln Lys Ala Lys
100 105 110
Asp Asp Ala Val Lys Gln Leu Thr Gly Gly Ile Glu Leu Leu Phe Lys
115 120 125
Lys Asn Lys Val Thr Tyr Tyr Lys Gly Asn Gly Ser Phe Glu Asp Glu
130 135 140
Thr Lys Ile Arg Val Thr Pro Val Asp Gly Leu Glu Gly Thr Val Lys
145 150 155 160
Glu Asp His Ile Leu Asp Val Lys Asn Ile Ile Val Ala Thr Gly Ser
165 170 175
Glu Val Thr Pro Phe Pro Gly Ile Glu Ile Asp Glu Glu Lys Ile Val
180 185 190
Ser Ser Thr Gly Ala Leu Ser Leu Lys Glu Ile Pro Lys Arg Leu Thr
195 200 205
Ile Ile Gly Gly Gly Ile Ile Gly Leu Glu Met Gly Ser Val Tyr Ser
210 215 220
Arg Leu Gly Ser Lys Val Thr Val Val Glu Phe Gln Pro Gln Ile Gly
225 230 235 240
Ala Ser Met Asp Gly Glu Val Ala Lys Ala Thr Gln Lys Phe Leu Lys
245 250 255
Lys Gln Gly Leu Asp Phe Lys Leu Ser Thr Lys Val Ile Ser Ala Lys
260 265 270
Arg Asn Asp Asp Lys Asn Val Val Glu Ile Val Val Glu Asp Thr Lys
275 280 285
Thr Asn Lys Gln Glu Asn Leu Glu Ala Glu Val Leu Leu Val Ala Val
290 295 300
Gly Arg Arg Pro Tyr Ile Ala Gly Leu Gly Ala Glu Lys Ile Gly Leu
305 310 315 320
Glu Val Asp Lys Arg Gly Arg Leu Val Ile Asp Asp Gln Phe Asn Ser
325 330 335
Lys Phe Pro His Ile Lys Val Val Gly Asp Val Thr Phe Gly Pro Met
340 345 350
Leu Ala His Lys Ala Glu Glu Glu Gly Ile Ala Ala Val Glu Met Leu
355 360 365
Lys Thr Gly His Gly His Val Asn Tyr Asn Asn Ile Pro Ser Val Met
370 375 380
Tyr Ser His Pro Glu Val Ala Trp Val Gly Lys Thr Glu Glu Gln Leu
385 390 395 400
Lys Glu Ala Gly Ile Asp Tyr Lys Ile Gly Lys Phe Pro Phe Ala Ala
405 410 415
Asn Ser Arg Ala Lys Thr Asn Gln Asp Thr Glu Gly Phe Val Lys Ile
420 425 430
Leu Ile Asp Ser Lys Thr Glu Arg Ile Leu Gly Ala His Ile Ile Gly
435 440 445
Pro Asn Ala Gly Glu Met Ile Ala Glu Ala Gly Leu Ala Leu Glu Tyr
450 455 460
Gly Ala Ser Ala Glu Asp Val Ala Arg Val Cys His Ala His Pro Thr
465 470 475 480
Leu Ser Glu Ala Phe Lys Glu Ala Asn Met Ala Ala Tyr Asp Lys Ala
485 490 495
Ile His Cys
<210> 67
<211> 887
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 67
Met Ser Glu Arg Phe Pro Asn Asp Val Asp Pro Ile Glu Thr Arg Asp
1 5 10 15
Trp Leu Gln Ala Ile Glu Ser Val Ile Arg Glu Glu Gly Val Glu Arg
20 25 30
Ala Gln Tyr Leu Ile Asp Gln Leu Leu Ala Glu Ala Arg Lys Gly Gly
35 40 45
Val Asn Val Ala Ala Gly Thr Gly Ile Ser Asn Tyr Ile Asn Thr Ile
50 55 60
Pro Val Glu Glu Gln Pro Glu Tyr Pro Gly Asn Leu Glu Leu Glu Arg
65 70 75 80
Arg Ile Arg Ser Ala Ile Arg Trp Asn Ala Ile Met Thr Val Leu Arg
85 90 95
Ala Ser Lys Lys Asp Leu Glu Leu Gly Gly His Met Ala Ser Phe Gln
100 105 110
Ser Ser Ala Thr Ile Tyr Asp Val Cys Phe Asn His Phe Phe Arg Ala
115 120 125
Arg Asn Glu Gln Asp Gly Gly Asp Leu Val Tyr Phe Gln Gly His Ile
130 135 140
Ser Pro Gly Val Tyr Ala Arg Ala Phe Leu Glu Gly Arg Leu Thr Gln
145 150 155 160
Glu Gln Leu Asp Asn Phe Arg Gln Glu Val His Gly Asn Gly Leu Ser
165 170 175
Ser Tyr Pro His Pro Lys Leu Met Pro Glu Phe Trp Gln Phe Pro Thr
180 185 190
Val Ser Met Gly Leu Gly Pro Ile Gly Ala Ile Tyr Gln Ala Lys Phe
195 200 205
Leu Lys Tyr Leu Glu His Arg Gly Leu Lys Asp Thr Ser Lys Gln Thr
210 215 220
Val Tyr Ala Phe Leu Gly Asp Gly Glu Met Asp Glu Pro Glu Ser Lys
225 230 235 240
Gly Ala Ile Thr Ile Ala Thr Arg Glu Lys Leu Asp Asn Leu Val Phe
245 250 255
Val Ile Asn Cys Asn Leu Gln Arg Leu Asp Gly Pro Val Thr Gly Asn
260 265 270
Gly Lys Ile Ile Asn Glu Leu Glu Gly Ile Phe Glu Gly Ala Gly Trp
275 280 285
Asn Val Ile Lys Val Met Trp Gly Ser Arg Trp Asp Glu Leu Leu Arg
290 295 300
Lys Asp Thr Ser Gly Lys Leu Ile Gln Leu Met Asn Glu Thr Val Asp
305 310 315 320
Gly Asp Tyr Gln Thr Phe Lys Ser Lys Asp Gly Ala Tyr Val Arg Glu
325 330 335
His Phe Phe Gly Lys Tyr Pro Glu Thr Ala Ala Leu Val Ala Asp Trp
340 345 350
Thr Asp Glu Gln Ile Trp Ala Leu Asn Arg Gly Gly His Asp Pro Lys
355 360 365
Lys Ile Tyr Ala Ala Phe Lys Lys Ala Gln Glu Thr Lys Gly Lys Ala
370 375 380
Thr Val Ile Leu Ala His Thr Ile Lys Gly Tyr Gly Met Gly Asp Ala
385 390 395 400
Ala Glu Gly Lys Asn Ile Ala His Gln Val Lys Lys Met Asn Met Asp
405 410 415
Gly Val Arg His Ile Arg Asp Arg Phe Asn Val Pro Val Ser Asp Ala
420 425 430
Asp Ile Glu Lys Leu Pro Tyr Ile Thr Phe Pro Glu Gly Ser Glu Glu
435 440 445
His Thr Tyr Leu His Ala Gln Arg Gln Lys Leu His Gly Tyr Leu Pro
450 455 460
Ser Arg Gln Pro Asn Phe Thr Glu Lys Leu Glu Leu Pro Ser Leu Gln
465 470 475 480
Asp Phe Gly Ala Leu Leu Glu Glu Gln Ser Lys Glu Ile Ser Thr Thr
485 490 495
Ile Ala Phe Val Arg Ala Leu Asn Val Met Leu Lys Asn Lys Ser Ile
500 505 510
Lys Asp Arg Leu Val Pro Ile Ile Ala Asp Glu Ala Arg Thr Phe Gly
515 520 525
Met Glu Gly Leu Phe Arg Gln Ile Gly Ile Tyr Ser Pro Asn Gly Gln
530 535 540
Gln Tyr Thr Pro Gln Asp Arg Glu Gln Val Ala Tyr Tyr Lys Glu Asp
545 550 555 560
Glu Lys Gly Gln Ile Leu Gln Glu Gly Ile Asn Glu Leu Gly Ala Gly
565 570 575
Cys Ser Trp Leu Ala Ala Ala Thr Ser Tyr Ser Thr Asn Asn Leu Pro
580 585 590
Met Ile Pro Phe Tyr Ile Tyr Tyr Ser Met Phe Gly Phe Gln Arg Ile
595 600 605
Gly Asp Leu Cys Trp Ala Ala Gly Asp Gln Gln Ala Arg Gly Phe Leu
610 615 620
Ile Gly Gly Thr Ser Gly Arg Thr Thr Leu Asn Gly Glu Gly Leu Gln
625 630 635 640
His Glu Asp Gly His Ser His Ile Gln Ser Leu Thr Ile Pro Asn Cys
645 650 655
Ile Ser Tyr Asp Pro Ala Tyr Ala Tyr Glu Val Ala Val Ile Met His
660 665 670
Asp Gly Leu Glu Arg Met Tyr Gly Glu Lys Gln Glu Asn Val Tyr Tyr
675 680 685
Tyr Ile Thr Thr Leu Asn Glu Asn Tyr His Met Pro Ala Met Pro Glu
690 695 700
Gly Ala Glu Glu Gly Ile Arg Lys Gly Ile Tyr Lys Leu Glu Thr Ile
705 710 715 720
Glu Gly Ser Lys Gly Lys Val Gln Leu Leu Gly Ser Gly Ser Ile Leu
725 730 735
Arg His Val Arg Glu Ala Ala Glu Ile Leu Ala Lys Asp Tyr Gly Val
740 745 750
Gly Ser Asp Val Tyr Ser Val Thr Ser Phe Thr Glu Leu Ala Arg Asp
755 760 765
Gly Gln Asp Cys Glu Arg Trp Asn Met Leu His Pro Leu Glu Thr Pro
770 775 780
Arg Val Pro Tyr Ile Ala Gln Val Met Asn Asp Ala Pro Ala Val Ala
785 790 795 800
Ser Thr Asp Tyr Met Lys Leu Phe Ala Glu Gln Val Arg Thr Tyr Val
805 810 815
Pro Ala Asp Asp Tyr Arg Val Leu Gly Thr Asp Gly Phe Gly Arg Ser
820 825 830
Asp Ser Arg Glu Asn Leu Arg His His Phe Glu Val Asp Ala Ser Tyr
835 840 845
Val Val Val Ala Ala Leu Gly Glu Leu Ala Lys Arg Gly Glu Ile Asp
850 855 860
Lys Lys Val Val Ala Asp Ala Ile Ala Lys Phe Asn Ile Asp Ala Asp
865 870 875 880
Lys Val Asn Pro Arg Leu Ala
885
<210> 68
<211> 630
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 68
Met Ala Ile Glu Ile Lys Val Pro Asp Ile Gly Ala Asp Glu Val Glu
1 5 10 15
Ile Thr Glu Ile Leu Val Lys Val Gly Asp Lys Val Glu Ala Glu Gln
20 25 30
Ser Leu Ile Thr Val Glu Gly Asp Lys Ala Ser Met Glu Val Pro Ser
35 40 45
Pro Gln Ala Gly Ile Val Lys Glu Ile Lys Val Ser Val Gly Asp Lys
50 55 60
Thr Gln Thr Gly Ala Leu Ile Met Ile Phe Asp Ser Ala Asp Gly Ala
65 70 75 80
Ala Asp Ala Ala Pro Ala Gln Ala Glu Glu Lys Lys Glu Ala Ala Pro
85 90 95
Ala Ala Ala Pro Ala Ala Ala Ala Ala Lys Asp Val Asn Val Pro Asp
100 105 110
Ile Gly Ser Asp Glu Val Glu Val Thr Glu Ile Leu Val Lys Val Gly
115 120 125
Asp Lys Val Glu Ala Glu Gln Ser Leu Ile Thr Val Glu Gly Asp Lys
130 135 140
Ala Ser Met Glu Val Pro Ala Pro Phe Ala Gly Thr Val Lys Glu Ile
145 150 155 160
Lys Val Asn Val Gly Asp Lys Val Ser Thr Gly Ser Leu Ile Met Val
165 170 175
Phe Glu Val Ala Gly Glu Ala Gly Ala Ala Ala Pro Ala Ala Lys Gln
180 185 190
Glu Ala Ala Pro Ala Ala Ala Pro Ala Pro Ala Ala Gly Val Lys Glu
195 200 205
Val Asn Val Pro Asp Ile Gly Gly Asp Glu Val Glu Val Thr Glu Val
210 215 220
Met Val Lys Val Gly Asp Lys Val Ala Ala Glu Gln Ser Leu Ile Thr
225 230 235 240
Val Glu Gly Asp Lys Ala Ser Met Glu Val Pro Ala Pro Phe Ala Gly
245 250 255
Val Val Lys Glu Leu Lys Val Asn Val Gly Asp Lys Val Lys Thr Gly
260 265 270
Ser Leu Ile Met Ile Phe Glu Val Glu Gly Ala Ala Pro Ala Ala Ala
275 280 285
Pro Ala Lys Gln Glu Ala Ala Ala Pro Ala Pro Ala Ala Lys Ala Glu
290 295 300
Ala Pro Ala Ala Ala Pro Ala Ala Lys Ala Glu Gly Lys Ser Glu Phe
305 310 315 320
Ala Glu Asn Asp Ala Tyr Val His Ala Thr Pro Leu Ile Arg Arg Leu
325 330 335
Ala Arg Glu Phe Gly Val Asn Leu Ala Lys Val Lys Gly Thr Gly Arg
340 345 350
Lys Gly Arg Ile Leu Arg Glu Asp Val Gln Ala Tyr Val Lys Glu Ala
355 360 365
Ile Lys Arg Ala Glu Ala Ala Pro Ala Ala Thr Gly Gly Gly Ile Pro
370 375 380
Gly Met Leu Pro Trp Pro Lys Val Asp Phe Ser Lys Phe Gly Glu Ile
385 390 395 400
Glu Glu Val Glu Leu Gly Arg Ile Gln Lys Ile Ser Gly Ala Asn Leu
405 410 415
Ser Arg Asn Trp Val Met Ile Pro His Val Thr His Phe Asp Lys Thr
420 425 430
Asp Ile Thr Glu Leu Glu Ala Phe Arg Lys Gln Gln Asn Glu Glu Ala
435 440 445
Ala Lys Arg Lys Leu Asp Val Lys Ile Thr Pro Val Val Phe Ile Met
450 455 460
Lys Ala Val Ala Ala Ala Leu Glu Gln Met Pro Arg Phe Asn Ser Ser
465 470 475 480
Leu Ser Glu Asp Gly Gln Arg Leu Thr Leu Lys Lys Tyr Ile Asn Ile
485 490 495
Gly Val Ala Val Asp Thr Pro Asn Gly Leu Val Val Pro Val Phe Lys
500 505 510
Asp Val Asn Lys Lys Gly Ile Ile Glu Leu Ser Arg Glu Leu Met Thr
515 520 525
Ile Ser Lys Lys Ala Arg Asp Gly Lys Leu Thr Ala Gly Glu Met Gln
530 535 540
Gly Gly Cys Phe Thr Ile Ser Ser Ile Gly Gly Leu Gly Thr Thr His
545 550 555 560
Phe Ala Pro Ile Val Asn Ala Pro Glu Val Ala Ile Leu Gly Val Ser
565 570 575
Lys Ser Ala Met Glu Pro Val Trp Asn Gly Lys Glu Phe Val Pro Arg
580 585 590
Leu Met Leu Pro Ile Ser Leu Ser Phe Asp His Arg Val Ile Asp Gly
595 600 605
Ala Asp Gly Ala Arg Phe Ile Thr Ile Ile Asn Asn Thr Leu Ser Asp
610 615 620
Ile Arg Arg Leu Val Met
625 630
<210> 69
<211> 474
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 69
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> 70
<211> 371
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 70
Met Ala Ala Lys Thr Lys Lys Ala Ile Val Asp Ser Lys Lys Gln Phe
1 5 10 15
Asp Ala Ile Lys Lys Gln Phe Glu Thr Phe Gln Ile Leu Asn Glu Lys
20 25 30
Gly Glu Val Val Asn Glu Ala Ala Met Pro Asp Leu Thr Asp Asp Gln
35 40 45
Leu Lys Glu Leu Met Arg Arg Met Val Phe Thr Arg Val Leu Asp Gln
50 55 60
Arg Ser Ile Ser Leu Asn Arg Gln Gly Arg Leu Gly Phe Tyr Ala Pro
65 70 75 80
Thr Ala Gly Gln Glu Ala Ser Gln Ile Ala Thr His Phe Ala Leu Glu
85 90 95
Lys Glu Asp Phe Val Leu Pro Gly Tyr Arg Asp Val Pro Gln Leu Ile
100 105 110
Trp His Gly Leu Pro Leu Tyr Gln Ala Phe Leu Phe Ser Arg Gly His
115 120 125
Phe Arg Gly Asn Gln Met Pro Asp Asp Val Asn Ala Leu Ser Pro Gln
130 135 140
Ile Ile Ile Gly Ala Gln Tyr Ile Gln Thr Ala Gly Val Ala Leu Gly
145 150 155 160
Leu Lys Lys Arg Gly Lys Lys Ala Val Ala Ile Thr Tyr Thr Gly Asp
165 170 175
Gly Gly Ala Ser Gln Gly Asp Phe Tyr Glu Gly Ile Asn Phe Ala Gly
180 185 190
Ala Tyr Lys Ala Pro Ala Ile Phe Val Val Gln Asn Asn Arg Tyr Ala
195 200 205
Ile Ser Thr Pro Val Glu Lys Gln Ser Ala Ala Glu Thr Ile Ala Gln
210 215 220
Lys Ala Val Ala Ala Gly Ile Val Gly Val Gln Val Asp Gly Met Asp
225 230 235 240
Pro Leu Ala Val Tyr Ala Ala Thr Ala Glu Ala Arg Glu Arg Ala Ile
245 250 255
Asn Gly Glu Gly Pro Thr Leu Ile Glu Thr Leu Thr Phe Arg Tyr Gly
260 265 270
Pro His Thr Met Ala Gly Asp Asp Pro Thr Lys Tyr Arg Thr Lys Glu
275 280 285
Ile Glu Asn Glu Trp Glu Gln Lys Asp Pro Leu Val Arg Phe Arg Ala
290 295 300
Phe Leu Glu Asn Lys Gly Leu Trp Ser Glu Glu Glu Glu Ala Lys Val
305 310 315 320
Ile Glu Asp Ala Lys Glu Glu Ile Lys Gln Ala Ile Lys Lys Ala Asp
325 330 335
Ala Glu Pro Lys Gln Lys Val Thr Asp Leu Met Lys Ile Met Tyr Glu
340 345 350
Lys Met Pro His Asn Leu Glu Glu Gln Phe Glu Ile Tyr Thr Gln Lys
355 360 365
Glu Ser Lys
370
<210> 71
<211> 325
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 71
Met Ala Gln Met Thr Met Ile Gln Ala Ile Thr Asp Ala Leu Arg Thr
1 5 10 15
Glu Leu Lys Asn Asp Glu Asn Val Leu Val Phe Gly Glu Asp Val Gly
20 25 30
Val Asn Gly Gly Val Phe Arg Ala Thr Glu Gly Leu Gln Lys Glu Phe
35 40 45
Gly Glu Asp Arg Val Phe Asp Thr Pro Leu Ala Glu Ser Gly Ile Gly
50 55 60
Gly Leu Ala Leu Gly Leu Gly Leu Asn Gly Phe Arg Pro Val Met Glu
65 70 75 80
Ile Gln Phe Phe Gly Phe Val Tyr Glu Val Met Asp Ser Val Ser Gly
85 90 95
Gln Met Ala Arg Met Arg Tyr Arg Ser Gly Gly Arg Trp Thr Ser Pro
100 105 110
Val Thr Ile Arg Ser Pro Phe Gly Gly Gly Val His Thr Pro Glu Leu
115 120 125
His Ala Asp Ser Leu Glu Gly Leu Val Ala Gln Gln Pro Gly Ile Lys
130 135 140
Val Val Ile Pro Ser Thr Pro Tyr Asp Ala Lys Gly Leu Leu Ile Ser
145 150 155 160
Ala Ile Arg Asp Asn Asp Pro Val Val Phe Leu Glu His Met Lys Leu
165 170 175
Tyr Arg Ser Phe Arg Gln Glu Val Pro Glu Glu Glu Tyr Thr Ile Glu
180 185 190
Leu Gly Lys Ala Asp Val Lys Arg Glu Gly Thr Asp Leu Ser Ile Ile
195 200 205
Thr Tyr Gly Ala Met Val His Glu Ser Leu Lys Ala Ala Asp Glu Leu
210 215 220
Glu Lys Asp Gly Ile Ser Ala Glu Val Val Asp Leu Arg Thr Val Ser
225 230 235 240
Pro Leu Asp Ile Asp Thr Ile Ile Ala Ser Val Glu Lys Thr Gly Arg
245 250 255
Ala Ile Val Val Gln Glu Ala Gln Lys Gln Ala Gly Ile Ala Ala Asn
260 265 270
Val Val Ala Glu Ile Asn Asp Arg Ala Ile Leu Ser Leu Glu Ala Pro
275 280 285
Val Leu Arg Val Ala Ala Pro Asp Thr Val Phe Pro Phe Ser Gln Ala
290 295 300
Glu Ser Val Trp Leu Pro Asn His Lys Asp Val Leu Glu Thr Ala Arg
305 310 315 320
Lys Val Leu Glu Phe
325
<210> 72
<211> 442
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 72
Met Ala Phe Glu Phe Lys Leu Pro Asp Ile Gly Glu Gly Ile His Glu
1 5 10 15
Gly Glu Ile Val Lys Trp Phe Val Lys Pro Asn Asp Glu Val Asp Glu
20 25 30
Asp Asp Val Leu Ala Glu Val Gln Asn Asp Lys Ala Val Val Glu Ile
35 40 45
Pro Ser Pro Val Lys Gly Lys Val Leu Glu Leu Lys Val Glu Glu Gly
50 55 60
Thr Val Ala Thr Val Gly Gln Thr Ile Ile Thr Phe Asp Ala Pro Gly
65 70 75 80
Tyr Glu Asp Leu Gln Phe Lys Gly Ser Asp Glu Ser Asp Asp Ala Lys
85 90 95
Thr Glu Ala Gln Val Gln Ser Thr Ala Glu Ala Gly Gln Asp Val Ala
100 105 110
Lys Glu Glu Gln Ala Gln Glu Pro Ala Lys Ala Thr Gly Ala Gly Gln
115 120 125
Gln Asp Gln Ala Glu Val Asp Pro Asn Lys Arg Val Ile Ala Met Pro
130 135 140
Ser Val Arg Lys Tyr Ala Arg Glu Lys Gly Val Asp Ile Arg Lys Val
145 150 155 160
Thr Gly Ser Gly Asn Asn Gly Arg Val Val Lys Glu Asp Ile Asp Ser
165 170 175
Phe Val Asn Gly Gly Ala Gln Glu Ala Ala Pro Gln Glu Thr Ala Ala
180 185 190
Pro Gln Glu Thr Ala Ala Lys Pro Ala Ala Ala Pro Ala Pro Glu Gly
195 200 205
Glu Phe Pro Glu Thr Arg Glu Lys Met Ser Gly Ile Arg Lys Ala Ile
210 215 220
Ala Lys Ala Met Val Asn Ser Lys His Thr Ala Pro His Val Thr Leu
225 230 235 240
Met Asp Glu Val Asp Val Thr Asn Leu Val Ala His Arg Lys Gln Phe
245 250 255
Lys Gln Val Ala Ala Asp Gln Gly Ile Lys Leu Thr Tyr Leu Pro Tyr
260 265 270
Val Val Lys Ala Leu Thr Ser Ala Leu Lys Lys Phe Pro Val Leu Asn
275 280 285
Thr Ser Ile Asp Asp Lys Thr Asp Glu Val Ile Gln Lys His Tyr Phe
290 295 300
Asn Ile Gly Ile Ala Ala Asp Thr Glu Lys Gly Leu Leu Val Pro Val
305 310 315 320
Val Lys Asn Ala Asp Arg Lys Ser Val Phe Glu Ile Ser Asp Glu Ile
325 330 335
Asn Gly Leu Ala Thr Lys Ala Arg Glu Gly Lys Leu Ala Pro Ala Glu
340 345 350
Met Lys Gly Ala Ser Cys Thr Ile Thr Asn Ile Gly Ser Ala Gly Gly
355 360 365
Gln Trp Phe Thr Pro Val Ile Asn His Pro Glu Val Ala Ile Leu Gly
370 375 380
Ile Gly Arg Ile Ala Glu Lys Ala Ile Val Arg Asp Gly Glu Ile Val
385 390 395 400
Ala Ala Pro Val Leu Ala Leu Ser Leu Ser Phe Asp His Arg Met Ile
405 410 415
Asp Gly Ala Thr Ala Gln Asn Ala Leu Asn His Ile Lys Arg Leu Leu
420 425 430
Asn Asp Pro Gln Leu Ile Leu Met Glu Ala
435 440
<210> 73
<211> 470
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 73
Met Val Val Gly Asp Phe Pro Ile Glu Thr Asp Thr Leu Val Ile Gly
1 5 10 15
Ala Gly Pro Gly Gly Tyr Val Ala Ala Ile Arg Ala Ala Gln Leu Gly
20 25 30
Gln Lys Val Thr Val Val Glu Lys Ala Thr Leu Gly Gly Val Cys Leu
35 40 45
Asn Val Gly Cys Ile Pro Ser Lys Ala Leu Ile Asn Ala Gly His Arg
50 55 60
Tyr Glu Asn Ala Lys His Ser Asp Asp Met Gly Ile Thr Ala Glu Asn
65 70 75 80
Val Thr Val Asp Phe Thr Lys Val Gln Glu Trp Lys Ala Ser Val Val
85 90 95
Asn Lys Leu Thr Gly Gly Val Ala Gly Leu Leu Lys Gly Asn Lys Val
100 105 110
Asp Val Val Lys Gly Glu Ala Tyr Phe Val Asp Ser Asn Ser Val Arg
115 120 125
Val Met Asp Glu Asn Ser Ala Gln Thr Tyr Thr Phe Lys Asn Ala Ile
130 135 140
Ile Ala Thr Gly Ser Arg Pro Ile Glu Leu Pro Asn Phe Lys Tyr Ser
145 150 155 160
Glu Arg Val Leu Asn Ser Thr Gly Ala Leu Ala Leu Lys Glu Ile Pro
165 170 175
Lys Lys Leu Val Val Ile Gly Gly Gly Tyr Ile Gly Thr Glu Leu Gly
180 185 190
Thr Ala Tyr Ala Asn Phe Gly Thr Glu Leu Val Ile Leu Glu Gly Gly
195 200 205
Asp Glu Ile Leu Pro Gly Phe Glu Lys Gln Met Ser Ser Leu Val Thr
210 215 220
Arg Arg Leu Lys Lys Lys Gly Asn Val Glu Ile His Thr Asn Ala Met
225 230 235 240
Ala Lys Gly Val Glu Glu Arg Pro Asp Gly Val Thr Val Thr Phe Glu
245 250 255
Val Lys Gly Glu Glu Lys Thr Val Asp Ala Asp Tyr Val Leu Ile Thr
260 265 270
Val Gly Arg Arg Pro Asn Thr Asp Glu Leu Gly Leu Glu Gln Val Gly
275 280 285
Ile Glu Met Thr Asp Arg Gly Ile Val Lys Thr Asp Lys Gln Cys Arg
290 295 300
Thr Asn Val Pro Asn Ile Tyr Ala Ile Gly Asp Ile Ile Glu Gly Pro
305 310 315 320
Pro Leu Ala His Lys Ala Ser Tyr Glu Gly Lys Ile Ala Ala Glu Ala
325 330 335
Ile Ala Gly Glu Pro Ala Glu Ile Asp Tyr Leu Gly Ile Pro Ala Val
340 345 350
Val Phe Ser Glu Pro Glu Leu Ala Ser Val Gly Tyr Thr Glu Ala Gln
355 360 365
Ala Lys Glu Glu Gly Leu Asp Ile Val Ala Ala Lys Phe Pro Phe Ala
370 375 380
Ala Asn Gly Arg Ala Leu Ser Leu Asn Glu Thr Asp Gly Phe Met Lys
385 390 395 400
Leu Ile Thr Arg Lys Glu Asp Gly Leu Val Ile Gly Ala Gln Ile Ala
405 410 415
Gly Ala Ser Ala Ser Asp Met Ile Ser Glu Leu Ser Leu Ala Ile Glu
420 425 430
Gly Gly Met Thr Ala Glu Asp Ile Ala Met Thr Ile His Ala His Pro
435 440 445
Thr Leu Gly Glu Ile Thr Met Glu Ala Ala Glu Val Ala Ile Gly Ser
450 455 460
Pro Ile His Ile Val Lys
465 470
<210> 74
<211> 2123
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 74
Met Arg Ser Ile Arg Lys Trp Ala Tyr Glu Thr Phe Asn Asp Glu Lys
1 5 10 15
Ile Ile Gln Phe Val Val Met Ala Thr Pro Asp Asp Leu His Ala Asn
20 25 30
Ser Glu Tyr Ile Arg Met Ala Asp Gln Tyr Val Gln Val Pro Gly Gly
35 40 45
Thr Asn Asn Asn Asn Tyr Ala Asn Ile Asp Leu Ile Leu Asp Val Ala
50 55 60
Glu Gln Thr Asp Val Asp Ala Val Trp Ala Gly Trp Gly His Ala Ser
65 70 75 80
Glu Asn Pro Cys Leu Pro Glu Leu Leu Ala Ser Ser Gln Arg Lys Ile
85 90 95
Leu Phe Ile Gly Pro Pro Gly Arg Ala Met Arg Ser Leu Gly Asp Lys
100 105 110
Ile Ser Ser Thr Ile Val Ala Gln Ser Ala Lys Ile Pro Cys Ile Pro
115 120 125
Trp Ser Gly Ser His Ile Asp Thr Ile His Ile Asp Asn Lys Thr Asn
130 135 140
Phe Val Ser Val Pro Asp Asp Val Tyr Val Arg Gly Cys Cys Ser Ser
145 150 155 160
Pro Glu Asp Ala Leu Glu Lys Ala Lys Leu Ile Gly Phe Pro Val Met
165 170 175
Ile Lys Ala Ser Glu Gly Gly Gly Gly Lys Gly Ile Arg Arg Val Asp
180 185 190
Asn Glu Asp Asp Phe Ile Ala Leu Tyr Arg Gln Ala Val Asn Glu Thr
195 200 205
Pro Gly Ser Pro Met Phe Val Met Lys Val Val Thr Asp Ala Arg His
210 215 220
Leu Glu Val Gln Leu Leu Ala Asp Gln Tyr Gly Thr Asn Ile Thr Leu
225 230 235 240
Phe Gly Arg Asp Cys Ser Ile Gln Arg Arg His Gln Lys Ile Ile Glu
245 250 255
Glu Ala Pro Val Thr Ile Thr Lys Pro Glu Thr Phe Gln Arg Met Glu
260 265 270
Arg Ala Ala Ile Arg Leu Gly Glu Leu Val Gly Tyr Val Ser Ala Gly
275 280 285
Thr Val Glu Tyr Leu Tyr Ser Pro Lys Asp Asp Lys Phe Tyr Phe Leu
290 295 300
Glu Leu Asn Pro Arg Leu Gln Val Glu His Pro Thr Thr Glu Met Ile
305 310 315 320
Ser Gly Val Asn Leu Pro Ala Thr Gln Leu Gln Ile Ala Met Gly Ile
325 330 335
Pro Met His Met Ile Ser Asp Ile Arg Lys Leu Tyr Gly Leu Asp Pro
340 345 350
Thr Gly Thr Ser Tyr Ile Asp Phe Lys Asn Leu Lys Arg Pro Ser Pro
355 360 365
Lys Gly His Cys Ile Ser Cys Arg Ile Thr Ser Glu Asp Pro Asn Glu
370 375 380
Gly Phe Lys Pro Ser Thr Gly Lys Ile His Glu Leu Asn Phe Arg Ser
385 390 395 400
Ser Ser Asn Val Trp Gly Tyr Phe Ser Val Gly Asn Asn Gly Ala Ile
405 410 415
His Ser Phe Ser Asp Ser Gln Phe Gly His Ile Phe Ala Val Gly Asn
420 425 430
Asp Arg Gln Asp Ala Lys Gln Asn Met Val Leu Ala Leu Lys Asp Phe
435 440 445
Ser Ile Arg Gly Glu Phe Lys Thr Pro Ile Glu Tyr Leu Ile Glu Leu
450 455 460
Leu Glu Thr Arg Asp Phe Glu Ser Asn Asn Ile Ser Thr Gly Trp Leu
465 470 475 480
Asp Asp Leu Ile Leu Lys Asn Leu Ser Ser Asp Ser Lys Leu Asp Pro
485 490 495
Thr Leu Ala Ile Ile Cys Gly Ala Ala Met Lys Ala Tyr Val Phe Thr
500 505 510
Glu Lys Val Arg Asn Lys Tyr Leu Glu Leu Leu Arg Arg Gly Gln Val
515 520 525
Pro Pro Lys Asp Phe Leu Lys Thr Lys Phe Pro Val Asp Phe Ile Phe
530 535 540
Asp Asn Asn Arg Tyr Leu Phe Asn Val Ala Gln Ser Ser Glu Glu Gln
545 550 555 560
Phe Ile Leu Ser Ile Asn Lys Ser Gln Cys Glu Val Asn Val Gln Lys
565 570 575
Leu Ser Ser Asp Cys Leu Leu Ile Ser Val Asp Gly Lys Cys His Thr
580 585 590
Val Tyr Trp Lys Asp Asp Ile Arg Gly Thr Arg Leu Ser Ile Asp Ser
595 600 605
Asn Thr Ile Phe Leu Glu Ala Glu Leu Asn Pro Thr Gln Val Ile Ser
610 615 620
Pro Thr Pro Gly Lys Leu Val Lys Tyr Leu Val Arg Ser Gly Asp His
625 630 635 640
Val Phe Ala Gly Gln Gln Tyr Ala Glu Ile Glu Ile Met Lys Met Gln
645 650 655
Met Pro Leu Val Ala Lys Ser Asp Gly Val Ile Glu Leu Leu Arg Gln
660 665 670
Pro Gly Ser Ile Ile Glu Ala Gly Asp Val Ile Ala Lys Leu Thr Leu
675 680 685
Asp Ser Pro Ser Lys Ala Asn Glu Ser Ser Leu Tyr Arg Gly Glu Leu
690 695 700
Pro Val Leu Gly Pro Pro Leu Ile Glu Gly Ser Arg Pro Asn His Lys
705 710 715 720
Leu Arg Val Leu Ile Asn Arg Leu Glu Asn Ile Leu Asn Gly Tyr His
725 730 735
Glu Asn Ser Gly Ile Glu Thr Thr Leu Lys Glu Leu Ile Lys Ile Leu
740 745 750
Arg Asp Gly Arg Leu Pro Tyr Ser Glu Trp Asp Ser Gln Ile Ser Thr
755 760 765
Val Arg Asn Arg Leu Pro Arg Gln Leu Asn Glu Gly Leu Gly Asn Leu
770 775 780
Val Lys Lys Ser Val Ser Phe Pro Ala Lys Glu Leu His Lys Leu Met
785 790 795 800
Lys Arg Tyr Leu Glu Glu Asn Thr Asn Asp His Val Val Tyr Val Ala
805 810 815
Leu Gln Pro Leu Leu Lys Ile Ser Glu Arg Tyr Ser Glu Gly Leu Ala
820 825 830
Asn His Glu Cys Glu Ile Phe Leu Lys Leu Ile Lys Lys Tyr Tyr Ala
835 840 845
Val Glu Lys Ile Phe Glu Asn His Asp Ile His Glu Glu Arg Asn Leu
850 855 860
Leu Asn Leu Arg Arg Lys Asp Leu Thr Asn Leu Lys Lys Ile Leu Cys
865 870 875 880
Ile Ser Leu Ser His Ala Asn Val Val Ala Lys Asn Lys Leu Val Thr
885 890 895
Ala Ile Leu His Glu Tyr Glu Pro Leu Cys Gln Asp Ser Ser Lys Met
900 905 910
Ser Leu Lys Phe Arg Ala Val Ile His Asp Leu Ala Ser Leu Glu Ser
915 920 925
Lys Trp Ala Lys Glu Val Ala Val Lys Ala Arg Ser Val Leu Leu Arg
930 935 940
Gly Ile Phe Pro Pro Ile Lys Lys Arg Lys Glu His Ile Lys Thr Leu
945 950 955 960
Leu Gln Leu His Ile Lys Asp Thr Gly Ala Glu Asn Ile His Ser Arg
965 970 975
Asn Ile Tyr Ser Cys Met Arg Asp Phe Gly Asn Leu Ile His Ser Asn
980 985 990
Leu Ile Gln Leu Gln Asp Leu Phe Phe Phe Phe Gly His Gln Asp Thr
995 1000 1005
Ala Leu Ser Ser Ile Ala Ser Glu Ile Tyr Ala Arg Tyr Ala Tyr
1010 1015 1020
Gly Asn Tyr Gln Leu Lys Ser Ile Lys Ile His Lys Gly Ala Pro
1025 1030 1035
Asp Leu Leu Met Ser Trp Gln Phe Ser Ser Leu Arg Asn Tyr Leu
1040 1045 1050
Val Asn Ser Asp Gly Glu Ser Asp Glu Phe Thr Lys Leu Ser Lys
1055 1060 1065
Pro Pro Ser Thr Ser Gly Lys Ser Ser Ala Asn Ser Phe Gly Leu
1070 1075 1080
Leu Val Asn Met Arg Ala Leu Glu Ser Leu Glu Lys Thr Leu Asp
1085 1090 1095
Glu Val Tyr Glu Gln Ile His Ile Pro Glu Glu Arg Leu Ser Ser
1100 1105 1110
Gly Glu Asn Ser Leu Ile Val Asn Ile Leu Ser Pro Ile Arg Tyr
1115 1120 1125
Arg Ser Glu Asn Asp Leu Ile Lys Thr Leu Lys Ile Lys Leu His
1130 1135 1140
Glu Asn Glu Arg Gly Leu Ser Lys Leu Lys Val Asn Arg Ile Thr
1145 1150 1155
Phe Ala Phe Ile Ala Ala Asn Ala Pro Ala Val Lys Phe Tyr Ser
1160 1165 1170
Phe Asp Gly Thr Thr Tyr Asp Glu Ile Ser Gln Ile Arg Asn Met
1175 1180 1185
Asp Pro Ser Tyr Glu Ala Pro Leu Glu Leu Gly Lys Met Ser Asn
1190 1195 1200
Tyr Lys Ile Arg Ser Leu Pro Thr Tyr Asp Ser Ser Ile Arg Ile
1205 1210 1215
Phe Glu Gly Ile Ser Lys Phe Thr Pro Leu Asp Lys Arg Phe Phe
1220 1225 1230
Val Arg Lys Ile Ile Asn Ser Phe Met Tyr Asn Asp Gln Lys Thr
1235 1240 1245
Thr Glu Glu Asn Leu Lys Ala Glu Ile Asn Ala Gln Val Val Tyr
1250 1255 1260
Met Leu Glu His Leu Gly Ala Val Asp Ile Ser Asn Ser Asp Leu
1265 1270 1275
Asn His Ile Phe Leu Ser Phe Asn Thr Val Leu Asn Ile Pro Val
1280 1285 1290
His Arg Leu Glu Glu Ile Val Ser Thr Ile Leu Lys Thr His Glu
1295 1300 1305
Thr Arg Leu Phe Gln Glu Arg Ile Thr Asp Val Glu Ile Cys Ile
1310 1315 1320
Ser Val Glu Cys Leu Glu Thr Lys Lys Pro Ala Pro Leu Arg Leu
1325 1330 1335
Leu Ile Ser Asn Lys Ser Gly Tyr Val Val Lys Ile Glu Thr Tyr
1340 1345 1350
Tyr Glu Lys Ile Gly Lys Asn Gly Asn Leu Ile Leu Glu Pro Cys
1355 1360 1365
Ser Glu Gln Ser His Tyr Ser Gln Lys Ser Leu Ser Leu Pro Tyr
1370 1375 1380
Ser Val Lys Asp Trp Leu Gln Pro Lys Arg Tyr Lys Ala Gln Phe
1385 1390 1395
Met Gly Thr Thr Tyr Val Tyr Asp Phe Pro Gly Leu Phe His Gln
1400 1405 1410
Ala Ala Ile Gln Gln Trp Lys Arg Tyr Phe Pro Lys His Lys Leu
1415 1420 1425
Asn Asp Ser Phe Phe Ser Trp Val Glu Leu Ile Glu Gln Asn Gly
1430 1435 1440
Asn Leu Ile Lys Val Asn Arg Glu Pro Gly Leu Asn Asn Ile Gly
1445 1450 1455
Met Val Ala Phe Glu Ile Met Val Gln Thr Pro Glu Tyr Pro Glu
1460 1465 1470
Gly Arg Asn Met Ile Val Ile Ser Asn Asp Ile Thr Tyr Asn Ile
1475 1480 1485
Gly Ser Phe Gly Pro Arg Glu Asp Leu Phe Phe Asp Arg Val Thr
1490 1495 1500
Asn Tyr Ala Arg Glu Arg Gly Ile Pro Arg Ile Tyr Leu Ala Ala
1505 1510 1515
Asn Ser Gly Ala Lys Leu Gly Ile Ala Glu Glu Leu Ile Pro Leu
1520 1525 1530
Phe Arg Val Ala Trp Asn Asp Pro Ser Asp Pro Thr Lys Gly Phe
1535 1540 1545
Gln Tyr Leu Tyr Leu Ala Pro Lys Asp Met Gln Leu Leu Lys Asp
1550 1555 1560
Ser Gly Lys Gly Asn Ser Val Val Val Glu His Lys Met Val Tyr
1565 1570 1575
Gly Glu Glu Arg Tyr Ile Ile Lys Ala Ile Val Gly Phe Glu Glu
1580 1585 1590
Gly Leu Gly Val Glu Cys Leu Gln Gly Ser Gly Leu Ile Ala Gly
1595 1600 1605
Ala Thr Ser Lys Ala Tyr Arg Asp Ile Phe Thr Ile Thr Ala Val
1610 1615 1620
Thr Cys Arg Ser Val Gly Ile Gly Ser Tyr Leu Val Arg Leu Gly
1625 1630 1635
Gln Arg Thr Ile Gln Val Glu Asp Lys Pro Ile Ile Leu Thr Gly
1640 1645 1650
Ala Ser Ala Ile Asn Lys Val Leu Gly Thr Asp Ile Tyr Thr Ser
1655 1660 1665
Asn Leu Gln Ile Gly Gly Thr Gln Ile Met Tyr Lys Asn Gly Ile
1670 1675 1680
Ala His Leu Thr Ala Ser Asn Asp Met Lys Ala Ile Glu Lys Ile
1685 1690 1695
Met Thr Trp Leu Ser Tyr Val Pro Ala Lys Arg Asp Met Ser Pro
1700 1705 1710
Pro Leu Leu Glu Thr Met Asp Arg Trp Asp Arg Asp Val Asp Phe
1715 1720 1725
Lys Pro Ala Lys Gln Val Pro Tyr Glu Ala Arg Trp Leu Ile Glu
1730 1735 1740
Gly Lys Trp Asp Ser Asn Asn Asn Phe Gln Ser Gly Leu Phe Asp
1745 1750 1755
Lys Asp Ser Phe Phe Glu Thr Leu Ser Gly Trp Ala Lys Gly Val
1760 1765 1770
Ile Val Gly Arg Ala Arg Leu Gly Gly Ile Pro Val Gly Val Ile
1775 1780 1785
Ala Val Glu Thr Lys Thr Ile Glu Glu Ile Ile Pro Ala Asp Pro
1790 1795 1800
Ala Asn Leu Asp Ser Ser Glu Phe Ser Val Lys Glu Ala Gly Gln
1805 1810 1815
Val Trp Tyr Pro Asn Ser Ala Phe Lys Thr Ala Gln Thr Ile Asn
1820 1825 1830
Asp Phe Asn Tyr Gly Glu Gln Leu Pro Leu Ile Ile Leu Ala Asn
1835 1840 1845
Trp Arg Gly Phe Ser Gly Gly Gln Arg Asp Met Tyr Asn Glu Val
1850 1855 1860
Leu Lys Tyr Gly Ser Phe Ile Val Asp Ala Leu Val Asp Tyr Lys
1865 1870 1875
Gln Pro Ile Leu Ile Tyr Ile Pro Pro Phe Gly Glu Leu Arg Gly
1880 1885 1890
Gly Ser Trp Val Val Ile Asp Pro Thr Ile Asn Pro Glu Gln Met
1895 1900 1905
Glu Met Tyr Ala Asp Val Glu Ser Arg Gly Gly Val Leu Glu Pro
1910 1915 1920
Asp Gly Val Val Ser Ile Lys Tyr Arg Lys Glu Lys Met Ile Glu
1925 1930 1935
Thr Met Ile Arg Leu Asp Ser Thr Tyr Gly His Leu Arg Arg Thr
1940 1945 1950
Leu Thr Glu Lys Lys Leu Ser Leu Glu Lys Gln Asn Asp Leu Thr
1955 1960 1965
Lys Arg Leu Lys Ile Arg Glu Arg Gln Leu Ile Pro Ile Tyr Asn
1970 1975 1980
Gln Ile Ser Ile Gln Phe Ala Asp Leu His Asp Arg Ser Thr Arg
1985 1990 1995
Met Leu Val Lys Gly Val Ile Arg Asn Glu Leu Glu Trp Lys Lys
2000 2005 2010
Ser Arg Arg Phe Leu Tyr Trp Arg Leu Arg Arg Arg Leu Asn Glu
2015 2020 2025
Gly Gln Val Ile Lys Arg Leu Gln Lys Lys Thr Cys Asp Asn Lys
2030 2035 2040
Thr Lys Met Lys Tyr Asp Asp Leu Leu Lys Ile Val Gln Ser Trp
2045 2050 2055
Tyr Asn Asp Leu Asp Val Asn Asp Asp Arg Ala Val Val Glu Phe
2060 2065 2070
Ile Glu Arg Asn Ser Lys Lys Ile Asp Lys Asn Ile Glu Glu Phe
2075 2080 2085
Glu Ile Ser Leu Leu Ile Asp Glu Leu Lys Lys Lys Phe Glu Asp
2090 2095 2100
Arg Arg Gly Asn Ile Val Leu Glu Glu Leu Thr Arg Leu Val Asp
2105 2110 2115
Ser Lys Arg Lys Arg
2120
<210> 75
<211> 319
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 75
Met Ser Leu Asn Phe Leu Asp Phe Glu Gln Pro Ile Ala Glu Leu Glu
1 5 10 15
Ala Lys Ile Asp Ser Leu Thr Ala Val Ser Arg Gln Asp Glu Lys Leu
20 25 30
Asp Ile Asn Ile Asp Glu Glu Val His Arg Leu Arg Glu Lys Ser Val
35 40 45
Glu Leu Thr Arg Lys Ile Phe Ala Asp Leu Gly Ala Trp Gln Ile Ala
50 55 60
Gln Leu Ala Arg His Pro Gln Arg Pro Tyr Thr Leu Asp Tyr Val Arg
65 70 75 80
Leu Ala Phe Asp Glu Phe Asp Glu Leu Ala Gly Asp Arg Ala Tyr Ala
85 90 95
Asp Asp Lys Ala Ile Val Gly Gly Ile Ala Arg Leu Asp Gly Arg Pro
100 105 110
Val Met Ile Ile Gly His Gln Lys Gly Arg Glu Thr Lys Glu Lys Ile
115 120 125
Arg Arg Asn Phe Gly Met Pro Ala Pro Glu Gly Tyr Arg Lys Ala Leu
130 135 140
Arg Leu Met Gln Met Ala Glu Arg Phe Lys Met Pro Ile Ile Thr Phe
145 150 155 160
Ile Asp Thr Pro Gly Ala Tyr Pro Gly Val Gly Ala Glu Glu Arg Gly
165 170 175
Gln Ser Glu Ala Ile Ala Arg Asn Leu Arg Glu Met Ser Arg Leu Gly
180 185 190
Val Pro Val Val Cys Thr Val Ile Gly Glu Gly Gly Ser Gly Gly Ala
195 200 205
Leu Ala Ile Gly Val Gly Asp Lys Val Asn Met Leu Gln Tyr Ser Thr
210 215 220
Tyr Ser Val Ile Ser Pro Glu Gly Cys Ala Ser Ile Leu Trp Lys Ser
225 230 235 240
Ala Asp Lys Ala Pro Leu Ala Ala Glu Ala Met Gly Ile Ile Ala Pro
245 250 255
Arg Leu Lys Glu Leu Lys Leu Ile Asp Ser Ile Ile Pro Glu Pro Leu
260 265 270
Gly Gly Ala His Arg Asn Pro Glu Ala Met Ala Ala Ser Leu Lys Ala
275 280 285
Gln Leu Leu Ala Asp Leu Ala Asp Leu Asp Val Leu Ser Thr Glu Asp
290 295 300
Leu Lys Asn Arg Arg Tyr Gln Arg Leu Met Ser Tyr Gly Tyr Ala
305 310 315
<210> 76
<211> 156
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 76
Met Asp Ile Arg Lys Ile Lys Lys Leu Ile Glu Leu Val Glu Glu Ser
1 5 10 15
Gly Ile Ser Glu Leu Glu Ile Ser Glu Gly Glu Glu Ser Val Arg Ile
20 25 30
Ser Arg Ala Ala Pro Ala Ala Ser Phe Pro Val Met Gln Gln Ala Tyr
35 40 45
Ala Ala Pro Met Met Gln Gln Pro Ala Gln Ser Asn Ala Ala Ala Pro
50 55 60
Ala Thr Val Pro Ser Met Glu Ala Pro Ala Ala Ala Glu Ile Ser Gly
65 70 75 80
His Ile Val Arg Ser Pro Met Val Gly Thr Phe Tyr Arg Thr Pro Ser
85 90 95
Pro Asp Ala Lys Ala Phe Ile Glu Val Gly Gln Lys Val Asn Val Gly
100 105 110
Asp Thr Leu Cys Ile Val Glu Ala Met Lys Met Met Asn Gln Ile Glu
115 120 125
Ala Asp Lys Ser Gly Thr Val Lys Ala Ile Leu Val Glu Ser Gly Gln
130 135 140
Pro Val Glu Phe Asp Glu Pro Leu Val Val Ile Glu
145 150 155
<210> 77
<211> 449
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 77
Met Leu Asp Lys Ile Val Ile Ala Asn Arg Gly Glu Ile Ala Leu Arg
1 5 10 15
Ile Leu Arg Ala Cys Lys Glu Leu Gly Ile Lys Thr Val Ala Val His
20 25 30
Ser Ser Ala Asp Arg Asp Leu Lys His Val Leu Leu Ala Asp Glu Thr
35 40 45
Val Cys Ile Gly Pro Ala Pro Ser Val Lys Ser Tyr Leu Asn Ile Pro
50 55 60
Ala Ile Ile Ser Ala Ala Glu Ile Thr Gly Ala Val Ala Ile His Pro
65 70 75 80
Gly Tyr Gly Phe Leu Ser Glu Asn Ala Asn Phe Ala Glu Gln Val Glu
85 90 95
Arg Ser Gly Phe Ile Phe Ile Gly Pro Lys Ala Glu Thr Ile Arg Leu
100 105 110
Met Gly Asp Lys Val Ser Ala Ile Ala Ala Met Lys Lys Ala Gly Val
115 120 125
Pro Cys Val Pro Gly Ser Asp Gly Pro Leu Gly Asp Asp Met Asp Lys
130 135 140
Asn Arg Ala Ile Ala Lys Arg Ile Gly Tyr Pro Val Ile Ile Lys Ala
145 150 155 160
Ser Gly Gly Gly Gly Gly Arg Gly Met Arg Val Val Arg Gly Asp Ala
165 170 175
Glu Leu Ala Gln Ser Ile Ser Met Thr Arg Ala Glu Ala Lys Ala Ala
180 185 190
Phe Ser Asn Asp Met Val Tyr Met Glu Lys Tyr Leu Glu Asn Pro Arg
195 200 205
His Val Glu Ile Gln Val Leu Ala Asp Gly Gln Gly Asn Ala Ile Tyr
210 215 220
Leu Ala Glu Arg Asp Cys Ser Met Gln Arg Arg His Gln Lys Val Val
225 230 235 240
Glu Glu Ala Pro Ala Pro Gly Ile Thr Pro Glu Leu Arg Arg Tyr Ile
245 250 255
Gly Glu Arg Cys Ala Lys Ala Cys Val Asp Ile Gly Tyr Arg Gly Ala
260 265 270
Gly Thr Phe Glu Phe Leu Phe Glu Asn Gly Glu Phe Tyr Phe Ile Glu
275 280 285
Met Asn Thr Arg Ile Gln Val Glu His Pro Val Thr Glu Met Ile Thr
290 295 300
Gly Val Asp Leu Ile Lys Glu Gln Leu Arg Ile Ala Ala Gly Gln Pro
305 310 315 320
Leu Ser Ile Lys Gln Glu Glu Val His Val Arg Gly His Ala Val Glu
325 330 335
Cys Arg Ile Asn Ala Glu Asp Pro Asn Thr Phe Leu Pro Ser Pro Gly
340 345 350
Lys Ile Thr Arg Phe His Ala Pro Gly Gly Phe Gly Val Arg Trp Glu
355 360 365
Ser His Ile Tyr Ala Gly Tyr Thr Val Pro Pro Tyr Tyr Asp Ser Met
370 375 380
Ile Gly Lys Leu Ile Cys Tyr Gly Glu Asn Arg Asp Val Ala Ile Ala
385 390 395 400
Arg Met Lys Asn Ala Leu Gln Glu Leu Ile Ile Asp Gly Ile Lys Thr
405 410 415
Asn Val Asp Leu Gln Ile Arg Ile Met Asn Asp Glu Asn Phe Gln His
420 425 430
Gly Gly Thr Asn Ile His Tyr Leu Glu Lys Lys Leu Gly Leu Gln Glu
435 440 445
Lys
<210> 78
<211> 304
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 78
Met Ser Trp Ile Glu Arg Ile Lys Ser Asn Ile Thr Pro Thr Arg Lys
1 5 10 15
Ala Ser Ile Pro Glu Gly Val Trp Thr Lys Cys Asp Ser Cys Gly Gln
20 25 30
Val Leu Tyr Arg Ala Glu Leu Glu Arg Asn Leu Glu Val Cys Pro Lys
35 40 45
Cys Asp His His Met Arg Met Thr Ala Arg Asn Arg Leu His Ser Leu
50 55 60
Leu Asp Glu Gly Ser Leu Val Glu Leu Gly Ser Glu Leu Glu Pro Lys
65 70 75 80
Asp Val Leu Lys Phe Arg Asp Ser Lys Lys Tyr Lys Asp Arg Leu Ala
85 90 95
Ser Ala Gln Lys Glu Thr Gly Glu Lys Asp Ala Leu Val Val Met Lys
100 105 110
Gly Thr Leu Tyr Gly Met Pro Val Val Ala Ala Ala Phe Glu Phe Ala
115 120 125
Phe Met Gly Gly Ser Met Gly Ser Val Val Gly Ala Arg Phe Val Arg
130 135 140
Ala Val Glu Gln Ala Leu Glu Asp Asn Cys Pro Leu Ile Cys Phe Ser
145 150 155 160
Ala Ser Gly Gly Ala Arg Met Gln Glu Ala Leu Met Ser Leu Met Gln
165 170 175
Met Ala Lys Thr Ser Ala Ala Leu Ala Lys Met Gln Glu Arg Gly Leu
180 185 190
Pro Tyr Ile Ser Val Leu Thr Asp Pro Thr Met Gly Gly Val Ser Ala
195 200 205
Ser Phe Ala Met Leu Gly Asp Leu Asn Ile Ala Glu Pro Lys Ala Leu
210 215 220
Ile Gly Phe Ala Gly Pro Arg Val Ile Glu Gln Thr Val Arg Glu Lys
225 230 235 240
Leu Pro Pro Gly Phe Gln Arg Ser Glu Phe Leu Ile Glu Lys Gly Ala
245 250 255
Ile Asp Met Ile Val Arg Arg Pro Glu Met Arg Leu Lys Leu Ala Ser
260 265 270
Ile Leu Ala Lys Leu Met Asn Leu Pro Ala Pro Asn Pro Glu Ala Pro
275 280 285
Arg Glu Gly Val Val Val Pro Pro Val Pro Asp Gln Glu Pro Glu Ala
290 295 300
<210> 79
<211> 282
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 79
Met Glu Glu Thr Ala Ile Pro Gln Ser Leu Thr Pro Trp Asp Arg Val
1 5 10 15
Gln Leu Ala Arg His Pro Gln Arg Pro His Thr Leu Asp Tyr Ile Ala
20 25 30
Ala Leu Cys Glu Asp Phe Val Glu Leu His Gly Asp Arg Arg Phe Gly
35 40 45
Asp Asp Pro Ala Met Val Gly Gly Met Ala Thr Phe Ala Gly Gln Thr
50 55 60
Val Met Val Ile Gly His Gln Lys Gly Asn Asp Thr Arg Glu Asn Met
65 70 75 80
Arg Arg Asn Phe Gly Met Pro His Pro Glu Gly Tyr Arg Lys Ala Gln
85 90 95
Arg Leu Met Arg His Ala Glu Lys Phe Gly Leu Pro Val Ile Cys Phe
100 105 110
Val Asp Thr Pro Ala Ala Asp Pro Thr Lys Ser Ser Glu Glu Arg Gly
115 120 125
Gln Ala Asn Ala Ile Ala Glu Ser Ile Met Leu Met Thr Thr Leu Arg
130 135 140
Val Pro Ser Ile Ala Val Val Ile Gly Glu Gly Gly Ser Gly Gly Ala
145 150 155 160
Leu Ala Ile Ser Val Ala Asp Arg Ile Leu Met Gln Glu Asn Ala Ile
165 170 175
Tyr Ser Val Ala Pro Pro Glu Ala Ala Ala Ser Ile Leu Trp Arg Asp
180 185 190
Ala Ala Lys Ala Pro Glu Ala Ala Arg Ala Leu Lys Leu Thr Ala Ala
195 200 205
Asp Leu Tyr Asp Leu Arg Ile Ile Asp Glu Val Ile Pro Glu Pro Pro
210 215 220
Gly Gly Ala His Ala Asp Arg Leu Thr Ala Ile Thr Thr Val Gly Glu
225 230 235 240
Arg Leu Arg Val His Leu Ala Asp Leu Gln Gln Arg Asp Ile Asp Thr
245 250 255
Leu Leu Arg Glu Arg Tyr Arg Lys Tyr Arg Ser Met Gly Gln Tyr Gln
260 265 270
Glu Gln Gln Met Asp Phe Phe Gly Arg Met
275 280
<210> 80
<211> 180
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 80
Met Met Leu Trp Gly Ala Met Lys Asp Glu Thr Thr Glu Leu Pro Ala
1 5 10 15
Asp Gln Pro Asp Pro Phe Gly Leu Ala Ala Val Arg Val Leu Leu Gln
20 25 30
Met Leu Glu Gln Ser Asp Val Tyr Glu Ile Thr Ile Glu Asn Gly Asn
35 40 45
Ala Lys Leu His Val Lys Arg Gly Gln Pro Gly Gly Val Ile Tyr Ser
50 55 60
Ala Pro Leu Pro Thr Ala Pro Val Pro Ser Pro Ser Leu Pro Ala Thr
65 70 75 80
Pro Val Thr Pro Phe Val Gln Pro Pro Pro Ala Pro Glu Gly Pro Pro
85 90 95
Val Glu Met Pro Ala Gly His Thr Ile Thr Ala Pro Met Val Gly Thr
100 105 110
Phe Tyr Ala Ala Pro Ser Pro Arg Asp Arg Pro Phe Val Gln Glu Gly
115 120 125
Asp Glu Val Arg Val Gly Asp Thr Val Gly Ile Val Glu Ala Met Lys
130 135 140
Met Met Asn Glu Ile Glu Ser Asp Val Ala Gly Arg Val Ala Arg Ile
145 150 155 160
Leu Val Lys Asn Gly Gln Pro Val Glu Tyr Gly Gln Pro Leu Met Val
165 170 175
Ile Glu Pro Leu
180
<210> 81
<211> 455
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 81
Met Ile Arg Lys Val Leu Val Ala Asn Arg Gly Glu Ile Ala Val Arg
1 5 10 15
Ile Ile Arg Ala Cys Gln Glu Leu Gly Ile Arg Thr Val Val Ala Tyr
20 25 30
Ser Thr Ala Asp Arg Asp Ser Leu Ala Val Arg Leu Ala Asp Glu Ala
35 40 45
Val Cys Ile Gly Pro Pro Pro Ala Ala Lys Ser Tyr Leu Asn Ala Pro
50 55 60
Ala Leu Ile Ser Ala Ala Leu Val Ser Gly Cys Asp Ala Ile His Pro
65 70 75 80
Gly Tyr Gly Phe Leu Ser Glu Asn Pro Tyr Phe Ala Glu Met Cys Ala
85 90 95
Asp Cys Lys Leu Thr Phe Ile Gly Pro Pro Pro Glu Pro Ile Arg Leu
100 105 110
Met Gly Asp Lys Ala Ile Gly Arg Glu Thr Met Arg Lys Ala Gly Val
115 120 125
Pro Thr Val Pro Gly Ser Asp Gly Glu Val Arg Ser Leu Glu Glu Ala
130 135 140
Ile Asp Val Ala Arg Gln Ile Gly Tyr Pro Val Leu Leu Lys Pro Ser
145 150 155 160
Gly Gly Gly Gly Gly Arg Gly Met Arg Val Ala Tyr Asp Glu Ala Asp
165 170 175
Leu Gln Arg Ala Phe Pro Thr Ala Arg Ala Glu Ala Glu Ala Ala Phe
180 185 190
Gly Asn Gly Ala Leu Leu Leu Glu Lys Tyr Leu Thr Arg Val Arg His
195 200 205
Val Glu Ile Gln Val Leu Ala Asp Gln Tyr Gly His Ala Ile His Leu
210 215 220
Gly Glu Arg Asp Cys Ser Ala Gln Arg Arg His Gln Lys Ile Val Glu
225 230 235 240
Glu Ala Pro Ser Pro Ala Val Thr Pro Glu Leu Arg Glu Arg Met Gly
245 250 255
Ala Asp Ala Val Arg Gly Ile Lys Ser Ile Gly Tyr Val Asn Ala Gly
260 265 270
Thr Leu Glu Phe Leu Leu Asp Gln Asp Gly Asn Tyr Tyr Phe Ile Glu
275 280 285
Met Asn Thr Arg Ile Gln Val Glu His Pro Val Thr Glu Gln Val Thr
290 295 300
Gly Ile Asp Leu Val Arg Trp Gln Leu Leu Ile Ala Ser Gly Glu Arg
305 310 315 320
Leu Thr Leu Arg Gln Glu Asp Ile Lys Ile Thr Arg His Ala Ile Glu
325 330 335
Cys Arg Ile Asn Ala Glu Asp Pro Glu Arg Asp Phe Leu Pro Ala Ser
340 345 350
Gly Glu Val Glu Phe Tyr Leu Pro Pro Gly Gly Pro Gly Val Arg Val
355 360 365
Asp Ser His Leu Tyr Ser Gly Tyr Thr Pro Pro Gly Thr Tyr Asp Ser
370 375 380
Leu Leu Ala Lys Ile Ile Thr Phe Gly Asp Thr Arg Asp Glu Ala Leu
385 390 395 400
Asn Arg Met Arg Arg Ala Leu Asn Glu Cys Val Ile Thr Gly Ile Lys
405 410 415
Thr Thr Ile Pro Phe Gln Leu Ala Leu Ile Asp Asp Pro Glu Phe Arg
420 425 430
Ala Gly Arg Ile His Thr Gly Tyr Val Ala Glu Leu Leu Arg Gln Trp
435 440 445
Lys Glu Thr Leu Asn Pro Val
450 455
<210> 82
<211> 305
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 82
Met Lys Glu Phe Phe Arg Leu Ser Arg Lys Gly Phe Thr Gly Arg Glu
1 5 10 15
Asp Gln Asp Ser Ala Gln Ile Pro Asp Asp Leu Trp Val Lys Cys Ser
20 25 30
Ser Cys Arg Glu Leu Ile Tyr Lys Lys Gln Leu Asn Asp Asn Leu Lys
35 40 45
Val Cys Pro Lys Cys Gly His His Met Arg Leu Ser Ala His Glu Trp
50 55 60
Leu Gly Leu Leu Asp Val Gly Ser Phe Arg Glu Met Asp Ala Asn Leu
65 70 75 80
Leu Pro Thr Asp Pro Leu Gly Phe Val Thr Asp Glu Glu Ser Tyr Ala
85 90 95
Ala Lys Leu Ala Lys Thr Gln Gln Arg Thr Gly Met Ala Asp Ala Val
100 105 110
Ile Ala Gly Ile Gly Ala Ile Ser Asn Met Gln Ile Cys Val Ala Val
115 120 125
Ala Asp Phe Ser Phe Met Gly Ala Ser Met Gly Ser Val Tyr Gly Glu
130 135 140
Lys Met Ala Arg Ser Ala Glu Arg Ala Ala Glu Leu Gly Val Pro Leu
145 150 155 160
Leu Thr Ile Asn Thr Ser Gly Gly Ala Arg Gln Gln Glu Gly Val Ile
165 170 175
Gly Leu Met Gln Met Ala Lys Val Thr Met Ala Leu Thr Arg Leu Ala
180 185 190
Asp Ala Gly Gln Pro His Ile Ala Leu Leu Val Asp Pro Cys Tyr Gly
195 200 205
Gly Val Thr Ala Ser Tyr Pro Ser Val Ala Asp Ile Ile Ile Ala Glu
210 215 220
Pro Gly Ala Asn Ile Gly Phe Ala Gly Lys Arg Leu Ile Glu Gln Ile
225 230 235 240
Met Arg Gln Lys Leu Pro Ala Gly Phe Gln Thr Ala Glu Phe Met Leu
245 250 255
Glu His Gly Met Ile Asp Met Val Val Pro Arg Ser Glu Met Arg Asp
260 265 270
Thr Leu Ala Arg Ile Leu Arg Leu Tyr Arg Gln Arg Ser Thr Ser Pro
275 280 285
Ala Lys Ala Glu Leu Ala Gly Arg Arg Ala Thr Leu Pro Gln Pro Ile
290 295 300
Met
305
<210> 83
<211> 378
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 83
Met Ile Ile Gly Val Pro Lys Glu Ile Lys Asn Asn Glu Asn Arg Val
1 5 10 15
Ala Leu Thr Pro Gly Gly Val Ser Gln Leu Ile Ser Asn Gly His Arg
20 25 30
Val Leu Val Glu Thr Gly Ala Gly Leu Gly Ser Gly Phe Glu Asn Glu
35 40 45
Ala Tyr Glu Ser Ala Gly Ala Glu Ile Ile Ala Asp Pro Lys Gln Val
50 55 60
Trp Asp Ala Glu Met Val Met Lys Val Lys Glu Pro Leu Pro Glu Glu
65 70 75 80
Tyr Val Tyr Phe Arg Lys Gly Leu Val Leu Phe Thr Tyr Leu His Leu
85 90 95
Ala Ala Glu Pro Glu Leu Ala Gln Ala Leu Lys Asp Lys Gly Val Thr
100 105 110
Ala Ile Ala Tyr Glu Thr Val Ser Glu Gly Arg Thr Leu Pro Leu Leu
115 120 125
Thr Pro Met Ser Glu Val Ala Gly Arg Met Ala Ala Gln Ile Gly Ala
130 135 140
Gln Phe Leu Glu Lys Pro Lys Gly Gly Lys Gly Ile Leu Leu Ala Gly
145 150 155 160
Val Pro Gly Val Ser Arg Gly Lys Val Thr Ile Ile Gly Gly Gly Val
165 170 175
Val Gly Thr Asn Ala Ala Lys Met Ala Val Gly Leu Gly Ala Asp Val
180 185 190
Thr Ile Ile Asp Leu Asn Ala Asp Arg Leu Arg Gln Leu Asp Asp Ile
195 200 205
Phe Gly His Gln Ile Lys Thr Leu Ile Ser Asn Pro Val Asn Ile Ala
210 215 220
Asp Ala Val Ala Glu Ala Asp Leu Leu Ile Cys Ala Val Leu Ile Pro
225 230 235 240
Gly Ala Lys Ala Pro Thr Leu Val Thr Glu Glu Met Val Lys Gln Met
245 250 255
Lys Pro Gly Ser Val Ile Val Asp Val Ala Ile Asp Gln Gly Gly Ile
260 265 270
Val Glu Thr Val Asp His Ile Thr Thr His Asp Gln Pro Thr Tyr Glu
275 280 285
Lys His Gly Val Val His Tyr Ala Val Ala Asn Met Pro Gly Ala Val
290 295 300
Pro Arg Thr Ser Thr Ile Ala Leu Thr Asn Val Thr Val Pro Tyr Ala
305 310 315 320
Leu Gln Ile Ala Asn Lys Gly Ala Val Lys Ala Leu Ala Asp Asn Thr
325 330 335
Ala Leu Arg Ala Gly Leu Asn Thr Ala Asn Gly His Val Thr Tyr Glu
340 345 350
Ala Val Ala Arg Asp Leu Gly Tyr Glu Tyr Val Pro Ala Glu Lys Ala
355 360 365
Leu Gln Asp Glu Ser Ser Val Ala Gly Ala
370 375
<210> 84
<211> 505
<212> PRT
<213> 粟酒裂殖酵母(Schizosaccharomyces pombe)
<400> 84
Met Phe Thr Asp Tyr Pro Asn Asp Ile Asn Cys Glu Ser Pro Arg Met
1 5 10 15
Ser Asp Leu Asp Gly Phe Cys Gln Asn Ala Phe Ser Asp Leu Asn Ser
20 25 30
Leu Asn Gln Gln Val Phe Lys Ala Asn Tyr Ala Val Arg Gly Ala Leu
35 40 45
Ala Ile Leu Ala Asp Glu Ile Gln Asp Asp Leu Leu Glu Asn Pro Ser
50 55 60
Ser Tyr Pro Phe Ser Glu Ile Val Tyr Ala Asn Ile Gly Asn Pro Gln
65 70 75 80
Gln Met Gly Gln Ser Pro Ile Thr Phe Val Arg Gln Val Leu Ser Leu
85 90 95
Cys Gln Tyr Pro Thr Leu Leu Asp His Ala Glu Glu Lys Trp Phe Gln
100 105 110
Asn Leu Phe Pro Thr Asp Val Val Gln Arg Ser Lys Met Leu Leu Lys
115 120 125
Glu Ser Gly Ser Leu Gly Ala Tyr Ser Ala Ser Gln Gly Ile Pro Leu
130 135 140
Val Arg Arg His Val Ala Asp Phe Ile Arg Ala Arg Asp Gly Phe Asp
145 150 155 160
Cys Glu Pro Ser Asp Ile Tyr Leu Thr Ser Gly Ala Ser His Ala Ala
165 170 175
Arg Leu Ile Met Thr Leu Ile Ile Ala Arg Pro Thr Asp Gly Val Met
180 185 190
Val Pro Ala Pro Gln Tyr Pro Leu Tyr Gly Ala Gln Ile Asp Leu Met
195 200 205
Ser Gly Ser Met Val Ser Tyr Ser Leu Ser Glu Glu Asn Asn Trp Asp
210 215 220
Ile Asp Phe Asp Gln Phe Lys Lys Ser Phe Asp Glu Ala Ser Lys Lys
225 230 235 240
Gly Ile Asn Val Arg Leu Cys Val Val Ile Asn Pro Gly Asn Pro Thr
245 250 255
Gly Ala Cys Ile Ser Glu Asn Ser Met Glu Lys Val Leu Arg Phe Ala
260 265 270
Lys Ala Lys Gly Ile Val Leu Leu Ala Asp Glu Val Tyr Gln Asn Asn
275 280 285
Ile Tyr Gln Asn Lys Phe His Ser Phe Arg Arg Lys Leu Gly Glu Leu
290 295 300
Arg Glu Lys Glu Pro Asp Asn His Trp Asp Gln Val Ser Leu Ile Ser
305 310 315 320
Val Asn Ser Val Ser Lys Gly Gln Phe Gly Glu Cys Gly Gln Arg Gly
325 330 335
Gly Tyr Leu Asp Val Val Asn Ile Pro Glu Pro Ala Lys Asp Gln Ile
340 345 350
Leu Lys Leu Ala Thr Ile Asp Ile Cys Pro Pro Val Ala Gly Gln Leu
355 360 365
Leu Val Asp Met Leu Val Asn Pro Pro Lys Pro Gly Asp Pro Ser Tyr
370 375 380
Asp Leu Phe Ile Lys Glu Val Asp Glu Ile His Glu Ala Leu Arg Leu
385 390 395 400
Gln Cys Arg Gln Leu Tyr Glu Gly Thr Lys Arg Met Lys Arg Val Ser
405 410 415
Cys Leu Glu Pro His Gly Ala Met Tyr Leu His Pro Ser Val Ser Leu
420 425 430
Pro Glu Lys Leu Ile Thr Thr Ala Lys Ala Gln Lys Ile Gln Pro Asp
435 440 445
Glu Phe Tyr Ala Ile Glu Leu Leu Lys Arg Ser Gly Ile Cys Val Val
450 455 460
Pro Gly Ser Gly Phe Gly Gln Pro Glu Gly Asp Tyr His Ile Arg Ile
465 470 475 480
Thr Phe Leu Ala Lys Gly Thr Glu Tyr Ile Glu Arg Phe Val Lys Ala
485 490 495
His Asn Glu Ile Met Asp Leu Tyr Glu
500 505
<210> 85
<211> 507
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 85
Met Thr Met Thr His Gln Gln Asp Leu Lys Gly Val Phe Thr Ala Lys
1 5 10 15
Asp Leu Asp Phe Lys Pro Ala Gly Lys Ile Thr Lys Lys Asp Leu Asn
20 25 30
Thr Gly Val Thr Lys Ala Glu Tyr Ala Val Arg Gly Ala Ile Pro Thr
35 40 45
Arg Ala Asp Glu Leu Lys Glu Glu Leu Lys Lys Asn Pro Glu Val Leu
50 55 60
Pro Phe Asp Asp Ile Ile Asn Ala Asn Ile Gly Asn Pro Gln Gln Leu
65 70 75 80
Asp Gln Lys Pro Leu Thr Phe Thr Arg Gln Val Leu Ala Ile Leu Glu
85 90 95
Tyr Pro Glu Ile Leu Arg Val Gly His Asn Glu Leu Ala Ser Leu Asn
100 105 110
Leu Phe Ser Arg Asp Ala Leu Glu Arg Ala Glu Arg Leu Leu Asn Asp
115 120 125
Ile Gly Gly Ser Ile Gly Ala Tyr Ser His Ser Gln Gly Val Pro Gly
130 135 140
Ile Arg Gln Thr Val Ala Asp Phe Ile Thr Arg Arg Asp Gly Gly Glu
145 150 155 160
Pro Ala Thr Pro Glu Asp Ile Tyr Leu Thr Thr Gly Ala Ser Ser Ala
165 170 175
Ala Thr Ser Leu Leu Ser Leu Leu Cys Lys Asp Ser Gln Thr Gly Leu
180 185 190
Leu Ile Pro Ile Pro Gln Tyr Pro Leu Tyr Thr Ala Ser Ala Ser Leu
195 200 205
Phe Asn Ala Gln Val Leu Pro Tyr Tyr Leu Asp Glu Glu Ser Asn Trp
210 215 220
Ser Thr Asn Ser Asp Glu Ile Glu Lys Val Val Gln Asp Ala Leu Lys
225 230 235 240
Lys Gln Ile Arg Pro Ser Val Leu Ile Val Ile Asn Pro Gly Asn Pro
245 250 255
Thr Gly Ala Val Leu Ser Glu Glu Thr Ile Ala Arg Ile Cys Leu Ile
260 265 270
Ala Ala Lys Tyr Gly Ile Thr Ile Ile Ser Asp Glu Val Tyr Gln Glu
275 280 285
Asn Ile Phe Asn Asp Val Lys Phe His Ser Met Lys Lys Val Leu Arg
290 295 300
Lys Leu Gln His Leu Tyr Pro Gly Lys Phe Asp Asn Val Gln Leu Ala
305 310 315 320
Ser Leu His Ser Ile Ser Lys Gly Phe Met Asp Glu Cys Gly Gln Arg
325 330 335
Gly Gly Tyr Met Glu Ile Ile Gly Phe Ser Gln Glu Ile Arg Asp Ala
340 345 350
Leu Phe Lys Leu Met Ser Ile Ser Ile Cys Ser Val Val Thr Gly Gln
355 360 365
Ala Val Val Asp Leu Met Val Lys Pro Pro Gln Pro Gly Asp Glu Ser
370 375 380
Tyr Glu Gln Asp His Asp Glu Arg Leu Lys Ile Phe His Glu Met Arg
385 390 395 400
Thr Arg Ala Asn Leu Leu Tyr Glu Thr Phe Lys Glu Leu Glu Gly Ile
405 410 415
Glu Cys Gln Lys Pro Gln Gly Ala Met Tyr Leu Phe Pro Arg Leu Val
420 425 430
Leu Pro Lys Lys Ala Leu Cys Glu Ser Glu Arg Leu Gly Ile Glu Pro
435 440 445
Asp Glu Phe Tyr Cys Thr Ser Leu Leu Glu Ser Thr Gly Ile Cys Thr
450 455 460
Val Pro Gly Ser Gly Phe Gly Gln Arg Pro Gly Thr Tyr His Val Arg
465 470 475 480
Thr Thr Phe Leu Ala Pro Gly Thr Lys Trp Ile Gln Asp Trp Lys Glu
485 490 495
Phe His Gln Asp Phe Phe Ser Lys Tyr Arg Asn
500 505
<210> 86
<211> 471
<212> PRT
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 86
Met Lys Asn Lys Trp Tyr Lys Pro Lys Arg His Trp Lys Glu Ile Glu
1 5 10 15
Leu Trp Lys Asp Val Pro Glu Glu Lys Trp Asn Asp Trp Leu Trp Gln
20 25 30
Leu Thr His Thr Val Arg Thr Leu Asp Asp Leu Lys Lys Val Ile Asn
35 40 45
Leu Thr Glu Asp Glu Glu Glu Gly Val Arg Ile Ser Thr Lys Thr Ile
50 55 60
Pro Leu Asn Ile Thr Pro Tyr Tyr Ala Ser Leu Met Asp Pro Asp Asn
65 70 75 80
Pro Arg Cys Pro Val Arg Met Gln Ser Val Pro Leu Ser Glu Glu Met
85 90 95
His Lys Thr Lys Tyr Asp Leu Glu Asp Pro Leu His Glu Asp Glu Asp
100 105 110
Ser Pro Val Pro Gly Leu Thr His Arg Tyr Pro Asp Arg Val Leu Phe
115 120 125
Leu Val Thr Asn Gln Cys Ser Met Tyr Cys Arg Tyr Cys Thr Arg Arg
130 135 140
Arg Phe Ser Gly Gln Ile Gly Met Gly Val Pro Lys Lys Gln Leu Asp
145 150 155 160
Ala Ala Ile Ala Tyr Ile Arg Glu Thr Pro Glu Ile Arg Asp Cys Leu
165 170 175
Ile Ser Gly Gly Asp Gly Leu Leu Ile Asn Asp Gln Ile Leu Glu Tyr
180 185 190
Ile Leu Lys Glu Leu Arg Ser Ile Pro His Leu Glu Val Ile Arg Ile
195 200 205
Gly Thr Arg Ala Pro Val Val Phe Pro Gln Arg Ile Thr Asp His Leu
210 215 220
Cys Glu Ile Leu Lys Lys Tyr His Pro Val Trp Leu Asn Thr His Phe
225 230 235 240
Asn Thr Ser Ile Glu Met Thr Glu Glu Ser Val Glu Ala Cys Glu Lys
245 250 255
Leu Val Asn Ala Gly Val Pro Val Gly Asn Gln Ala Val Val Leu Ala
260 265 270
Gly Ile Asn Asp Ser Val Pro Ile Met Lys Lys Leu Met His Asp Leu
275 280 285
Val Lys Ile Arg Val Arg Pro Tyr Tyr Ile Tyr Gln Cys Asp Leu Ser
290 295 300
Glu Gly Ile Gly His Phe Arg Ala Pro Val Ser Lys Gly Leu Glu Ile
305 310 315 320
Ile Glu Gly Leu Arg Gly His Thr Ser Gly Tyr Ala Val Pro Thr Phe
325 330 335
Val Val Asp Ala Pro Gly Gly Gly Gly Lys Ile Ala Leu Gln Pro Asn
340 345 350
Tyr Val Leu Ser Gln Ser Pro Asp Lys Val Ile Leu Arg Asn Phe Glu
355 360 365
Gly Val Ile Thr Ser Tyr Pro Glu Pro Glu Asn Tyr Ile Pro Asn Gln
370 375 380
Ala Asp Ala Tyr Phe Glu Ser Val Phe Pro Glu Thr Ala Asp Lys Lys
385 390 395 400
Glu Pro Ile Gly Leu Ser Ala Ile Phe Ala Asp Lys Glu Val Ser Phe
405 410 415
Thr Pro Glu Asn Val Asp Arg Ile Lys Arg Arg Glu Ala Tyr Ile Ala
420 425 430
Asn Pro Glu His Glu Thr Leu Lys Asp Arg Arg Glu Lys Arg Asp Gln
435 440 445
Leu Lys Glu Lys Lys Phe Leu Ala Gln Gln Lys Lys Gln Lys Glu Thr
450 455 460
Glu Cys Gly Gly Asp Ser Ser
465 470
<210> 87
<211> 416
<212> PRT
<213> 牙龈卟啉单胞菌(Porphyromonas gingivalis)
<400> 87
Met Ala Glu Ser Arg Arg Lys Tyr Tyr Phe Pro Asp Val Thr Asp Glu
1 5 10 15
Gln Trp Asn Asp Trp His Trp Gln Val Leu Asn Arg Ile Glu Thr Leu
20 25 30
Asp Gln Leu Lys Lys Tyr Val Thr Leu Thr Ala Glu Glu Glu Glu Gly
35 40 45
Val Lys Glu Ser Leu Lys Val Leu Arg Met Ala Ile Thr Pro Tyr Tyr
50 55 60
Leu Ser Leu Ile Asp Pro Glu Asn Pro Asn Cys Pro Ile Arg Lys Gln
65 70 75 80
Ala Ile Pro Thr His Gln Glu Leu Val Arg Ala Pro Glu Asp Gln Val
85 90 95
Asp Pro Leu Ser Glu Asp Glu Asp Ser Pro Val Pro Gly Leu Thr His
100 105 110
Arg Tyr Pro Asp Arg Val Leu Phe Leu Ile Thr Asp Lys Cys Ser Met
115 120 125
Tyr Cys Arg His Cys Thr Arg Arg Arg Phe Ala Gly Gln Lys Asp Ala
130 135 140
Ser Ser Pro Ser Glu Arg Ile Asp Arg Cys Ile Asp Tyr Ile Ala Asn
145 150 155 160
Thr Pro Thr Val Arg Asp Val Leu Leu Ser Gly Gly Asp Ala Leu Leu
165 170 175
Val Ser Asp Glu Arg Leu Glu Tyr Ile Leu Lys Arg Leu Arg Glu Ile
180 185 190
Pro His Val Glu Ile Val Arg Ile Gly Ser Arg Thr Pro Val Val Leu
195 200 205
Pro Gln Arg Ile Thr Pro Gln Leu Val Asp Met Leu Lys Lys Tyr His
210 215 220
Pro Val Trp Leu Asn Thr His Phe Asn His Pro Asn Glu Val Thr Glu
225 230 235 240
Glu Ala Val Glu Ala Cys Glu Arg Met Ala Asn Ala Gly Ile Pro Leu
245 250 255
Gly Asn Gln Thr Val Leu Leu Arg Gly Ile Asn Asp Cys Thr His Val
260 265 270
Met Lys Arg Leu Val His Leu Leu Val Lys Met Arg Val Arg Pro Tyr
275 280 285
Tyr Ile Tyr Val Cys Asp Leu Ser Leu Gly Ile Gly His Phe Arg Thr
290 295 300
Pro Val Ser Lys Gly Ile Glu Ile Ile Glu Asn Leu Arg Gly His Thr
305 310 315 320
Ser Gly Tyr Ala Val Pro Thr Phe Val Val Asp Ala Pro Gly Gly Gly
325 330 335
Gly Lys Ile Pro Val Met Pro Asn Tyr Val Val Ser Gln Ser Pro Arg
340 345 350
His Val Val Leu Arg Asn Tyr Glu Gly Val Ile Thr Thr Tyr Thr Glu
355 360 365
Pro Glu Asn Tyr His Glu Glu Cys Asp Cys Glu Asp Cys Arg Ala Gly
370 375 380
Lys His Lys Glu Gly Val Ala Ala Leu Ser Gly Gly Gln Gln Leu Ala
385 390 395 400
Ile Glu Pro Ser Asp Leu Ala Arg Lys Lys Arg Lys Phe Asp Lys Asn
405 410 415
<210> 88
<211> 425
<212> PRT
<213> 具核梭杆菌(Fusobacterium nucleatum)
<400> 88
Met Asn Thr Val Asn Thr Arg Lys Lys Phe Phe Pro Asn Val Thr Asp
1 5 10 15
Glu Glu Trp Asn Asp Trp Thr Trp Gln Val Lys Asn Arg Ile Glu Lys
20 25 30
Ile Asp Asp Leu Lys Lys Tyr Val Glu Leu Ser Ala Glu Glu Glu Glu
35 40 45
Gly Val Val Arg Thr Leu Glu Thr Leu Arg Met Ala Ile Thr Pro Tyr
50 55 60
Tyr Phe Ser Leu Ile Asp Met Asn Ser Asp Arg Cys Pro Ile Arg Lys
65 70 75 80
Gln Ala Ile Pro Thr Ile Gln Glu Ile His Gln Ser Asp Ala Asp Leu
85 90 95
Leu Asp Pro Leu His Glu Asp Glu Asp Ser Pro Val Pro Gly Leu Thr
100 105 110
His Arg Tyr Pro Asp Arg Val Leu Leu Leu Ile Thr Asp Met Cys Ser
115 120 125
Met Tyr Cys Arg His Cys Thr Arg Arg Arg Phe Ala Gly Ser Ser Asp
130 135 140
Asp Ala Met Pro Met Asp Arg Ile Asp Lys Ala Ile Glu Tyr Ile Ala
145 150 155 160
Lys Thr Pro Gln Val Arg Asp Val Leu Leu Ser Gly Gly Asp Ala Leu
165 170 175
Leu Val Ser Asp Lys Lys Leu Glu Ser Ile Ile Gln Lys Leu Arg Ala
180 185 190
Ile Pro His Val Glu Ile Ile Arg Ile Gly Ser Arg Thr Pro Val Val
195 200 205
Leu Pro Gln Arg Ile Thr Pro Glu Leu Cys Asn Met Leu Lys Lys Tyr
210 215 220
His Pro Ile Trp Leu Asn Thr His Phe Asn His Pro Gln Glu Val Thr
225 230 235 240
Pro Glu Ala Lys Lys Ala Cys Glu Met Leu Ala Asp Ala Gly Val Pro
245 250 255
Leu Gly Asn Gln Thr Val Leu Leu Arg Gly Ile Asn Asp Ser Val Pro
260 265 270
Val Met Lys Arg Leu Val His Asp Leu Val Met Met Arg Val Arg Pro
275 280 285
Tyr Tyr Ile Tyr Gln Cys Asp Leu Ser Met Gly Leu Glu His Phe Arg
290 295 300
Thr Pro Val Ser Lys Gly Ile Glu Ile Ile Glu Gly Leu Arg Gly His
305 310 315 320
Thr Ser Gly Tyr Ala Val Pro Thr Phe Val Val Asp Ala Pro Gly Gly
325 330 335
Gly Gly Lys Thr Pro Val Met Pro Gln Tyr Val Ile Ser Gln Ser Pro
340 345 350
His Arg Val Val Leu Arg Asn Phe Glu Gly Val Ile Thr Thr Tyr Thr
355 360 365
Glu Pro Glu Asn Tyr Thr His Glu Pro Cys Tyr Asp Glu Glu Lys Phe
370 375 380
Glu Lys Met Tyr Glu Ile Ser Gly Val Tyr Met Leu Asp Glu Gly Leu
385 390 395 400
Lys Met Ser Leu Glu Pro Ser His Leu Ala Arg His Glu Arg Asn Lys
405 410 415
Lys Arg Ala Glu Ala Glu Gly Lys Lys
420 425
<210> 89
<211> 517
<212> PRT
<213> 埃氏巨球形菌(Megasphaera elsdenii)
<400> 89
Met Arg Lys Val Glu Ile Ile Thr Ala Glu Gln Ala Ala Gln Leu Val
1 5 10 15
Lys Asp Asn Asp Thr Ile Thr Ser Ile Gly Phe Val Ser Ser Ala His
20 25 30
Pro Glu Ala Leu Thr Lys Ala Leu Glu Lys Arg Phe Leu Asp Thr Asn
35 40 45
Thr Pro Gln Asn Leu Thr Tyr Ile Tyr Ala Gly Ser Gln Gly Lys Arg
50 55 60
Asp Gly Arg Ala Ala Glu His Leu Ala His Thr Gly Leu Leu Lys Arg
65 70 75 80
Ala Ile Ile Gly His Trp Gln Thr Val Pro Ala Ile Gly Lys Leu Ala
85 90 95
Val Glu Asn Lys Ile Glu Ala Tyr Asn Phe Ser Gln Gly Thr Leu Val
100 105 110
His Trp Phe Arg Ala Leu Ala Gly His Lys Leu Gly Val Phe Thr Asp
115 120 125
Ile Gly Leu Glu Thr Phe Leu Asp Pro Arg Gln Leu Gly Gly Lys Leu
130 135 140
Asn Asp Val Thr Lys Glu Asp Leu Val Lys Leu Ile Glu Val Asp Gly
145 150 155 160
His Glu Gln Leu Phe Tyr Pro Thr Phe Pro Val Asn Val Ala Phe Leu
165 170 175
Arg Gly Thr Tyr Ala Asp Glu Ser Gly Asn Ile Thr Met Asp Glu Glu
180 185 190
Ile Gly Pro Phe Glu Ser Thr Ser Val Ala Gln Ala Val His Asn Cys
195 200 205
Gly Gly Lys Val Val Val Gln Val Lys Asp Val Val Ala His Gly Ser
210 215 220
Leu Asp Pro Arg Met Val Lys Ile Pro Gly Ile Tyr Val Asp Tyr Val
225 230 235 240
Val Val Ala Ala Pro Glu Asp His Gln Gln Thr Tyr Asp Cys Glu Tyr
245 250 255
Asp Pro Ser Leu Ser Gly Glu His Arg Ala Pro Glu Gly Ala Thr Asp
260 265 270
Ala Ala Leu Pro Met Ser Ala Lys Lys Ile Ile Gly Arg Arg Gly Ala
275 280 285
Leu Glu Leu Thr Glu Asn Ala Val Val Asn Leu Gly Val Gly Ala Pro
290 295 300
Glu Tyr Val Ala Ser Val Ala Gly Glu Glu Gly Ile Ala Asp Thr Ile
305 310 315 320
Thr Leu Thr Val Glu Gly Gly Ala Ile Gly Gly Val Pro Gln Gly Gly
325 330 335
Ala Arg Phe Gly Ser Ser Arg Asn Ala Asp Ala Ile Ile Asp His Thr
340 345 350
Tyr Gln Phe Asp Phe Tyr Asp Gly Gly Gly Leu Asp Ile Ala Tyr Leu
355 360 365
Gly Leu Ala Gln Cys Asp Gly Ser Gly Asn Ile Asn Val Ser Lys Phe
370 375 380
Gly Thr Asn Val Ala Gly Cys Gly Gly Phe Pro Asn Ile Ser Gln Gln
385 390 395 400
Thr Pro Asn Val Tyr Phe Cys Gly Thr Phe Thr Ala Gly Gly Leu Lys
405 410 415
Ile Ala Val Glu Asp Gly Lys Val Lys Ile Leu Gln Glu Gly Lys Ala
420 425 430
Lys Lys Phe Ile Lys Ala Val Asp Gln Ile Thr Phe Asn Gly Ser Tyr
435 440 445
Ala Ala Arg Asn Gly Lys His Val Leu Tyr Ile Thr Glu Arg Cys Val
450 455 460
Phe Glu Leu Thr Lys Glu Gly Leu Lys Leu Ile Glu Val Ala Pro Gly
465 470 475 480
Ile Asp Ile Glu Lys Asp Ile Leu Ala His Met Asp Phe Lys Pro Ile
485 490 495
Ile Asp Asn Pro Lys Leu Met Asp Ala Arg Leu Phe Gln Asp Gly Pro
500 505 510
Met Gly Leu Lys Lys
515
<210> 90
<211> 145
<212> PRT
<213> 丙酸梭菌(Clostridium propionicum)
<400> 90
Met Val Gly Lys Lys Val Val His His Leu Met Met Ser Ala Lys Asp
1 5 10 15
Ala His Tyr Thr Gly Asn Leu Val Asn Gly Ala Arg Ile Val Asn Gln
20 25 30
Trp Gly Asp Val Gly Thr Glu Leu Met Val Tyr Val Asp Gly Asp Ile
35 40 45
Ser Leu Phe Leu Gly Tyr Lys Asp Ile Glu Phe Thr Ala Pro Val Tyr
50 55 60
Val Gly Asp Phe Met Glu Tyr His Gly Trp Ile Glu Lys Val Gly Asn
65 70 75 80
Gln Ser Tyr Thr Cys Lys Phe Glu Ala Trp Lys Val Ala Thr Met Val
85 90 95
Asp Ile Thr Asn Pro Gln Asp Thr Arg Ala Thr Ala Cys Glu Pro Pro
100 105 110
Val Leu Cys Gly Arg Ala Thr Gly Ser Leu Phe Ile Ala Lys Lys Asp
115 120 125
Gln Arg Gly Pro Gln Glu Ser Ser Phe Lys Glu Arg Lys His Pro Gly
130 135 140
Glu
145
<210> 91
<211> 260
<212> PRT
<213> 大豆疫霉菌(Phytophthora sojae)
<400> 91
Met Ala Ala Glu Tyr Glu Ser Ile Leu Thr Glu Val Arg Gly Lys Val
1 5 10 15
Ala Ile Ile Thr Leu Asn Arg Pro Lys Ala Leu Asn Ala Leu Cys Ser
20 25 30
Pro Leu Ile Glu Glu Leu Asn Gly Ala Ala His Ala Phe Asp Ala Asp
35 40 45
Pro Ser Ile Gly Ala Ile Val Ile Thr Gly Ser Gly Ser Lys Ala Phe
50 55 60
Ala Ala Gly Ala Asp Ile Lys Glu Met Ala Thr Lys Thr Phe Val Asp
65 70 75 80
Ala Tyr Lys Ser Asn Met Phe Ala Asn Trp Gly Asp Ile Thr Lys Val
85 90 95
Ser Lys Pro Val Ile Ala Ala Val Asn Gly Tyr Ala Leu Gly Gly Gly
100 105 110
Cys Glu Leu Ala Met Leu Cys Asp Leu Ile Ile Ala Gly Asp Ser Ala
115 120 125
Lys Phe Gly Gln Pro Glu Ile Thr Leu Gly Thr Ile Pro Gly Cys Gly
130 135 140
Gly Thr Gln Arg Leu Ile Arg Ala Val Gly Lys Ser Lys Ala Met Glu
145 150 155 160
Met Ile Leu Thr Gly Asn Met Ile Asp Ala Gln Gln Ala Glu Arg Asp
165 170 175
Gly Leu Val Ala Arg Val Val Pro Ala Asp Gln Leu Leu Asp Glu Ala
180 185 190
Leu Lys Thr Ala Asn Lys Ile Ala Ser Phe Ser Gln Pro Val Val Lys
195 200 205
Met Ala Lys Glu Ala Val Asn Ala Ala Tyr Glu Gln Ser Leu Gln Glu
210 215 220
Gly Leu Lys Tyr Glu Ser Arg Leu Phe Trp Ser Ser Phe Ala Thr Lys
225 230 235 240
Asp Gln Lys Glu Gly Met Ala Ala Phe Val Glu Lys Arg Lys Ala Asp
245 250 255
Phe Lys Asp Glu
260
<210> 92
<211> 258
<212> PRT
<213> 橙色绿屈挠菌(Chloroflexus aurantiacus)
<400> 92
Met Ser Glu Glu Ser Leu Val Leu Ser Thr Ile Glu Gly Pro Ile Ala
1 5 10 15
Ile Leu Thr Leu Asn Arg Pro Gln Ala Leu Asn Ala Leu Ser Pro Ala
20 25 30
Leu Ile Asp Asp Leu Ile Arg His Leu Glu Ala Cys Asp Ala Asp Asp
35 40 45
Thr Ile Arg Val Ile Ile Ile Thr Gly Ala Gly Arg Ala Phe Ala Ala
50 55 60
Gly Ala Asp Ile Lys Ala Met Ala Asn Ala Thr Pro Ile Asp Met Leu
65 70 75 80
Thr Ser Gly Met Ile Ala Arg Trp Ala Arg Ile Ala Ala Val Arg Lys
85 90 95
Pro Val Ile Ala Ala Val Asn Gly Tyr Ala Leu Gly Gly Gly Cys Glu
100 105 110
Leu Ala Met Met Cys Asp Ile Ile Ile Ala Ser Glu Asn Ala Gln Phe
115 120 125
Gly Gln Pro Glu Ile Asn Leu Gly Ile Ile Pro Gly Ala Gly Gly Thr
130 135 140
Gln Arg Leu Thr Arg Ala Leu Gly Pro Tyr Arg Ala Met Glu Leu Ile
145 150 155 160
Leu Thr Gly Ala Thr Ile Ser Ala Gln Glu Ala Leu Ala His Gly Leu
165 170 175
Val Cys Arg Val Cys Pro Pro Glu Ser Leu Leu Asp Glu Ala Arg Arg
180 185 190
Ile Ala Gln Thr Ile Ala Thr Lys Ser Pro Leu Ala Val Gln Leu Ala
195 200 205
Lys Glu Ala Val Arg Met Ala Ala Glu Thr Thr Val Arg Glu Gly Leu
210 215 220
Ala Ile Glu Leu Arg Asn Phe Tyr Leu Leu Phe Ala Ser Ala Asp Gln
225 230 235 240
Lys Glu Gly Met Gln Ala Phe Ile Glu Lys Arg Ala Pro Asn Phe Ser
245 250 255
Gly Arg
<210> 93
<211> 288
<212> PRT
<213> 深红红螺菌(Rhodospirillum rubrum)
<400> 93
Met Ala Ala Ala Pro Gly Pro Pro Ala Ala Arg Pro Arg Ala Ala Arg
1 5 10 15
Gln Ser Arg Met Ile Leu Pro Pro Leu Arg Glu Gln Ala Gln Met Ala
20 25 30
Tyr Glu Asn Ile Leu Val Glu Thr Asn Gly Lys Val Gly Ile Val Thr
35 40 45
Leu Asn Arg Pro Lys Ala Leu Asn Ala Leu Ser Ala Gly Leu Val Arg
50 55 60
Asp Leu Gly Ala Ala Leu Asp Ala Phe Glu Ala Asp Val Asn Val His
65 70 75 80
Val Ile Val Leu Thr Gly Ser Asp Lys Ala Phe Ala Ala Gly Ala Asp
85 90 95
Ile Lys Glu Met Ala Glu Lys Ser Tyr Met Asp Ala Tyr Leu Glu Asp
100 105 110
Phe Ile Thr Lys Gly Trp Glu Arg Val Thr Thr Cys Arg Lys Pro Ile
115 120 125
Ile Ala Ala Val Ala Gly Phe Ala Leu Gly Gly Gly Cys Glu Met Ala
130 135 140
Met Met Cys Asp Phe Ile Ile Ala Ala Gln Asn Ala Lys Phe Gly Gln
145 150 155 160
Pro Glu Ile Asn Leu Gly Thr Leu Pro Gly Ala Gly Gly Thr Gln Arg
165 170 175
Leu Thr Arg Phe Val Gly Lys Ser Lys Ala Met Asp Met Cys Leu Thr
180 185 190
Gly Arg Met Met Asp Ala Asp Glu Ala Trp Lys Cys Gly Leu Val Ser
195 200 205
Arg Ile Val Pro Val Asp Asp Leu Lys Asp Glu Val Leu Lys Ile Ala
210 215 220
Glu Ala Ile Ala Asp Lys Ser Leu Pro Ile Thr Met Met Val Lys Glu
225 230 235 240
Ala Val Asn Ala Ala Tyr Glu Thr Thr Leu Ala Gln Gly Val Arg Phe
245 250 255
Glu Arg Arg Leu Phe Gln Ala Ser Phe Ala Thr Asp Asp Gln Lys Glu
260 265 270
Gly Met Asn Ala Phe Ile Glu Lys Arg Gln Pro Ser Phe Thr Asp Arg
275 280 285
<210> 94
<211> 258
<212> PRT
<213> 荚膜红细菌(Rhodobacter capsulatus)
<400> 94
Met Ser Tyr Gln Thr Leu Ile Val Glu Ile Ala Asp Gly Val Ala Leu
1 5 10 15
Ile Arg Leu Asn Arg Pro Glu Ala Leu Asn Ala Leu Asn Ser Gln Leu
20 25 30
Leu Gly Glu Leu Ala Ala Ala Leu Ser Thr Leu Asp Ala Asp Pro Ala
35 40 45
Val Arg Cys Phe Val Leu Thr Gly Ser Asp Lys Ala Phe Ala Ala Gly
50 55 60
Ala Asp Ile Lys Glu Met Ala Asp Lys Ser Phe Val Asp Met Leu Lys
65 70 75 80
Leu Asp Phe Phe Gly Thr Glu Gly Asp Ala Ile Leu Arg Thr Arg Lys
85 90 95
Pro Val Ile Ala Ala Val Ala Gly Tyr Ala Leu Gly Gly Gly Cys Glu
100 105 110
Leu Ala Met Met Cys Asp Phe Ile Leu Cys Ala Glu Asn Ala Lys Phe
115 120 125
Gly Gln Pro Glu Ile Asn Leu Gly Val Val Ala Gly Ile Gly Gly Thr
130 135 140
Gln Arg Leu Thr Arg Phe Val Gly Lys Ser Lys Ser Met Glu Met His
145 150 155 160
Leu Thr Gly Arg Phe Met Asp Ala Ala Glu Ala Glu Arg Ser Gly Leu
165 170 175
Val Ser Arg Val Leu Pro Leu Ala Asp Leu Leu Pro Glu Ala Leu Ala
180 185 190
Thr Ala Arg Lys Ile Ala Glu Lys Ser Ala Ile Ala Thr Met Val Ala
195 200 205
Lys Asp Cys Val Asn Arg Ala Tyr Glu Thr Thr Leu Arg Glu Gly Val
210 215 220
Leu Tyr Glu Arg Arg Val Phe His Ala Leu Phe Ala Thr Glu Asp Gln
225 230 235 240
Lys Glu Gly Met Ala Ala Phe Ala Glu Lys Arg Pro Ala Lys Phe Ala
245 250 255
Asp Lys
<210> 95
<211> 290
<212> PRT
<213> 智人(Homo sapiens)
<400> 95
Met Ala Ala Leu Arg Val Leu Leu Ser Cys Ala Arg Gly Pro Leu Arg
1 5 10 15
Pro Pro Val Arg Cys Pro Ala Trp Arg Pro Phe Ala Ser Gly Ala Asn
20 25 30
Phe Glu Tyr Ile Ile Ala Glu Lys Arg Gly Lys Asn Asn Thr Val Gly
35 40 45
Leu Ile Gln Leu Asn Arg Pro Lys Ala Leu Asn Ala Leu Cys Asp Gly
50 55 60
Leu Ile Asp Glu Leu Asn Gln Ala Leu Lys Ile Phe Glu Glu Asp Pro
65 70 75 80
Ala Val Gly Ala Ile Val Leu Thr Gly Gly Asp Lys Ala Phe Ala Ala
85 90 95
Gly Ala Asp Ile Lys Glu Met Gln Asn Leu Ser Phe Gln Asp Cys Tyr
100 105 110
Ser Ser Lys Phe Leu Lys His Trp Asp His Leu Thr Gln Val Lys Lys
115 120 125
Pro Val Ile Ala Ala Val Asn Gly Tyr Ala Phe Gly Gly Gly Cys Glu
130 135 140
Leu Ala Met Met Cys Asp Ile Ile Tyr Ala Gly Glu Lys Ala Gln Phe
145 150 155 160
Ala Gln Pro Glu Ile Leu Ile Gly Thr Ile Pro Gly Ala Gly Gly Thr
165 170 175
Gln Arg Leu Thr Arg Ala Val Gly Lys Ser Leu Ala Met Glu Met Val
180 185 190
Leu Thr Gly Asp Arg Ile Ser Ala Gln Asp Ala Lys Gln Ala Gly Leu
195 200 205
Val Ser Lys Ile Cys Pro Val Glu Thr Leu Val Glu Glu Ala Ile Gln
210 215 220
Cys Ala Glu Lys Ile Ala Ser Asn Ser Lys Ile Val Val Ala Met Ala
225 230 235 240
Lys Glu Ser Val Asn Ala Ala Phe Glu Met Thr Leu Thr Glu Gly Ser
245 250 255
Lys Leu Glu Lys Lys Leu Phe Tyr Ser Thr Phe Ala Thr Asp Asp Arg
260 265 270
Lys Glu Gly Met Thr Ala Phe Val Glu Lys Arg Lys Ala Asn Phe Lys
275 280 285
Asp Gln
290
<210> 96
<211> 399
<212> PRT
<213> 荧光假单胞菌(Pseudomonas fluorescens)
<400> 96
Met Arg Val Val Leu Cys Lys Phe Ala Leu Leu Arg Ser His Asp Gly
1 5 10 15
Ser Ser Ala Asn Ile Leu Ile Lys Asn Asn Lys Ser Arg Glu Leu Ser
20 25 30
Met Thr Ala Gln Val Ser Thr Glu Ala Ser His Ala Ala Ile Leu Gln
35 40 45
Asp Glu Val Leu Ala Glu Val Arg Asn His Ile Gly His Leu Thr Leu
50 55 60
Asn Arg Pro Ala Gly Leu Asn Ala Leu Thr Leu Gln Met Val Arg Ser
65 70 75 80
Leu Thr Ser Gln Leu Gln Ala Trp Ser Asp Asp Pro Gln Val Tyr Ala
85 90 95
Val Val Leu Arg Gly Ala Gly Glu Lys Ala Phe Cys Ala Gly Gly Asp
100 105 110
Ile Arg Ser Leu Tyr Asp Ser Phe Lys Asn Gly Asp Thr Leu His Gln
115 120 125
Asp Phe Phe Val Glu Glu Tyr Ala Leu Asp Leu Ala Ile His His Tyr
130 135 140
Arg Lys Pro Val Leu Ala Leu Met Asp Gly Phe Val Leu Gly Gly Gly
145 150 155 160
Met Gly Leu Val Gln Gly Ala Asp Leu Arg Val Val Thr Glu Arg Ser
165 170 175
Arg Leu Ala Met Pro Glu Val Ala Ile Gly Tyr Phe Pro Asp Val Gly
180 185 190
Gly Ser Tyr Phe Leu Pro Arg Ile Pro Gly Glu Leu Gly Ile Tyr Leu
195 200 205
Gly Val Thr Gly Val Gln Ile Arg Ala Ala Asp Ala Leu Tyr Cys Gly
210 215 220
Leu Ala Asp Trp Tyr Leu Asp Ser His Lys Leu Ala Asp Leu Asp Gln
225 230 235 240
Lys Leu Asp Asn Leu Arg Trp His Asp Ser Pro Leu Lys Asp Leu Gln
245 250 255
Gly Ala Leu Ala Arg Leu Ala Val Gln Gln Leu Pro Asp Ala Pro Leu
260 265 270
Ala Ala Leu Arg Pro Ala Ile Asp His Phe Phe Ala Leu Pro Asp Val
275 280 285
Pro Ser Ile Val Glu Gln Leu Gln Gln Val Thr Val Ala Asp Ser His
290 295 300
Glu Trp Ala Leu Asn Thr Val Ser Leu Met Gln Thr Arg Ser Pro Leu
305 310 315 320
Ala Met Ala Val Thr Leu Glu Met Leu Arg Arg Gly Arg Arg Leu Ser
325 330 335
Leu Glu Gln Cys Phe Ala Leu Glu Leu His Leu Asp Arg Gln Trp Phe
340 345 350
Glu Arg Gly Asp Leu Ile Glu Gly Val Arg Ala Leu Ile Ile Asp Lys
355 360 365
Asp Lys Ser Pro Arg Trp Asn Pro Pro Thr Leu His Gly Leu Ala Leu
370 375 380
Asn His Val Glu Ser Phe Phe His His Phe Glu Lys Val Val Lys
385 390 395
<210> 97
<211> 351
<212> PRT
<213> 荧光假单胞菌(Pseudomonas fluorescens)
<400> 97
Met Thr Glu Gln Val Leu Phe Ser Val Ser Glu Asn Gly Val Ala Thr
1 5 10 15
Ile Thr Leu Asn Arg Pro Lys Ala Leu Asn Ser Leu Ser Tyr Asp Met
20 25 30
Leu Gln Pro Ile Gly Gln Lys Leu Lys Glu Trp Glu His Asp Glu Arg
35 40 45
Ile Ala Leu Ile Val Leu Lys Gly Ala Gly Thr Lys Gly Phe Cys Ala
50 55 60
Gly Gly Asp Ile Lys Thr Leu Tyr Glu Ala Arg Ser Asn Glu Val Ala
65 70 75 80
Leu Gln His Ala Glu Arg Phe Phe Glu Glu Glu Tyr Glu Ile Asp Thr
85 90 95
Tyr Ile Tyr Gln Tyr Thr Lys Pro Ile Ile Ala Cys Leu Asp Gly Ile
100 105 110
Val Met Gly Gly Gly Val Gly Leu Thr Asn Gly Ala Lys Tyr Arg Ile
115 120 125
Val Thr Glu Arg Thr Lys Trp Ala Met Pro Glu Met Asn Ile Gly Phe
130 135 140
Phe Pro Asp Val Gly Ala Ala Tyr Phe Leu Asn Lys Ala Pro Gly Tyr
145 150 155 160
Thr Gly Arg Phe Val Ala Leu Thr Ala Ser Ile Leu Lys Ala Ser Asp
165 170 175
Val Leu Phe Ile Asn Ala Ala Asp Tyr Phe Met Thr Ser Asp Ser Leu
180 185 190
Pro Glu Phe Leu Thr Glu Leu Glu Ser Val Asn Trp His Lys Glu Asp
195 200 205
Asp Val His Thr Asn Leu Lys Glu Val Ile Arg Thr Phe Ala Thr Ala
210 215 220
Pro Asn Leu Glu Ser Glu Leu Ala Pro Ser Leu Glu Val Ile Asn Ser
225 230 235 240
His Phe Ala Phe Asp Thr Ile Glu Glu Ile Ile His Ser Leu Glu Lys
245 250 255
Asp Glu Ser Ser Phe Ala Leu Lys Thr Lys Glu Ile Leu Leu Ser Lys
260 265 270
Ser Pro Ile Ser Leu Lys Val Thr Leu Lys Gln Phe Ile Asp Gly Gln
275 280 285
Asp Lys Ser Val Glu Glu Cys Phe Ala Thr Asp Leu Ile Leu Ala Lys
290 295 300
Asn Phe Met Arg His Glu Asp Phe Phe Glu Gly Val Arg Ser Val Val
305 310 315 320
Val Asp Lys Asp Gln Asn Pro Asn Tyr Lys Tyr Lys Gln Leu Ser Asp
325 330 335
Val Ser Glu Glu Asp Val Asn Arg Phe Phe Asn Leu Leu Asn Ala
340 345 350
<210> 98
<211> 381
<212> PRT
<213> 智人(Homo sapiens)
<400> 98
Met Trp Arg Leu Met Ser Arg Phe Asn Ala Phe Lys Arg Thr Asn Thr
1 5 10 15
Ile Leu His His Leu Arg Met Ser Lys His Thr Asp Ala Ala Glu Glu
20 25 30
Val Leu Leu Glu Lys Lys Gly Cys Ala Gly Val Ile Thr Leu Asn Arg
35 40 45
Pro Lys Phe Leu Asn Ala Leu Thr Leu Asn Met Ile Arg Gln Ile Tyr
50 55 60
Pro Gln Leu Lys Lys Trp Glu Gln Asp Pro Glu Thr Phe Val Ile Ile
65 70 75 80
Ile Lys Gly Ala Gly Gly Lys Ala Phe Cys Ala Gly Gly Asp Ile Arg
85 90 95
Val Ile Ser Glu Ala Glu Lys Ala Lys Gln Lys Ile Ala Pro Val Phe
100 105 110
Phe Arg Glu Glu Tyr Met Leu Asn Asn Ala Val Gly Ser Cys Gln Lys
115 120 125
Pro Tyr Val Ala Leu Ile His Gly Ile Thr Met Gly Gly Gly Val Gly
130 135 140
Leu Ser Val His Gly Gln Phe Arg Val Ala Thr Glu Lys Cys Leu Phe
145 150 155 160
Ala Met Pro Glu Thr Ala Ile Gly Leu Phe Pro Asp Val Gly Gly Gly
165 170 175
Tyr Phe Phe Ala Thr Thr Pro Arg Lys Thr Trp Leu Leu Pro Cys Ile
180 185 190
Asn Gly Phe Arg Leu Lys Gly Arg Asp Val Tyr Arg Ala Gly Ile Ala
195 200 205
Thr His Phe Val Asp Ser Glu Lys Leu Ala Met Leu Glu Glu Asp Leu
210 215 220
Leu Ala Leu Lys Ser Pro Ser Lys Glu Asn Ile Ala Ser Val Leu Glu
225 230 235 240
Asn Tyr His Thr Glu Ser Lys Ile Asp Arg Asp Lys Ser Phe Ile Leu
245 250 255
Glu Glu His Met Asp Lys Ile Asn Ser Cys Phe Ser Ala Asn Thr Val
260 265 270
Glu Glu Ile Ile Glu Asn Leu Gln Gln Asp Gly Ser Ser Phe Ala Leu
275 280 285
Glu Gln Leu Lys Val Ile Asn Lys Met Ser Pro Thr Ser Leu Lys Ile
290 295 300
Thr Leu Arg Gln Leu Met Glu Gly Ser Ser Lys Thr Leu Gln Glu Val
305 310 315 320
Leu Thr Met Glu Tyr Arg Leu Ser Gln Ala Cys Met Arg Gly His Asp
325 330 335
Phe His Glu Gly Val Arg Ala Val Leu Ile Asp Lys Asp Gln Ser Pro
340 345 350
Lys Trp Lys Pro Ala Asp Leu Lys Glu Val Thr Glu Glu Asp Leu Asn
355 360 365
Asn His Phe Lys Ser Leu Gly Ser Ser Asp Leu Lys Phe
370 375 380
<210> 99
<211> 261
<212> PRT
<213> 埃氏巨球形菌(Megasphaera elsdenii)
<400> 99
Val Lys Thr Val Tyr Thr Leu Gly Ile Asp Val Gly Ser Ser Ser Ser
1 5 10 15
Lys Ala Val Ile Leu Glu Asp Gly Lys Lys Ile Val Ala His Ala Val
20 25 30
Val Glu Ile Gly Thr Gly Ser Thr Gly Pro Glu Arg Val Leu Asp Glu
35 40 45
Val Phe Lys Asp Thr Asn Leu Lys Ile Glu Asp Met Ala Asn Ile Ile
50 55 60
Ala Thr Gly Tyr Gly Arg Phe Asn Val Asp Cys Ala Lys Gly Glu Val
65 70 75 80
Ser Glu Ile Thr Cys His Ala Lys Gly Ala Leu Phe Glu Cys Pro Gly
85 90 95
Thr Thr Thr Ile Leu Asp Ile Gly Gly Gln Asp Val Lys Ser Ile Lys
100 105 110
Leu Asn Gly Gln Gly Leu Val Met Gln Phe Ala Met Asn Asp Lys Cys
115 120 125
Ala Ala Gly Thr Gly Arg Phe Leu Asp Val Met Ser Lys Val Leu Glu
130 135 140
Ile Pro Met Ser Glu Met Gly Asp Trp Tyr Phe Lys Ser Lys His Pro
145 150 155 160
Ala Ala Val Ser Ser Thr Cys Thr Val Phe Ala Glu Ser Glu Val Ile
165 170 175
Ser Leu Leu Ser Lys Asn Val Pro Lys Glu Asp Ile Val Ala Gly Val
180 185 190
His Gln Ser Ile Ala Ala Lys Ala Cys Ala Leu Val Arg Arg Val Gly
195 200 205
Val Gly Glu Asp Leu Thr Met Thr Gly Gly Gly Ser Arg Asp Pro Gly
210 215 220
Val Val Asp Ala Val Ser Lys Glu Leu Gly Ile Pro Val Arg Val Ala
225 230 235 240
Leu His Pro Gln Ala Val Gly Ala Leu Gly Ala Ala Leu Ile Ala Tyr
245 250 255
Asp Lys Ile Lys Lys
260
<210> 100
<211> 428
<212> PRT
<213> 埃氏巨球形菌(Megasphaera elsdenii)
<400> 100
Met Ser Glu Glu Lys Thr Val Asp Ile Glu Ser Met Ser Ser Lys Glu
1 5 10 15
Ala Leu Gly Tyr Phe Leu Pro Lys Val Asp Glu Asp Ala Arg Lys Ala
20 25 30
Lys Lys Glu Gly Arg Leu Val Cys Trp Ser Ala Ser Val Ala Pro Pro
35 40 45
Glu Phe Cys Thr Ala Met Asp Ile Ala Ile Val Tyr Pro Glu Thr His
50 55 60
Ala Ala Gly Ile Gly Ala Arg His Gly Ala Pro Ala Met Leu Glu Val
65 70 75 80
Ala Glu Asn Lys Gly Tyr Asn Gln Asp Ile Cys Ser Tyr Cys Arg Val
85 90 95
Asn Met Gly Tyr Met Glu Leu Leu Lys Gln Gln Ala Leu Thr Gly Glu
100 105 110
Thr Pro Glu Val Leu Lys Asn Ser Pro Ala Ser Pro Ile Pro Leu Pro
115 120 125
Asp Val Val Leu Thr Cys Asn Asn Ile Cys Asn Thr Leu Leu Lys Trp
130 135 140
Tyr Glu Asn Leu Ala Lys Glu Leu Asn Val Pro Leu Ile Asn Ile Asp
145 150 155 160
Val Pro Phe Asn His Glu Phe Pro Val Thr Lys His Ala Lys Gln Tyr
165 170 175
Ile Val Gly Glu Phe Lys His Ala Ile Lys Gln Leu Glu Asp Leu Cys
180 185 190
Gly Arg Pro Phe Asp Tyr Asp Lys Phe Phe Glu Val Gln Lys Gln Thr
195 200 205
Gln Arg Ser Ile Ala Ala Trp Asn Lys Ile Ala Thr Tyr Phe Gln Tyr
210 215 220
Lys Pro Ser Pro Leu Asn Gly Phe Asp Leu Phe Asn Tyr Met Gly Leu
225 230 235 240
Ala Val Ala Ala Arg Ser Leu Asn Tyr Ser Glu Ile Thr Phe Asn Lys
245 250 255
Phe Leu Lys Glu Leu Asp Glu Lys Val Ala Asn Lys Lys Trp Ala Phe
260 265 270
Gly Glu Asn Glu Lys Ser Arg Val Thr Trp Glu Gly Ile Ala Val Trp
275 280 285
Ile Ala Leu Gly His Thr Phe Lys Glu Leu Lys Gly Gln Gly Ala Leu
290 295 300
Met Thr Gly Ser Ala Tyr Pro Gly Met Trp Asp Val Ser Tyr Glu Pro
305 310 315 320
Gly Asp Leu Glu Ser Met Ala Glu Ala Tyr Ser Arg Thr Tyr Ile Asn
325 330 335
Cys Cys Leu Glu Gln Arg Gly Ala Val Leu Glu Lys Val Val Arg Asp
340 345 350
Gly Lys Cys Asp Gly Leu Ile Met His Gln Asn Arg Ser Cys Lys Asn
355 360 365
Met Ser Leu Leu Asn Asn Glu Gly Gly Gln Arg Ile Gln Lys Asn Leu
370 375 380
Gly Val Pro Tyr Val Ile Phe Asp Gly Asp Gln Thr Asp Ala Arg Asn
385 390 395 400
Phe Ser Glu Ala Gln Phe Asp Thr Arg Val Glu Ala Leu Ala Glu Met
405 410 415
Met Ala Asp Lys Lys Ala Asn Glu Gly Gly Asn His
420 425
<210> 101
<211> 372
<212> PRT
<213> 埃氏巨球形菌(Megasphaera elsdenii)
<400> 101
Met Ser Gln Ile Asp Glu Leu Ile Ser Lys Leu Gln Glu Val Ser Asn
1 5 10 15
His Pro Gln Lys Thr Val Leu Asn Tyr Lys Lys Gln Gly Lys Gly Leu
20 25 30
Val Gly Met Met Pro Tyr Tyr Ala Pro Glu Glu Ile Val Tyr Ala Ala
35 40 45
Gly Tyr Leu Pro Val Gly Met Phe Gly Ser Gln Asn Pro Gln Ile Ser
50 55 60
Ala Ala Arg Thr Tyr Leu Pro Pro Phe Ala Cys Ser Leu Met Gln Ala
65 70 75 80
Asp Met Glu Leu Gln Leu Asn Gly Thr Tyr Asp Cys Leu Asp Ala Val
85 90 95
Ile Phe Ser Val Pro Cys Asp Thr Leu Arg Cys Met Ser Gln Lys Trp
100 105 110
His Gly Lys Ala Pro Val Ile Val Phe Thr Gln Pro Gln Asn Arg Lys
115 120 125
Ile Arg Pro Ala Val Asp Phe Leu Lys Ala Glu Tyr Glu His Val Arg
130 135 140
Thr Glu Leu Gly Arg Ile Leu Asn Val Lys Ile Ser Asp Leu Ala Ile
145 150 155 160
Gln Glu Ala Ile Lys Val Tyr Asn Glu Asn Arg Gln Val Met Arg Glu
165 170 175
Phe Cys Asp Val Ala Ala Gln Tyr Pro Gln Ile Phe Thr Pro Ile Lys
180 185 190
Arg His Asp Val Ile Lys Ala Arg Trp Phe Met Asp Lys Ala Glu His
195 200 205
Thr Ala Leu Val Arg Glu Leu Ile Asp Ala Val Lys Lys Glu Pro Val
210 215 220
Gln Pro Trp Asn Gly Lys Lys Val Ile Leu Ser Gly Ile Met Ala Glu
225 230 235 240
Pro Asp Glu Phe Leu Asp Ile Phe Ser Glu Phe Asn Ile Ala Val Val
245 250 255
Ala Asp Asp Leu Ala Gln Glu Ser Arg Gln Phe Arg Thr Asp Val Pro
260 265 270
Ser Gly Ile Asp Pro Leu Glu Gln Leu Ala Gln Gln Trp Gln Asp Phe
275 280 285
Asp Gly Cys Pro Leu Ala Leu Asn Glu Asp Lys Pro Arg Gly Gln Met
290 295 300
Leu Ile Asp Met Thr Lys Lys Tyr Asn Ala Asp Ala Val Val Ile Cys
305 310 315 320
Met Met Arg Phe Cys Asp Pro Glu Glu Phe Asp Tyr Pro Ile Tyr Lys
325 330 335
Pro Glu Phe Glu Ala Ala Gly Val Arg Tyr Thr Val Leu Asp Leu Asp
340 345 350
Ile Glu Ser Pro Ser Leu Glu Gln Leu Arg Thr Arg Ile Gln Ala Phe
355 360 365
Ser Glu Ile Leu
370
<210> 102
<211> 519
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 102
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> 103
<211> 495
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 103
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> 104
<211> 495
<212> PRT
<213> 肺炎克雷伯氏菌(Klebsiella pneumoniae)
<400> 104
Met Asn Phe Gln His Leu Ala Tyr Trp Gln Glu Lys Ala Lys Asn Leu
1 5 10 15
Ala Ile Glu Thr Arg Leu Phe Ile Asn Gly Glu Tyr Cys Ala Ala Ala
20 25 30
Asp Asn Thr Thr Phe Glu Thr Ile Asp Pro Ala Ala Gln Gln Thr Leu
35 40 45
Ala Gln Val Ala Arg Gly Lys Lys Ala Asp Val Glu Arg Ala Val Lys
50 55 60
Ala Ala Arg Gln Ala Phe Asp Asn Gly Asp Trp Ser Gln Ala Ser Pro
65 70 75 80
Ala Gln Arg Lys Ala Ile Leu Thr Arg Phe Ala Asp Leu Met Glu Ala
85 90 95
His Arg 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 Leu Asp Lys Val Tyr Gly Glu Val Ala Pro
130 135 140
Thr Gly Ser Asn Glu Leu Ala Met Ile Val Arg Glu Pro Ile Gly Val
145 150 155 160
Ile Ala Ala Val Val Pro Trp Asn Phe Pro Leu Leu Leu Ala 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 Thr Ala Leu Arg Leu Ala Gly Leu Ala Lys
195 200 205
Glu Ala Gly Leu Pro Asp Gly Val Leu Asn Val Val Ser Gly Phe Gly
210 215 220
His Glu Ala Gly Gln Ala Leu Ala Leu His Pro Asp Val Glu Val Ile
225 230 235 240
Thr 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 Val Arg
275 280 285
Ala Thr Ala Gly 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
Arg Leu Lys Ala Glu Ala Gln His Trp Gln Pro Gly Asn Pro Leu Asp
325 330 335
Pro Asp Thr Thr Met Gly Met Leu Ile Asp Asn Thr His Ala Asp Asn
340 345 350
Val His Ser Phe Ile Arg Gly Gly Glu Ser Gln Ser Thr Leu Phe Leu
355 360 365
Asp Gly Arg Lys Asn Pro Trp Pro Ala Ala Val Gly Pro Thr Ile Phe
370 375 380
Val Asp Val Asp Pro Ala Ser Thr Leu Ser Arg Glu Glu Ile Phe Gly
385 390 395 400
Pro Val Leu Val Val Thr Arg Phe Lys Ser Glu Glu Glu Ala Leu Lys
405 410 415
Leu Ala Asn Asp Ser Asp 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 Ala Leu Glu Ser
485 490 495
<210> 105
<211> 1053
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 105
atgtcttacg aaatcccaca aacacaaaag gcctgtgtct tttacgaaaa cggcggccca 60
atcacataca aggacattcc agttccaaag ccaaaaccta ctgagatttt agtcaaggtt 120
ctgtactctg gtgtctgcca caccgacttg cacgcatgga agggtgactg gcctctagct 180
accaagttgc cattggttgg tggtcacgaa ggtgccggtg ttgttgttgc caagggtgaa 240
aacgtcacct cttttgagat tggtgattac gcaggtatca agtggttgaa tggttcatgt 300
atgggttgtg aattctgtga acaaggtgct gaaccaaact gtcctaaggc cgacttgagt 360
ggttacaccc acgacggttc cttccaacag tatgctactg ctgacgctat tcaagctgca 420
cacatctcca aggaaaccga cttggctggt gttgctccaa tcttgtgtgc aggtgtcact 480
gtctacaagg ctttaaagac tgcagacctt agagcaggtg aatgggtttg tatttccggt 540
gcagctggtg gtttaggttc tcttgctatt caatatgcaa aggctatggg tctgagagtt 600
gttggtattg acggtggtga cgaaaagaag gaattgtgta aatcccttgg tgctgaagca 660
tttattgatt tcacaaagac caaggatatc gtcaaggctg tccaagaggc aaccaatggt 720
ggtccacatg gtgtcatcaa tgtctctgtc tctgaagctg caatttctca atcttgtgaa 780
tacgttagac ctctaggtaa ggttgttctt gttggtttac cagcaggcgc acaagtcaaa 840
actggtgtct ttgaagccgt tgtcaagtct attgaaatta agggttctta tgtcggtaac 900
agaaaggata ccgccgaagc acttgacttc tacactagag gcttggtcaa gtctccattc 960
aagattgtcg gtttatccga attgccaaaa gtctttgaac tcatggaaca gggtaagatt 1020
ttaggtagaa tggtcttaga cacctccaaa taa 1053
<210> 106
<211> 350
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 106
Met Ser Tyr Glu Ile Pro Gln Thr Gln Lys Ala Cys Val Phe Tyr Glu
1 5 10 15
Asn Gly Gly Pro Ile Thr Tyr Lys Asp Ile Pro Val Pro Lys Pro Lys
20 25 30
Pro Thr Glu Ile Leu Val Lys Val Leu Tyr Ser Gly Val Cys His Thr
35 40 45
Asp Leu His Ala Trp Lys Gly Asp Trp Pro Leu Ala Thr Lys Leu Pro
50 55 60
Leu Val Gly Gly His Glu Gly Ala Gly Val Val Val Ala Lys Gly Glu
65 70 75 80
Asn Val Thr Ser Phe Glu Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu
85 90 95
Asn Gly Ser Cys Met Gly Cys Glu Phe Cys Glu Gln Gly Ala Glu Pro
100 105 110
Asn Cys Pro Lys Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe
115 120 125
Gln Gln Tyr Ala Thr Ala Asp Ala Ile Gln Ala Ala His Ile Ser Lys
130 135 140
Glu Thr Asp Leu Ala Gly Val Ala Pro Ile Leu Cys Ala Gly Val Thr
145 150 155 160
Val Tyr Lys Ala Leu Lys Thr Ala Asp Leu Arg Ala Gly Glu Trp Val
165 170 175
Cys Ile Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Ile Gln Tyr
180 185 190
Ala Lys Ala Met Gly Leu Arg Val Val Gly Ile Asp Gly Gly Asp Glu
195 200 205
Lys Lys Glu Leu Cys Lys Ser Leu Gly Ala Glu Ala Phe Ile Asp Phe
210 215 220
Thr Lys Thr Lys Asp Ile Val Lys Ala Val Gln Glu Ala Thr Asn Gly
225 230 235 240
Gly Pro His Gly Val Ile Asn Val Ser Val Ser Glu Ala Ala Ile Ser
245 250 255
Gln Ser Cys Glu Tyr Val Arg Pro Leu Gly Lys Val Val Leu Val Gly
260 265 270
Leu Pro Ala Gly Ala Gln Val Lys Thr Gly Val Phe Glu Ala Val Val
275 280 285
Lys Ser Ile Glu Ile Lys Gly Ser Tyr Val Gly Asn Arg Lys Asp Thr
290 295 300
Ala Glu Ala Leu Asp Phe Tyr Thr Arg Gly Leu Val Lys Ser Pro Phe
305 310 315 320
Lys Ile Val Gly Leu Ser Glu Leu Pro Lys Val Phe Glu Leu Met Glu
325 330 335
Gln Gly Lys Ile Leu Gly Arg Met Val Leu Asp Thr Ser Lys
340 345 350
<210> 107
<211> 1131
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 107
atgtttgcat caaccttcag aagtcaagct gtaagagctg caagatttac tagattccaa 60
tccacttttg ccattcctga gaagcaaatg ggtgttatct ttgaaactca tggtggtcct 120
ttacaataca aggaaattcc agttccaaaa ccaaaaccaa ctgaaatttt aatcaatgtt 180
aaatactctg gtgtctgcca taccgattta cacgcatgga aaggtgactg gccattacca 240
gcaaagttac ccctagttgg tggtcacgaa ggtgcgggca ttgttgttgc gaaaggttct 300
gcagttacca actttgagat tggcgattat gctggtatta agtggttaaa cggttcatgt 360
atgtcatgtg aattctgtga acaaggtgat gaatctaact gtgaacatgc cgatttgagt 420
ggttatactc atgatggttc tttccaacaa tatgccactg ctgacgctat tcaagctgca 480
aagatcccaa agggtaccga cttatctgaa gttgcgccaa ttttatgtgc tggtgttact 540
gtctataaag ctttgaaaac tgctgattta agagcaggtc aatgggttgc gatttctggt 600
gccgctggtg gtctaggttc tcttgctgtc caatatgcaa aggcaatggg tctaagagtt 660
ttaggtatcg atggtggtga aggtaaaaag gaactttttg aacaatgtgg tggtgatgtg 720
tttatcgatt tcaccagata cccaagagat gcacctgaaa agatggttgc tgatattaag 780
gctgcaacta acggtttggg tccacacggt gttatcaatg tctctgtctc cccagctgct 840
atctctcaat catgtgacta tgttagagca actggtaagg ttgtccttgt cggtatgcca 900
tctggtgctg tctgtaagtc tgatgtcttc actcatgttg ttaaatcctt acaaattaaa 960
ggttcttatg ttggtaacag agcagatacc agagaagctt tggaattctt taatgaaggt 1020
aaggtcagat ctccaatcaa ggttgtccca ttatctactt tacctgaaat ttacgaattg 1080
atggagcaag gtaagatttt aggtagatac gttgttgata cttctaaata a 1131
<210> 108
<211> 376
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 108
Met Phe Ala Ser Thr Phe Arg Ser Gln Ala Val Arg Ala Ala Arg Phe
1 5 10 15
Thr Arg Phe Gln Ser Thr Phe Ala Ile Pro Glu Lys Gln Met Gly Val
20 25 30
Ile Phe Glu Thr His Gly Gly Pro Leu Gln Tyr Lys Glu Ile Pro Val
35 40 45
Pro Lys Pro Lys Pro Thr Glu Ile Leu Ile Asn Val Lys Tyr Ser Gly
50 55 60
Val Cys His Thr Asp Leu His Ala Trp Lys Gly Asp Trp Pro Leu Pro
65 70 75 80
Ala Lys Leu Pro Leu Val Gly Gly His Glu Gly Ala Gly Ile Val Val
85 90 95
Ala Lys Gly Ser Ala Val Thr Asn Phe Glu Ile Gly Asp Tyr Ala Gly
100 105 110
Ile Lys Trp Leu Asn Gly Ser Cys Met Ser Cys Glu Phe Cys Glu Gln
115 120 125
Gly Asp Glu Ser Asn Cys Glu His Ala Asp Leu Ser Gly Tyr Thr His
130 135 140
Asp Gly Ser Phe Gln Gln Tyr Ala Thr Ala Asp Ala Ile Gln Ala Ala
145 150 155 160
Lys Ile Pro Lys Gly Thr Asp Leu Ser Glu Val Ala Pro Ile Leu Cys
165 170 175
Ala Gly Val Thr Val Tyr Lys Ala Leu Lys Thr Ala Asp Leu Arg Ala
180 185 190
Gly Gln Trp Val Ala Ile Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu
195 200 205
Ala Val Gln Tyr Ala Lys Ala Met Gly Leu Arg Val Leu Gly Ile Asp
210 215 220
Gly Gly Glu Gly Lys Lys Glu Leu Phe Glu Gln Cys Gly Gly Asp Val
225 230 235 240
Phe Ile Asp Phe Thr Arg Tyr Pro Arg Asp Ala Pro Glu Lys Met Val
245 250 255
Ala Asp Ile Lys Ala Ala Thr Asn Gly Leu Gly Pro His Gly Val Ile
260 265 270
Asn Val Ser Val Ser Pro Ala Ala Ile Ser Gln Ser Cys Asp Tyr Val
275 280 285
Arg Ala Thr Gly Lys Val Val Leu Val Gly Met Pro Ser Gly Ala Val
290 295 300
Cys Lys Ser Asp Val Phe Thr His Val Val Lys Ser Leu Gln Ile Lys
305 310 315 320
Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu Ala Leu Glu Phe
325 330 335
Phe Asn Glu Gly Lys Val Arg Ser Pro Ile Lys Val Val Pro Leu Ser
340 345 350
Thr Leu Pro Glu Ile Tyr Glu Leu Met Glu Gln Gly Lys Ile Leu Gly
355 360 365
Arg Tyr Val Val Asp Thr Ser Lys
370 375
<210> 109
<211> 1134
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 109
atgttatcca agaccatcac tgctgcattg aggggcaata caactcgtac tgcattcaga 60
atcaatgcca ttagaagttt agcgatccca gctattccag agacacaaaa gggtgttatc 120
ttttatgaga acggaggtga actattttac aaggacattc cagttccaaa gccaaagcca 180
aatgagattt tggtgaatgt caagtattct ggtgtttgtc ataccgattt acacgcatgg 240
aaaggtgact ggcctttggc gaccaagttg ccattggttg gtggacatga aggtgccgga 300
gttgttgttg ctaaggggga caatgtcacc aactttgaaa ttggcgatta tgccggtatc 360
aagtggttga atggttcatg tatggggtgt gaattttgcc aacaaggtgc agagccaaac 420
tgtccacagg ccgacttgag tggttacacc catgacgggt cctttcaaca atatgccact 480
gccgatgctg ttcaggcagc caagattcct cagggcactg atttggctca agttgcgcca 540
attttatgtg caggtattac tgtctataag gctttaaaga ctgcagaatt aagaccaggt 600
caatgggttg ccatttctgg tgctgctgga ggtttaggtt ctcttgctgt tcaatatgcc 660
aaggccatgg gtttgagagt tttgggtatt gatggtggtg aggagaaggg caagtttgca 720
aagtctcttg gagctgaagt tttcattgat ttcaccaaat ccaaggacat tgtcaaggat 780
atccaagagg ccaccaatgg tggtccacat ggtgtcatta atgtttctgt ttctccagct 840
gctatttctc aaagtaccca gtatgtcaga accttgggta aggttgtcct tgttggatta 900
ccagcgcatg ctgtatgcga gtcttcggtt ttcgaccatg ttgtcaagtc gattcaaatt 960
agaggctctt atgttggtaa cagggaagat actagtgagg ctattgattt tttcaccagg 1020
ggtttagtga agtcaccaat taagattgtt ggtttgagtg agttgccaaa gatctatgaa 1080
ttgatggagc aaggtaagat tttaggcaga tatgttgttg acacttcgaa atga 1134
<210> 110
<211> 377
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 110
Met Leu Ser Lys Thr Ile Thr Ala Ala Leu Arg Gly Asn Thr Thr Arg
1 5 10 15
Thr Ala Phe Arg Ile Asn Ala Ile Arg Ser Leu Ala Ile Pro Ala Ile
20 25 30
Pro Glu Thr Gln Lys Gly Val Ile Phe Tyr Glu Asn Gly Gly Glu Leu
35 40 45
Phe Tyr Lys Asp Ile Pro Val Pro Lys Pro Lys Pro Asn Glu Ile Leu
50 55 60
Val Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu His Ala Trp
65 70 75 80
Lys Gly Asp Trp Pro Leu Ala Thr Lys Leu Pro Leu Val Gly Gly His
85 90 95
Glu Gly Ala Gly Val Val Val Ala Lys Gly Asp Asn Val Thr Asn Phe
100 105 110
Glu Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly Ser Cys Met
115 120 125
Gly Cys Glu Phe Cys Gln Gln Gly Ala Glu Pro Asn Cys Pro Gln Ala
130 135 140
Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Gln Tyr Ala Thr
145 150 155 160
Ala Asp Ala Val Gln Ala Ala Lys Ile Pro Gln Gly Thr Asp Leu Ala
165 170 175
Gln Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr Lys Ala Leu
180 185 190
Lys Thr Ala Glu Leu Arg Pro Gly Gln Trp Val Ala Ile Ser Gly Ala
195 200 205
Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys Ala Met Gly
210 215 220
Leu Arg Val Leu Gly Ile Asp Gly Gly Glu Glu Lys Gly Lys Phe Ala
225 230 235 240
Lys Ser Leu Gly Ala Glu Val Phe Ile Asp Phe Thr Lys Ser Lys Asp
245 250 255
Ile Val Lys Asp Ile Gln Glu Ala Thr Asn Gly Gly Pro His Gly Val
260 265 270
Ile Asn Val Ser Val Ser Pro Ala Ala Ile Ser Gln Ser Thr Gln Tyr
275 280 285
Val Arg Thr Leu Gly Lys Val Val Leu Val Gly Leu Pro Ala His Ala
290 295 300
Val Cys Glu Ser Ser Val Phe Asp His Val Val Lys Ser Ile Gln Ile
305 310 315 320
Arg Gly Ser Tyr Val Gly Asn Arg Glu Asp Thr Ser Glu Ala Ile Asp
325 330 335
Phe Phe Thr Arg Gly Leu Val Lys Ser Pro Ile Lys Ile Val Gly Leu
340 345 350
Ser Glu Leu Pro Lys Ile Tyr Glu Leu Met Glu Gln Gly Lys Ile Leu
355 360 365
Gly Arg Tyr Val Val Asp Thr Ser Lys
370 375
<210> 111
<211> 1347
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 111
atgtctcctt cacaaattaa cgttgacaac ttatctaatt ggactgaaga attcaaatct 60
gacgccaaga ctcaaatcgg gggttctgta ttgcaacatt ccaacattga tgaggtcttg 120
attaacagag atgcagaaat cgccaacaag catatcttca accacaagat tgaaattgaa 180
ggtctacctg tcatggatca gaaggcttct ggtagatgtt ggttgtttgc atcgactaac 240
ttgatgcgtg ttactgcaat gaagaaatac aatttgaagg aaatcaagct ttccccatcg 300
tatttgtttt tctatgacaa attggaaaga gcaaactatt tccttgaaca aatcatcgac 360
actcataagg aaccaatcga ttcaagattg gttcaatatt tcctgaccaa tccagttgaa 420
gatggtggtc aattcaccat gatggcacaa attgctacca aatacggtgt tgttcctgat 480
caagtctacc cagattcttt caacacaacc acttcgagga ttatgaacag attagtcaac 540
cacagattac gttcttatgc aatgacttta cgtaacgctc tagatgaagg taaagatgta 600
atgtccttga agaatgagat gcaaaaagaa atttatcgtt tgctaacaat gttccttggt 660
aacccaccaa agccaaacga agagtttgtc tgggaattca ccgataaaga tggtaaatat 720
gaatctatta aaactacacc attaaaatat gcaactgaag ttttggattt ccatgctcca 780
gaatatgttt ccttgttaaa tgacccaaga aataagtata acaagatggt tcaagttgaa 840
agattaggta atgttgctgg tggcgaacca gttgcatact taaacttaga aattgaaaag 900
ttatctcaag ctgttgttaa cagaatcaaa aataacaaac cagttttctt tggtaccgat 960
acacctaaat ttatggataa aagtagaggt attatggata tcaatttatg ggactatgag 1020
ttattaggtt atgatgtccg taccatgtca aagaaggaaa gagttgtttt tggtgattct 1080
ttaatgaccc acgctatgtt gattactgca gtgcacgttg atgaaaatgg caaacctgtc 1140
agatacagag tcgaaaacag ttggggtacc aagagtggtc aagaaggtta ttacacaatg 1200
acccaagaat attttgaaga gtacgtttat caagtagtca ttgaaaagag tgaatttgct 1260
gccctaaacc tcgatgtttc cattctggag gataaagaac cagtcgtctt gccaccttat 1320
gaccctatgg gtgcacttgc tttataa 1347
<210> 112
<211> 448
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 112
Met Ser Pro Ser Gln Ile Asn Val Asp Asn Leu Ser Asn Trp Thr Glu
1 5 10 15
Glu Phe Lys Ser Asp Ala Lys Thr Gln Ile Gly Gly Ser Val Leu Gln
20 25 30
His Ser Asn Ile Asp Glu Val Leu Ile Asn Arg Asp Ala Glu Ile Ala
35 40 45
Asn Lys His Ile Phe Asn His Lys Ile Glu Ile Glu Gly Leu Pro Val
50 55 60
Met Asp Gln Lys Ala Ser Gly Arg Cys Trp Leu Phe Ala Ser Thr Asn
65 70 75 80
Leu Met Arg Val Thr Ala Met Lys Lys Tyr Asn Leu Lys Glu Ile Lys
85 90 95
Leu Ser Pro Ser Tyr Leu Phe Phe Tyr Asp Lys Leu Glu Arg Ala Asn
100 105 110
Tyr Phe Leu Glu Gln Ile Ile Asp Thr His Lys Glu Pro Ile Asp Ser
115 120 125
Arg Leu Val Gln Tyr Phe Leu Thr Asn Pro Val Glu Asp Gly Gly Gln
130 135 140
Phe Thr Met Met Ala Gln Ile Ala Thr Lys Tyr Gly Val Val Pro Asp
145 150 155 160
Gln Val Tyr Pro Asp Ser Phe Asn Thr Thr Thr Ser Arg Ile Met Asn
165 170 175
Arg Leu Val Asn His Arg Leu Arg Ser Tyr Ala Met Thr Leu Arg Asn
180 185 190
Ala Leu Asp Glu Gly Lys Asp Val Met Ser Leu Lys Asn Glu Met Gln
195 200 205
Lys Glu Ile Tyr Arg Leu Leu Thr Met Phe Leu Gly Asn Pro Pro Lys
210 215 220
Pro Asn Glu Glu Phe Val Trp Glu Phe Thr Asp Lys Asp Gly Lys Tyr
225 230 235 240
Glu Ser Ile Lys Thr Thr Pro Leu Lys Tyr Ala Thr Glu Val Leu Asp
245 250 255
Phe His Ala Pro Glu Tyr Val Ser Leu Leu Asn Asp Pro Arg Asn Lys
260 265 270
Tyr Asn Lys Met Val Gln Val Glu Arg Leu Gly Asn Val Ala Gly Gly
275 280 285
Glu Pro Val Ala Tyr Leu Asn Leu Glu Ile Glu Lys Leu Ser Gln Ala
290 295 300
Val Val Asn Arg Ile Lys Asn Asn Lys Pro Val Phe Phe Gly Thr Asp
305 310 315 320
Thr Pro Lys Phe Met Asp Lys Ser Arg Gly Ile Met Asp Ile Asn Leu
325 330 335
Trp Asp Tyr Glu Leu Leu Gly Tyr Asp Val Arg Thr Met Ser Lys Lys
340 345 350
Glu Arg Val Val Phe Gly Asp Ser Leu Met Thr His Ala Met Leu Ile
355 360 365
Thr Ala Val His Val Asp Glu Asn Gly Lys Pro Val Arg Tyr Arg Val
370 375 380
Glu Asn Ser Trp Gly Thr Lys Ser Gly Gln Glu Gly Tyr Tyr Thr Met
385 390 395 400
Thr Gln Glu Tyr Phe Glu Glu Tyr Val Tyr Gln Val Val Ile Glu Lys
405 410 415
Ser Glu Phe Ala Ala Leu Asn Leu Asp Val Ser Ile Leu Glu Asp Lys
420 425 430
Glu Pro Val Val Leu Pro Pro Tyr Asp Pro Met Gly Ala Leu Ala Leu
435 440 445
<210> 113
<211> 1737
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 113
atgttactca gatcactaaa ctcttctgct cgttgtgtca aacaaacaac cagaacaaag 60
gttaggtatc tcagccacgt cagtggtgca agcatggcga aacctacatt gaagaacaac 120
tcgagagaat ccaacaaatc cagaaactat ctaattgctg ctgtgacagc attggctgta 180
tcaacctcaa ttggagttgc cgtacatgtg aaggacccct tgtataacga tgctaccggc 240
agtgattctc cgagaagtat atctgttgac gagtttgtca agcataattc acaaaacgac 300
tgttggattg caatcaatgg caaggtttat gatttcactg attttattcc aaaccatcca 360
ggtggggtac ctccattagt taatcatgct ggttatgatg gtactaaact ttatgagaaa 420
ttgcatccaa aaggtacaat tgagaaattc ttgccaaagg ataagtttct gggtgtgtta 480
gatggtgaag cgccaaaatt ggaagcagac tatttggtgg acgatgatga acaagagaga 540
ctggattatt tgaacaactt acctcctttg tcatctattc agaatgttta tgatttcgaa 600
tacttggcca agaagatttt acctaaagat gcctgggcat attattcttg tggtgccgat 660
gatgaaatca caatgagaga aaaccattat gcttatcaaa gagtttattt cagaccaaga 720
atttgtgttg atgtcaagga agttgatact tcttatgaaa tgttaggcac taaaacctct 780
gttccttttt atgtatctgc caccgctttg gctaaattag gccatcctga tggtgaatgc 840
tcaattgcta gaggcgctgg taaggaaggt gtcgttcaaa tgatttcgac cctttcctca 900
atgtcattag atgaaattgc cgctgctaga attccaggtg caacccaatg gttccaatta 960
tacattaatg aggatagaaa tgtcgctaaa ggtctggtca aacatgcaga agacttgggt 1020
atgaaggcta tctttataac tgttgatgct ccttctctag gtaacagaga aaaggataaa 1080
agattaaagt ttgttaatga caccgatgtc gatttgggtg attccgcaga tcgaaacagt 1140
ggtgcttcaa aggcactatc ttcgttcatt gatgcttctg tctcttggaa tgacgtcaaa 1200
gcggtcaagt cgtggactaa attgcctgtc ttagttaaag gtgttcaaac agttgaagac 1260
gttattgaag cttacgatgc tggttgtcaa ggtgttgttt tgtcaaacca cggtggtagg 1320
caactagata ctgctcctcc tccaatcgaa ttattagctg aaactgttcc aactttgaag 1380
agattgggta aattaagacc agattttgaa attttaattg acggtggtgt caaaagaggt 1440
accgatattt tgaaagcagt cgcaatcggt ggccaagatg tcagagtttc agttggtatg 1500
ggtagacctt tcttatatgc caactcttgc tatggtgaag caggtgttag aaaattaatt 1560
caaaatctaa aggatgaatt agaaatggat atgagattgt tgggtgtcac taaaatggac 1620
cagctatctt cgaaacatgt cgatactaaa cgtttgattg gtagagatgc gatcaactat 1680
ttgtatgata atgtatacag cccaatcgaa accgttaaat tcaacaatga agattga 1737
<210> 114
<211> 578
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 114
Met Leu Leu Arg Ser Leu Asn Ser Ser Ala Arg Cys Val Lys Gln Thr
1 5 10 15
Thr Arg Thr Lys Val Arg Tyr Leu Ser His Val Ser Gly Ala Ser Met
20 25 30
Ala Lys Pro Thr Leu Lys Asn Asn Ser Arg Glu Ser Asn Lys Ser Arg
35 40 45
Asn Tyr Leu Ile Ala Ala Val Thr Ala Leu Ala Val Ser Thr Ser Ile
50 55 60
Gly Val Ala Val His Val Lys Asp Pro Leu Tyr Asn Asp Ala Thr Gly
65 70 75 80
Ser Asp Ser Pro Arg Ser Ile Ser Val Asp Glu Phe Val Lys His Asn
85 90 95
Ser Gln Asn Asp Cys Trp Ile Ala Ile Asn Gly Lys Val Tyr Asp Phe
100 105 110
Thr Asp Phe Ile Pro Asn His Pro Gly Gly Val Pro Pro Leu Val Asn
115 120 125
His Ala Gly Tyr Asp Gly Thr Lys Leu Tyr Glu Lys Leu His Pro Lys
130 135 140
Gly Thr Ile Glu Lys Phe Leu Pro Lys Asp Lys Phe Leu Gly Val Leu
145 150 155 160
Asp Gly Glu Ala Pro Lys Leu Glu Ala Asp Tyr Leu Val Asp Asp Asp
165 170 175
Glu Gln Glu Arg Leu Asp Tyr Leu Asn Asn Leu Pro Pro Leu Ser Ser
180 185 190
Ile Gln Asn Val Tyr Asp Phe Glu Tyr Leu Ala Lys Lys Ile Leu Pro
195 200 205
Lys Asp Ala Trp Ala Tyr Tyr Ser Cys Gly Ala Asp Asp Glu Ile Thr
210 215 220
Met Arg Glu Asn His Tyr Ala Tyr Gln Arg Val Tyr Phe Arg Pro Arg
225 230 235 240
Ile Cys Val Asp Val Lys Glu Val Asp Thr Ser Tyr Glu Met Leu Gly
245 250 255
Thr Lys Thr Ser Val Pro Phe Tyr Val Ser Ala Thr Ala Leu Ala Lys
260 265 270
Leu Gly His Pro Asp Gly Glu Cys Ser Ile Ala Arg Gly Ala Gly Lys
275 280 285
Glu Gly Val Val Gln Met Ile Ser Thr Leu Ser Ser Met Ser Leu Asp
290 295 300
Glu Ile Ala Ala Ala Arg Ile Pro Gly Ala Thr Gln Trp Phe Gln Leu
305 310 315 320
Tyr Ile Asn Glu Asp Arg Asn Val Ala Lys Gly Leu Val Lys His Ala
325 330 335
Glu Asp Leu Gly Met Lys Ala Ile Phe Ile Thr Val Asp Ala Pro Ser
340 345 350
Leu Gly Asn Arg Glu Lys Asp Lys Arg Leu Lys Phe Val Asn Asp Thr
355 360 365
Asp Val Asp Leu Gly Asp Ser Ala Asp Arg Asn Ser Gly Ala Ser Lys
370 375 380
Ala Leu Ser Ser Phe Ile Asp Ala Ser Val Ser Trp Asn Asp Val Lys
385 390 395 400
Ala Val Lys Ser Trp Thr Lys Leu Pro Val Leu Val Lys Gly Val Gln
405 410 415
Thr Val Glu Asp Val Ile Glu Ala Tyr Asp Ala Gly Cys Gln Gly Val
420 425 430
Val Leu Ser Asn His Gly Gly Arg Gln Leu Asp Thr Ala Pro Pro Pro
435 440 445
Ile Glu Leu Leu Ala Glu Thr Val Pro Thr Leu Lys Arg Leu Gly Lys
450 455 460
Leu Arg Pro Asp Phe Glu Ile Leu Ile Asp Gly Gly Val Lys Arg Gly
465 470 475 480
Thr Asp Ile Leu Lys Ala Val Ala Ile Gly Gly Gln Asp Val Arg Val
485 490 495
Ser Val Gly Met Gly Arg Pro Phe Leu Tyr Ala Asn Ser Cys Tyr Gly
500 505 510
Glu Ala Gly Val Arg Lys Leu Ile Gln Asn Leu Lys Asp Glu Leu Glu
515 520 525
Met Asp Met Arg Leu Leu Gly Val Thr Lys Met Asp Gln Leu Ser Ser
530 535 540
Lys His Val Asp Thr Lys Arg Leu Ile Gly Arg Asp Ala Ile Asn Tyr
545 550 555 560
Leu Tyr Asp Asn Val Tyr Ser Pro Ile Glu Thr Val Lys Phe Asn Asn
565 570 575
Glu Asp
<210> 115
<211> 1698
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<220>
<221> 尚未归类的特征(misc_feature)
<222> (941)..(941)
<223> N= A, C, T, or G
<220>
<221> 尚未归类的特征(misc_feature)
<222> (943)..(943)
<223> N= A, C, T, or G
<400> 115
atgttaagat cccagttcaa aaacattttg aaaaatgtta acaagaacca ttctctaagg 60
agaactttta cttccagcac ctcaaaggct ggaaaaaatg cttcatacaa tgccaagatt 120
atatctgcaa ccgtggcctc gattgttgca gcagctggct cttatatgtt ggtccagcct 180
tcactagcta atgatgaggc acagtctgct aatccaacta ggaagatctc tgttgacgaa 240
tttgttaaac acaaccatgc cgatgattgt tggatcactg ttaacggtaa cgtctatgac 300
ttgactgatt tcatttcaat gcatccaggt ggtactaccc cattgattca aaatgcaggt 360
cacgacgcaa ctgaaattta caacaagatt catccaaagg gtacaatcga gaacttctta 420
ccaaaggaaa agcaattggg tgttttggat ggtgaagctc ctaaaatcga agttgtgctt 480
gacgaaaagg agaaacacag attggagttg ttgaatcatc tccctgctct ttccagaatt 540
caaaacattt atgatttcga acatattgct tctagagttt tgagcgacca agcatggaac 600
tactattcat gtggtgccga agatgaaatc accttgaggg aaaatcatta tgcttaccaa 660
agaatctact ttaagccaaa atgttgtgtc aatgttgcag aagttgatac ctctcatgaa 720
attttaggta caaaagcttc tgttcctttc tacgtttccg cagccgcttc tgcaaagttg 780
gggcacgagg atggtgaatg ttccattgct agaggtgcag gtaaggaagg cgttattcaa 840
atgatttctt ccttctcttc caactctttg gaggaaattg cagaatccag aattcctggt 900
gcaacacaat ggtttcaatt atacgttaat gaagacaagg ntnttgtgaa gaagacttta 960
aaaagggccg aaaacttggg tatgaaggcc atctttgtca ctgtggacgc tgctagtaga 1020
ggtaatagag aaaaagacat tacaatgaga attaccgaag atacagatga gttaatagac 1080
gattcttctg ttagagctgg ttctacctct ggtgcattgc cagctttcat tgacaagagg 1140
ctgacttggg atgaagttaa ggatatcatt tcatggacca agttaccagt tttgctgaag 1200
ggtgttcaaa gaactgatga tattgagaag gcaattgata ttggttgtaa gggtgttgtc 1260
ttgtccaatc atggtggtag acaattagat acttctcctc ctccaataga agttatggct 1320
gaatctgttc caatcctaaa gcaaaagggt aaactggatc caaatttcag tattttcgtt 1380
gatggtggtg ttagaagagg tacagatatt ttgaaagctt tggctattgg tggcagagac 1440
tgtaaagttg ctgttggtct gggtagacct ttcctttatg caaatactgg ttatggtgaa 1500
aagggtgtca gaaaggccgt gcaaattcta agagaagaat taaaggctga catgagaatg 1560
ttgggcgtta cctctttgaa cgagctagac gactcttaca ttgacaccag aagattacta 1620
ggtagagatg ctgttaacca catatacaac aacaactact acccaatgtc taagattcaa 1680
ttcaaaaacg aaaaataa 1698
<210> 116
<211> 565
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<220>
<221> 尚未归类的特征(misc_feature)
<222> (314)..(314)
<223> Xaa = Asp, Gly, Ala, or Val
<220>
<221> 尚未归类的特征(misc_feature)
<222> (315)..(315)
<223> Xaa = Ile, Val, Leu, or Phe
<400> 116
Met Leu Arg Ser Gln Phe Lys Asn Ile Leu Lys Asn Val Asn Lys Asn
1 5 10 15
His Ser Leu Arg Arg Thr Phe Thr Ser Ser Thr Ser Lys Ala Gly Lys
20 25 30
Asn Ala Ser Tyr Asn Ala Lys Ile Ile Ser Ala Thr Val Ala Ser Ile
35 40 45
Val Ala Ala Ala Gly Ser Tyr Met Leu Val Gln Pro Ser Leu Ala Asn
50 55 60
Asp Glu Ala Gln Ser Ala Asn Pro Thr Arg Lys Ile Ser Val Asp Glu
65 70 75 80
Phe Val Lys His Asn His Ala Asp Asp Cys Trp Ile Thr Val Asn Gly
85 90 95
Asn Val Tyr Asp Leu Thr Asp Phe Ile Ser Met His Pro Gly Gly Thr
100 105 110
Thr Pro Leu Ile Gln Asn Ala Gly His Asp Ala Thr Glu Ile Tyr Asn
115 120 125
Lys Ile His Pro Lys Gly Thr Ile Glu Asn Phe Leu Pro Lys Glu Lys
130 135 140
Gln Leu Gly Val Leu Asp Gly Glu Ala Pro Lys Ile Glu Val Val Leu
145 150 155 160
Asp Glu Lys Glu Lys His Arg Leu Glu Leu Leu Asn His Leu Pro Ala
165 170 175
Leu Ser Arg Ile Gln Asn Ile Tyr Asp Phe Glu His Ile Ala Ser Arg
180 185 190
Val Leu Ser Asp Gln Ala Trp Asn Tyr Tyr Ser Cys Gly Ala Glu Asp
195 200 205
Glu Ile Thr Leu Arg Glu Asn His Tyr Ala Tyr Gln Arg Ile Tyr Phe
210 215 220
Lys Pro Lys Cys Cys Val Asn Val Ala Glu Val Asp Thr Ser His Glu
225 230 235 240
Ile Leu Gly Thr Lys Ala Ser Val Pro Phe Tyr Val Ser Ala Ala Ala
245 250 255
Ser Ala Lys Leu Gly His Glu Asp Gly Glu Cys Ser Ile Ala Arg Gly
260 265 270
Ala Gly Lys Glu Gly Val Ile Gln Met Ile Ser Ser Phe Ser Ser Asn
275 280 285
Ser Leu Glu Glu Ile Ala Glu Ser Arg Ile Pro Gly Ala Thr Gln Trp
290 295 300
Phe Gln Leu Tyr Val Asn Glu Asp Lys Xaa Xaa Val Lys Lys Thr Leu
305 310 315 320
Lys Arg Ala Glu Asn Leu Gly Met Lys Ala Ile Phe Val Thr Val Asp
325 330 335
Ala Ala Ser Arg Gly Asn Arg Glu Lys Asp Ile Thr Met Arg Ile Thr
340 345 350
Glu Asp Thr Asp Glu Leu Ile Asp Asp Ser Ser Val Arg Ala Gly Ser
355 360 365
Thr Ser Gly Ala Leu Pro Ala Phe Ile Asp Lys Arg Leu Thr Trp Asp
370 375 380
Glu Val Lys Asp Ile Ile Ser Trp Thr Lys Leu Pro Val Leu Leu Lys
385 390 395 400
Gly Val Gln Arg Thr Asp Asp Ile Glu Lys Ala Ile Asp Ile Gly Cys
405 410 415
Lys Gly Val Val Leu Ser Asn His Gly Gly Arg Gln Leu Asp Thr Ser
420 425 430
Pro Pro Pro Ile Glu Val Met Ala Glu Ser Val Pro Ile Leu Lys Gln
435 440 445
Lys Gly Lys Leu Asp Pro Asn Phe Ser Ile Phe Val Asp Gly Gly Val
450 455 460
Arg Arg Gly Thr Asp Ile Leu Lys Ala Leu Ala Ile Gly Gly Arg Asp
465 470 475 480
Cys Lys Val Ala Val Gly Leu Gly Arg Pro Phe Leu Tyr Ala Asn Thr
485 490 495
Gly Tyr Gly Glu Lys Gly Val Arg Lys Ala Val Gln Ile Leu Arg Glu
500 505 510
Glu Leu Lys Ala Asp Met Arg Met Leu Gly Val Thr Ser Leu Asn Glu
515 520 525
Leu Asp Asp Ser Tyr Ile Asp Thr Arg Arg Leu Leu Gly Arg Asp Ala
530 535 540
Val Asn His Ile Tyr Asn Asn Asn Tyr Tyr Pro Met Ser Lys Ile Gln
545 550 555 560
Phe Lys Asn Glu Lys
565
<210> 117
<211> 1167
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 117
atggtgtccc ctgctgaaag attatctact attgcgtcca caatcaagcc aaacagaaaa 60
gattctacat cattacaacc agaagactat ccggaacatc cgttcaaggt gacggttgtt 120
ggttccggta actgggggtg tacaattgcc aaggttatag cggaaaacac cgttgagaga 180
cctcgtcaat ttcaaagaga tgttaatatg tgggtctatg aagaattgat tgaaggcgaa 240
aagttgactg aaatcataaa taccaaacac gaaaacgtca agtacttgcc aggtatcaag 300
ttgccagtta acgttgttgc agttccagac attgttgagg cttgtgcagg ctcagacttg 360
attgtcttta atattcctca ccaattttta ccaagaattt tatcccaatt aaagggtaag 420
gtgaatccaa aggctagagc aatttcttgt ttgaaaggtt tggatgtcaa tcctaatgga 480
tgtaagttgc tctccactgt tattactgaa gagttgggta tttattgtgg tgccttatca 540
ggtgctaatt tagctcctga agttgcacaa tgtaaatggt cggaaacaac tgttgcatat 600
acaattccgg acgatttcag aggtaaaggc aaggatattg accatcaaat tctaaagagt 660
ttgttccata gaccttattt ccatgttcgt gttattagtg atgttgcagg tatttccatt 720
gccggtgcac tcaagaatgt cgttgctatg gctgctggat ttgtcgaagg tttaggttgg 780
ggtgataatg caaaggctgc agtcatgaga ataggtttgg tggaaaccat tcaatttgcc 840
aagacttttt tcgatggctg tcatgctgca acctttactc atgaatctgc aggtgttgcc 900
gacctaatca ctacctgtgc cggcggccgt aacgttagag ttggtagata tatggcacaa 960
cattctgtct ctgcaacgga ggctgaagaa aagttgttga atggccaatc ctgtcaaggt 1020
atccacacaa ctagggaagt ttacgagttc ctctccaaca tgggcaggac agatgagttc 1080
ccactattta ccaccaccta ccgtatcatc tacgaaaact tcccaattga gaagctgcca 1140
gaatgccttg aacctgtgga agattaa 1167
<210> 118
<211> 388
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 118
Met Val Ser Pro Ala Glu Arg Leu Ser Thr Ile Ala Ser Thr Ile Lys
1 5 10 15
Pro Asn Arg Lys Asp Ser Thr Ser Leu Gln Pro Glu Asp Tyr Pro Glu
20 25 30
His Pro Phe Lys Val Thr Val Val Gly Ser Gly Asn Trp Gly Cys Thr
35 40 45
Ile Ala Lys Val Ile Ala Glu Asn Thr Val Glu Arg Pro Arg Gln Phe
50 55 60
Gln Arg Asp Val Asn Met Trp Val Tyr Glu Glu Leu Ile Glu Gly Glu
65 70 75 80
Lys Leu Thr Glu Ile Ile Asn Thr Lys His Glu Asn Val Lys Tyr Leu
85 90 95
Pro Gly Ile Lys Leu Pro Val Asn Val Val Ala Val Pro Asp Ile Val
100 105 110
Glu Ala Cys Ala Gly Ser Asp Leu Ile Val Phe Asn Ile Pro His Gln
115 120 125
Phe Leu Pro Arg Ile Leu Ser Gln Leu Lys Gly Lys Val Asn Pro Lys
130 135 140
Ala Arg Ala Ile Ser Cys Leu Lys Gly Leu Asp Val Asn Pro Asn Gly
145 150 155 160
Cys Lys Leu Leu Ser Thr Val Ile Thr Glu Glu Leu Gly Ile Tyr Cys
165 170 175
Gly Ala Leu Ser Gly Ala Asn Leu Ala Pro Glu Val Ala Gln Cys Lys
180 185 190
Trp Ser Glu Thr Thr Val Ala Tyr Thr Ile Pro Asp Asp Phe Arg Gly
195 200 205
Lys Gly Lys Asp Ile Asp His Gln Ile Leu Lys Ser Leu Phe His Arg
210 215 220
Pro Tyr Phe His Val Arg Val Ile Ser Asp Val Ala Gly Ile Ser Ile
225 230 235 240
Ala Gly Ala Leu Lys Asn Val Val Ala Met Ala Ala Gly Phe Val Glu
245 250 255
Gly Leu Gly Trp Gly Asp Asn Ala Lys Ala Ala Val Met Arg Ile Gly
260 265 270
Leu Val Glu Thr Ile Gln Phe Ala Lys Thr Phe Phe Asp Gly Cys His
275 280 285
Ala Ala Thr Phe Thr His Glu Ser Ala Gly Val Ala Asp Leu Ile Thr
290 295 300
Thr Cys Ala Gly Gly Arg Asn Val Arg Val Gly Arg Tyr Met Ala Gln
305 310 315 320
His Ser Val Ser Ala Thr Glu Ala Glu Glu Lys Leu Leu Asn Gly Gln
325 330 335
Ser Cys Gln Gly Ile His Thr Thr Arg Glu Val Tyr Glu Phe Leu Ser
340 345 350
Asn Met Gly Arg Thr Asp Glu Phe Pro Leu Phe Thr Thr Thr Tyr Arg
355 360 365
Ile Ile Tyr Glu Asn Phe Pro Ile Glu Lys Leu Pro Glu Cys Leu Glu
370 375 380
Pro Val Glu Asp
385
<210> 119
<211> 1566
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 119
atgttgtccc tctctaaaca gtcaagaaac tttttcaaat tgaactattt ttcagtcacc 60
caaatagcaa aaatgtctgc aacttccgtc actttcccaa ttatcaacga aacttaccaa 120
cagccaaccg ggcttttcat caacaatgaa tttgttagtg caaagtcagg taagactttt 180
gatgttaaca ccccaattga tgagtctctc atttgtaaag tccaacaggc cgatgctgaa 240
gatgttgaaa ttgccgttca agcagcatct aaagcttaca agacttggag atttacaccg 300
ccaaatgaaa gaggcagata cttgaacaaa ttggccgatt tgatggacga aaagagagac 360
ttacttgcca aaattgaatc ccttgataat ggtaaggcct tacattgtgc aaaattcgat 420
gtcaatcttg tcattgaata tttcagatac tgtgcaggtt actgtgataa aatcgatggt 480
agaacaatta caaccgatgt agaacatttt acctacacta gaaaggaacc tttaggtgtc 540
tgtggtgcaa ttacaccttg gaacttccca ttgctgatgt ttgcttggaa aatcggcccg 600
gctttagcaa ccggtaatac cattatcttg aagcctgcca gtgcaacacc tctatcaaac 660
ctctttactt gtaccttgat caaggaggcg ggcattccag ccggtgttgt taatgttgtt 720
ccaggttccg gtagaggctg tggtaactcc attttacaac atcctaaaat taagaaggtt 780
gcgtttaccg gatctacaga agttggtaaa actgttatga aggaatgtgc taattccatc 840
aaaaaggtta ctctcgaatt gggtggtaag tctccaaaca ttgttttcaa agactgtaac 900
gttgaacaaa ccattcaaaa tttgattact ggtattttct tcaatggtgg tgaagtctgt 960
tgtgctggtt ctagaattta cattgaagca accgatgaga aatggtatac tgaattcttg 1020
accaaattca aggagactgt tgaaaaatta aagattggta acccatttga agagggtgtt 1080
ttccaaggtg cacaaaccac tccagatcaa ttccaaactg tcttggacta catcaccgct 1140
gctaacgaat ccagcttgaa actattaact ggtggtaaaa gaattggcaa taagggatac 1200
tttgttgagc caactatctt ctacgatgtt cctcaaaatt ccaagttaac tcaagaagaa 1260
atctttggtc cagttgctgt tgttttacct ttcaagtcca ctgaagaatt gattgaaaag 1320
gcaaatgatt ccgattttgg cttaggttcc ggtattcaca ctgaagattt caacaaggca 1380
atttgggttt ccgaaaggct tgaagcaggt tctgtttgga tcaacactta caatgatttc 1440
cacccagctg ctccattcgg tggttacaag gaatccggta ttggcagaga aatgggtatt 1500
gaagctttcg acaactatac tcaaaccaag ttagttagag ctagagttaa caagccagct 1560
ttttag 1566
<210> 120
<211> 521
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 120
Met Leu Ser Leu Ser Lys Gln Ser Arg Asn Phe Phe Lys Leu Asn Tyr
1 5 10 15
Phe Ser Val Thr Gln Ile Ala Lys Met Ser Ala Thr Ser Val Thr Phe
20 25 30
Pro Ile Ile Asn Glu Thr Tyr Gln Gln Pro Thr Gly Leu Phe Ile Asn
35 40 45
Asn Glu Phe Val Ser Ala Lys Ser Gly Lys Thr Phe Asp Val Asn Thr
50 55 60
Pro Ile Asp Glu Ser Leu Ile Cys Lys Val Gln Gln Ala Asp Ala Glu
65 70 75 80
Asp Val Glu Ile Ala Val Gln Ala Ala Ser Lys Ala Tyr Lys Thr Trp
85 90 95
Arg Phe Thr Pro Pro Asn Glu Arg Gly Arg Tyr Leu Asn Lys Leu Ala
100 105 110
Asp Leu Met Asp Glu Lys Arg Asp Leu Leu Ala Lys Ile Glu Ser Leu
115 120 125
Asp Asn Gly Lys Ala Leu His Cys Ala Lys Phe Asp Val Asn Leu Val
130 135 140
Ile Glu Tyr Phe Arg Tyr Cys Ala Gly Tyr Cys Asp Lys Ile Asp Gly
145 150 155 160
Arg Thr Ile Thr Thr Asp Val Glu His Phe Thr Tyr Thr Arg Lys Glu
165 170 175
Pro Leu Gly Val Cys Gly Ala Ile Thr Pro Trp Asn Phe Pro Leu Leu
180 185 190
Met Phe Ala Trp Lys Ile Gly Pro Ala Leu Ala Thr Gly Asn Thr Ile
195 200 205
Ile Leu Lys Pro Ala Ser Ala Thr Pro Leu Ser Asn Leu Phe Thr Cys
210 215 220
Thr Leu Ile Lys Glu Ala Gly Ile Pro Ala Gly Val Val Asn Val Val
225 230 235 240
Pro Gly Ser Gly Arg Gly Cys Gly Asn Ser Ile Leu Gln His Pro Lys
245 250 255
Ile Lys Lys Val Ala Phe Thr Gly Ser Thr Glu Val Gly Lys Thr Val
260 265 270
Met Lys Glu Cys Ala Asn Ser Ile Lys Lys Val Thr Leu Glu Leu Gly
275 280 285
Gly Lys Ser Pro Asn Ile Val Phe Lys Asp Cys Asn Val Glu Gln Thr
290 295 300
Ile Gln Asn Leu Ile Thr Gly Ile Phe Phe Asn Gly Gly Glu Val Cys
305 310 315 320
Cys Ala Gly Ser Arg Ile Tyr Ile Glu Ala Thr Asp Glu Lys Trp Tyr
325 330 335
Thr Glu Phe Leu Thr Lys Phe Lys Glu Thr Val Glu Lys Leu Lys Ile
340 345 350
Gly Asn Pro Phe Glu Glu Gly Val Phe Gln Gly Ala Gln Thr Thr Pro
355 360 365
Asp Gln Phe Gln Thr Val Leu Asp Tyr Ile Thr Ala Ala Asn Glu Ser
370 375 380
Ser Leu Lys Leu Leu Thr Gly Gly Lys Arg Ile Gly Asn Lys Gly Tyr
385 390 395 400
Phe Val Glu Pro Thr Ile Phe Tyr Asp Val Pro Gln Asn Ser Lys Leu
405 410 415
Thr Gln Glu Glu Ile Phe Gly Pro Val Ala Val Val Leu Pro Phe Lys
420 425 430
Ser Thr Glu Glu Leu Ile Glu Lys Ala Asn Asp Ser Asp Phe Gly Leu
435 440 445
Gly Ser Gly Ile His Thr Glu Asp Phe Asn Lys Ala Ile Trp Val Ser
450 455 460
Glu Arg Leu Glu Ala Gly Ser Val Trp Ile Asn Thr Tyr Asn Asp Phe
465 470 475 480
His Pro Ala Ala Pro Phe Gly Gly Tyr Lys Glu Ser Gly Ile Gly Arg
485 490 495
Glu Met Gly Ile Glu Ala Phe Asp Asn Tyr Thr Gln Thr Lys Leu Val
500 505 510
Arg Ala Arg Val Asn Lys Pro Ala Phe
515 520
<210> 121
<211> 1506
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 121
atgtcagcac tgttcagaac cattgagact ccaaacggta aaaccctgga acaaccactg 60
ggtctcttca tcgacaatga gtgggtgaaa acaaaccgta cttttgagac cattaatccg 120
tccacaggtg aggcgatctg tcatgtttac cgtgctgggg tccaggaggt gaacgacgct 180
gtcgaagctg caaatagagc atttagaaac gaatcttggt caggtctaac tggttctcaa 240
cgtggcgatt tactgtatcg catgtaccaa gttatcaaaa gagacgccga gagcattgca 300
tcgattgagt ccatggataa tggtaaaccg tatgctgcag aatgcctaga tggagattta 360
ggtgaagctg ctgacgtttt caaatattat gccggttggg ccgacaagat caccggtgaa 420
ctcattggct cgagtgtatt gggtaagaat aagatgtgtt atgtcgagcc tacaccactg 480
ggtgccgttg gcggtatagt cccttggaat ttcccgttta ccatgatggc atggaaaatt 540
gccccggcac tggcgacggg ttgtacagtg gttatgaagt caagtgaagt cacaccgttg 600
acggcattat ggtatggcaa gattgcactt gaagtgggtc tacctaaagg tgtacttaac 660
atcctctccg gttttggatc ggatgttgga tcggccatgg cttcacatcc aaagttggct 720
aagatagcgt tcactggctc aactgcaact ggtaaaaaaa tcatggaagc agcaggtggt 780
tccaacttga aaaaggttac actagagtgt ggtggtaaat ctccttacat tgtttttgat 840
gatgctgact tagaattggc agtagaatgg gcatattggg gtatttggta taacaaaggt 900
gaggtttgta cttcaacttc gagatttttg attcaggaag acatttacga taagtttgtt 960
gagagttttg ttgagttgac caagacgaga gcaatcactg ctgatccgtt tgatgataga 1020
tgcactatcg ggcctttggt ttctagctca cagtacgaaa aagtcaaaaa gtacgttgaa 1080
ataggtaaaa atgaaggagc aaagctacta actggcaaat tcatcgacgg gccaggctat 1140
ttctgtgagc catttatctt cagtgaatgc actgacgata tgacaatcat gaaagaggaa 1200
atctttggcc ctgttgtggg gattactaaa ttctcaacgg ttaaagaggc gatcgagaga 1260
gccaatgcta cgacttacgg tttaggagct gcgttgtttt cctctaacat aacaaaggca 1320
cattctgtgg ctgccaagtt ggaggctgga atggtgtgga tcaattctaa tggtgattct 1380
gatatccaca ttccatttgg tggttccaaa atgagtggta taggtaggga gttggggcca 1440
tacgcactag acttgtttac tgagaaaaag gcagttcatg tcaacttatc gcttccggtc 1500
aagtga 1506
<210> 122
<211> 501
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 122
Met Ser Ala Leu Phe Arg Thr Ile Glu Thr Pro Asn Gly Lys Thr Leu
1 5 10 15
Glu Gln Pro Leu Gly Leu Phe Ile Asp Asn Glu Trp Val Lys Thr Asn
20 25 30
Arg Thr Phe Glu Thr Ile Asn Pro Ser Thr Gly Glu Ala Ile Cys His
35 40 45
Val Tyr Arg Ala Gly Val Gln Glu Val Asn Asp Ala Val Glu Ala Ala
50 55 60
Asn Arg Ala Phe Arg Asn Glu Ser Trp Ser Gly Leu Thr Gly Ser Gln
65 70 75 80
Arg Gly Asp Leu Leu Tyr Arg Met Tyr Gln Val Ile Lys Arg Asp Ala
85 90 95
Glu Ser Ile Ala Ser Ile Glu Ser Met Asp Asn Gly Lys Pro Tyr Ala
100 105 110
Ala Glu Cys Leu Asp Gly Asp Leu Gly Glu Ala Ala Asp Val Phe Lys
115 120 125
Tyr Tyr Ala Gly Trp Ala Asp Lys Ile Thr Gly Glu Leu Ile Gly Ser
130 135 140
Ser Val Leu Gly Lys Asn Lys Met Cys Tyr Val Glu Pro Thr Pro Leu
145 150 155 160
Gly Ala Val Gly Gly Ile Val Pro Trp Asn Phe Pro Phe Thr Met Met
165 170 175
Ala Trp Lys Ile Ala Pro Ala Leu Ala Thr Gly Cys Thr Val Val Met
180 185 190
Lys Ser Ser Glu Val Thr Pro Leu Thr Ala Leu Trp Tyr Gly Lys Ile
195 200 205
Ala Leu Glu Val Gly Leu Pro Lys Gly Val Leu Asn Ile Leu Ser Gly
210 215 220
Phe Gly Ser Asp Val Gly Ser Ala Met Ala Ser His Pro Lys Leu Ala
225 230 235 240
Lys Ile Ala Phe Thr Gly Ser Thr Ala Thr Gly Lys Lys Ile Met Glu
245 250 255
Ala Ala Gly Gly Ser Asn Leu Lys Lys Val Thr Leu Glu Cys Gly Gly
260 265 270
Lys Ser Pro Tyr Ile Val Phe Asp Asp Ala Asp Leu Glu Leu Ala Val
275 280 285
Glu Trp Ala Tyr Trp Gly Ile Trp Tyr Asn Lys Gly Glu Val Cys Thr
290 295 300
Ser Thr Ser Arg Phe Leu Ile Gln Glu Asp Ile Tyr Asp Lys Phe Val
305 310 315 320
Glu Ser Phe Val Glu Leu Thr Lys Thr Arg Ala Ile Thr Ala Asp Pro
325 330 335
Phe Asp Asp Arg Cys Thr Ile Gly Pro Leu Val Ser Ser Ser Gln Tyr
340 345 350
Glu Lys Val Lys Lys Tyr Val Glu Ile Gly Lys Asn Glu Gly Ala Lys
355 360 365
Leu Leu Thr Gly Lys Phe Ile Asp Gly Pro Gly Tyr Phe Cys Glu Pro
370 375 380
Phe Ile Phe Ser Glu Cys Thr Asp Asp Met Thr Ile Met Lys Glu Glu
385 390 395 400
Ile Phe Gly Pro Val Val Gly Ile Thr Lys Phe Ser Thr Val Lys Glu
405 410 415
Ala Ile Glu Arg Ala Asn Ala Thr Thr Tyr Gly Leu Gly Ala Ala Leu
420 425 430
Phe Ser Ser Asn Ile Thr Lys Ala His Ser Val Ala Ala Lys Leu Glu
435 440 445
Ala Gly Met Val Trp Ile Asn Ser Asn Gly Asp Ser Asp Ile His Ile
450 455 460
Pro Phe Gly Gly Ser Lys Met Ser Gly Ile Gly Arg Glu Leu Gly Pro
465 470 475 480
Tyr Ala Leu Asp Leu Phe Thr Glu Lys Lys Ala Val His Val Asn Leu
485 490 495
Ser Leu Pro Val Lys
500
<210> 123
<211> 1506
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 123
atggctcttc cacttgcaac tacaatctcc ttatcaagcg gcaaaacatt agaacagcca 60
attggtttat ttattgataa tgaatttgtc aatccaattt ctgtttctaa tgcaagaaca 120
ctaacaacct tcaacccaag cacaggtgag ccaataaccg atgttcattg tgcctcagct 180
gcagatgttg atgttgcggt aaatgctgca aacaaggcaa tggaaacatg gaaagacatt 240
gatcctactg ttcgtgtcga acttttacta aaattggcca gcttagttga cgagcattcc 300
caagcaattg ctgaaattga agcactagac tcgggtaaac cattgtactc gaatgcactg 360
gcggatgttc aatcggttgc tgagtactta aggtactgtg ccggttgggc ggataaatta 420
cacggtacgc aaattccaat aaactctaag gtaatggcta ttacaaaacg tgtaccctta 480
gttgtcggct gcatcattcc atggaactac ccaatttcaa tggcctcctg gaagttctgt 540
ccagcattgg ctgccggatg tactattgta atgaagtcaa gtgagataac cccgttatcg 600
ttactttatt ttgcgaattt ggtcaaatta gcaggtttcc ctaagggtgt ttttaatgtc 660
gtctctggat ttggtgatga tgttggctca gcgctttcaa atcacccaaa gttgggtaag 720
attgcattta caggctcgac cttgaccggg caaaaggtga tggcggatgc tgccagatca 780
aatttgaaaa gcgtatcttt ggaatgtggt ggtaaatctc cacttattgt cttcgaagat 840
gcagaattgg atgaatgcgt taaatgggca agttttggtg tcatgtataa caccggacaa 900
aattgtactg ccaattctcg tattattgtg catgataagg tttatgatca atttatcgaa 960
aagttcctgt ctcaactcaa ggaagattgg aaaatgggag atgtcatgaa tgaaaagact 1020
acattgggac cacttgtcag ccaacaacaa tatgagcgtg ttcagtcgta tattgatata 1080
ggtgtcaaag aaggggctac actgattcaa ccgcttaagg agagcactcc atcaaatgga 1140
ttctacatct ctcctactgt ttttactaac gttaaggaag atatgagaat tgttaaggag 1200
gaaatatttg gtcctgtcgt aactatctcc aaattctcaa ctgaggaaga ggcaatttca 1260
aaggcgaatg atacaattta tggcttagct gcaatgttat ttactactaa ttttgaacgt 1320
gccaacagag ttgctgataa gctggaagct ggcagtgtgt acattaatag ctctaacaac 1380
gagagtacca aagttccatt tggaggaatg aagatgagtg gtattggaag agagttgggg 1440
caagaagcat ttaatttgta cactgttaca aagagtattt attatagtta tggtgctaag 1500
ctttaa 1506
<210> 124
<211> 501
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 124
Met Ala Leu Pro Leu Ala Thr Thr Ile Ser Leu Ser Ser Gly Lys Thr
1 5 10 15
Leu Glu Gln Pro Ile Gly Leu Phe Ile Asp Asn Glu Phe Val Asn Pro
20 25 30
Ile Ser Val Ser Asn Ala Arg Thr Leu Thr Thr Phe Asn Pro Ser Thr
35 40 45
Gly Glu Pro Ile Thr Asp Val His Cys Ala Ser Ala Ala Asp Val Asp
50 55 60
Val Ala Val Asn Ala Ala Asn Lys Ala Met Glu Thr Trp Lys Asp Ile
65 70 75 80
Asp Pro Thr Val Arg Val Glu Leu Leu Leu Lys Leu Ala Ser Leu Val
85 90 95
Asp Glu His Ser Gln Ala Ile Ala Glu Ile Glu Ala Leu Asp Ser Gly
100 105 110
Lys Pro Leu Tyr Ser Asn Ala Leu Ala Asp Val Gln Ser Val Ala Glu
115 120 125
Tyr Leu Arg Tyr Cys Ala Gly Trp Ala Asp Lys Leu His Gly Thr Gln
130 135 140
Ile Pro Ile Asn Ser Lys Val Met Ala Ile Thr Lys Arg Val Pro Leu
145 150 155 160
Val Val Gly Cys Ile Ile Pro Trp Asn Tyr Pro Ile Ser Met Ala Ser
165 170 175
Trp Lys Phe Cys Pro Ala Leu Ala Ala Gly Cys Thr Ile Val Met Lys
180 185 190
Ser Ser Glu Ile Thr Pro Leu Ser Leu Leu Tyr Phe Ala Asn Leu Val
195 200 205
Lys Leu Ala Gly Phe Pro Lys Gly Val Phe Asn Val Val Ser Gly Phe
210 215 220
Gly Asp Asp Val Gly Ser Ala Leu Ser Asn His Pro Lys Leu Gly Lys
225 230 235 240
Ile Ala Phe Thr Gly Ser Thr Leu Thr Gly Gln Lys Val Met Ala Asp
245 250 255
Ala Ala Arg Ser Asn Leu Lys Ser Val Ser Leu Glu Cys Gly Gly Lys
260 265 270
Ser Pro Leu Ile Val Phe Glu Asp Ala Glu Leu Asp Glu Cys Val Lys
275 280 285
Trp Ala Ser Phe Gly Val Met Tyr Asn Thr Gly Gln Asn Cys Thr Ala
290 295 300
Asn Ser Arg Ile Ile Val His Asp Lys Val Tyr Asp Gln Phe Ile Glu
305 310 315 320
Lys Phe Leu Ser Gln Leu Lys Glu Asp Trp Lys Met Gly Asp Val Met
325 330 335
Asn Glu Lys Thr Thr Leu Gly Pro Leu Val Ser Gln Gln Gln Tyr Glu
340 345 350
Arg Val Gln Ser Tyr Ile Asp Ile Gly Val Lys Glu Gly Ala Thr Leu
355 360 365
Ile Gln Pro Leu Lys Glu Ser Thr Pro Ser Asn Gly Phe Tyr Ile Ser
370 375 380
Pro Thr Val Phe Thr Asn Val Lys Glu Asp Met Arg Ile Val Lys Glu
385 390 395 400
Glu Ile Phe Gly Pro Val Val Thr Ile Ser Lys Phe Ser Thr Glu Glu
405 410 415
Glu Ala Ile Ser Lys Ala Asn Asp Thr Ile Tyr Gly Leu Ala Ala Met
420 425 430
Leu Phe Thr Thr Asn Phe Glu Arg Ala Asn Arg Val Ala Asp Lys Leu
435 440 445
Glu Ala Gly Ser Val Tyr Ile Asn Ser Ser Asn Asn Glu Ser Thr Lys
450 455 460
Val Pro Phe Gly Gly Met Lys Met Ser Gly Ile Gly Arg Glu Leu Gly
465 470 475 480
Gln Glu Ala Phe Asn Leu Tyr Thr Val Thr Lys Ser Ile Tyr Tyr Ser
485 490 495
Tyr Gly Ala Lys Leu
500
<210> 125
<211> 1544
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<220>
<221> 尚未归类的特征(misc_feature)
<222> (1319)..(1319)
<223> N= A, C, G, OR T
<400> 125
atgatgggcg caactacagc aaaaattagt attccaaatg gtaacaaata cgagcaacct 60
acaggtttgt tcatcaatgg tgagtttgtt gcttcaagtg atggtaaaac tgcagaagtt 120
gagaatccag gcaatggaaa cattgtatgt tctgtccact tagcttctat tgaggatatt 180
aataccgccg tagaagctgc tgaagatgca tttttcaaaa ggtgggccac catcagtggt 240
aaagccaagg gagaatactt gagtaagatt gccgatctaa tcgttaaata ttctgatcaa 300
ttggcagatc tagaggctat tgaatcaggt aagccaaagg acaccaatgc aatctttgat 360
gttttacatt cggctgatgt tttcagatac tatgctggca aggctgtcac tgcacaaagc 420
ggcaagacta tcgagtccga actctccaaa tttacataca cagtttacga gccctatggt 480
gtttgtgccg ctatcatcgc atggaacttc ccaatgagca catttgcgtg gaaagttgcg 540
gcatgtttag ctgctggtaa tacaatggtt gtcaaaactt ccgagctgac tccgttatct 600
gcattgttca tgtgtaagat tttccaagaa gcagatctac ctgctggagt tataaacgtc 660
acatgtggtt taggttctgt tgcaggtgtt cgattgagtg aacatgaaaa ggttcagaaa 720
atttcgttta ctggctccac tggcgttggt aagttgatcc aagaatccgc agcaaagtct 780
aacttaaagt attgtacgct tgaatgtggt ggtaagtctc cgttagtgat ttacgaggat 840
gcagatcttg agcaagcagt gaagtgggct gcctttggta tttttttcaa caaaggtgaa 900
atttgcacag cctcttccag aatatatgtt caagaatcag tctatgacaa atttttgact 960
atgtacaagg atcatgtgga agaagcctat gttcaaggag aacagtttgc cactggtgtt 1020
aacgttgggc ctactgtctg caaagcccaa caagagaaaa tactggccta cattgaaagt 1080
gccaagcaag aaggtggtag aattatcact ggtggtaaaa taccatctta cacgaacaaa 1140
aatggttact atctcgaacc aacaattatt gcagattgta accaggatat gaaggtagtc 1200
agggaagaga ttttcggacc agtcgttact gtatccaaat tcactagtga tgaagaagcc 1260
atcaaattaa gcaatgattc cgaatatggc ttggcagcat atttattcac tystsrasna 1320
ssrgttyrgy taaaatyrth thraaggacc tcgttagatc tcagaattat atcagaaaag 1380
tgcaaagcgg acaggtcttt gtcaacttca cctttgcggc tgatttcagg ttgccatttg 1440
gcggatataa gatgagtggt aacggaagag agcttggtga tgaaggactg agtgctttcc 1500
agcaagtcaa agcagtacac attaatctca ctgggaagtt gtaa 1544
<210> 126
<211> 503
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 126
Met Met Gly Ala Thr Thr Ala Lys Ile Ser Ile Pro Asn Gly Asn Lys
1 5 10 15
Tyr Glu Gln Pro Thr Gly Leu Phe Ile Asn Gly Glu Phe Val Ala Ser
20 25 30
Ser Asp Gly Lys Thr Ala Glu Val Glu Asn Pro Gly Asn Gly Asn Ile
35 40 45
Val Cys Ser Val His Leu Ala Ser Ile Glu Asp Ile Asn Thr Ala Val
50 55 60
Glu Ala Ala Glu Asp Ala Phe Phe Lys Arg Trp Ala Thr Ile Ser Gly
65 70 75 80
Lys Ala Lys Gly Glu Tyr Leu Ser Lys Ile Ala Asp Leu Ile Val Lys
85 90 95
Tyr Ser Asp Gln Leu Ala Asp Leu Glu Ala Ile Glu Ser Gly Lys Pro
100 105 110
Lys Asp Thr Asn Ala Ile Phe Asp Val Leu His Ser Ala Asp Val Phe
115 120 125
Arg Tyr Tyr Ala Gly Lys Ala Val Thr Ala Gln Ser Gly Lys Thr Ile
130 135 140
Glu Ser Glu Leu Ser Lys Phe Thr Tyr Thr Val Tyr Glu Pro Tyr Gly
145 150 155 160
Val Cys Ala Ala Ile Ile Ala Trp Asn Phe Pro Met Ser Thr Phe Ala
165 170 175
Trp Lys Val Ala Ala Cys Leu Ala Ala Gly Asn Thr Met Val Val Lys
180 185 190
Thr Ser Glu Leu Thr Pro Leu Ser Ala Leu Phe Met Cys Lys Ile Phe
195 200 205
Gln Glu Ala Asp Leu Pro Ala Gly Val Ile Asn Val Thr Cys Gly Leu
210 215 220
Gly Ser Val Ala Gly Val Arg Leu Ser Glu His Glu Lys Val Gln Lys
225 230 235 240
Ile Ser Phe Thr Gly Ser Thr Gly Val Gly Lys Leu Ile Gln Glu Ser
245 250 255
Ala Ala Lys Ser Asn Leu Lys Tyr Cys Thr Leu Glu Cys Gly Gly Lys
260 265 270
Ser Pro Leu Val Ile Tyr Glu Asp Ala Asp Leu Glu Gln Ala Val Lys
275 280 285
Trp Ala Ala Phe Gly Ile Phe Phe Asn Lys Gly Glu Ile Cys Thr Ala
290 295 300
Ser Ser Arg Ile Tyr Val Gln Glu Ser Val Tyr Asp Lys Phe Leu Thr
305 310 315 320
Met Tyr Lys Asp His Val Glu Glu Ala Tyr Val Gln Gly Glu Gln Phe
325 330 335
Ala Thr Gly Val Asn Val Gly Pro Thr Val Cys Lys Ala Gln Gln Glu
340 345 350
Lys Ile Leu Ala Tyr Ile Glu Ser Ala Lys Gln Glu Gly Gly Arg Ile
355 360 365
Ile Thr Gly Gly Lys Ile Pro Ser Tyr Thr Asn Lys Asn Gly Tyr Tyr
370 375 380
Leu Glu Pro Thr Ile Ile Ala Asp Cys Asn Gln Asp Met Lys Val Val
385 390 395 400
Arg Glu Glu Ile Phe Gly Pro Val Val Thr Val Ser Lys Phe Thr Ser
405 410 415
Asp Glu Glu Ala Ile Lys Leu Ser Asn Asp Ser Glu Tyr Gly Leu Ala
420 425 430
Ala Tyr Leu Phe Thr Lys Asp Leu Val Arg Ser Gln Asn Tyr Ile Arg
435 440 445
Lys Val Gln Ser Gly Gln Val Phe Val Asn Phe Thr Phe Ala Ala Asp
450 455 460
Phe Arg Leu Pro Phe Gly Gly Tyr Lys Met Ser Gly Asn Gly Arg Glu
465 470 475 480
Leu Gly Asp Glu Gly Leu Ser Ala Phe Gln Gln Val Lys Ala Val His
485 490 495
Ile Asn Leu Thr Gly Lys Leu
500
<210> 127
<211> 1716
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<220>
<221> 尚未归类的特征(misc_feature)
<222> (655)..(655)
<223> n is a, c, g, or t
<400> 127
atggctccaa ctgctgttga tatccataac gagtacaaac agaatgtttc caacgaacag 60
gaaattcctt tcaacaaaac tgaaagaaag tcatcgattg catctaaatt aggactgaat 120
ccagacgcta agattcacta caattctgct gttcctatat tatacgaaga tggtttaaag 180
gaaaaaggta caaccatttc ctcttctggt gcattgattg cattctctgg ttccaaaaca 240
ggtagatctc caaaggacaa aagaattgtc gatgaagaga cttcaacaga caacatctgg 300
tggggtccag tcaataagaa ggttgatgaa aacacttgga atatctcgaa atctagagcg 360
attgattatt tgagaacaag agagaaggtt tacattatcg atgcttttgc tggttgggat 420
ccaagataca gaattaaggt tagaattgtc tgtgctagag cttaccatgc tttgttcatg 480
aagaatatgt taattagacc aacaacggaa gaattaaaga actttggtga gcctgatttc 540
accatttgga atgcaggtca attccctgct aatgtttaca ctaagggtat gacttcttca 600
acttctgttg aaataaattt caagtctthr ysgymtthrs rsrthrsrva gtasnhyssr 660
atggaaatgg ttatcctagg tactgaatac gcaggtgaaa tgaagaaagg tatctttacc 720
gttatgttct acttgatgcc aatcagacac aaggttttaa ctttacactc ttctgcaaat 780
caaggtaaaa aggatggtga tgtcacatta ttctttggtt tatctggtac aggtaaaaca 840
accttgtctg cagatcctca tagagaattg attggtgatg atgaacattg ctggtctgat 900
catggtgttt tcaacattga aggtggatgt tatgctaagt gtttggactt atctgctgaa 960
agagaacctg agattttcaa tgcaattagg tttggatctg tcttggagaa tgttgtctat 1020
gatccagttg atagaactgt tgactattcc gctgctaatg tcactgaaaa tactagatgt 1080
gcttatccta tcgactttat tccttctgct aagatcccat gtctggcaga ttctcatcca 1140
aagaatattg ttcttttaac ttgtgatgca agaggtgttt tgccacctgt ctccaagcta 1200
actaatgcac aagtcatgta tcactttatc tctggttaca cctccaagat ggcaggtacc 1260
gaagttggtg tcactgaacc agaagcaacc ttctctgcat gttttggtca acctttctta 1320
gttttacatc caatgaaata cgcacaacaa ctctctgata aaatggctga acattcttcc 1380
accgcttggt tattgaatac cggttggact ggtcaatctt atgttaaagg tggtaagaga 1440
tgtccattga agtatactag agcaatttta gatgctattc actctggtga gcttgcaaaa 1500
caggaattcg aaacataccc tactttcggt ttacaagttc caaaaacttg tccaggtgtc 1560
ccagaaagtg ttctgaaccc atctaaacac tgggctactg gtgaagctga tttcaaggct 1620
gaagtcacta acttggctaa attatttgct gagaactttg aaaagtattc tgcagaatgt 1680
actgcagaag ttgttgctgc tggtcctgct ttataa 1716
<210> 128
<211> 560
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 128
Met Ala Pro Thr Ala Val Asp Ile His Asn Glu Tyr Lys Gln Asn Val
1 5 10 15
Ser Asn Glu Gln Glu Ile Pro Phe Asn Lys Thr Glu Arg Lys Ser Ser
20 25 30
Ile Ala Ser Lys Leu Gly Leu Asn Pro Asp Ala Lys Ile His Tyr Asn
35 40 45
Ser Ala Val Pro Ile Leu Tyr Glu Asp Gly Leu Lys Glu Lys Gly Thr
50 55 60
Thr Ile Ser Ser Ser Gly Ala Leu Ile Ala Phe Ser Gly Ser Lys Thr
65 70 75 80
Gly Arg Ser Pro Lys Asp Lys Arg Ile Val Asp Glu Glu Thr Ser Thr
85 90 95
Asp Asn Ile Trp Trp Gly Pro Val Asn Lys Lys Val Asp Glu Asn Thr
100 105 110
Trp Asn Ile Ser Lys Ser Arg Ala Ile Asp Tyr Leu Arg Thr Arg Glu
115 120 125
Lys Val Tyr Ile Ile Asp Ala Phe Ala Gly Trp Asp Pro Arg Tyr Arg
130 135 140
Ile Lys Val Arg Ile Val Cys Ala Arg Ala Tyr His Ala Leu Phe Met
145 150 155 160
Lys Asn Met Leu Ile Arg Pro Thr Thr Glu Glu Leu Lys Asn Phe Gly
165 170 175
Glu Pro Asp Phe Thr Ile Trp Asn Ala Gly Gln Phe Pro Ala Asn Val
180 185 190
Tyr Thr Lys Gly Met Thr Ser Ser Thr Ser Val Glu Ile Asn Phe Lys
195 200 205
Ser Met Glu Met Val Ile Leu Gly Thr Glu Tyr Ala Gly Glu Met Lys
210 215 220
Lys Gly Ile Phe Thr Val Met Phe Tyr Leu Met Pro Ile Arg His Lys
225 230 235 240
Val Leu Thr Leu His Ser Ser Ala Asn Gln Gly Lys Lys Asp Gly Asp
245 250 255
Val Thr Leu Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr Thr Leu Ser
260 265 270
Ala Asp Pro His Arg Glu Leu Ile Gly Asp Asp Glu His Cys Trp Ser
275 280 285
Asp His Gly Val Phe Asn Ile Glu Gly Gly Cys Tyr Ala Lys Cys Leu
290 295 300
Asp Leu Ser Ala Glu Arg Glu Pro Glu Ile Phe Asn Ala Ile Arg Phe
305 310 315 320
Gly Ser Val Leu Glu Asn Val Val Tyr Asp Pro Val Asp Arg Thr Val
325 330 335
Asp Tyr Ser Ala Ala Asn Val Thr Glu Asn Thr Arg Cys Ala Tyr Pro
340 345 350
Ile Asp Phe Ile Pro Ser Ala Lys Ile Pro Cys Leu Ala Asp Ser His
355 360 365
Pro Lys Asn Ile Val Leu Leu Thr Cys Asp Ala Arg Gly Val Leu Pro
370 375 380
Pro Val Ser Lys Leu Thr Asn Ala Gln Val Met Tyr His Phe Ile Ser
385 390 395 400
Gly Tyr Thr Ser Lys Met Ala Gly Thr Glu Val Gly Val Thr Glu Pro
405 410 415
Glu Ala Thr Phe Ser Ala Cys Phe Gly Gln Pro Phe Leu Val Leu His
420 425 430
Pro Met Lys Tyr Ala Gln Gln Leu Ser Asp Lys Met Ala Glu His Ser
435 440 445
Ser Thr Ala Trp Leu Leu Asn Thr Gly Trp Thr Gly Gln Ser Tyr Val
450 455 460
Lys Gly Gly Lys Arg Cys Pro Leu Lys Tyr Thr Arg Ala Ile Leu Asp
465 470 475 480
Ala Ile His Ser Gly Glu Leu Ala Lys Gln Glu Phe Glu Thr Tyr Pro
485 490 495
Thr Phe Gly Leu Gln Val Pro Lys Thr Cys Pro Gly Val Pro Glu Ser
500 505 510
Val Leu Asn Pro Ser Lys His Trp Ala Thr Gly Glu Ala Asp Phe Lys
515 520 525
Ala Glu Val Thr Asn Leu Ala Lys Leu Phe Ala Glu Asn Phe Glu Lys
530 535 540
Tyr Ser Ala Glu Cys Thr Ala Glu Val Val Ala Ala Gly Pro Ala Leu
545 550 555 560
<210> 129
<211> 267
<212> PRT
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 129
Met Ser Gln Gly Arg Lys Ala Ala Glu Arg Leu Ala Lys Lys Thr Val
1 5 10 15
Leu Ile Thr Gly Ala Ser Ala Gly Ile Gly Lys Ala Thr Ala Leu Glu
20 25 30
Tyr Leu Glu Ala Ser Asn Gly Asp Met Lys Leu Ile Leu Ala Ala Arg
35 40 45
Arg Leu Glu Lys Leu Glu Glu Leu Lys Lys Thr Ile Asp Gln Glu Phe
50 55 60
Pro Asn Ala Lys Val His Val Ala Gln Leu Asp Ile Thr Gln Ala Glu
65 70 75 80
Lys Ile Lys Pro Phe Ile Glu Asn Leu Pro Gln Glu Phe Lys Asp Ile
85 90 95
Asp Ile Leu Val Asn Asn Ala Gly Lys Ala Leu Gly Ser Asp Arg Val
100 105 110
Gly Gln Ile Ala Thr Glu Asp Ile Gln Asp Val Phe Asp Thr Asn Val
115 120 125
Thr Ala Leu Ile Asn Ile Thr Gln Ala Val Leu Pro Ile Phe Gln Ala
130 135 140
Lys Asn Ser Gly Asp Ile Val Asn Leu Gly Ser Ile Ala Gly Arg Asp
145 150 155 160
Ala Tyr Pro Thr Gly Ser Ile Tyr Cys Ala Ser Lys Phe Ala Val Gly
165 170 175
Ala Phe Thr Asp Ser Leu Arg Lys Glu Leu Ile Asn Thr Lys Ile Arg
180 185 190
Val Ile Leu Ile Ala Pro Gly Leu Val Glu Thr Glu Phe Ser Leu Val
195 200 205
Arg Tyr Arg Gly Asn Glu Glu Gln Ala Lys Asn Val Tyr Lys Asp Thr
210 215 220
Thr Pro Leu Met Ala Asp Asp Val Ala Asp Leu Ile Val Tyr Ala Thr
225 230 235 240
Ser Arg Lys Gln Asn Thr Val Ile Ala Asp Thr Leu Ile Phe Pro Thr
245 250 255
Asn Gln Ala Ser Pro His His Ile Phe Arg Gly
260 265
<210> 130
<211> 420
<212> DNA
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 130
atgttaagaa ccatgttcaa atctaagatt cacagagcaa ctgttactca agcagatctc 60
cattatgttg gttccgttac tattgatgca gacttgttag acgcagcaga cttgttgcca 120
ggtgaattgg ttcacatcgt tgacattacg aacggtgcta gattggaaac ttacgtcatt 180
gaaggtgaac gtggttccgg tgttgttggt atcaatggtg ctgccgctca tttagttcat 240
cctggtgatc ttgttatcat tatctcctat gcacaagttt cagatgcaga agcacgtgca 300
ttgcgtccaa gagttgttca cgttgacaga gacaatagag ttgttgcgct tggtgcggat 360
ccagccgaac cagtcccagg ttccgaccaa gctagatccc cacaagctgt tactgcataa 420
<210> 131
<211> 384
<212> DNA
<213> 丙酮丁醇梭菌(Clostridium acetobutylicum)
<400> 131
atgcacttga acatgttgaa gtccaagatc cacagagcta ccgtcgttca agcagacttg 60
aactacgtcg gttccatcac catcgacaga aacttgatgg acaaggcaaa catcttggaa 120
tacgaaaagg tcgagatcgc aaacatcaac aacggtgcaa gattcgaaac ctacgtcatc 180
gctggtgagg ctggttccgg tatcatctgt ttgaacggtg ctgctgcaag atgtgcacaa 240
gcgggtgaca aggttatcat catgtgttac tgttccttga ccccagaaga agcttccgag 300
cacagaccaa aggtcgtttt cgtcaacgac gacaactcca tctccaacgt caccgaatac 360
gagaagcacg gcaccatcgg ttaa 384
<210> 132
<211> 354
<212> DNA
<213> 幽门螺杆菌(Helicobacter pylori)
<400> 132
atgaccttcg agatgttgta ctccaagatc cacagagcaa ccatcaccga cgcaaacttg 60
aactacatcg gctccatcac catcgacgag gacttggcta agttggctaa gttgagagag 120
ggtatgaagg tcgaaatcgt cgacgtcaac aacggcgaga gattctccac ctacgtcatc 180
ttgggtaaga agagaggtga aatctgcgtc aacggtgcag cagccagaaa ggtcgctatc 240
ggtgacgtcg tcatcatctt ggcttacgca tccatgaacg aggacgagat caacgctcac 300
aagccatcca tcgtcttggt cgacgaaaag aacgaaatct tggaaaaggg ttaa 354
<210> 133
<211> 117
<212> PRT
<213> 幽门螺杆菌(Helicobacter pylori)
<400> 133
Met Thr Phe Glu Met Leu Tyr Ser Lys Ile His Arg Ala Thr Ile Thr
1 5 10 15
Asp Ala Asn Leu Asn Tyr Ile Gly Ser Ile Thr Ile Asp Glu Asp Leu
20 25 30
Ala Lys Leu Ala Lys Leu Arg Glu Gly Met Lys Val Glu Ile Val Asp
35 40 45
Val Asn Asn Gly Glu Arg Phe Ser Thr Tyr Val Ile Leu Gly Lys Lys
50 55 60
Arg Gly Glu Ile Cys Val Asn Gly Ala Ala Ala Arg Lys Val Ala Ile
65 70 75 80
Gly Asp Val Val Ile Ile Leu Ala Tyr Ala Ser Met Asn Glu Asp Glu
85 90 95
Ile Asn Ala His Lys Pro Ser Ile Val Leu Val Asp Glu Lys Asn Glu
100 105 110
Ile Leu Glu Lys Gly
115
<210> 134
<211> 387
<212> DNA
<213> 芽孢杆菌属物种TS25(Bacillus sp. TS25)
<400> 134
atgtacagaa ccatgatgaa gtccaagttg cacagagcga ccgtcaccga agcaaacttg 60
aactacgtcg gttccatcac catcgaccaa gacttgatgg aagctgcaga catcttggaa 120
aacgagaagg tccaaatcgt caacaacaac aacggtgcta gattcgaaac ctacgtcatc 180
gctggtccaa gaggctccgg caccatctgt ttgaacggtg cagcagcaag attggtccaa 240
ccaggtgaca ccgttatcat catctcctac gcaatgttgg aagaagccga ggctagaaag 300
caccaacctg tcgtcgtttt gttgaaccca gacaacacca tccaagaatt gatcagagaa 360
acccacggtg ctaccgctac cgtctaa 387
<210> 135
<211> 128
<212> PRT
<213> 芽孢杆菌属物种TS25(Bacillus sp. TS25)
<400> 135
Met Tyr Arg Thr Met Met Lys Ser Lys Leu His Arg Ala Thr Val Thr
1 5 10 15
Glu Ala Asn Leu Asn Tyr Val Gly Ser Ile Thr Ile Asp Gln Asp Leu
20 25 30
Met Glu Ala Ala Asp Ile Leu Glu Asn Glu Lys Val Gln Ile Val Asn
35 40 45
Asn Asn Asn Gly Ala Arg Phe Glu Thr Tyr Val Ile Ala Gly Pro Arg
50 55 60
Gly Ser Gly Thr Ile Cys Leu Asn Gly Ala Ala Ala Arg Leu Val Gln
65 70 75 80
Pro Gly Asp Thr Val Ile Ile Ile Ser Tyr Ala Met Leu Glu Glu Ala
85 90 95
Glu Ala Arg Lys His Gln Pro Val Val Val Leu Leu Asn Pro Asp Asn
100 105 110
Thr Ile Gln Glu Leu Ile Arg Glu Thr His Gly Ala Thr Ala Thr Val
115 120 125
<210> 136
<211> 411
<212> DNA
<213> 谷氨酸棒杆菌(Corynebacterium glutamicum)
<400> 136
atgttgagaa ccatcttggg ttccaaaatc cacagagcta ccgttaccca agcagacttg 60
gactacgttg gttcagtcac catcgatgca gacttggttc atgccgcagg tttgatcgaa 120
ggtgaaaagg ttgccatcgt cgacatcacc aatggcgcta gattggaaac ctacgttatc 180
gttggtgatg ctggcaccgg taacatctgt atcaacggtg cagcagcaca cttgatcaac 240
ccaggtgatt tggttatcat catgtcctac ttgcaagcta ccgatgcaga ggccaaggct 300
tacgaaccaa agatcgttca cgtcgacgca gacaacagaa tcgttgcttt gggtaacgac 360
ttggcagaag ctttgcctgg ttccggtttg ttgacctcaa gatctatcta a 411
<210> 137
<211> 136
<212> PRT
<213> 谷氨酸棒杆菌(Corynebacterium glutamicum)
<400> 137
Met Leu Arg Thr Ile Leu Gly Ser Lys Ile His Arg Ala Thr Val Thr
1 5 10 15
Gln Ala Asp Leu Asp Tyr Val Gly Ser Val Thr Ile Asp Ala Asp Leu
20 25 30
Val His Ala Ala Gly Leu Ile Glu Gly Glu Lys Val Ala Ile Val Asp
35 40 45
Ile Thr Asn Gly Ala Arg Leu Glu Thr Tyr Val Ile Val Gly Asp Ala
50 55 60
Gly Thr Gly Asn Ile Cys Ile Asn Gly Ala Ala Ala His Leu Ile Asn
65 70 75 80
Pro Gly Asp Leu Val Ile Ile Met Ser Tyr Leu Gln Ala Thr Asp Ala
85 90 95
Glu Ala Lys Ala Tyr Glu Pro Lys Ile Val His Val Asp Ala Asp Asn
100 105 110
Arg Ile Val Ala Leu Gly Asn Asp Leu Ala Glu Ala Leu Pro Gly Ser
115 120 125
Gly Leu Leu Thr Ser Arg Ser Ile
130 135
<210> 138
<211> 384
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 138
atgtacagaa ccttgatgtc cgctaagttg cacagagcta gagtcaccga agcaaacttg 60
aactacgttg gttccgtcac catcgacgag gacttgttgg acgcagtcgg tatgatggca 120
aacgaaaagg tccaaatcgt caacaacaac aacggtgcta gattggaaac ctacatcatc 180
cctggtgaga gaggttccgg tgtcgtctgt ttgaacggtg ctgcagctag attggtccaa 240
gttggtgacg tcgttatcat cgtttcctac gccatgatgt ccgaagaaga agcaaagacc 300
cacaagccaa aggtcgctgt tttgaacgaa agaaacgaaa tcgaagagat gttgggtcaa 360
gaaccagcta gaaccatctt gtaa 384
<210> 139
<211> 127
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 139
Met Tyr Arg Thr Leu Met Ser Ala Lys Leu His Arg Ala Arg Val Thr
1 5 10 15
Glu Ala Asn Leu Asn Tyr Val Gly Ser Val Thr Ile Asp Glu Asp Leu
20 25 30
Leu Asp Ala Val Gly Met Met Ala Asn Glu Lys Val Gln Ile Val Asn
35 40 45
Asn Asn Asn Gly Ala Arg Leu Glu Thr Tyr Ile Ile Pro Gly Glu Arg
50 55 60
Gly Ser Gly Val Val Cys Leu Asn Gly Ala Ala Ala Arg Leu Val Gln
65 70 75 80
Val Gly Asp Val Val Ile Ile Val Ser Tyr Ala Met Met Ser Glu Glu
85 90 95
Glu Ala Lys Thr His Lys Pro Lys Val Ala Val Leu Asn Glu Arg Asn
100 105 110
Glu Ile Glu Glu Met Leu Gly Gln Glu Pro Ala Arg Thr Ile Leu
115 120 125
<210> 140
<211> 1356
<212> DNA
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 140
atgacaccac agccaaaccc acaagtcggt gcagcagtca aagctgcaga tagagcacac 60
gtcttccact cttggtctgc acaagaattg atcgatccat tggctgttgc tggtgcagag 120
ggttcctact tctgggatta cgatggtaga cgttaccttg acttcacctc cggcttagtc 180
ttcaccaaca tcggttacca acacccaaag gttgtcgcag ctatccaaga acaagccgca 240
tctttgacta catttgcccc agctttcgca gttgaagcaa gatccgaagc tgcaagattg 300
atcgctgagc gtactccagg tgatttagac aaaatcttct tcaccaacgg tggcgcagat 360
gctatcgagc acgctgttcg tatggcaaga atccacgctg gtagaccaaa ggtcttatcc 420
gcatacagat cataccacgg tggtacacaa caggcagtca acatcactgg tgatccaagg 480
agatgggcat ccgattccgc ttctgctggc gttgtccact tctgggctcc atacttatac 540
agatccagat tctacgccga aactgagcaa caagaatgtg agcgtgctct tgagcacttg 600
gaaactacta tcgccttcga aggtccaggt actattgccg ctatcgtttt ggaaactgtc 660
ccaggtactg ctggtatcat ggttcctcca ccaggttact tagcaggtgt tagagaattg 720
tgtgacaaat acggtatcgt cttcgtcttg gatgaagtca tggctggttt cggcagaact 780
ggcgaatggt tcgctgcaga cttattcgat gttaccccag acttgatgac cttcgctaag 840
ggtgtcaact caggttacgt tccattgggt ggtgttgcta tctccggcaa aatcgcagag 900
actttcggta agagagctta cccaggtggt ttgacgtact ccggtcaccc tcttgcttgc 960
gcagccgctg ttgctactat caacgttatg gcagaagaag gtgtcgttga aaacgctgca 1020
aacttgggtg ctagagttat cgaaccaggt ttgagagaac ttgcagagag acacccatca 1080
gttggtgaag ttagaggtgt tggtatgttc tgggctttgg aattggttaa ggatagagaa 1140
accagagaac ctttggtccc atacaatgcc gctggtgaag cgaacgcacc aatggctgct 1200
ttcggtgcag ctgcaaaggc aaacggtttg tggccattca tcaacatgaa cagaacccac 1260
gttgttcctc cttgtaacgt taccgaagcc gaagctaaag aaggcttggc agcattggat 1320
gcagctttat ctgttgcaga tgagtacact gtctaa 1356
<210> 141
<211> 1416
<212> DNA
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 141
atgtccatct gtgagcaata ctacccagaa gaaccaacca agccaaccgt caagaccgaa 60
tccatccctg gtccagaatc ccaaaagcaa ttgaaggaat tgggtgaagt tttcgacacc 120
agaccagctt acttcttggc agactacgaa aagtccttgg gtaactacat caccgacgtc 180
gacggtaaca cctacttgga cttgtacgct caaatctcct ccatcgcctt gggttacaac 240
aacccagcat tgatcaaagc cgctcaatcc ccagaaatga tcagagcatt ggttgacaga 300
ccagccttgg gtaacttccc ttccaaggac ttggacaaga tcttgaagca aatcttgaag 360
tccgctccaa agggtcagga ccacgtctgg tccggtttgt ccggtgcaga cgcaaacgaa 420
ttggctttca aggctgcatt catctactac agagctaagc aaagaggtta cgacgcagac 480
ttctccgaaa aggaaaactt gtccgttatg gacaacgacg caccaggtgc tccacacttg 540
gcagttttgt ccttcaagag agctttccac ggtagattgt tcgcatccgg ttccactacc 600
tgttccaagc caatccacaa gttggacttc cctgctttcc actggccaca cgccgaatac 660
ccatcctacc agtacccttt ggacgaaaac tccgacgcaa acagaaagga ggacgaccac 720
tgcttggcaa tcgttgaaga attgatcaag acctggtcca tccctgttgc tgcgttgatc 780
atcgaaccaa tccaatccga gggtggtgac aaccacgcct ccaagtactt cttgcaaaag 840
ttgagagaca tcaccttgaa gtacaacgtc gtctacatca tcgacgaagt ccaaactggc 900
gttggcgcaa ccggtaagtt gtggtgtcac gaatacgcag acatccagcc tccagtcgac 960
ttggtcacct tctccaagaa gttccaatcc gctggctact tcttccacga ccctaagttc 1020
atcccaaaca agccatacag acaattcaac acctggtgtg gtgaaccagc aagaatgatc 1080
atcgcaggtg caatcggtca agagatctcc gacaagaagt tgaccgaaca atgctccaga 1140
gtcggtgact acttgttcaa gaagttggaa ggtttgcaaa agaagtaccc agagaacttc 1200
caaaacttga gaggtaaggg tagaggcacc ttcatcgctt gggacttgcc aactggcgag 1260
aagagagact tgttgttgaa gaagttgaag ttgaacggct gtaacgttgg tggctgtgct 1320
gttcacgcag tcagattgag accttccttg accttcgaag agaagcacgc agacatcttc 1380
atcgaagctt tggctaagtc cgtcaacgaa ttgtaa 1416
<210> 142
<211> 1428
<212> DNA
<213> 克鲁维酵母(Saccharomyces kluyveri)
<400> 142
atgccatcct actccgttgc agaattgtac tacccagacg aacctaccga acctaagatc 60
tccacctcct cctacccagg tccaaaggca aagcaagaat tggaaaagtt gtccaacgtc 120
ttcgacacca gagcagctta cttgttggca gactactaca agtcccgtgg taactacatc 180
gttgaccagg acggtaacgt cttgttggac gtttacgctc aaatctcctc catcgccttg 240
ggttacaaca acccagaaat cttgaaggtt gcaaagtccg acgcaatgtc cgttgcattg 300
gccaaccgtc cagcattggc ttgtttccca tccaacgact acggtcaatt gttggaagac 360
ggtttgttga aggcagcacc acaaggtcaa gacaagatct ggaccgcttt gtccggttcc 420
gacgcaaacg aaaccgcctt caaagcctgc ttcatgtacc aagctgcgaa gaagagaaac 480
ggtagatcct tctccaccga agaattggaa tccgttatgg acaaccaatt gccaggcacc 540
tccgaaatgg ttatctgttc cttcgaaaag ggtttccacg gtcgtttgtt cggttccttg 600
tccactacca gatccaagcc tatccacaag ttggacatcc cagctttcaa ctggcctaaa 660
gccccattcc cagacttgaa gtacccattg gaagaaaaca aggaagccaa caaggctgaa 720
gaatcctcct gtatcgaaaa gttctcccaa atcgttcaag agtggcaagg taagatcgct 780
gcagttatca tcgaaccaat ccagtccgag ggtggcgaca accacgcttc ctccgacttc 840
ttccaaaagt tgagagaaat caccatcgaa aacggtatct tgatgatcgt cgacgaagtt 900
caaaccggtg tcggtgctac cggcaagatg tgggcacacg aacactggaa cttgtccaac 960
cctccagact tggttacctt ctccaagaag ttccaagcag caggtttcta ctaccacgac 1020
ccaaagttgc aaccagacca gccattcaga cagttcaaca cctggtgtgg tgacccatcc 1080
aaggctttga tcgccaaggt tatctacgaa gaaatcgtta agcacgactt ggtcaccaga 1140
actgccgaag tcggtaacta cttgttcaac agattggaaa agttgttcga aggtaagaac 1200
tacatccaga acttgagagg taagggtcaa ggcacctaca tcgctttcga cttcggcacc 1260
tcctccgaga gagactcctt cttgtccaga ttgagatgta acggtgcaaa cgtcgctggt 1320
tgcggtgact ccgctgtcag attgagacca tccttgacct tcgaagagaa gcacgcagac 1380
gtcttggttt ccatcttcga caagaccttg agacaattgt acggctaa 1428
<210> 143
<211> 747
<212> DNA
<213> 大肠杆菌(Escherichia coli)
<400> 143
atgatcgttt tggtcaccgg tgcaaccgca ggtttcggcg aatgtatcac cagaagattc 60
atccagcagg gtcacaaggt tatcgctacc ggtagaagac aagagagatt gcaagaattg 120
aaggacgagt tgggtgacaa cttgtacatc gctcaattgg acgttagaaa cagagcagct 180
atcgaagaaa tgttggcatc cttgccagct gaatggtgca acatcgacat cttggtcaac 240
aacgctggtt tggcattggg tatggaacca gctcacaagg ctagtgttga ggactgggag 300
accatgatcg acaccaacaa caagggtttg gtctacatga ccagagcagt tttgcctggt 360
atggttgaaa gaaaccacgg tcacatcatc aacatcggtt ccaccgctgg ttcctggcca 420
tacgctggcg gtaacgtcta cggtgctacc aaggctttcg ttagacagtt ctccttgaac 480
ttgagaaccg acttgcacgg caccgctgtt agagttaccg acatcgaacc aggtttggtt 540
ggtggcaccg aattctccaa cgtcagattc aagggcgacg acggtaaggc tgaaaagacc 600
taccaaaaca ccgtcgcttt gaccccagaa gacgtttcag aggctgtttg gtgggtcagt 660
accttgccag cacacgtcaa catcaacacc ttggaaatga tgccagtcac ccaatcctac 720
gcaggtttga acgttcacag acaataa 747
<210> 144
<211> 804
<212> DNA
<213> 酿酒酵母(Saccharomyces cerevisiae)
<400> 144
atgtcccaag gtagaaaggc agcagaaaga ttggcaaaga agaccgtctt gatcaccggt 60
gcgtccgctg gtatcggtaa ggctaccgcg ttggagtact tggaagcatc caacggtgac 120
atgaagttga tcttggcagc aagaagattg gagaagttgg aagaattgaa gaagaccatc 180
gaccaagaat tcccaaacgc taaggtccac gttgcacaat tggacatcac ccaagcagag 240
aagatcaagc cattcatcga aaacttgcca caagaattca aggacatcga catcttggtc 300
aacaacgctg gtaaggcgtt gggttccgac agagttggtc aaatcgcaac cgaagacatc 360
caagacgtct tcgacaccaa cgtcaccgct ttgatcaaca tcacccaagc tgttttgcca 420
atcttccaag cgaagaactc cggtgacatc gtcaacttgg gttccatcgc tggtagagac 480
gcatacccaa ccggctccat ctactgcgcc tccaagttcg ctgtcggtgc tttcaccgac 540
tccttgagaa aggaattgat caacaccaag atcagagtca tcttgattgc ccctggtttg 600
gtcgaaaccg aattctcctt ggttagatac agaggtaacg aagaacaagc aaagaacgtt 660
tacaaggaca ctaccccatt gatggccgac gacgttgcag acttgatcgt ttacgctacc 720
tccagaaagc aaaacaccgt tatcgcagac accttgatct tcccaaccaa ccaagcatcc 780
ccacaccaca tcttcagagg ttaa 804
<210> 145
<211> 420
<212> DNA
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 145
atgcttagaa ccatgttcaa atccaagatc cacagagcaa ccgtcactca agcagatttg 60
cattacgttg gttctgttac tattgacgca gacttacttg atgcagccga tttacttcct 120
ggtgagcttg ttcatattgt tgatatcacc aatggtgcgc gtcttgaaac ctatgtcatt 180
gagggtgaac gtggttccgg tgtcgtcggt atcaatggcg cagctgcaca cctcgtccat 240
ccaggtgacc tggtcatcat catttcttat gcacaggtct ccgatgctga agcccgtgcc 300
ttaagaccaa gagtcgttca cgtcgacaga gataacagag ttgtcgcttt aggtgcagac 360
ccagcagagc cagttccagg ttccgatcaa gctagatcac cacaggctgt taccgcctaa 420
<210> 146
<211> 420
<212> DNA
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 146
atgttaagaa ctatgtttaa gtccaagatt cacagagcta ccgtcaccca agcagatttg 60
cattacgtcg gttccgttac cattgatgca gatttactcg atgcggcaga cttgttacct 120
ggtgaactag ttcacattgt tgacattacc aatggtgcta gattggaaac ttacgtcatt 180
gaaggtgaaa gaggtagtgg tgtcgttggt atcaatggtg cagctgctca cttagttcac 240
ccaggtgact tagtcatcat catttcatat gcacaagttt ccgatgcgga agctagagca 300
ttaagaccaa gagttgttca tgttgataga gacaacagag tcgttgcact tggtgcagat 360
cctgctgaac cagttccagg ttcagatcaa gctaggtctc cacaagcagt tactgcataa 420
<210> 147
<211> 420
<212> DNA
<213> 除虫链霉菌(Streptomyces avermitilis)
<400> 147
atgttaagaa ctatgttcaa gagtaagatt catagggcta ccgttaccca ggcagatcta 60
cattatgttg gttctgttac cattgacgca gatttgttgg atgcagcaga cttgttacca 120
ggtgaactcg ttcacattgt cgatatcacc aacggtgcca gactggaaac ttatgtcatt 180
gaaggtgaga gaggcagtgg tgttgtcggc attaacggtg ccgcagcaca cttggttcat 240
ccaggtgact tggtcatcat catttcttac gcacaagtct ccgatgccga agctagagca 300
ttgagaccta gagtcgttca cgttgacaga gacaatagag ttgttgctct tggtgcagat 360
ccagctgagc cagttccagg ttccgatcaa gcgagatctc ctcaagctgt tactgcataa 420
<210> 148
<211> 384
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 148
atgtatagaa ccttgatgag tgcaaagctt cacagagcaa gagtcactga agcaaacttg 60
aactacgttg gttctgttac tattgacgaa gacttacttg atgcagtcgg tatgatggca 120
aacgagaaag ttcaaattgt caacaataac aatggtgcgc gtcttgaaac ctatatcatt 180
cctggtgaac gtggttccgg tgtcgtctgc ttgaatggcg cagctgcaag gctcgtccaa 240
gttggtgacg tcgtcatcat tgtttcttat gcaatgatgt ccgaagaaga agccaaaaca 300
cataagccaa aggtcgctgt cctcaatgaa agaaacgaaa ttgaggaaat gttaggtcag 360
gaaccagcga gaaccatctt ataa 384
<210> 149
<211> 384
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 149
atgtatagaa ccttgatgag tgcaaagctt cacagagcaa gagtcactga agcaaacttg 60
aactacgttg gttctgttac tattgacgaa gacttacttg atgcagtcgg tatgatggca 120
aacgagaaag ttcaaattgt caacaataac aatggtgcgc gtcttgaaac ctatatcatt 180
cctggtgaac gtggttccgg tgtcgtctgc ttgaatggcg cagctgcaag gctcgtccaa 240
gttggtgacg tcgtcatcat tgtttcttat gcaatgatgt ccgaagaaga agccaaaaca 300
cataagccaa aggtcgctgt cctcaatgaa agaaacgaaa ttgaggaaat gttaggtcag 360
gaaccagcga gaaccatctt ataa 384
<210> 150
<211> 384
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 150
atgtacagaa cgttaatgtc tgccaagtta cacagagcga gagttactga agcaaatctc 60
aactatgttg gttcagttac cattgatgag gatctattgg atgctgttgg tatgatggca 120
aatgaaaagg ttcaaatcgt caataacaac aatggtgcta gattagaaac ttacatcatt 180
cctggtgaaa gaggttcagg tgttgtttgc ttaaacggtg ctgccgcaag attagttcaa 240
gtcggtgatg ttgttatcat cgtctcttat gctatgatga gtgaggaaga agctaagact 300
cataagccta aggttgccgt tctcaatgag agaaacgaaa tcgaagaaat gcttggccaa 360
gaacctgcca gaaccatcct gtaa 384
<210> 151
<211> 384
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 151
atgtatagaa cattgatgtc tgcaaagttg catagggctc gtgttactga ggcaaatcta 60
aactacgtcg gttccgtcac aatcgatgaa gatctactcg atgccgttgg tatgatggcc 120
aatgaaaaag ttcaaattgt caacaacaac aacggtgcaa gattggagac ctatatcatt 180
cctggtgaaa gaggttcagg tgttgtttgt ctgaacggtg ctgctgcgag gttggtccaa 240
gtcggtgatg ttgtcattat cgtttcttat gcaatgatgt cagaagaaga agctaagacc 300
cataagccaa aggttgctgt tttgaacgaa agaaatgaga ttgaggaaat gttaggtcaa 360
gaaccagcaa gaacaatctt ataa 384
<210> 152
<211> 945
<212> DNA
<213> 勤奋生金球菌(Metallosphaera sedula)
<400> 152
atgaccgaaa aggtctccgt cgtcggtgca ggcgtcatcg gtgtcggttg ggctaccttg 60
ttcgcttcca agggttactc cgtctccttg tacaccgaaa agaaggaaac cttggacaag 120
ggtatcgaaa agttgcgtaa ctacgtccaa gtcatgaaga acaactccca aatcaccgaa 180
gacgtcaaca ccgtcatctc cagagtttcc cctactacca acttggacga agccgttaga 240
ggtgcaaact tcgtcatcga ggctgtcatc gaagactacg acgctaagaa gaagatcttc 300
ggttacttgg actccgtctt ggacaaggaa gttatcttgg catcctccac ctccggtttg 360
ttgatcaccg aagtccaaaa ggctatgtcc aagcacccag aaagagctgt catcgcccac 420
ccatggaacc caccacactt gttgcctttg gtcgaaatcg ttccaggtga gaagacctcc 480
atggaagtcg ttgagagaac caagtccttg atggaaaagt tggacagaat cgtcgtcgtt 540
ttgaagaagg aaatcccagg tttcatcggt aacagattgg cattcgcttt gttcagagaa 600
gctgtctact tggttgacga gggtgtcgca accgtcgagg acatcgacaa ggttatgact 660
gccgctatcg gcttgagatg ggccttcatg ggtccattct tgacctacca cttgggtggt 720
ggtgagggtg gtttggaata cttcttcaac agaggtttcg gttacggtgc caacgaatgg 780
atgcacacct tggctaagta cgacaagttc ccatacaccg gtgtcaccaa ggccatccag 840
caaatgaagg aatactcctt catcaagggt aagaccttcc aagagatctc caagtggaga 900
gacgaaaagt tgttgaaggt ctacaagttg gtctgggaaa agtaa 945
<210> 153
<211> 1728
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 153
atgactgaca aaatctccct aggtacttat ctgtttgaaa agttaaagga agcaggctct 60
tattccatct ttggtgttcc tggtgatttc aatttggcat tgttggacca cgtcaaggaa 120
gttgaaggca ttagatgggt cggtaacgct aacgagttga atgccggcta cgaagctgat 180
ggttatgcaa gaatcaatgg atttgcatcc ctaatcacca cctttggtgt cggtgaattg 240
tctgccgtca atgccattgc aggttcttat gctgaacacg tcccattgat ccatattgtt 300
ggtatgcctt ccttgtctgc tatgaagaac aacttgttgt tacaccatac cttgggtgac 360
acaagattcg acaacttcac cgaaatgtca aagaaaatca gtgcaaaggt tgaaattgtt 420
tacgatttgg aatcagctcc aaaattaatt aataacttga ttgaaaccgc ttatcacaca 480
aagagaccag tctacttggg acttccttcc aactttgctg atgaattggt tccagcggca 540
ttagttaagg aaaacaagtt acatttagaa gaacctctaa acaaccccgt tgctgaagaa 600
gaattcattc ataacgttgt tgaaatggtc aagaaggcag aaaaaccaat cattctcgtt 660
gacgcttgtg ctgcaagaca taacatttct aaggaagtga gagagttggc taaattgact 720
aaattccctg tcttcaccac cccaatgggt aaatctactg ttgatgaaga tgatgaagaa 780
ttctttggct tatacttggg ttctctatct gctccagatg ttaaggacat tgttggccca 840
accgattgta tcttatcctt aggtggttta ccttctgatt tcaacaccgg ttccttctca 900
tatggttaca ccactaagaa tgtcgttgaa ttccattcca actactgtaa attcaaatct 960
gcaacttatg aaaacttgat gatgaagggc gcagtccaaa gattgatcag cgaattgaag 1020
aatattaagt attccaatgt ctcaacttta tctccaccaa aatctaaatt tgcttacgaa 1080
tctgcaaagg ttgctccaga aggtatcatc actcaagatt acctgtggaa gagattatct 1140
tacttcttaa agccaagaga tatcattgtc actgaaactg gtacttcctc ctttggtgtc 1200
ttggctaccc acttaccaag agattcaaag tctatctccc aagtcttatg gggttccatt 1260
ggtttctcct taccagctgc agttggtgct gcatttgctg ctgaagatgc acacaaacaa 1320
actggcgaac aagaaagaag aactgttttg tttattggtg atggttcttt acaattgact 1380
gtccaatcaa tctcagatgc tgcaagatgg aacatcaagc catacatctt catcttaaac 1440
aacagaggtt acactatcga aaagttgatc cacggtcgtc atgaggacta caaccaaatt 1500
caaccatggg atcaccaatt gttattgaag ctctttgctg acaagaccca atatgaaaac 1560
catgttgtta aatccgctaa ggacttggac gctttgatga aggatgaagc attcaacaag 1620
gaagataaga ttagagtcat tgaattattc ttggatgaat tcgatgctcc agaaatcttg 1680
gttgctcaag ctaaattatc tgatgaaatc aactctaaag ccgcttaa 1728
<210> 154
<211> 575
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 154
Met Thr Asp Lys Ile Ser Leu Gly Thr Tyr Leu Phe Glu Lys Leu Lys
1 5 10 15
Glu Ala Gly Ser Tyr Ser Ile Phe Gly Val Pro Gly Asp Phe Asn Leu
20 25 30
Ala Leu Leu Asp His Val Lys Glu Val Glu Gly Ile Arg Trp Val Gly
35 40 45
Asn Ala Asn Glu Leu Asn Ala Gly Tyr Glu Ala Asp Gly Tyr Ala Arg
50 55 60
Ile Asn Gly Phe Ala Ser Leu Ile Thr Thr Phe Gly Val Gly Glu Leu
65 70 75 80
Ser Ala Val Asn Ala Ile Ala Gly Ser Tyr Ala Glu His Val Pro Leu
85 90 95
Ile His Ile Val Gly Met Pro Ser Leu Ser Ala Met Lys Asn Asn Leu
100 105 110
Leu Leu His His Thr Leu Gly Asp Thr Arg Phe Asp Asn Phe Thr Glu
115 120 125
Met Ser Lys Lys Ile Ser Ala Lys Val Glu Ile Val Tyr Asp Leu Glu
130 135 140
Ser Ala Pro Lys Leu Ile Asn Asn Leu Ile Glu Thr Ala Tyr His Thr
145 150 155 160
Lys Arg Pro Val Tyr Leu Gly Leu Pro Ser Asn Phe Ala Asp Glu Leu
165 170 175
Val Pro Ala Ala Leu Val Lys Glu Asn Lys Leu His Leu Glu Glu Pro
180 185 190
Leu Asn Asn Pro Val Ala Glu Glu Glu Phe Ile His Asn Val Val Glu
195 200 205
Met Val Lys Lys Ala Glu Lys Pro Ile Ile Leu Val Asp Ala Cys Ala
210 215 220
Ala Arg His Asn Ile Ser Lys Glu Val Arg Glu Leu Ala Lys Leu Thr
225 230 235 240
Lys Phe Pro Val Phe Thr Thr Pro Met Gly Lys Ser Thr Val Asp Glu
245 250 255
Asp Asp Glu Glu Phe Phe Gly Leu Tyr Leu Gly Ser Leu Ser Ala Pro
260 265 270
Asp Val Lys Asp Ile Val Gly Pro Thr Asp Cys Ile Leu Ser Leu Gly
275 280 285
Gly Leu Pro Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Gly Tyr Thr
290 295 300
Thr Lys Asn Val Val Glu Phe His Ser Asn Tyr Cys Lys Phe Lys Ser
305 310 315 320
Ala Thr Tyr Glu Asn Leu Met Met Lys Gly Ala Val Gln Arg Leu Ile
325 330 335
Ser Glu Leu Lys Asn Ile Lys Tyr Ser Asn Val Ser Thr Leu Ser Pro
340 345 350
Pro Lys Ser Lys Phe Ala Tyr Glu Ser Ala Lys Val Ala Pro Glu Gly
355 360 365
Ile Ile Thr Gln Asp Tyr Leu Trp Lys Arg Leu Ser Tyr Phe Leu Lys
370 375 380
Pro Arg Asp Ile Ile Val Thr Glu Thr Gly Thr Ser Ser Phe Gly Val
385 390 395 400
Leu Ala Thr His Leu Pro Arg Asp Ser Lys Ser Ile Ser Gln Val Leu
405 410 415
Trp Gly Ser Ile Gly Phe Ser Leu Pro Ala Ala Val Gly Ala Ala Phe
420 425 430
Ala Ala Glu Asp Ala His Lys Gln Thr Gly Glu Gln Glu Arg Arg Thr
435 440 445
Val Leu Phe Ile Gly Asp Gly Ser Leu Gln Leu Thr Val Gln Ser Ile
450 455 460
Ser Asp Ala Ala Arg Trp Asn Ile Lys Pro Tyr Ile Phe Ile Leu Asn
465 470 475 480
Asn Arg Gly Tyr Thr Ile Glu Lys Leu Ile His Gly Arg His Glu Asp
485 490 495
Tyr Asn Gln Ile Gln Pro Trp Asp His Gln Leu Leu Leu Lys Leu Phe
500 505 510
Ala Asp Lys Thr Gln Tyr Glu Asn His Val Val Lys Ser Ala Lys Asp
515 520 525
Leu Asp Ala Leu Met Lys Asp Glu Ala Phe Asn Lys Glu Asp Lys Ile
530 535 540
Arg Val Ile Glu Leu Phe Leu Asp Glu Phe Asp Ala Pro Glu Ile Leu
545 550 555 560
Val Ala Gln Ala Lys Leu Ser Asp Glu Ile Asn Ser Lys Ala Ala
565 570 575
<210> 155
<211> 1167
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 155
atggtgtccc ctgctgaaag attatctact attgcgtcca caatcaagcc aaacagaaaa 60
gattctacat cattacaacc agaagactat ccggaacatc cgttcaaggt gacggttgtt 120
ggttccggta actgggggtg tacaattgcc aaggttatag cggaaaacac cgttgagaga 180
cctcgtcaat ttcaaagaga tgttaatatg tgggtctatg aagaattgat tgaaggcgaa 240
aagttgactg aaatcataaa taccaaacac gaaaacgtca agtacttgcc aggtatcaag 300
ttgccagtta acgttgttgc agttccagac attgttgagg cttgtgcagg ctcagacttg 360
attgtcttta atattcctca ccaattttta ccaagaattt tatcccaatt aaagggtaag 420
gtgaatccaa aggctagagc aatttcttgt ttgaaaggtt tggatgtcaa tcctaatgga 480
tgtaagttgc tctccactgt tattactgaa gagttgggta tttattgtgg tgccttatca 540
ggtgctaatt tagctcctga agttgcacaa tgtaaatggt cggaaacaac tgttgcatat 600
acaattccgg acgatttcag aggtaaaggc aaggatattg accatcaaat tctaaagagt 660
ttgttccata gaccttattt ccatgttcgt gttattagtg atgttgcagg tatttccatt 720
gccggtgcac tcaagaatgt cgttgctatg gctgctggat ttgtcgaagg tttaggttgg 780
ggtgataatg caaaggctgc agtcatgaga ataggtttgg tggaaaccat tcaatttgcc 840
aagacttttt tcgatggctg tcatgctgca acctttactc atgaatctgc aggtgttgcc 900
gacctaatca ctacctgtgc cggcggccgt aacgttagag ttggtagata tatggcacaa 960
cattctgtct ctgcaacgga ggctgaagaa aagttgttga atggccaatc ctgtcaaggt 1020
atccacacaa ctagggaagt ttacgagttc ctctccaaca tgggcaggac agatgagttc 1080
ccactattta ccaccaccta ccgtatcatc tacgaaaact tcccaattga gaagctgcca 1140
gaatgccttg aacctgtgga agattaa 1167
<210> 156
<211> 388
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 156
Met Val Ser Pro Ala Glu Arg Leu Ser Thr Ile Ala Ser Thr Ile Lys
1 5 10 15
Pro Asn Arg Lys Asp Ser Thr Ser Leu Gln Pro Glu Asp Tyr Pro Glu
20 25 30
His Pro Phe Lys Val Thr Val Val Gly Ser Gly Asn Trp Gly Cys Thr
35 40 45
Ile Ala Lys Val Ile Ala Glu Asn Thr Val Glu Arg Pro Arg Gln Phe
50 55 60
Gln Arg Asp Val Asn Met Trp Val Tyr Glu Glu Leu Ile Glu Gly Glu
65 70 75 80
Lys Leu Thr Glu Ile Ile Asn Thr Lys His Glu Asn Val Lys Tyr Leu
85 90 95
Pro Gly Ile Lys Leu Pro Val Asn Val Val Ala Val Pro Asp Ile Val
100 105 110
Glu Ala Cys Ala Gly Ser Asp Leu Ile Val Phe Asn Ile Pro His Gln
115 120 125
Phe Leu Pro Arg Ile Leu Ser Gln Leu Lys Gly Lys Val Asn Pro Lys
130 135 140
Ala Arg Ala Ile Ser Cys Leu Lys Gly Leu Asp Val Asn Pro Asn Gly
145 150 155 160
Cys Lys Leu Leu Ser Thr Val Ile Thr Glu Glu Leu Gly Ile Tyr Cys
165 170 175
Gly Ala Leu Ser Gly Ala Asn Leu Ala Pro Glu Val Ala Gln Cys Lys
180 185 190
Trp Ser Glu Thr Thr Val Ala Tyr Thr Ile Pro Asp Asp Phe Arg Gly
195 200 205
Lys Gly Lys Asp Ile Asp His Gln Ile Leu Lys Ser Leu Phe His Arg
210 215 220
Pro Tyr Phe His Val Arg Val Ile Ser Asp Val Ala Gly Ile Ser Ile
225 230 235 240
Ala Gly Ala Leu Lys Asn Val Val Ala Met Ala Ala Gly Phe Val Glu
245 250 255
Gly Leu Gly Trp Gly Asp Asn Ala Lys Ala Ala Val Met Arg Ile Gly
260 265 270
Leu Val Glu Thr Ile Gln Phe Ala Lys Thr Phe Phe Asp Gly Cys His
275 280 285
Ala Ala Thr Phe Thr His Glu Ser Ala Gly Val Ala Asp Leu Ile Thr
290 295 300
Thr Cys Ala Gly Gly Arg Asn Val Arg Val Gly Arg Tyr Met Ala Gln
305 310 315 320
His Ser Val Ser Ala Thr Glu Ala Glu Glu Lys Leu Leu Asn Gly Gln
325 330 335
Ser Cys Gln Gly Ile His Thr Thr Arg Glu Val Tyr Glu Phe Leu Ser
340 345 350
Asn Met Gly Arg Thr Asp Glu Phe Pro Leu Phe Thr Thr Thr Tyr Arg
355 360 365
Ile Ile Tyr Glu Asn Phe Pro Ile Glu Lys Leu Pro Glu Cys Leu Glu
370 375 380
Pro Val Glu Asp
385
<210> 157
<211> 1683
<212> DNA
<213> 埃及伊蚊(Aedes aegypti)
<400> 157
atgccagcca acggaatgtt cgacgtcgct ctgcaggtca tcgacgactc caacgtgtct 60
agtggatcgg acagtgctgg tgtgtctgaa gatgaagatg ttcaactgtt ttgcagtaaa 120
gggaatacca tagtcccgaa gccgttgaaa aaatccatat cgaaaatcaa ggatgaagag 180
ttcagcaaaa ccgccaaagc aaacgagaaa cgatacgcaa gtcttccgag ccgtgaacat 240
caccagcaat tcttgaccga cttcctgtcg gaagtgctga acaatgccgt tttcaacgct 300
accgaacggg ctaacaaagt tctgaactgg gtggatccgg agcagctcaa gcgaaccttg 360
gacctggaac tgaaggacga gcccgattca catgagaagc tgctggaact gaccagggcc 420
accataaagc actcggtcaa aaccggacat ccctacttca tgaaccaact gttctcgtcc 480
gtcgatccgt acgggttcgc tggacagatc cttaccgatg cgttgaaccc cagtgtctac 540
acgttcgaag tgtctccggt gttcgtcctg atggaggaag tggtgctcaa agaaatgaga 600
accatcgttg gctacccgga cggaaccgga gatggcattt tctgcccagg tggttcgatg 660
gctaacggct attccattag ctgtgcccgc ttcaaacaca tgcccgatgt caagacaaaa 720
ggattacatt cacttccgcg cttggtaatc ttcacatctg aagacgctca ttattcggtg 780
aaaaaattgg catcgttcat gggcatcggc tcggacaacg tgtacccaat ccatacggac 840
gccatcggca agatcagagt ggatcatcta gagtcggaaa ttctgcgcgc caaatcggag 900
ggagccgtgc cgtttatggt gtcggccacc gcaggaacaa cagtgatcgg agcgtttgat 960
ccgctggagc agatcgcaga tctgtgcaaa aagtacaacc tctggatgca cgtggatgcc 1020
gcctggggtg gtggtgcact tatgtccaag aagtaccgat cgctgcttaa aggaatcgaa 1080
cgatcggact cggtcacctg gaacccacac aaattgctcg ccgctccgca gcaatgctcc 1140
accttcctga cccgccacga gggaatccta tcggagtgcc actcgaccaa cgcgacctat 1200
ctcttccaga aggacaaatt ctacgacacc cagtacgaca ccggcgataa gcacatccag 1260
tgtggtcgcc gtgccgacgt cctcaaattt tggttcatgt ggcgtgccaa gggcacttcc 1320
gggctggaac agcacatcga caaggtgttc gagaatgcgg agcacttcac cagcagtatt 1380
aaatcgcgag aaggttttga aatggtcgtc gagaatcccg agtgtaccaa cgtgtgcttc 1440
tggtacgtgc cacctggatt gcggaacgta ccacgtgaca gcgcagagtt caccgaacgg 1500
ctgcacaagg ttgcccccaa ggtcaaggaa cgcatgatgc gggaaggttc gatgatgatc 1560
acataccaac cgatccacga taagcccaac ttcttccgat tggtcctgca gaactcggcc 1620
ctggacaaat cggacatgaa ctacatcatc gatgaaatcg aacgacttgc tgcagatttg 1680
taa 1683
<210> 158
<211> 1683
<212> DNA
<213> 埃及伊蚊(Aedes aegypti)
<400> 158
atgccagcca acggtatgtt cgacgttgca ttgcaagtta tcgacgactc caacgtctcc 60
tccggttccg actccgctgg tgtctccgaa gacgaagacg tccaattgtt ctgttctaag 120
ggcaacacca tcgtcccaaa gccattgaag aagtccatct ccaagatcaa ggacgaagaa 180
ttctccaaga ccgctaaggc aaacgaaaag agatacgcct ccttgccatc cagagagcat 240
caccagcaat tcttgaccga cttcttgtcc gaagttttga acaacgcagt tttcaacgct 300
accgaaagag caaacaaggt cttgaactgg gttgacccag aacaattgaa gagaaccttg 360
gacttggaat tgaaggacga accagactcc cacgaaaagt tgttggaatt gaccagagca 420
accatcaagc actccgttaa gaccggtcac ccatacttca tgaaccaatt gttctcctcc 480
gtcgacccat acggtttcgc tggtcaaatc ttgaccgacg ctttgaaccc atccgtctac 540
accttcgaag tttccccagt cttcgttttg atggaagaag tcgttttgaa ggaaatgaga 600
accatcgttg gttacccaga cggcaccggt gacggtatct tctgtccagg tggttccatg 660
gcaaacggtt actccatctc ctgtgcaaga ttcaagcaca tgccagacgt taagaccaag 720
ggtttgcact ccttgccaag attggtcatc ttcacctccg aagacgccca ctactccgtc 780
aagaagttgg cttccttcat gggtatcggc tccgacaacg tctacccaat ccacaccgac 840
gcaatcggca agatcagagt cgaccacttg gaatccgaaa tcttgagagc caagtccgaa 900
ggtgcggttc ctttcatggt ctccgctact gctggcacta ccgttatcgg tgcattcgac 960
ccattggaac aaatcgcaga cttgtgcaag aagtacaact tgtggatgca cgtcgacgct 1020
gcttggggtg gtggtgcttt gatgtccaag aagtacagat ccttgttgaa gggtatcgaa 1080
agatccgact ccgttacctg gaacccacac aagttgttag ccgcacctca acaatgttcc 1140
accttcttga ccagacacga aggtatcttg tccgaatgtc actccaccaa cgcaacctac 1200
ttgttccaaa aggacaagtt ctacgacacc caatacgaca ccggtgacaa gcacatccaa 1260
tgtggtagaa gggcagacgt cttgaagttc tggttcatgt ggcgtgccaa gggcacctcc 1320
ggtttggaac aacacatcga caaggttttc gaaaacgctg aacacttcac ctcctccatc 1380
aagtccagag aaggtttcga aatggttgtt gagaacccag aatgtaccaa cgtctgtttc 1440
tggtacgtcc caccaggttt gagaaacgtc cctagagact ccgctgaatt caccgaaaga 1500
ttgcacaagg tcgcaccaaa ggtcaaggaa agaatgatga gagaaggttc catgatgatc 1560
acctaccaac ctatccacga caagccaaac ttcttccgtt tggttttgca aaactccgca 1620
ttggacaagt ccgacatgaa ctacatcatc gacgagatcg agagattggc agcagacttg 1680
taa 1683
<210> 159
<211> 1683
<212> DNA
<213> 埃及伊蚊(Aedes aegypti)
<400> 159
atgccagcca acggtatgtt cgacgttgct ttgcaagtta tcgacgactc caacgtttcc 60
tccggttccg actccgcagg tgtttccgaa gacgaagacg tccaattgtt ctgctccaag 120
ggtaacacca tcgtcccaaa gccattgaag aagtccatct ccaagatcaa ggacgaggaa 180
ttctccaaga ccgctaaggc aaacgaaaag agatacgcgt ccttgccttc cagagaacat 240
caccagcaat tcttgaccga cttcttgtcc gaagtcttga acaacgcagt tttcaacgct 300
accgaaagag caaacaaggt cttgaactgg gttgacccag agcaattgaa gagaaccttg 360
gacttggaat tgaaggacga accagactcc cacgaaaagt tgttggaatt gaccagggca 420
accatcaagc actccgtcaa gaccggtcac ccttacttca tgaaccaatt gttctcctcc 480
gtcgacccat acggtttcgc tggtcaaatc ttgaccgacg ctttgaaccc atccgtttac 540
accttcgaag tctccccagt tttcgtcttg atggaagagg ttgtcttgaa ggagatgaga 600
accatcgtcg gttacccaga cggcaccggt gacggcatct tctgtccagg tggttccatg 660
gcaaacggtt actccatctc ctgtgcaaga ttcaagcaca tgccagacgt caagaccaag 720
ggtttgcact ccttgccaag attggtcatc ttcacctccg aagacgctca ctactccgtc 780
aagaagttgg cttccttcat gggtatcggt tccgacaacg tctacccaat ccacaccgac 840
gcaatcggta agatcagagt tgaccacttg gaatccgaaa tcttgagagc taagtccgaa 900
ggtgcagtcc ctttcatggt ctccgcaacc gcaggcacta ccgtcatcgg tgcattcgac 960
cctttggagc aaatcgcaga cttgtgtaag aagtacaact tgtggatgca cgttgacgct 1020
gcttggggtg gtggcgcatt gatgtccaag aagtacagat ccttgttgaa gggtatcgag 1080
agatccgact ccgtcacctg gaaccctcac aagttgttag ccgctccaca gcaatgttcc 1140
accttcttga ccagacacga aggtatcttg tccgaatgtc actccaccaa cgcaacctac 1200
ttgttccaaa aggacaagtt ctacgacacc caatacgaca ccggtgacaa gcacatccaa 1260
tgcggtagaa gggcagacgt cttgaagttc tggttcatgt ggcgtgcaaa gggcacctcc 1320
ggtttggagc aacacatcga caaggtcttc gaaaacgccg aacacttcac ctcctccatc 1380
aagtcccgtg aaggtttcga gatggtcgtc gaaaacccag agtgcaccaa cgtctgtttc 1440
tggtacgtcc ctccaggttt gagaaacgtc ccacgtgact ccgctgaatt caccgagaga 1500
ttgcacaagg ttgccccaaa ggtcaaggaa cgtatgatga gagagggttc catgatgatc 1560
acctaccaac caatccacga caagccaaac ttcttcagat tggttttgca gaactccgca 1620
ttggacaagt ccgacatgaa ctacatcatc gacgagatcg aaagattggc tgccgacttg 1680
taa 1683
<210> 160
<211> 1683
<212> DNA
<213> 埃及伊蚊(Aedes aegypti)
<400> 160
atgcctgcaa acggtatgtt cgacgtcgca ttgcaagtta tcgacgactc caacgtctcc 60
tccggttccg actccgctgg tgtctccgaa gacgaagacg tccagttgtt ctgttccaag 120
ggtaacacca tcgtccctaa gccattgaag aagtccatct ccaagatcaa ggacgaagaa 180
ttctccaaga ccgcaaaggc aaacgaaaag agatacgctt ccttgccatc cagagagcat 240
caccagcaat tcttgaccga cttcttgtcc gaagttttga acaacgcagt cttcaacgct 300
accgagagag ccaacaaggt cttgaactgg gtcgacccag aacaattgaa gagaaccttg 360
gacttggaat tgaaggacga accagactcc cacgaaaagt tgttggagtt gaccagagct 420
accatcaagc actccgtcaa gaccggtcac ccttacttca tgaaccaatt gttctcctcc 480
gtcgacccat acggtttcgc aggccaaatc ttgaccgacg ctttgaaccc ttccgtctac 540
accttcgaag tctccccagt tttcgtcttg atggaagaag tcgttttgaa ggagatgaga 600
accatcgttg gttacccaga cggcaccggt gacggcatct tctgtccagg cggttccatg 660
gcaaacggtt actccatctc ctgtgcacgt ttcaagcaca tgccagacgt caagaccaag 720
ggtttgcact ccttgccaag gttggtcatc ttcacctccg aagacgccca ctactccgtt 780
aagaagttgg catccttcat gggtatcggt tccgacaacg tctacccaat ccacaccgac 840
gcaatcggta agatcagggt tgaccacttg gaatccgaaa tcttgagagc taagtccgaa 900
ggtgctgtcc cattcatggt ctccgcaacc gctggcacta ccgtcatcgg tgccttcgac 960
cctttggaac aaatcgccga cttgtgcaag aagtacaact tgtggatgca cgttgatgct 1020
gcgtggggtg gtggtgcgtt gatgtccaag aagtacagat ccttgttgaa gggtatcgaa 1080
cgttccgact ccgtcacctg gaacccacac aagttgttag cggcaccaca acagtgctcc 1140
accttcttga ccagacacga gggtatcttg tccgaatgcc actccaccaa cgctacctac 1200
ttgttccaga aggacaagtt ctacgacacc caatacgaca ctggcgacaa gcacatccag 1260
tgtggtagaa gagcagacgt tttgaagttc tggttcatgt ggagagctaa gggcacctcc 1320
ggtttggaac aacacatcga caaggtcttc gaaaacgccg aacacttcac ctcctccatc 1380
aagtccagag aaggtttcga aatggttgtc gaaaacccag aatgtaccaa cgtttgtttc 1440
tggtacgttc caccaggttt gaggaacgtc ccaagagact ccgcagagtt caccgaaaga 1500
ttgcacaagg tcgcacctaa ggttaaggaa agaatgatga gagagggttc catgatgatc 1560
acctaccaac ctatccacga caagccaaac ttcttcaggt tggtcttgca aaactccgcg 1620
ttggacaagt ccgacatgaa ctacatcatc gacgagatcg aaagattggc agcagacttg 1680
taa 1683
<210> 161
<211> 1683
<212> DNA
<213> 埃及伊蚊(Aedes aegypti)
<400> 161
atgcctgcaa acggtatgtt cgacgtcgct ttgcaagtca tcgacgactc caacgtctcc 60
tccggttccg actccgcagg cgtctccgaa gacgaagacg ttcaattgtt ctgttccaag 120
ggtaacacca tcgttccaaa gccattgaag aagtccatct ccaagatcaa ggacgaagaa 180
ttctccaaga ccgctaaggc aaacgagaag agatacgctt ccttgccatc cagagaacat 240
caccaacaat tcttgaccga cttcttgtcc gaagttttga acaacgctgt cttcaacgct 300
accgaaagag ccaacaaggt cttgaactgg gtcgacccag aacaattgaa gagaaccttg 360
gacttggaat tgaaggacga gccagactcc cacgagaagt tgttggaatt gaccagagct 420
accatcaagc actccgtcaa gaccggtcac ccatacttca tgaaccagtt gttctcctcc 480
gtcgacccat acggtttcgc tggtcaaatc ttgaccgacg cattgaaccc ttccgtttac 540
accttcgaag tttcccctgt cttcgtcttg atggaagaag tcgtcttgaa ggaaatgaga 600
accatcgtcg gttacccaga cggcactggc gacggtatct tctgtcctgg tggttccatg 660
gccaacggtt actccatctc ctgtgctaga ttcaagcaca tgccagacgt taagaccaag 720
ggtttgcact ccttgcctag attggtcatc ttcacctccg aagacgctca ctactccgtt 780
aagaagttgg cctccttcat gggtatcggt tccgacaacg tctacccaat ccacaccgac 840
gcaatcggta agatcagagt cgaccacttg gagtccgaaa tcttgagagc aaagtccgaa 900
ggcgcagtcc cattcatggt ctccgcgacc gcaggcacta ccgtcatcgg tgctttcgac 960
ccattggaac agatcgcaga cttgtgtaag aagtacaact tgtggatgca cgtcgatgcc 1020
gcttggggtg gtggcgcttt gatgtccaag aagtacagat ccttgttgaa gggcatcgag 1080
agatccgact ccgttacctg gaacccacac aagttgttgg ctgctccaca acaatgttcc 1140
accttcttga ccagacacga gggtatcttg tccgaatgtc actccaccaa cgctacctac 1200
ttgttccaaa aggacaagtt ctacgacacc caatacgaca ccggtgacaa gcacatccaa 1260
tgtggtagaa gggcagacgt cttgaagttc tggttcatgt ggagggcaaa gggcacctcc 1320
ggtttggagc aacacatcga caaggtcttc gaaaacgctg aacacttcac ctcctccatc 1380
aagtccagag aaggcttcga aatggtcgtt gaaaaccctg aatgtaccaa cgtctgcttc 1440
tggtacgtcc caccaggttt gaggaacgtc cctagagact ccgctgaatt caccgaacgt 1500
ttgcacaagg tcgcaccaaa ggttaaggaa aggatgatga gagaaggttc catgatgatc 1560
acctaccaac caatccacga caagccaaac ttcttcagat tggttttgca aaactccgca 1620
ttggacaagt ccgacatgaa ctacatcatc gacgaaatcg aaagattggc agcagacttg 1680
taa 1683
<210> 162
<211> 560
<212> PRT
<213> 埃及伊蚊(Aedes aegypti)
<400> 162
Met Pro Ala Asn Gly Met Phe Asp Val Ala Leu Gln Val Ile Asp Asp
1 5 10 15
Ser Asn Val Ser Ser Gly Ser Asp Ser Ala Gly Val Ser Glu Asp Glu
20 25 30
Asp Val Gln Leu Phe Cys Ser Lys Gly Asn Thr Ile Val Pro Lys Pro
35 40 45
Leu Lys Lys Ser Ile Ser Lys Ile Lys Asp Glu Glu Phe Ser Lys Thr
50 55 60
Ala Lys Ala Asn Glu Lys Arg Tyr Ala Ser Leu Pro Ser Arg Glu His
65 70 75 80
His Gln Gln Phe Leu Thr Asp Phe Leu Ser Glu Val Leu Asn Asn Ala
85 90 95
Val Phe Asn Ala Thr Glu Arg Ala Asn Lys Val Leu Asn Trp Val Asp
100 105 110
Pro Glu Gln Leu Lys Arg Thr Leu Asp Leu Glu Leu Lys Asp Glu Pro
115 120 125
Asp Ser His Glu Lys Leu Leu Glu Leu Thr Arg Ala Thr Ile Lys His
130 135 140
Ser Val Lys Thr Gly His Pro Tyr Phe Met Asn Gln Leu Phe Ser Ser
145 150 155 160
Val Asp Pro Tyr Gly Phe Ala Gly Gln Ile Leu Thr Asp Ala Leu Asn
165 170 175
Pro Ser Val Tyr Thr Phe Glu Val Ser Pro Val Phe Val Leu Met Glu
180 185 190
Glu Val Val Leu Lys Glu Met Arg Thr Ile Val Gly Tyr Pro Asp Gly
195 200 205
Thr Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser Met Ala Asn Gly Tyr
210 215 220
Ser Ile Ser Cys Ala Arg Phe Lys His Met Pro Asp Val Lys Thr Lys
225 230 235 240
Gly Leu His Ser Leu Pro Arg Leu Val Ile Phe Thr Ser Glu Asp Ala
245 250 255
His Tyr Ser Val Lys Lys Leu Ala Ser Phe Met Gly Ile Gly Ser Asp
260 265 270
Asn Val Tyr Pro Ile His Thr Asp Ala Ile Gly Lys Ile Arg Val Asp
275 280 285
His Leu Glu Ser Glu Ile Leu Arg Ala Lys Ser Glu Gly Ala Val Pro
290 295 300
Phe Met Val Ser Ala Thr Ala Gly Thr Thr Val Ile Gly Ala Phe Asp
305 310 315 320
Pro Leu Glu Gln Ile Ala Asp Leu Cys Lys Lys Tyr Asn Leu Trp Met
325 330 335
His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr
340 345 350
Arg Ser Leu Leu Lys Gly Ile Glu Arg Ser Asp Ser Val Thr Trp Asn
355 360 365
Pro His Lys Leu Leu Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Thr
370 375 380
Arg His Glu Gly Ile Leu Ser Glu Cys His Ser Thr Asn Ala Thr Tyr
385 390 395 400
Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Gln Tyr Asp Thr Gly Asp
405 410 415
Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe
420 425 430
Met Trp Arg Ala Lys Gly Thr Ser Gly Leu Glu Gln His Ile Asp Lys
435 440 445
Val Phe Glu Asn Ala Glu His Phe Thr Ser Ser Ile Lys Ser Arg Glu
450 455 460
Gly Phe Glu Met Val Val Glu Asn Pro Glu Cys Thr Asn Val Cys Phe
465 470 475 480
Trp Tyr Val Pro Pro Gly Leu Arg Asn Val Pro Arg Asp Ser Ala Glu
485 490 495
Phe Thr Glu Arg Leu His Lys Val Ala Pro Lys Val Lys Glu Arg Met
500 505 510
Met Arg Glu Gly Ser Met Met Ile Thr Tyr Gln Pro Ile His Asp Lys
515 520 525
Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Ala Leu Asp Lys Ser
530 535 540
Asp Met Asn Tyr Ile Ile Asp Glu Ile Glu Arg Leu Ala Ala Asp Leu
545 550 555 560
<210> 163
<211> 563
<212> PRT
<213> 云南致倦库蚊(Culex quinquefasciatus)
<400> 163
Met Pro Thr Asn Gly Met Phe Asp Val Ala Leu Gln Val Ile Glu Asp
1 5 10 15
Ala Asn Leu Ser Ser Gly Ser Asp Ser Ala Gly Val Ser Glu Asp Glu
20 25 30
Asp Val Gln Leu Phe Cys Thr Thr Gly Asn Val Val Ser Ser Lys Pro
35 40 45
Leu Lys Lys Pro Ser Leu Lys Pro Val Thr Thr Val Lys Asp Glu Asp
50 55 60
Gln Asn Lys Met Lys Thr Asn Ala Lys Arg Tyr Ala Ser Leu Pro Asn
65 70 75 80
Arg Glu Gln His Gln Arg Phe Leu Thr Asp Phe Leu Ser Glu Val Leu
85 90 95
Asn Asn Ala Ile Phe Asn Ala Thr Asp Arg Ser Asn Lys Val Leu Asn
100 105 110
Trp Val Asp Pro Glu Glu Leu Lys Arg Ser Ile Asp Leu Ser Leu Lys
115 120 125
Ala Glu Pro Asp Ser Asp Glu Lys Leu Leu Glu Leu Ala Arg Ala Thr
130 135 140
Ile Asp His Ser Val Lys Thr Gly His Pro Tyr Phe Met Asn Gln Leu
145 150 155 160
Phe Ser Ser Val Asp Val Tyr Gly Phe Ala Gly Gln Cys Leu Thr Asp
165 170 175
Ala Leu Asn Pro Ser Val Tyr Thr Phe Glu Val Ser Pro Val Phe Val
180 185 190
Leu Met Glu Glu Val Val Leu Lys Glu Met Arg Thr Ile Val Gly Phe
195 200 205
Pro Gly Gly Val Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser Met Ala
210 215 220
Asn Gly Tyr Ala Ile Ser Cys Ala Arg Phe Lys His Met Pro Asp Val
225 230 235 240
Lys Thr Lys Gly Leu His Ser Leu Pro Arg Leu Val Ile Phe Thr Ser
245 250 255
Glu Asp Ala His Tyr Ser Ile Lys Lys Leu Ala Ser Phe Met Gly Ile
260 265 270
Gly Ser Asp Asn Val Tyr Pro Ile Arg Thr Asp Ala Val Gly Lys Ile
275 280 285
Gln Pro Asp His Leu Glu Ala Glu Ile Leu Arg Ala Lys Ser Glu Gly
290 295 300
Ala Leu Pro Phe Met Val Ser Ala Thr Ala Gly Thr Thr Val Ile Gly
305 310 315 320
Ala Phe Asp Pro Leu Glu Gln Ile Ala Asp Leu Cys Gln Lys Tyr Asn
325 330 335
Leu Trp Met His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser
340 345 350
Lys Lys Tyr Arg Thr Leu Leu Lys Gly Val Glu Arg Ala Asp Ser Val
355 360 365
Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln Gln Cys Ser Thr
370 375 380
Phe Leu Thr Arg His Glu Gly Ile Leu Ser Gly Cys His Ser Thr Asn
385 390 395 400
Ala Thr Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Gln Tyr Asp
405 410 415
Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val Leu Lys
420 425 430
Phe Trp Phe Met Trp Arg Ala Lys Gly Thr Ser Gly Phe Glu Gln His
435 440 445
Ile Asp Lys Val Phe Glu Asn Ala Glu Tyr Phe Thr Asn Ser Ile Lys
450 455 460
Ala Arg Pro Gly Phe Glu Met Val Ile Glu Asn Pro Glu Cys Thr Asn
465 470 475 480
Val Cys Phe Trp Tyr Val Pro Pro Gly Leu Arg Gln Val Pro Arg Asp
485 490 495
Ser Ala Glu Phe Gly Glu Arg Leu His Lys Val Ala Pro Lys Val Lys
500 505 510
Glu Arg Met Met Arg Glu Gly Ser Met Met Ile Thr Tyr Gln Pro Ile
515 520 525
His Asp Lys Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly Leu
530 535 540
Asp Lys Ser Asp Met Asn Tyr Ile Ile Asp Glu Ile Glu Arg Leu Ala
545 550 555 560
Ser Asp Leu
<210> 164
<211> 567
<212> PRT
<213> 冈比亚按蚊(Anopheles gambiae)
<400> 164
Met Pro Ala Asn Gly Val Cys Ser Val Gly Leu Glu Val Ile Glu Asp
1 5 10 15
Asn Ala Thr Tyr Ala Ser Gly Ser Asp Ser Ala Gly Val Ser Glu Asp
20 25 30
Glu Asp Val Gln Gln Leu Phe Val Ser Gly Ala Asp Arg Val Thr Ser
35 40 45
Val Leu Pro Lys Lys Ser Asp Ile Arg Lys Ala Ser Gln Val Asp Glu
50 55 60
Gln Ala Ala Ala Ala Ala Ala Ala Ala Val Ser Glu Lys Arg Tyr Ala
65 70 75 80
Ser Leu Pro Asn Arg Glu Gln His Gln Gln Phe Leu Thr Gln Phe Leu
85 90 95
Thr Glu Val Leu Asn Ser Ala Val Phe Asn Ala Thr Asp Arg Ala Asn
100 105 110
Lys Val Leu Asn Trp Val Asp Pro Glu Glu Leu Gln Arg Thr Leu Asp
115 120 125
Leu Ala Leu Lys Asp Glu Pro Asp Thr His Glu Lys Leu Leu Glu Leu
130 135 140
Thr Arg Ala Thr Ile Arg His Ser Val Lys Thr Gly His Pro Tyr Phe
145 150 155 160
Met Asn Gln Leu Phe Ser Ser Val Asp Pro Tyr Gly Phe Ala Gly Gln
165 170 175
Val Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser
180 185 190
Pro Val Phe Val Leu Met Glu Glu Val Val Leu Arg Glu Met Arg Thr
195 200 205
Ile Val Gly Tyr Pro Asp Gly Glu Gly Asp Gly Ile Phe Ala Pro Gly
210 215 220
Gly Ser Met Ala Asn Gly Tyr Ala Ile Ser Cys Ala Arg His Lys Phe
225 230 235 240
Met Pro Asp Ile Lys Thr Lys Gly Leu His Ala Leu Pro Arg Leu Val
245 250 255
Ile Phe Thr Ser Glu Asp Ala His Tyr Ser Val Lys Lys Leu Ala Ser
260 265 270
Phe Met Gly Ile Gly Ser Asp Asn Val Tyr Ala Ile Lys Thr Asp Asn
275 280 285
Val Gly Lys Ile Arg Val Glu His Leu Glu Ser Glu Ile Leu Arg Ala
290 295 300
Lys Ser Glu Gly Ala Leu Pro Phe Met Val Ser Ala Thr Ala Gly Thr
305 310 315 320
Thr Val Ile Gly Ala Phe Asp Pro Leu Glu Gln Ile Ala Asp Leu Cys
325 330 335
Ala Lys Tyr Asn Leu Trp Met His Val Asp Ala Ala Trp Gly Gly Gly
340 345 350
Ala Leu Met Ser Lys Lys Tyr Arg Thr Leu Leu Lys Gly Ile Glu Arg
355 360 365
Ser Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln
370 375 380
Gln Cys Ser Thr Leu Leu Thr Arg His Arg Asn Ile Leu Ala Glu Ala
385 390 395 400
His Ser Thr Asn Ala Thr Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp
405 410 415
Thr Arg Tyr Asp Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg Ala
420 425 430
Asp Val Leu Lys Phe Trp Phe Met Trp Arg Ala Lys Gly Thr Ala Gly
435 440 445
Phe Glu Ala His Ile Asp Lys Val Phe Glu Asn Ala Glu His Phe Thr
450 455 460
Ser Ser Ile Lys Ala Arg Pro Gly Phe Glu Met Val Ile Glu Gln Pro
465 470 475 480
Glu Cys Thr Asn Val Cys Phe Trp Tyr Val Pro Pro Gly Leu Arg Gly
485 490 495
Val Pro Arg Asp Ser Ala Glu Tyr Arg Asp Arg Leu His Lys Val Ala
500 505 510
Pro Lys Val Lys Glu Arg Met Met Lys Asp Gly Ser Met Met Ile Thr
515 520 525
Tyr Gln Pro Ile His Asp Lys Pro Asn Phe Phe Arg Leu Val Leu Gln
530 535 540
Asn Ser Ser Leu Asp Lys Ser Asp Met Asn Tyr Ile Ile Asp Glu Ile
545 550 555 560
Glu Arg Leu Gly Lys Asp Leu
565
<210> 165
<211> 540
<212> PRT
<213> 赤拟谷盗(Tribolium castaneum)
<400> 165
Met Pro Ala Thr Gly Glu Asp Gln Asp Leu Val Gln Asp Leu Ile Glu
1 5 10 15
Glu Pro Ala Thr Phe Ser Asp Ala Val Leu Ser Ser Asp Glu Glu Leu
20 25 30
Phe His Gln Lys Cys Pro Lys Pro Ala Pro Ile Tyr Ser Pro Val Ser
35 40 45
Lys Pro Val Ser Phe Glu Ser Leu Pro Asn Arg Arg Leu His Glu Glu
50 55 60
Phe Leu Arg Ser Ser Val Asp Val Leu Leu Gln Glu Ala Val Phe Glu
65 70 75 80
Gly Thr Asn Arg Lys Asn Arg Val Leu Gln Trp Arg Glu Pro Glu Glu
85 90 95
Leu Arg Arg Leu Met Asp Phe Gly Val Arg Ser Ala Pro Ser Thr His
100 105 110
Glu Glu Leu Leu Glu Val Leu Lys Lys Val Val Thr Tyr Ser Val Lys
115 120 125
Thr Gly His Pro Tyr Phe Val Asn Gln Leu Phe Ser Ala Val Asp Pro
130 135 140
Tyr Gly Leu Val Ala Gln Trp Ala Thr Asp Ala Leu Asn Pro Ser Val
145 150 155 160
Tyr Thr Tyr Glu Val Ser Pro Val Phe Val Leu Met Glu Glu Val Val
165 170 175
Leu Arg Glu Met Arg Ala Ile Val Gly Phe Glu Gly Gly Lys Gly Asp
180 185 190
Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser
195 200 205
Cys Ala Arg Tyr Arg Phe Met Pro Asp Ile Lys Lys Lys Gly Leu His
210 215 220
Ser Leu Pro Arg Leu Val Leu Phe Thr Ser Glu Asp Ala His Tyr Ser
225 230 235 240
Ile Lys Lys Leu Ala Ser Phe Gln Gly Ile Gly Thr Asp Asn Val Tyr
245 250 255
Leu Ile Arg Thr Asp Ala Arg Gly Arg Met Asp Val Ser His Leu Val
260 265 270
Glu Glu Ile Glu Arg Ser Leu Arg Glu Gly Ala Ala Pro Phe Met Val
275 280 285
Ser Ala Thr Ala Gly Thr Thr Val Ile Gly Ala Phe Asp Pro Ile Glu
290 295 300
Lys Ile Ala Asp Val Cys Gln Lys Tyr Lys Leu Trp Leu His Val Asp
305 310 315 320
Ala Ala Trp Gly Gly Gly Ala Leu Val Ser Ala Lys His Arg His Leu
325 330 335
Leu Lys Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys
340 345 350
Leu Leu Thr Ala Pro Gln Gln Cys Ser Thr Leu Leu Leu Arg His Glu
355 360 365
Gly Val Leu Ala Glu Ala His Ser Thr Asn Ala Ala Tyr Leu Phe Gln
370 375 380
Lys Asp Lys Phe Tyr Asp Thr Lys Tyr Asp Thr Gly Asp Lys His Ile
385 390 395 400
Gln Cys Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys
405 410 415
Ala Lys Gly Thr Ser Gly Leu Glu Lys His Val Asp Lys Val Phe Glu
420 425 430
Asn Ala Arg Phe Phe Thr Asp Cys Ile Lys Asn Arg Glu Gly Phe Glu
435 440 445
Met Val Ile Ala Glu Pro Glu Tyr Thr Asn Ile Cys Phe Trp Tyr Val
450 455 460
Pro Lys Ser Leu Arg Gly Arg Lys Asp Glu Ala Asp Tyr Lys Asp Lys
465 470 475 480
Leu His Lys Val Ala Pro Arg Ile Lys Glu Arg Met Met Lys Glu Gly
485 490 495
Ser Met Met Val Thr Tyr Gln Ala Gln Lys Gly His Pro Asn Phe Phe
500 505 510
Arg Ile Val Phe Gln Asn Ser Gly Leu Asp Lys Ala Asp Met Val His
515 520 525
Leu Val Glu Glu Ile Glu Arg Leu Gly Ser Asp Leu
530 535 540
<210> 166
<211> 600
<212> PRT
<213> 红缘皮蠹(Attagenus smirnovi)
<220>
<221> 尚未归类的特征(misc_feature)
<222> (36)..(36)
<223> Xaa可以是任何天然存在的氨基酸
<400> 166
Met Val Thr Ile Trp Pro Thr Ser Pro Gln Gly Lys Val Ile Ala Ala
1 5 10 15
Arg Pro Ala Arg Asp Pro Ser Thr Ser Tyr Asn Gly Gly Phe Ala Ala
20 25 30
Gln Tyr Arg Xaa Phe Val Ile Gln Ser Cys Lys Gln Leu Asn Phe Ser
35 40 45
Ser Arg Gly Ile Thr Asn His Ala Arg Gln Arg Arg Ser Glu Lys Tyr
50 55 60
Asn Leu Glu Glu Asp Val Thr Asp Glu Ala Thr Ser Asn Ser Asp His
65 70 75 80
Ser Pro Ser Glu Asp Asp Asp Leu Tyr Val Arg His Thr Asn Gly Phe
85 90 95
Asn Ile Lys Pro Ser Lys Pro Ile Ile Glu Pro Arg Lys Phe Ala Thr
100 105 110
Phe Pro Ser Val Pro Asp Lys Glu His His Glu Asp Phe Ile Lys Ala
115 120 125
Cys Val Asp Ile Leu Leu Lys Glu Ala Val Phe Asp Gly Thr Asn Arg
130 135 140
Lys Asn Arg Val Leu Glu Trp His Ser Pro Glu Glu Leu Lys Lys Leu
145 150 155 160
Phe Asp Phe Asn Leu Arg Lys Ser Gly Ser Ser His Glu Glu Leu Thr
165 170 175
Arg Leu Ile Lys Asp Thr Ile His Tyr Ser Val Lys Thr Gly His Pro
180 185 190
Tyr Phe Val Asn Gln Leu Phe Ser Ser Val Asp Leu Tyr Gly Leu Val
195 200 205
Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu
210 215 220
Val Ser Pro Val Phe Thr Leu Met Glu Glu Thr Val Leu Arg Glu Met
225 230 235 240
Arg Thr Ile Val Gly Phe Glu Asp Gly Lys Gly Asp Gly Ile Phe Cys
245 250 255
Pro Gly Gly Ser Met Ala Asn Gly Tyr Ala Ile Ser Cys Ala Arg Tyr
260 265 270
Lys Phe Lys Pro Asp Ile Lys Asp Thr Gly Leu His Gly Leu Pro Arg
275 280 285
Leu Val Leu Phe Thr Ser Glu Asp Ala His Tyr Ser Ile Lys Lys Met
290 295 300
Ala Ser Leu Leu Gly Ile Gly Ser Asn Asn Val Tyr Leu Ile Lys Thr
305 310 315 320
Asp Glu Leu Gly Arg Met Ser Val Pro His Leu Val Glu Gln Ile Glu
325 330 335
Arg Val His Lys Glu Gly Gly Ala Pro Phe Met Val Ser Ala Thr Ala
340 345 350
Gly Thr Thr Val Leu Gly Ala Phe Asp Pro Ile Gln Glu Leu Ala Asp
355 360 365
Val Cys Glu Arg Tyr Asn Leu Trp Leu His Val Asp Ala Ala Trp Gly
370 375 380
Gly Gly Ala Leu Ile Ser Gln Arg His Arg His Leu Leu Thr Gly Ile
385 390 395 400
Asn Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Thr Ala
405 410 415
Pro Gln Gln Cys Ser Thr Leu Leu Leu Arg His Glu Gly Ile Leu Ala
420 425 430
Gly Ala His Ser Ala Asn Ala Ala Tyr Leu Phe Gln Lys Asp Lys Phe
435 440 445
Tyr Asp Thr Arg Tyr Asp Thr Gly Asp Lys His Val Gln Cys Gly Arg
450 455 460
Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys Gly Thr
465 470 475 480
Ile Gly Phe Glu Gln His Ile Asp Lys Val Phe Asp Asn Ala Lys Phe
485 490 495
Phe Ser Asp Asn Ile Arg His Arg Pro Gly Phe Arg Met Val Leu Glu
500 505 510
Asp Pro Glu Cys Thr Asn Ile Cys Phe Trp Tyr Val Pro Pro Ser Met
515 520 525
Arg Gly Cys Glu Asp Gln Gln Asp Phe Asn Glu Arg Leu His Lys Val
530 535 540
Ala Pro Lys Ile Lys Glu Arg Met Met Lys Glu Gly Ser Met Met Val
545 550 555 560
Thr Tyr Gln Pro Gln Lys Ser Leu Pro Asn Phe Phe Arg Ile Val Phe
565 570 575
Gln Asn Ser Gly Leu Glu Arg Ser Asp Met Leu His Leu Ile Lys Glu
580 585 590
Phe Glu Arg Leu Gly His Asp Leu
595 600
<210> 167
<211> 537
<212> PRT
<213> 豌豆蚜(Acyrthosiphon pisum)
<400> 167
Met Pro Ile Val Met Pro Ala Ala Thr Val Pro Thr Asp Tyr Ala Thr
1 5 10 15
Ala Arg Pro Val Glu Leu Met Val Thr Ala Ser Thr Leu Asp Glu Thr
20 25 30
Pro Cys Gly Lys Gly Pro Met Met Glu Ser Leu Ser Ala Ala Val Cys
35 40 45
Gly Tyr Lys Ser Ala Pro Asn Ala Ala Asp His Glu Ala Phe Val Arg
50 55 60
Asp Ala Val Arg Leu Met Leu Glu Gln Ala Val Phe Arg Gly Thr Asp
65 70 75 80
Arg Arg Arg Pro Val Leu Asn Trp Lys Ser Pro Glu Glu Leu Gln Ala
85 90 95
Ala Phe Asp Phe Ala Leu Asp Arg Ser Pro Thr Thr His Gly His Leu
100 105 110
Leu Asn Leu Ile Glu Asp Thr Ile Glu His Ser Val Lys Thr Gly His
115 120 125
Pro Tyr Phe Ile Asn Gln Leu Phe Ser Ser Val Asp Pro Tyr Gly Leu
130 135 140
Ile Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Phe
145 150 155 160
Glu Val Ala Pro Val Met Thr Ile Met Glu Glu Thr Val Leu Thr Glu
165 170 175
Met Arg Lys Phe Leu Gly Tyr Pro Asp Gly Lys Gly Asp Gly Ile Phe
180 185 190
Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Asn Cys Ala Arg
195 200 205
Phe Ser Ala Phe Pro Glu Val Lys Thr Arg Gly Met His Gly Leu Pro
210 215 220
Arg Leu Val Val Tyr Thr Ser Ala Asp Ala His Tyr Ser Ile Lys Lys
225 230 235 240
Leu Cys Ala Phe Glu Gly Ile Gly Ser Asp Asn Leu Tyr Leu Ile Asn
245 250 255
Thr Asp Ala Lys Gly Lys Met Asp Val Ser His Leu Arg Gln Gln Ile
260 265 270
Gln Arg Thr Leu Glu Glu Lys Ala Val Pro Ile Met Val Ser Ala Thr
275 280 285
Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Pro Ile Ala Glu Ile Ala
290 295 300
Asp Val Cys His Glu Tyr Gly Ile Trp Leu His Val Asp Ala Ala Trp
305 310 315 320
Gly Gly Gly Ala Leu Val Ser Lys Lys His Lys His Leu Leu Thr Gly
325 330 335
Ile Asp Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys Met Leu Thr
340 345 350
Ala Pro Gln Gln Cys Ser Thr Phe Leu Thr Lys His Glu Arg Val Leu
355 360 365
Thr Glu Ser Asn Ser Ser Cys Ala Gln Tyr Leu Phe Gln Lys Asp Lys
370 375 380
Phe Tyr Asp Thr Thr Tyr Asp Thr Gly Asp Lys His Ile Gln Cys Gly
385 390 395 400
Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys Ala Lys Gly
405 410 415
Thr Asp Gly Leu Glu Ala His Val Asp Glu Asn Phe Asp Asn Ala Lys
420 425 430
Tyr Phe Thr Glu Met Ile Arg Asn Arg Ala Gly Phe Lys Leu Val Leu
435 440 445
Glu Glu Pro Glu Tyr Thr Asn Ile Thr Phe Trp Tyr Ile Pro Pro Ser
450 455 460
Leu Arg Gly Arg Gln Asn Glu Pro Asp Phe Lys Asn Lys Leu His Lys
465 470 475 480
Val Ala Pro Arg Ile Lys Glu Arg Met Met Lys Glu Gly Thr Met Met
485 490 495
Ile Thr Tyr Gln Pro Ser Asp Asp Leu Pro Asn Phe Phe Arg Leu Val
500 505 510
Leu Gln Asn Ser Ser Leu Asp Gln Asn Asp Met Asp Tyr Phe Val Asn
515 520 525
Glu Ile Glu Arg Leu Gly Ser Asp Leu
530 535
<210> 168
<211> 576
<212> PRT
<213> 瑟车利亚果蝇(Drosophila sechellia)
<400> 168
Met Leu Ala Ser Glu Asn Phe Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Thr Ser Gly Asp Asp Leu Ala Ser Val Thr
20 25 30
Pro Leu Thr Ala Thr Ala Ala Leu Val Ala Ser Thr Pro Ser Pro Ala
35 40 45
Asp Ser Thr Ser Ala Val Ala Phe Glu Gln Ala Ser Lys Met Leu Ala
50 55 60
Thr Ala Ala Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Ile Thr Ser
65 70 75 80
Thr Lys Asp Asp Leu Ser Ser Phe Val Ala Ser His Pro Ala Ala Glu
85 90 95
Phe Glu Gly Phe Ile Arg Ala Cys Val Asp Glu Ile Ile Lys Leu Ala
100 105 110
Val Phe Gln Gly Thr Asn Arg Ser Ser Lys Val Val Glu Trp His Glu
115 120 125
Pro Ala Glu Leu Arg Gln Leu Phe Asp Phe Gln Leu Arg Glu Gln Gly
130 135 140
Glu Ser Gln Asp Lys Leu Arg Glu Leu Leu Arg Glu Thr Ile Arg Phe
145 150 155 160
Ser Val Lys Thr Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly
165 170 175
Val Asp Pro Tyr Ala Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn
180 185 190
Pro Ser Val Tyr Thr Tyr Glu Val Ala Pro Leu Phe Thr Leu Met Glu
195 200 205
Glu Gln Val Leu Ala Glu Met Arg Arg Ile Val Gly Phe Pro Asn Gly
210 215 220
Gly Gln Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly
225 230 235 240
Tyr Ala Ile Ser Cys Ala Arg Tyr Arg His Ser Pro Glu Ser Lys Lys
245 250 255
Asn Gly Leu Phe Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp
260 265 270
Ala His Tyr Ser Val Glu Lys Leu Ala Met Phe Met Gly Phe Gly Ser
275 280 285
Glu His Val Arg Lys Ile Ala Thr Asn Glu Val Gly Lys Met Arg Leu
290 295 300
Ser Asp Leu Glu Glu Gln Val Lys Gln Cys Leu Glu Asn Gly Trp Gln
305 310 315 320
Pro Leu Met Val Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala Phe
325 330 335
Asp Asp Leu Ala Gly Ile Ser Glu Leu Cys Lys Lys Tyr Asn Met Trp
340 345 350
Met His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys
355 360 365
Tyr Arg His Leu Leu Asn Gly Ile Glu Arg Ala Asp Ser Val Thr Trp
370 375 380
Asn Pro His Lys Leu Leu Ala Ala Ser Gln Gln Cys Ser Thr Phe Leu
385 390 395 400
Thr Arg His Gln Gln Val Leu Ala Gln Cys His Ser Thr Asn Ala Thr
405 410 415
Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly
420 425 430
Asp Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val Phe Lys Phe Trp
435 440 445
Phe Met Trp Lys Ala Lys Gly Thr Gln Gly Leu Glu Ala His Val Glu
450 455 460
Lys Val Phe Arg Met Ala Glu Phe Phe Thr Ala Lys Val Arg Glu Arg
465 470 475 480
Pro Gly Phe Glu Leu Val Leu Glu Ser Pro Glu Cys Thr Asn Ile Ser
485 490 495
Phe Trp Tyr Val Pro Pro Gly Leu Arg Glu Met Glu Arg Asn Arg Glu
500 505 510
Phe Tyr Asp Arg Leu His Lys Val Ala Pro Lys Val Lys Glu Gly Met
515 520 525
Ile Lys Lys Gly Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Gln Leu
530 535 540
Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Cys Leu Glu Glu Ser
545 550 555 560
Asp Met Val Tyr Phe Leu Asp Glu Ile Glu Ser Leu Ala Gln Asn Leu
565 570 575
<210> 169
<211> 575
<212> PRT
<213> 黑腹果蝇(Drosophila melanogaster)
<400> 169
Met Leu Ala Ser Glu Asn Phe Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Thr Ser Gly Asp Asp Leu Ala Ser Val Ser
20 25 30
Pro Leu Thr Ala Thr Ala Ala Leu Val Ala Ser Thr Ser Ser Pro Ala
35 40 45
Asp Ser Thr Ser Thr Val Ala Phe Glu Gln Ala Ser Lys Met Leu Ala
50 55 60
Asn Ala Ala Asn Asn Asn Asn Asn Asn Asn Asn Asn Ile Thr Ser Thr
65 70 75 80
Lys Asp Asp Leu Ser Ser Phe Val Ala Ser His Pro Ala Ala Glu Phe
85 90 95
Glu Gly Phe Ile Arg Ala Cys Val Asp Glu Ile Ile Lys Leu Ala Val
100 105 110
Phe Gln Gly Thr Asn Arg Ser Ser Lys Val Val Glu Trp His Glu Pro
115 120 125
Ala Glu Leu Arg Gln Leu Phe Asp Phe Gln Leu Arg Glu Gln Gly Glu
130 135 140
Ser Gln Asp Lys Leu Arg Glu Leu Leu Arg Glu Thr Ile Arg Phe Ser
145 150 155 160
Val Lys Thr Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val
165 170 175
Asp Pro Tyr Ala Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro
180 185 190
Ser Val Tyr Thr Tyr Glu Val Ala Pro Leu Phe Thr Leu Met Glu Glu
195 200 205
Gln Val Leu Ala Glu Met Arg Arg Ile Val Gly Phe Pro Asn Gly Gly
210 215 220
Gln Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr
225 230 235 240
Ala Ile Ser Cys Ala Arg Tyr Arg His Ser Pro Glu Ser Lys Lys Asn
245 250 255
Gly Leu Phe Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala
260 265 270
His Tyr Ser Val Glu Lys Leu Ala Met Phe Met Gly Phe Gly Ser Asp
275 280 285
His Val Arg Lys Ile Ala Thr Asn Glu Val Gly Lys Met Arg Leu Ser
290 295 300
Asp Leu Glu Lys Gln Val Lys Leu Cys Leu Glu Asn Gly Trp Gln Pro
305 310 315 320
Leu Met Val Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp
325 330 335
Asp Leu Ala Gly Ile Ser Glu Val Cys Lys Lys Tyr Asn Met Trp Met
340 345 350
His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr
355 360 365
Arg His Leu Leu Asn Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn
370 375 380
Pro His Lys Leu Leu Ala Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr
385 390 395 400
Arg His Gln Gln Val Leu Ala Gln Cys His Ser Thr Asn Ala Thr Tyr
405 410 415
Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly Asp
420 425 430
Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe
435 440 445
Met Trp Lys Ala Lys Gly Thr Gln Gly Leu Glu Ala His Val Glu Lys
450 455 460
Val Phe Arg Met Ala Glu Phe Phe Thr Ala Lys Val Arg Glu Arg Pro
465 470 475 480
Gly Phe Glu Leu Val Leu Glu Ser Pro Glu Cys Thr Asn Ile Ser Phe
485 490 495
Trp Tyr Val Pro Pro Gly Leu Arg Glu Met Glu Arg Asn Arg Glu Phe
500 505 510
Tyr Asp Arg Leu His Lys Val Ala Pro Lys Val Lys Glu Gly Met Ile
515 520 525
Lys Lys Gly Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Gln Leu Pro
530 535 540
Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp
545 550 555 560
Met Val Tyr Phe Leu Asp Glu Ile Glu Ser Leu Ala Gln Asn Leu
565 570 575
<210> 170
<211> 508
<212> PRT
<213> 大斑蝶(Danaus plexippus)
<400> 170
Met Arg Val Asp Ser Lys Ile Ile Val Lys Lys Glu Thr Gln Asp Glu
1 5 10 15
Asn Leu Tyr Gln Ser Leu Ala Glu Arg Ser Lys His Glu Glu Phe Leu
20 25 30
Arg Lys Ala Val Asp Leu Leu Val Glu Arg Val Val Phe Gly Arg Ser
35 40 45
Thr Arg Ser Ser Lys Val Val Glu Trp Ala Ala Pro Asp Glu Ile Lys
50 55 60
Lys Ala Ile Asp Leu Lys Pro Arg Leu Gly Pro Ala Ser His Asp Glu
65 70 75 80
Leu Leu Ala Phe Met Ala Asn Val Ala Arg Tyr Ser Val Asn Thr Gly
85 90 95
His Pro Tyr Phe Val Asn Gln Leu Phe Ser Ser Val Asp Pro Tyr Gly
100 105 110
Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr
115 120 125
Phe Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Glu Val Leu Arg
130 135 140
Glu Met Arg Lys Ile Val Gly Trp Pro Glu Gly Glu Gly Asp Gly Ile
145 150 155 160
Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys Ala
165 170 175
Arg His His Phe Tyr Pro Glu Val Lys Tyr Lys Gly Val His Ala Val
180 185 190
Pro Lys Leu Val Leu Phe Thr Ser Glu Leu Ala His Tyr Ser Thr Lys
195 200 205
Lys Met Ala Ala Phe Met Gly Ile Gly Ser Asp Asn Cys Val Asn Ile
210 215 220
Lys Thr Asp Asp Val Gly Lys Met Asn Ile Val Asp Leu Glu Met Lys
225 230 235 240
Ile Lys Ile Ala Ile Asp Asn Lys Cys Thr Pro Phe Met Val Thr Ala
245 250 255
Thr Ser Gly Thr Thr Val Phe Gly Ala Phe Asp Pro Leu Val Ala Ile
260 265 270
Ser Asp Leu Cys Lys Lys Tyr Asn Leu Trp Leu His Val Asp Ala Ala
275 280 285
Trp Gly Gly Gly Ala Leu Met Ser Lys Lys His Arg His Leu Leu Asn
290 295 300
Gly Ile Glu Leu Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
305 310 315 320
Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Thr Arg His Lys Lys Val
325 330 335
Leu Ser Glu Gly His Ser Ser Asn Ala Lys Tyr Leu Phe Gln Lys Asp
340 345 350
Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His Ile Gln Cys
355 360 365
Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys
370 375 380
Gly Thr Glu Gly Phe Glu Lys His Val Asp Lys Leu Phe Asp Asn Ala
385 390 395 400
Lys Tyr Phe Leu Asp His Ile Lys Gln Arg Glu Gly Phe Gln Leu Val
405 410 415
Ile Ala Glu Pro Gln Cys Thr Asn Ile Met Phe Trp Tyr Ile Pro Lys
420 425 430
Cys Leu Arg Gly Cys Glu Asn Asp Ala Asp Tyr Tyr Glu Arg Leu His
435 440 445
Lys Val Ala Pro Lys Ile Lys Glu Arg Met Ile Lys Glu Gly Ser Met
450 455 460
Met Val Thr Tyr Gln Pro Gln Gly Asp Leu Val Asn Phe Phe Arg Ile
465 470 475 480
Val Phe Gln Asn Ser Ala Leu Asp His Lys Asp Met Val Tyr Phe Ala
485 490 495
Asn Glu Phe Glu Arg Leu Gly Ser Asp Met Ile Val
500 505
<210> 171
<211> 570
<212> PRT
<213> 亚库巴果蝇(Drosophila yakuba)
<400> 171
Met Leu Ala Ser Glu Asn Phe Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Ser Gly Asp Asp Leu Ala Ser Ala Thr Pro
20 25 30
Leu Thr Ala Ala Ala Ala Leu Val Ala Thr Thr Ser Ser Pro Ala Asp
35 40 45
Ser Ser Ser Ala Val Ala Phe Glu Thr Ala Ser Lys Met Leu Ala Thr
50 55 60
Asn Asn Asn Asn Asn Asn Asn Ile Thr Ser Ser Lys Asp Asp Leu Ser
65 70 75 80
Ser Phe Val Ala Ser His Pro Ala Ala Glu Phe Glu Gly Phe Ile Arg
85 90 95
Ala Cys Val Asp Glu Ile Ile Lys Leu Ala Val Phe Gln Gly Thr Asn
100 105 110
Arg Ser Ser Lys Val Val Glu Trp His Glu Pro Ala Glu Leu Arg Gln
115 120 125
Leu Phe Asp Phe Gln Leu Arg Glu Lys Gly Glu Ser Gln Asp Lys Leu
130 135 140
Arg Glu Leu Leu Arg Glu Thr Ile Arg Phe Ser Val Lys Thr Gly His
145 150 155 160
Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro Tyr Ala Leu
165 170 175
Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr
180 185 190
Glu Val Ala Pro Leu Phe Thr Leu Met Glu Glu Gln Val Leu Ala Glu
195 200 205
Met Arg Arg Ile Val Gly Phe Pro Asn Gly Gly Gln Gly Asp Gly Ile
210 215 220
Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys Ala
225 230 235 240
Arg Tyr Arg His Ser Pro Glu Ser Lys Lys Asn Gly Leu Phe Asn Ala
245 250 255
Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala His Tyr Ser Val Glu
260 265 270
Lys Leu Ala Met Phe Met Gly Phe Gly Ser Glu His Val Arg Lys Ile
275 280 285
Ala Thr Asn Glu Val Gly Lys Met Arg Leu Ser Asp Leu Glu Glu Gln
290 295 300
Val Lys Gln Cys Leu Glu Asn Asn Trp Gln Pro Leu Met Val Ser Ala
305 310 315 320
Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Asp Leu Ala Gly Ile
325 330 335
Ser Glu Leu Cys Lys Lys Tyr Asn Met Trp Met His Val Asp Ala Ala
340 345 350
Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr Arg His Leu Leu Ser
355 360 365
Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
370 375 380
Ala Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr Arg His Gln Gln Val
385 390 395 400
Leu Ala Gln Cys His Ser Thr Asn Ala Thr Tyr Leu Phe Gln Lys Asp
405 410 415
Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly Asp Lys His Ile Gln Cys
420 425 430
Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys Ala Lys
435 440 445
Gly Thr Gln Gly Leu Glu Ala His Val Glu Lys Val Phe Arg Met Ala
450 455 460
Glu Phe Phe Thr Ala Lys Val Arg Glu Arg Pro Gly Phe Glu Leu Val
465 470 475 480
Leu Glu Ser Pro Glu Cys Thr Asn Ile Ser Phe Trp Tyr Val Pro Pro
485 490 495
Gly Leu Arg Glu Met Glu Arg Asn Arg Glu Phe Tyr Asp Arg Leu His
500 505 510
Lys Val Ala Pro Lys Val Lys Glu Gly Met Ile Lys Lys Gly Ser Met
515 520 525
Met Ile Thr Tyr Gln Pro Leu Arg Gln Leu Pro Asn Phe Phe Arg Leu
530 535 540
Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp Met Val Tyr Phe Leu
545 550 555 560
Asp Glu Ile Glu Ser Leu Ala Gln Asn Leu
565 570
<210> 172
<211> 572
<212> PRT
<213> 埃瑞克塔果蝇(Drosophila erecta)
<400> 172
Met Leu Ala Ser Glu Asn Phe Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Ser Gly Asp Asp Leu Ala Ser Val Thr Pro
20 25 30
Leu Thr Ala Ala Ala Ala Leu Val Ala Ser Thr Ser Ser Pro Ala Asp
35 40 45
Ser Ser Ser Ala Val Ala Phe Glu Thr Ala Ser Lys Met Leu Thr Thr
50 55 60
Thr Asn Ser Asn Asn Asn Asn Asn Asn Thr Thr Ser Ala Lys Asp Asp
65 70 75 80
Leu Ser Ser Phe Val Ala Ser His Pro Ala Ala Glu Phe Glu Gly Phe
85 90 95
Ile Arg Ala Cys Val Asp Glu Ile Ile Lys Leu Ala Val Phe Gln Gly
100 105 110
Thr Asn Arg Ser Ser Lys Val Val Glu Trp His Glu Pro Ala Glu Leu
115 120 125
Arg Gln Leu Phe Asp Phe Gln Leu Arg Glu Lys Gly Glu Ser Gln Asp
130 135 140
Lys Leu Arg Glu Leu Leu Arg Glu Thr Ile Arg Phe Ser Val Lys Thr
145 150 155 160
Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro Tyr
165 170 175
Ala Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr
180 185 190
Thr Tyr Glu Val Ala Pro Leu Phe Thr Leu Met Glu Glu Gln Val Leu
195 200 205
Ala Glu Met Arg Arg Ile Val Gly Phe Pro Asn Gly Gly Gln Gly Asp
210 215 220
Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser
225 230 235 240
Cys Ala Arg Tyr Arg His Ser Pro Glu Ser Lys Lys Asn Gly Leu Phe
245 250 255
Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala His Tyr Ser
260 265 270
Val Glu Lys Leu Ala Met Phe Met Gly Phe Gly Ser Glu His Val Arg
275 280 285
Lys Ile Ala Thr Asn Glu Val Gly Lys Met Arg Leu Ser Asp Leu Glu
290 295 300
Glu Gln Val Lys Gln Cys Leu Glu Asn Asp Trp Gln Pro Leu Met Val
305 310 315 320
Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Asp Leu Ala
325 330 335
Gly Ile Ser Asp Val Cys Lys Lys Tyr Asn Met Trp Met His Val Asp
340 345 350
Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr Arg His Leu
355 360 365
Leu Ser Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys
370 375 380
Leu Leu Ala Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr Arg His Gln
385 390 395 400
Gln Val Leu Ala Gln Cys His Ser Thr Asn Ala Thr Tyr Leu Phe Gln
405 410 415
Lys Asp Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly Asp Lys His Ile
420 425 430
Gln Cys Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys
435 440 445
Ala Lys Gly Thr Gln Gly Leu Glu Ala His Val Glu Lys Val Phe Arg
450 455 460
Met Ala Glu Phe Phe Thr Ala Lys Val Arg Glu Arg Pro Gly Phe Glu
465 470 475 480
Leu Val Leu Glu Ser Pro Glu Cys Thr Asn Ile Ser Phe Trp Tyr Val
485 490 495
Pro Pro Gly Leu Arg Glu Met Glu Arg Asn Arg Glu Phe Tyr Asp Arg
500 505 510
Leu His Lys Val Ala Pro Lys Val Lys Glu Gly Met Ile Lys Lys Gly
515 520 525
Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Gln Leu Pro Asn Phe Phe
530 535 540
Arg Leu Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp Met Val Tyr
545 550 555 560
Phe Leu Asp Glu Ile Glu Ser Leu Ala Gln Asn Leu
565 570
<210> 173
<211> 508
<212> PRT
<213> 柑橘凤蝶(Papilio xuthus)
<400> 173
Met Pro Ala Asp Ser Asn Leu Ile Val Ala Gly Glu Ala Ile Lys Glu
1 5 10 15
Ser Leu Phe Gln Ser Leu Pro Glu Arg Ser Lys His Glu Glu Phe Ile
20 25 30
Arg Arg Ala Val Asp Leu Leu Val Glu Arg Val Val Phe Gly Arg Ser
35 40 45
Gln Arg Ser Ala Lys Val Val Glu Trp Ala Ala Pro Asp Glu Ile Lys
50 55 60
Ser Val Ile Asp Leu Lys Pro Arg Glu Gly Pro Val Ser His Asp Glu
65 70 75 80
Leu Leu Ala Ile Met Ala Asp Val Ala Arg Tyr Ser Val Asn Thr Gly
85 90 95
His Pro Tyr Phe Val Asn Gln Leu Phe Ser Thr Val Asp Pro Tyr Gly
100 105 110
Leu Ile Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr
115 120 125
Tyr Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Glu Val Leu Arg
130 135 140
Glu Met Arg Ala Ile Val Gly Trp Lys Asp Gly Asp Gly Asp Gly Ile
145 150 155 160
Phe Cys Pro Gly Gly Ser Ile Ser Asn Gly Tyr Ala Ile Ser Cys Ala
165 170 175
Arg His His Phe Tyr Pro Asp Val Lys Ser Lys Gly Val Tyr Ala Val
180 185 190
Pro Lys Leu Val Leu Phe Thr Ser Glu Leu Ala His Tyr Ser Thr Lys
195 200 205
Lys Met Ala Cys Phe Met Gly Ile Gly Ser Asp Asn Cys Ile Met Ile
210 215 220
Lys Thr Asp Glu Leu Gly Lys Met Asp Val Gly Asp Leu Glu Ile Lys
225 230 235 240
Ile Ser Glu Ala Ile Asn Ser Gly Ser Thr Pro Phe Met Val Thr Ala
245 250 255
Thr Ala Gly Thr Thr Val Phe Gly Ala Phe Asp Pro Leu Ile Pro Ile
260 265 270
Ser Asp Leu Cys Lys Lys Tyr Asn Leu Trp Leu His Val Asp Ala Ala
275 280 285
Trp Gly Gly Gly Ala Leu Met Ser Lys Lys His Arg His Leu Leu Lys
290 295 300
Gly Ile Glu Leu Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
305 310 315 320
Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Val Arg His Lys Asn Val
325 330 335
Leu Lys Glu Gly His Ser Ser Asn Ala Lys Tyr Leu Phe Gln Lys Asp
340 345 350
Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His Ile Gln Cys
355 360 365
Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys
370 375 380
Gly Ser Asp Gly Phe Glu Lys His Ile Asp Lys Leu Phe Asp Asn Ala
385 390 395 400
Lys Tyr Phe Leu Asp His Ile Lys Gln Arg Ala Gly Phe Lys Leu Val
405 410 415
Leu Glu Asn Pro Glu Cys Thr Asn Ile Met Phe Trp Tyr Val Pro Asn
420 425 430
Cys Leu Arg Gly Cys Glu Asn Asp Pro Asn Tyr Arg Glu Arg Leu His
435 440 445
Lys Val Ala Pro Lys Ile Lys Glu Arg Met Ile Lys Glu Gly Ser Met
450 455 460
Met Val Thr Tyr Gln Pro Gln Gly Asn Leu Val Asn Phe Phe Arg Ile
465 470 475 480
Val Phe Gln Asn Ser Ala Leu Asp His Lys Asp Met Val Tyr Phe Ala
485 490 495
Asn Glu Phe Glu Arg Leu Gly Ser Asp Ile Ile Val
500 505
<210> 174
<211> 589
<212> PRT
<213> 黑翅果蝇(Drosophila persimilis)
<400> 174
Met Leu Ala Ser Glu Thr Phe Pro Ala His Arg Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Ser Gly Ser Ala Ser Val Asp Asp Leu Ala
20 25 30
Ser Val Asn Lys Thr Leu Thr Ser Ala Ala Thr Thr Ser Ser Ser Pro
35 40 45
Asp Thr His Ala Ala Val Asp Ile Ala Pro Thr Ala Ser Ser Val Glu
50 55 60
Phe Glu Thr Ala Arg Lys Met Leu Thr Asn Asn Ser Ser Ser Ser Ser
65 70 75 80
Asn Asn Ser Asn Asn Asn Asn Asn Asn Asn Asn Asn Asp Ala Lys Asp
85 90 95
Asp Ile Ser Gly Phe Val Ala Ser His Pro Ala Ala Pro Phe Glu Gly
100 105 110
Phe Ile Arg Ala Cys Val Asp Glu Ile Ile Lys Leu Ala Val Phe Gln
115 120 125
Gly Thr Asn Arg Ser Thr Lys Val Val Glu Trp His Glu Pro Ala Glu
130 135 140
Leu Arg Gln Leu Phe Asp Phe Gln Leu Arg Asp Lys Gly Glu Pro Gln
145 150 155 160
Glu Lys Leu Arg Glu Leu Leu Arg Glu Thr Ile Arg Phe Ser Val Lys
165 170 175
Thr Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro
180 185 190
Tyr Ala Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val
195 200 205
Tyr Thr Tyr Glu Val Ala Pro Leu Phe Thr Leu Met Glu Glu Gln Val
210 215 220
Leu Ala Glu Met Arg Arg Ile Val Gly Phe Pro Asn Gly Gly Lys Gly
225 230 235 240
Asp Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile
245 250 255
Ser Cys Ala Arg Tyr Thr Tyr Ala Pro Glu Ser Lys Lys Asn Gly Leu
260 265 270
Phe Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala His Tyr
275 280 285
Ser Val Glu Lys Leu Ala Met Phe Met Gly Phe Gly Ser Glu His Val
290 295 300
Val Lys Ile Ala Thr Asn Glu Val Gly Lys Met Arg Leu Ser Asp Leu
305 310 315 320
Glu Asp Gln Val Arg Arg Cys Leu Asp Asn Gly Trp Gln Pro Leu Met
325 330 335
Val Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Asp Leu
340 345 350
Thr Gly Ile Gly Asp Leu Cys Arg Lys Tyr Asn Met Trp Met His Val
355 360 365
Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr Arg His
370 375 380
Leu Leu Asn Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His
385 390 395 400
Lys Leu Leu Ala Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr Arg His
405 410 415
Gln Leu Val Leu Gly Gln Cys His Ser Thr Asn Ala Ala Tyr Leu Phe
420 425 430
Gln Lys Asp Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His
435 440 445
Ile Gln Cys Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp
450 455 460
Lys Ala Lys Gly Asn Leu Gly Leu Glu Ser His Val Glu Lys Val Phe
465 470 475 480
Arg Met Ala Glu Phe Phe Thr Ala Lys Val Arg Glu Arg Pro Gly Phe
485 490 495
Glu Leu Val Leu Glu Ser Pro Glu Cys Thr Asn Ile Ser Phe Trp Tyr
500 505 510
Val Pro Pro Ser Leu Arg Thr Met Glu Arg Asp Arg Glu Phe Tyr Asp
515 520 525
Lys Leu His Lys Val Ala Pro Lys Val Lys Glu Arg Met Ile Lys Lys
530 535 540
Gly Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Gln Leu Pro Asn Phe
545 550 555 560
Phe Arg Leu Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp Met Ile
565 570 575
Tyr Phe Leu Asp Glu Ile Glu Ser Leu Ala Lys Asn Leu
580 585
<210> 175
<211> 511
<212> PRT
<213> 家蚕(Bombyx mori)
<400> 175
Met Pro Ala Asp Ser Asp Leu Ile Val Ala Glu Arg Glu Asp Leu Gly
1 5 10 15
Ile Asp Arg Thr Leu Tyr Lys Ser Leu Ser Glu Arg Ser Lys His Glu
20 25 30
Asp Phe Ile Arg Arg Ala Val Asp Leu Leu Val Glu Arg Val Val Phe
35 40 45
Gly Arg Ser Leu Arg Thr Ser Lys Val Val Glu Trp Ala Val Pro Ser
50 55 60
Glu Ile Lys Lys Ala Ile Asp Leu Lys Pro Arg Asp Gly Pro Ile Ser
65 70 75 80
His Asp Glu Leu Leu Gly Leu Met Ala Asp Val Ala Arg Tyr Ser Val
85 90 95
Asn Thr Ala His Pro Tyr Phe Val Asn Gln Leu Tyr Ser Ser Val Asp
100 105 110
Pro Tyr Gly Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser
115 120 125
Val Tyr Thr Tyr Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Glu
130 135 140
Val Leu Lys Glu Met Arg Val Leu Val Gly Trp Lys Asp Gly Glu Gly
145 150 155 160
Asp Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile
165 170 175
Ser Cys Ala Arg Phe His Phe Tyr Pro Glu Ile Lys Thr Lys Gly Val
180 185 190
Tyr Ala Val Pro Lys Leu Thr Leu Tyr Thr Ser Glu Leu Ala His Tyr
195 200 205
Ser Thr Lys Lys Leu Ala Ala Phe Met Gly Ile Gly Asp Glu Asn Cys
210 215 220
Val Leu Ile Lys Thr Asp Lys Tyr Gly Lys Ile Asp Val Glu Asp Leu
225 230 235 240
Glu Ala Lys Ile Val Glu Gly Ile Glu Glu Gly Ala Ala Pro Phe Leu
245 250 255
Val Thr Ala Thr Ala Gly Thr Thr Val Phe Gly Ala Phe Asp Pro Leu
260 265 270
Val Pro Ile Ala Ala Leu Cys Lys Lys Tyr Asn Leu Trp Leu His Val
275 280 285
Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys His Arg His
290 295 300
Leu Leu Asn Gly Ile Glu Leu Ala Asp Ser Val Thr Trp Asn Pro His
305 310 315 320
Lys Leu Leu Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Ile Arg His
325 330 335
Lys Asn Val Leu Lys Glu Gly His Ser Cys Asn Ala Lys Tyr Leu Phe
340 345 350
Gln Lys Asp Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His
355 360 365
Ile Gln Cys Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp
370 375 380
Lys Ala Lys Gly Ser Asp Gly Phe Glu Asn His Ile Asp Thr Leu Phe
385 390 395 400
Asp Asn Ala Arg Phe Phe Leu Glu Gln Ile Arg Asn Arg Glu Gly Phe
405 410 415
Glu Leu Val Ile Glu Lys Pro Glu Cys Thr Asn Ile Met Phe Trp Tyr
420 425 430
Val Pro Arg Cys Leu Arg Gly Cys Glu Asn Glu Ser Asp Tyr Arg Glu
435 440 445
Arg Leu His Lys Val Ala Pro Lys Ile Lys Glu Leu Met Ile Lys Glu
450 455 460
Gly Ser Met Met Val Thr Tyr Gln Pro Gln Gly Asp Leu Val Asn Phe
465 470 475 480
Phe Arg Ile Val Phe Gln Asn Ser Ala Leu Asp His Lys Asp Met Ile
485 490 495
Tyr Phe Val Asn Glu Phe Glu Arg Leu Gly Arg Asp Ile Leu Val
500 505 510
<210> 176
<211> 578
<212> PRT
<213> 嗜凤梨果蝇(Drosophila ananassae)
<400> 176
Met Leu Ala Ser Lys Asn Tyr Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Ser Thr Ser Gly Asp Asp Leu Ala Ser Val Asn Thr
20 25 30
Leu Thr Ala Ser Ala Ala Ser Ala Ala Met Val Ala Thr Thr Ser Ser
35 40 45
Ser Ala Asp Thr Val Ala Val Asp Phe Glu Asn Ala Arg Arg Met Leu
50 55 60
Ala Thr Asn Gly Gly Ile Ala Ser Asn Gly Asn Asn Asn Asn Asn Val
65 70 75 80
Leu Asp Ser Lys Asp Ser Leu Ser Gly Phe Val Ala Ser His Pro Ala
85 90 95
Ala Gln Phe Asp Gly Phe Ile Arg Ala Cys Val Asp Glu Ile Ile Lys
100 105 110
Leu Ala Val Phe Gln Gly Thr Asn Arg Ser Ser Lys Val Val Glu Trp
115 120 125
His Glu Pro Ala Glu Leu Arg Gln Leu Phe Asp Phe Gln Leu Arg Glu
130 135 140
Lys Gly Glu Ser Gln Asp Lys Leu Arg Glu Leu Leu Arg Glu Thr Ile
145 150 155 160
Arg Phe Ser Val Lys Thr Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr
165 170 175
Ser Gly Val Asp Pro Tyr Ala Leu Val Gly Gln Trp Leu Thr Asp Ala
180 185 190
Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ala Pro Leu Phe Thr Leu
195 200 205
Met Glu Glu Gln Val Leu Ala Glu Met Arg Arg Ile Val Gly Phe Pro
210 215 220
Asn Gly Gly Gln Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala
225 230 235 240
Asn Gly Tyr Ala Ile Ser Cys Ala Arg Tyr Lys Tyr Thr Pro Glu Ser
245 250 255
Lys Lys Asn Gly Leu Phe Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser
260 265 270
Glu Asp Ala His Tyr Ser Val Glu Lys Leu Ala Met Phe Met Gly Phe
275 280 285
Gly Ser Glu His Val Arg Lys Ile Ala Thr Asn Glu Val Gly Lys Met
290 295 300
Arg Val Glu Asp Leu Glu Asn Gln Ile Lys Met Cys Leu Glu Asn Asn
305 310 315 320
Cys Gln Pro Leu Met Val Ser Ala Thr Ala Gly Thr Thr Val Leu Gly
325 330 335
Ala Phe Asp Asp Leu Val Gly Ile Ser Glu Leu Cys Lys Lys Tyr Asn
340 345 350
Met Trp Met His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser
355 360 365
Lys Lys Tyr Arg His Leu Leu Asn Gly Ile Glu Arg Ala Asp Ser Val
370 375 380
Thr Trp Asn Pro His Lys Leu Leu Ala Ala Ser Gln Gln Cys Ser Thr
385 390 395 400
Phe Leu Thr Pro His Gln Gln Ile Leu Ala Gln Cys His Ser Thr Asn
405 410 415
Ala Thr Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Ser Phe Asp
420 425 430
Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val Phe Lys
435 440 445
Phe Trp Phe Met Trp Lys Ala Lys Gly Ser Gln Gly Leu Glu Ala His
450 455 460
Val Glu Lys Val Phe Arg Met Ala Glu Phe Phe Thr Ala Lys Val Arg
465 470 475 480
Glu Arg Pro Gly Phe Glu Leu Val Leu Asp Gln Pro Glu Cys Thr Asn
485 490 495
Ile Ser Phe Trp Tyr Val Pro Pro Ser Leu Arg Gln Met Glu Arg Asn
500 505 510
Arg Glu Phe Tyr Asp Arg Leu His Lys Val Ala Pro Lys Val Lys Glu
515 520 525
Gly Met Ile Lys Lys Gly Ser Met Met Ile Thr Tyr Gln Pro Leu Arg
530 535 540
Gln Leu Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Cys Leu Glu
545 550 555 560
Glu Ser Asp Met Leu Tyr Phe Leu Asn Glu Ile Glu Ser Leu Ala Gln
565 570 575
Asn Leu
<210> 177
<211> 580
<212> PRT
<213> 漠海威果蝇(Drosophila mojavensis)
<400> 177
Met Leu Ala Ser Glu Asn Phe Gln Ala His Leu Tyr Asn Arg Ser Ser
1 5 10 15
Ile Tyr Lys Pro Tyr Asn Glu Glu Leu Ala Ser Met Ala Lys Gln Leu
20 25 30
Thr Thr Ala Thr Ala Asp Ala Ala Gly Ile Asp Ala Ala Gln Ala Ala
35 40 45
Val Asp Tyr Gly Ser Pro Ser Lys Gln Met Leu Gly Ser Asn Asn Gly
50 55 60
Ser Ser Ser Gly Ser Ser Asn Lys Thr Ser Ser Ala Asn Ser Asn Asn
65 70 75 80
Asn Asn Asn Asn Val Ala Asn Gly Leu Ser Ser Phe Val Ala Ser His
85 90 95
Pro Ser Ala Glu Phe Glu Gly Phe Ile Arg Ala Cys Val Asp Glu Ile
100 105 110
Ile Gln Leu Ala Val Leu Gln Gly Thr Asn Arg Ser Ser Lys Val Val
115 120 125
Glu Trp His Glu Pro Ala Glu Leu Arg Lys Leu Phe Asp Phe Glu Leu
130 135 140
Arg Asp Gln Pro Asp Ser Pro Asp Lys Leu Arg Gln Leu Leu Arg Glu
145 150 155 160
Thr Ile Arg Phe Ser Val Lys Thr Gly His Pro Tyr Phe Ile Asn Gln
165 170 175
Leu Tyr Ser Gly Val Asp Pro Tyr Ala Leu Ile Gly Gln Trp Leu Thr
180 185 190
Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ala Pro Val Phe
195 200 205
Thr Leu Met Glu Glu Gln Val Leu Gly Glu Met Arg Arg Ile Val Gly
210 215 220
Phe Pro Asn Asn Gly Gln Gly Asp Gly Ile Phe Cys Pro Gly Gly Ser
225 230 235 240
Ile Ala Asn Gly Tyr Ala Ile Ser Cys Ala Arg Tyr Gln Tyr Ala Pro
245 250 255
Glu Ser Lys Lys Asn Gly Leu Phe Asn Ala Lys Pro Leu Val Ile Phe
260 265 270
Thr Ser Glu Asp Ala His Tyr Ser Val Glu Lys Leu Ala Met Phe Met
275 280 285
Gly Phe Gly Ser Glu His Val Arg Lys Ile Ala Thr Asn Glu Leu Gly
290 295 300
Lys Met Arg Leu Ser Asp Leu Glu Gln Gln Ile Gln Phe Cys Leu Asp
305 310 315 320
Asn Asn Trp Gln Pro Leu Met Val Ser Ala Thr Ala Gly Thr Thr Val
325 330 335
Leu Gly Ala Phe Asp Asp Leu Leu Gly Ile Ser Glu Leu Cys Arg Lys
340 345 350
His Asn Met Trp Met His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu
355 360 365
Met Ser Lys Lys Tyr Arg Gln Leu Leu Asn Gly Ile Glu Arg Ala Asp
370 375 380
Ser Val Thr Trp Asn Pro His Lys Leu Leu Ser Ala Ser Gln Gln Cys
385 390 395 400
Ser Thr Phe Leu Thr Arg His Thr Gln Ile Leu Gly Gln Cys His Ser
405 410 415
Thr Asn Ala Ala Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr Ser
420 425 430
Phe Asp Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg Ala Asp Val
435 440 445
Phe Lys Phe Trp Phe Met Trp Lys Ala Lys Gly Thr Lys Gly Phe Glu
450 455 460
Ala His Val Glu Gln Val Phe Glu Met Ser Glu Tyr Phe Thr Asn Lys
465 470 475 480
Leu Arg Glu Arg Pro Gly Phe Glu Leu Val Leu Asp Lys Pro Glu Cys
485 490 495
Thr Asn Ile Thr Phe Trp Tyr Val Pro Pro Ser Leu Arg Gln Met Glu
500 505 510
Arg Asn Gln Glu Phe Tyr Asp Lys Leu His Lys Val Ala Pro Lys Ile
515 520 525
Lys Glu Ala Met Ile Lys Lys Gly Ser Met Met Ile Thr Tyr Gln Pro
530 535 540
Leu Arg Lys Leu Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Cys
545 550 555 560
Leu Asp Glu Ser Asp Met Leu Tyr Phe Leu Asn Glu Ile Glu Thr Leu
565 570 575
Gly Gln Lys Leu
580
<210> 178
<211> 588
<212> PRT
<213> 格瑞姆肖果蝇(Drosophila grimshawi)
<400> 178
Met Leu Ala Ser Lys Thr Phe Pro Thr His His Phe Lys Lys Ser Ile
1 5 10 15
Tyr Thr Thr Tyr Asn Gly Ala Ser Ala Pro Thr Asn Val Glu Asp Leu
20 25 30
Ala Asn Val Ala Lys Thr Leu Thr Thr Thr Thr Ser Ser Ser Ser Asp
35 40 45
Ser Thr Val Val Glu Ala Asn Thr Ser Pro Val Glu Phe Ser Thr Pro
50 55 60
Ser Lys Met Leu Ser Ser Thr Ser Thr Thr Thr Thr Thr Thr Thr Asn
65 70 75 80
Asn Asn Asn Asn Asn Asn Ser Asn Asn Asn Asn Asn Ile Val Asn Gly
85 90 95
Leu Ser Ser Phe Val Ala Ser His Pro Ala Ala Glu Phe Glu Gly Phe
100 105 110
Ile Arg Ala Cys Val Asp Glu Ile Ile His Leu Ala Val Phe Gln Gly
115 120 125
Thr Asp Arg Ala Ser Lys Val Val Glu Trp His Glu Pro Ala Glu Leu
130 135 140
Arg Lys Leu Phe Asp Phe Glu Leu Arg Glu Lys Gly Glu Ser Gln Glu
145 150 155 160
Lys Leu Arg Gln Leu Met Arg Glu Thr Ile Arg Tyr Ser Val Lys Thr
165 170 175
Gly His Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro Tyr
180 185 190
Ala Leu Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr
195 200 205
Thr Tyr Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Gln Val Leu
210 215 220
Ala Glu Met Arg Arg Ile Val Gly Phe Pro Asp Asn Gly His Gly Asp
225 230 235 240
Gly Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser
245 250 255
Cys Ala Arg Tyr Asn Tyr Ala Pro Glu Ser Lys Lys Asn Gly Leu Phe
260 265 270
Asn Ala Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala His Tyr Ser
275 280 285
Val Glu Lys Leu Ala Met Phe Met Gly Phe Gly Ser Glu Asn Val Arg
290 295 300
Lys Ile Ala Thr Asn Glu Val Gly Lys Met Arg Leu Ser Asp Leu Glu
305 310 315 320
Glu Gln Ile Gln Leu Cys Leu Asp Asn Asn Trp Gln Pro Leu Met Val
325 330 335
Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Asp Leu Val
340 345 350
Gly Ile Ser Glu Leu Cys Arg Lys His Asn Met Trp Met His Val Asp
355 360 365
Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr Arg His Leu
370 375 380
Leu Asn Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys
385 390 395 400
Leu Leu Ser Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr Arg His Ala
405 410 415
Gln Ile Leu Gly Gln Cys His Ser Thr Asn Ala Ala Tyr Leu Phe Gln
420 425 430
Lys Asp Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly Asp Lys His Ile
435 440 445
Gln Cys Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys
450 455 460
Ala Lys Gly Ser Lys Gly Phe Glu Ala His Val Glu Gln Val Phe Glu
465 470 475 480
Met Ser Glu Phe Phe Thr Ala Lys Leu Arg Glu Arg Pro Gly Phe Glu
485 490 495
Leu Val Leu Asp His Pro Glu Cys Thr Asn Ile Thr Phe Trp Tyr Val
500 505 510
Pro Pro Ser Leu Arg His Met Glu His Asn Gln Glu Phe Tyr Asp Lys
515 520 525
Leu His Lys Val Ala Pro Lys Ile Lys Glu Ala Met Ile Lys Lys Gly
530 535 540
Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Lys Leu Pro Asn Phe Phe
545 550 555 560
Arg Leu Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp Met Leu Tyr
565 570 575
Phe Ile Asn Glu Ile Glu Ser Leu Gly Gln Asn Leu
580 585
<210> 179
<211> 461
<212> PRT
<213> 桦尺蠖(Biston betularia)
<400> 179
Ser Gln Arg Ser Ala Lys Val Val Glu Trp Ala Ala Pro Glu Glu Ile
1 5 10 15
Lys Lys Ala Ile Asp Leu Lys Pro Arg Asp Gly Pro Ala Ser His Asp
20 25 30
Gln Leu Leu Gly Leu Met Ala Asp Val Ala Arg Tyr Ser Val Asn Thr
35 40 45
Gly His Pro Tyr Phe Val Asn Gln Leu Phe Ser Ser Val Asp Pro Tyr
50 55 60
Gly Leu Ile Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr
65 70 75 80
Thr Phe Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Glu Val Leu
85 90 95
Lys Glu Met Arg Ser Leu Val Gly Trp Lys Asn Gly Asp Gly Asp Gly
100 105 110
Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys
115 120 125
Ala Arg Phe Tyr Tyr Tyr Pro Asp Ile Lys Thr Lys Gly Val Tyr Ala
130 135 140
Val Pro Arg Leu Val Leu Phe Thr Ser Glu Leu Ala His Tyr Ser Thr
145 150 155 160
Lys Lys Met Ala Ala Phe Met Gly Ile Gly Ser Asp Asn Cys Ile Leu
165 170 175
Val Lys Ala Asp Lys Leu Gly Lys Met Asp Ala Glu Asp Leu Glu Val
180 185 190
Lys Ile Asn Glu Ala Leu Asp Asp Gly Ala Thr Pro Phe Leu Val Thr
195 200 205
Ala Thr Ala Gly Thr Thr Val Tyr Gly Ala Phe Asp Pro Leu Ala Gln
210 215 220
Ile Ser Ser Leu Cys Lys Lys Tyr Asn Leu Trp Leu His Val Asp Ala
225 230 235 240
Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys His Arg His Leu Leu
245 250 255
Thr Gly Ile Glu Leu Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu
260 265 270
Leu Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Ile Lys His Lys Asn
275 280 285
Val Leu Lys Asp Gly His Ser Ser Asn Ala Lys Tyr Leu Phe Gln Lys
290 295 300
Asp Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His Ile Gln
305 310 315 320
Cys Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala
325 330 335
Lys Gly Ser Glu Gly Phe Glu Gln His Ile Asp Thr Leu Phe Asp Asn
340 345 350
Ala Lys His Phe Val Tyr Leu Ile Arg Asn Arg Glu Gly Tyr Arg Leu
355 360 365
Val Ile Glu Glu Pro Glu Cys Thr Asn Ile Met Phe Trp Tyr Ile Pro
370 375 380
Lys Cys Leu Arg Gly Cys Glu Asn Glu Pro Asp Tyr Lys Glu Arg Leu
385 390 395 400
Asn Lys Val Ala Pro Lys Ile Lys Glu Arg Met Ile Lys Glu Gly Ser
405 410 415
Met Met Val Thr Tyr Gln Pro Gln Gly Asp Leu Ala Asn Phe Phe Arg
420 425 430
Ile Val Phe Gln Asn Ser Ala Leu Asp His Lys Asp Met Val Tyr Phe
435 440 445
Ala Asn Glu Phe Glu Arg Leu Gly Arg Asp Ile Val Val
450 455 460
<210> 180
<211> 583
<212> PRT
<213> 威尔斯托尼果蝇(Drosophila willistoni)
<400> 180
Met Leu Ala Ser Glu Asn Phe Pro Thr His His Phe Lys Glu Ser Ile
1 5 10 15
Phe Lys Pro Tyr Asn Ala Thr Thr Ser Ala Ala Asn Ala Ala Ala Ala
20 25 30
Ala Thr Val Glu Asp Leu Ala Asn Val Ala Lys Thr Leu Thr Ser Lys
35 40 45
Ser Thr Thr Ser Ser Ser Val Ala Ser Asp Ala Ala Ala Thr Val Ser
50 55 60
Leu Met Gly Ala Val Asp Ile Glu Thr Ala Arg Lys Met Leu Ala Asn
65 70 75 80
Asn Asn Val Asn Ile Asn Asn Asn Asn Asn Asn Asn Ile Asn Asn Asn
85 90 95
Asn Ser Lys Asp Thr Ala Glu Phe Glu Ser Phe Leu Arg Gly Cys Ile
100 105 110
Asp Glu Ile Ile Lys Leu Ala Val Val Glu Gly Thr Asn Arg Ser Ser
115 120 125
Lys Val Val Glu Trp His Glu Pro Ser Glu Leu Arg Gln Ile Phe Asp
130 135 140
Phe Gln Leu Arg Glu Lys Gly Glu Ser Gln Asp Lys Leu Arg Glu Leu
145 150 155 160
Leu Arg Glu Thr Ile Arg Phe Ser Val Lys Thr Gly His Pro Tyr Phe
165 170 175
Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro Tyr Ala Leu Val Gly Gln
180 185 190
Trp Leu Thr Asp Ser Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ala
195 200 205
Pro Val Phe Thr Leu Met Glu Glu Glu Val Leu Ala Glu Met Arg Arg
210 215 220
Ile Val Gly Phe Pro Asp Asn Gly Leu Gly Asp Gly Ile Phe Cys Pro
225 230 235 240
Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys Ala Arg Tyr Lys
245 250 255
Tyr Ala Pro Glu Ser Lys Lys Asn Gly Leu Phe Ser Gly Lys Pro Leu
260 265 270
Ile Ile Phe Thr Ser Glu Asp Ala His Tyr Ser Val Glu Lys Leu Ala
275 280 285
Met Phe Met Gly Phe Gly Ser Glu His Val Arg Lys Ile Ala Thr Asn
290 295 300
Glu Val Gly Lys Met Arg Leu Ser Asp Leu Glu Gln Gln Ile Gln Leu
305 310 315 320
Cys Leu Asp Asn Asn Trp Gln Pro Leu Met Val Ser Ala Thr Ala Gly
325 330 335
Thr Thr Val Leu Gly Ala Phe Asp Asp Leu Val Gly Ile Ser Glu Leu
340 345 350
Cys Arg Lys His Asn Met Trp Met His Val Asp Ala Ala Trp Gly Gly
355 360 365
Gly Ala Leu Met Ser Lys Lys Tyr Arg His Leu Leu Asn Gly Ile Glu
370 375 380
Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Ser
385 390 395 400
Gln Gln Cys Ser Thr Phe Leu Thr Arg His Gln Gln Ile Leu Gly Gln
405 410 415
Cys His Ser Thr Asn Ala Thr Tyr Leu Phe Gln Lys Asp Lys Phe Tyr
420 425 430
Asp Thr Ser Tyr Asp Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg
435 440 445
Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys Ala Lys Gly Ser Glu
450 455 460
Gly Leu Arg Ala His Val Glu Gln Val Phe Arg Met Ser Glu Tyr Phe
465 470 475 480
Thr Gln Gln Val Arg Glu Arg Pro Gly Phe Glu Leu Val Leu Glu Ser
485 490 495
Pro Glu Cys Thr Asn Ile Ser Phe Trp Tyr Ile Pro Pro Ser Leu Arg
500 505 510
His Met Glu Arg Asn Gln Glu Phe Tyr Asp Lys Leu His Lys Val Ala
515 520 525
Pro Lys Ile Lys Glu Gly Met Ile Lys Lys Gly Ser Met Met Ile Thr
530 535 540
Tyr Gln Pro Leu Arg Arg Leu Pro Asn Phe Phe Arg Leu Val Leu Gln
545 550 555 560
Asn Ser Cys Leu Glu Glu Ser Asp Met Leu Tyr Phe Leu Asn Glu Ile
565 570 575
Glu Ser Leu Gly His Gln Leu
580
<210> 181
<211> 548
<212> PRT
<213> 西方蜜蜂(Apis mellifera)
<400> 181
Met Pro Ala Asn Glu Glu Thr Leu Ser Met Thr Ser Asp Gln Ile Arg
1 5 10 15
Asn Asn Leu Leu Gly Lys Ala Cys Asp Asp Ser Met Gln Asn Asn Phe
20 25 30
Asp Cys Glu Ser Ser Gly Asp Asp Glu Glu Asp Tyr Gln Asn Asp Cys
35 40 45
Ser Arg Ser Ile Val Lys Glu Asn Phe Lys Glu Val Cys Asn Tyr Lys
50 55 60
Ser Leu Pro Val Arg Glu Ile His Lys Lys Phe Met Lys Ser Phe Val
65 70 75 80
Asn Leu Leu Leu Glu Glu Ala Val Phe Glu Gly Thr Leu Arg Arg Asn
85 90 95
Lys Val Val Glu Trp Ile Glu Pro Thr Thr Leu His Ser Ile Ile Asp
100 105 110
Leu Lys Leu Ser Asp Gln Gly Cys Ser Tyr Glu Thr Leu Leu Thr Leu
115 120 125
Ala His Asn Val Ile Lys Tyr Ser Val Lys Thr Gly His Pro Arg Phe
130 135 140
Ile Asn Gln Leu Tyr Ser Ser Val Asp Pro Tyr Gly Leu Leu Gly Gln
145 150 155 160
Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser
165 170 175
Pro Val Phe Ser Leu Met Glu Glu Glu Ile Leu Arg Glu Met Arg Lys
180 185 190
Ile Val Gly Trp Lys Asp Gly Arg Ser Glu Gly Ile Phe Cys Pro Gly
195 200 205
Gly Ser Ile Ala Asn Gly Tyr Ala Ile Asn Leu Ala Arg Tyr Tyr Lys
210 215 220
Phe Pro Gln Ser Lys Glu Leu Gly Leu Phe Asn Thr Gly Arg Leu Ile
225 230 235 240
Ile Phe Thr Ser Arg Asp Ala His Tyr Ser Val Lys Lys Leu Ser Ala
245 250 255
Phe Leu Gly Ile Gly Thr Glu Asn Val Tyr Glu Val Lys Thr Asp Asp
260 265 270
Lys Gly Lys Met Cys Ile Thr Asp Leu Lys Ile Gln Ile Lys Lys Ala
275 280 285
Leu Glu Glu Asp Ala Ile Pro Leu Met Val Ser Ala Thr Ala Gly Thr
290 295 300
Thr Val Leu Gly Ala Phe Asp Pro Leu Lys Asn Ile Ala Ala Ile Cys
305 310 315 320
Lys Asn Tyr Asn Leu Trp Phe His Val Asp Ala Ala Trp Gly Gly Gly
325 330 335
Ala Leu Met Ser Lys Lys Tyr Lys Tyr Leu Leu Asp Gly Ile Glu Leu
340 345 350
Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln
355 360 365
Gln Cys Ser Thr Leu Leu Leu Arg His Glu Gly Leu Leu Gln Asp Ala
370 375 380
His Gly Ser Lys Ala Ser Tyr Leu Phe Gln Pro Asp Lys Phe Tyr Asp
385 390 395 400
Thr Ser Phe Asp Ser Gly Asp Lys His Ile Gln Cys Gly Arg Arg Ala
405 410 415
Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys Gly Thr Arg Gly
420 425 430
Leu Glu Lys His Val Asp Arg Val Phe Lys Leu Ala Arg Tyr Phe Thr
435 440 445
Asn Tyr Ile Lys His Arg Glu Gly Phe Lys Leu Ile Leu Glu Pro Glu
450 455 460
Cys Thr Asn Val Cys Phe Trp Tyr Val Pro Pro Ser Lys Arg Gln Leu
465 470 475 480
Gln Asn Glu Glu Leu Leu Lys Ala Leu Gln Lys Ile Gly Pro Ala Val
485 490 495
Lys Glu Arg Met Met Lys Lys Gly Ser Met Leu Ile Thr Tyr Gln Pro
500 505 510
Leu Arg Glu Leu Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly
515 520 525
Leu Thr Glu Thr Asp Met Arg Phe Phe Ala Glu Glu Ile Glu Arg Leu
530 535 540
Ala Ile Asp Leu
545
<210> 182
<211> 586
<212> PRT
<213> 维尔利斯果蝇(Drosophila virilis)
<400> 182
Met Leu Ala Ser Glu Thr Phe Pro Thr His His Phe Lys Asn Ser Ile
1 5 10 15
Tyr Lys Pro Tyr Asn Gly Ala Ser Ser Ala Pro Asp Val Glu Asp Leu
20 25 30
Ala Ser Met Ala Lys Thr Leu Thr Thr Thr Thr Leu Ser Ser Ser Asp
35 40 45
Ala Ala Val Ile Asp Val Val Lys Thr Thr Val Glu Tyr Gly Asn Pro
50 55 60
Asn Lys Met Leu Asn Ser Ser Val Ser Ser Ser Ser Asn Ser Asn Asn
65 70 75 80
Lys Asn Asn Asn Ile Lys Asn Thr Asn Gly Asn Val Asn Gly Leu Ala
85 90 95
Ser Phe Val Ala Ser His Pro Ala Ala Glu Phe Glu Gly Phe Ile Arg
100 105 110
Ala Cys Val Asp Glu Ile Ile Lys Leu Ala Val Phe Gln Gly Thr Asn
115 120 125
Arg Ser Ser Lys Val Val Glu Trp His Glu Pro Ala Glu Leu Arg Lys
130 135 140
Leu Phe Asp Phe Glu Leu Arg Glu Lys Gly Glu Ser Pro Asp Lys Leu
145 150 155 160
Arg Gln Leu Leu Arg Glu Thr Ile Arg Phe Ser Val Lys Thr Gly His
165 170 175
Pro Tyr Phe Ile Asn Gln Leu Tyr Ser Gly Val Asp Pro Tyr Ala Leu
180 185 190
Val Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr
195 200 205
Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Gln Val Leu Ala Glu
210 215 220
Met Arg Arg Ile Val Gly Phe Pro Asn Asn Gly His Gly Asp Gly Ile
225 230 235 240
Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys Ala
245 250 255
Arg Tyr Lys Tyr Ala Pro Glu Ser Lys Lys Asn Gly Leu Phe Asn Ala
260 265 270
Lys Pro Leu Ile Ile Phe Thr Ser Glu Asp Ala His Tyr Ser Val Glu
275 280 285
Lys Leu Ala Met Phe Met Gly Phe Gly Ser Glu His Val Arg Lys Ile
290 295 300
Ala Thr Asn Glu Leu Gly Lys Met Arg Leu Ser Asp Leu Glu Asp Gln
305 310 315 320
Ile Gln Leu Cys Leu Asp Asn Asn Trp Gln Pro Leu Met Val Ser Ala
325 330 335
Thr Ala Gly Thr Thr Val Leu Gly Ala Phe Asp Asp Leu Val Gly Ile
340 345 350
Ser Glu Leu Cys Arg Lys His Asn Met Trp Met His Val Asp Ala Ala
355 360 365
Trp Gly Gly Gly Ala Leu Met Ser Lys Lys Tyr Arg Gln Leu Leu Asn
370 375 380
Gly Ile Glu Arg Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
385 390 395 400
Ser Ala Ser Gln Gln Cys Ser Thr Phe Leu Thr Pro His Ala Gln Ile
405 410 415
Leu Gly Gln Cys His Ser Thr Asn Ala Ala Tyr Leu Phe Gln Lys Asp
420 425 430
Lys Phe Tyr Asp Thr Ser Phe Asp Thr Gly Asp Lys His Ile Gln Cys
435 440 445
Gly Arg Arg Ala Asp Val Phe Lys Phe Trp Phe Met Trp Lys Ala Lys
450 455 460
Gly Ser Lys Gly Phe Glu Ala His Val Glu Gln Val Phe Glu Met Ser
465 470 475 480
Glu Tyr Phe Thr Ala Lys Leu Arg Glu Arg Pro Gly Phe Glu Leu Val
485 490 495
Leu Glu Lys Pro Glu Cys Thr Asn Ile Thr Phe Trp Tyr Val Pro Pro
500 505 510
Ser Leu Arg Gln Met Glu Arg Asn Gln Glu Phe Phe Asp Lys Leu His
515 520 525
Lys Val Ala Pro Lys Ile Lys Glu Ala Met Ile Lys Lys Gly Ser Met
530 535 540
Met Ile Thr Tyr Gln Pro Leu Arg Lys Leu Pro Asn Phe Phe Arg Leu
545 550 555 560
Val Leu Gln Asn Ser Cys Leu Glu Glu Ser Asp Met Leu Tyr Phe Leu
565 570 575
Asn Glu Ile Glu Asp Leu Gly Gln Asn Leu
580 585
<210> 183
<211> 547
<212> PRT
<213> 丽蝇蛹集金小蜂(Nasonia vitripennis)
<400> 183
Met Pro Ala His Glu Glu Thr Gln Leu Gln Glu Gly Ser Ala Ile Glu
1 5 10 15
Ala Leu Glu Arg Pro Arg Ser Ala Asp Ser Glu Lys His Ser Arg Leu
20 25 30
Asp Tyr Glu Arg Glu Glu Glu Ala His Gln Ile Gly Glu Asp Ser Asp
35 40 45
Glu Gly Tyr Ala Met Asp Asn Phe Asn Glu Glu Gly Phe Phe Cys Ser
50 55 60
Leu Pro Gly Arg Glu Ser His Glu Lys Phe Ile Arg Asp Ala Val Glu
65 70 75 80
Met Ile Leu Arg Glu Ala Val Phe Lys Gly Thr Ser Arg Lys Asn Arg
85 90 95
Val Val Glu Trp Ile Glu Pro Ala Thr Leu Pro Ser Lys Ile Asp Leu
100 105 110
Pro Pro Arg Lys Thr Gly Glu Ser His Glu Ala Leu Leu Arg Leu Leu
115 120 125
Asp Ser Val Ile Arg Tyr Ser Val Lys Thr Gly His Pro His Phe Val
130 135 140
Asn Gln Leu Tyr Ser Ser Val Asp Pro Tyr Gly Leu Val Gly Gln Trp
145 150 155 160
Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser Pro
165 170 175
Val Phe Ser Leu Met Glu Glu Ala Val Leu Lys Glu Met Arg Ala Ile
180 185 190
Val Gly Trp Gln Asn Gly Glu Gly Asp Gly Ile Phe Cys Pro Gly Gly
195 200 205
Ser Met Ala Asn Gly Tyr Ala Ile Asn Leu Ala Arg His Trp Met Phe
210 215 220
Pro Ile Val Lys Glu Gln Gly Leu Thr Ala Val Pro Arg Leu Val Val
225 230 235 240
Phe Thr Ser Glu Asp Ala His Tyr Ser Val Lys Lys Leu Ala Ala Phe
245 250 255
Leu Gly Ile Gly Ile Ala Asn Val Tyr Ser Val Lys Val Asp Glu Ser
260 265 270
Gly Lys Met Cys Val Ser Asp Leu Arg Ala Gln Ile Asp Arg Ala Ile
275 280 285
Gln Glu Gly Ala Arg Pro Leu Met Val Ser Ala Thr Ala Gly Thr Thr
290 295 300
Val Leu Gly Ala Phe Asp Pro Leu Arg Ser Ile Ala Glu Leu Cys Arg
305 310 315 320
Glu His Asn Met Trp Phe His Val Asp Ala Ala Trp Gly Gly Gly Ala
325 330 335
Leu Val Ser Pro Lys His Arg His Leu Leu Asp Gly Val Glu Leu Ala
340 345 350
Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln Gln
355 360 365
Cys Ser Thr Leu Leu Thr Arg His Lys Gly Leu Leu Gln Ser Ala His
370 375 380
Gly Cys Lys Ala Thr Tyr Leu Phe Gln Gln Asp Lys Phe Tyr Asp Thr
385 390 395 400
Ser Tyr Asp Phe Gly Asp Lys His Val Gln Cys Gly Arg Arg Ala Asp
405 410 415
Val Leu Lys Phe Trp Leu Met Trp Lys Ala Lys Gly Thr Asp Gly Leu
420 425 430
Glu Lys His Val Asp Arg Val Phe Gln Leu Ser Arg Tyr Phe Val Gly
435 440 445
Ile Ile Arg Asn Arg Pro Gly Trp Gln Leu Leu Phe Glu Pro Glu Cys
450 455 460
Thr Asn Val Cys Phe Arg Tyr Val Pro Pro Ser Lys Arg His Leu Asn
465 470 475 480
Gly Gln Asp Leu Phe Gln Ala Leu His Lys Val Ala Pro Leu Val Lys
485 490 495
Glu Arg Met Val Lys Thr Gly Ser Met Leu Ile Thr Tyr Gln Pro Ile
500 505 510
Arg Glu Gln Ala Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly Leu
515 520 525
Thr Glu Ala Asp Met His Phe Phe Val Glu Glu Ile Glu Arg Leu Ser
530 535 540
Glu Asp Leu
545
<210> 184
<211> 508
<212> PRT
<213> 偏瞳蔽眼蝶(Bicyclus anynana)
<400> 184
Met Pro Ala Asp Ser Asn Ile Ile Val Ala Val Glu Glu Lys Lys Glu
1 5 10 15
Asp Gly Leu Phe Gln Ser Leu Thr Glu Arg Ser Lys His Glu Asp Phe
20 25 30
Ile Arg Arg Ala Val Asp Leu Leu Val Glu Arg Val Val Phe Gly Arg
35 40 45
Ala Ser Arg Thr Ser Lys Val Val Glu Trp Ser Ala Pro Glu Asp Ile
50 55 60
Lys Gln Ala Ile Asp Leu Lys Val Arg Asp Gly Pro Ala Ser His Glu
65 70 75 80
Glu Leu Leu Ala Phe Met Ala Asp Val Ala Arg Tyr Ser Val Asn Thr
85 90 95
Ala His Pro Tyr Phe Val Asn Gln Leu Phe Ser Ser Val Asp Pro Tyr
100 105 110
Gly Leu Ile Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr
115 120 125
Thr Phe Glu Val Ala Pro Val Phe Thr Leu Met Glu Glu Glu Val Leu
130 135 140
Arg Glu Met Arg Ser Ile Val Gly Trp Ala Asp Gly Glu Gly Asp Gly
145 150 155 160
Ile Phe Cys Pro Gly Gly Ser Ile Ala Asn Gly Tyr Ala Ile Ser Cys
165 170 175
Ala Arg Ser Tyr Phe Tyr Pro Glu Ile Lys Asn Lys Gly Val Tyr Ala
180 185 190
Val Pro Lys Leu Val Ile Phe Thr Ser Glu Leu Ala His Tyr Ser Thr
195 200 205
Lys Lys Met Ala Val Phe Met Gly Ile Gly Ser Asp Asn Cys Ile Leu
210 215 220
Val Lys Ala Asp Glu Asn Gly Arg Met Asp Val Asn Asp Phe Glu Arg
225 230 235 240
Lys Ile Asn Glu Ala Ile Glu Ala Gly Ala Thr Pro Phe Leu Val Thr
245 250 255
Ser Thr Ser Gly Thr Thr Val Tyr Gly Ala Phe Asp Pro Ile Val Pro
260 265 270
Ile Ser Asn Ile Cys Lys Lys Tyr Asn Leu Trp Leu His Val Asp Ala
275 280 285
Ala Trp Gly Gly Gly Ala Leu Met Ser Arg Lys His Arg Asn Leu Leu
290 295 300
Asn Gly Ile Glu Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
305 310 315 320
Ala Ala Pro Gln Gln Cys Ser Thr Phe Leu Leu Lys His Lys Asn Val
325 330 335
Leu Lys Glu Ala His Ser Ser Asn Ala Gln Tyr Leu Phe Gln Lys Asp
340 345 350
Lys Phe Tyr Asp Thr Ser Tyr Asp Thr Gly Asp Lys His Ile Gln Cys
355 360 365
Gly Arg Arg Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys
370 375 380
Gly Ser Glu Gly Phe Glu Lys His Val Glu Lys Leu Phe Asp Asn Ala
385 390 395 400
Asn Tyr Phe Leu Glu His Ile Arg Gln Arg Glu Gly Phe Arg Leu Val
405 410 415
Ile Pro Lys Pro Glu Cys Thr Asn Ile Met Phe Trp Tyr Ile Pro Lys
420 425 430
Cys Leu Arg Ser Cys Glu Asn Glu Pro Asn Tyr Tyr Glu Arg Leu His
435 440 445
Lys Val Ala Pro Lys Ile Lys Glu Arg Met Ile Lys Glu Gly Ser Met
450 455 460
Met Val Thr Tyr Gln Pro Gln Gly Asn Leu Val Asn Phe Phe Arg Ile
465 470 475 480
Val Phe Gln Asn Ser Ala Leu Asp His Lys Asp Met Ile Tyr Phe Ala
485 490 495
Asn Glu Phe Glu Arg Leu Gly Ser Asp Ile Val Val
500 505
<210> 185
<211> 543
<212> PRT
<213> 印度跳蚁(Harpegnathos saltator)
<400> 185
Met Pro Ala Asn Glu Asp Thr Ser Asn Glu Ser Leu Glu Arg Ile Glu
1 5 10 15
Pro Gly Arg Ile Ser Pro Ser Leu Ser Gln Arg Lys Met Ser Gly Gly
20 25 30
Leu Gln Asn Leu Ala Ser Ser Leu Leu Pro Gly Ala Thr Val Asp Asp
35 40 45
Asp Leu Glu Ser Ser Arg Val Glu Glu Arg Asn Phe Arg Ser Ile Pro
50 55 60
Arg Arg Asp Ile His Glu Lys Leu Phe Arg Asp Phe Phe Glu Leu Val
65 70 75 80
Leu Gln Gln Ala Val Phe Gln Ser Thr Ser Gly Lys Glu Arg Val Val
85 90 95
Glu Trp Met Asn Pro Ser Asp Leu Arg Ser Val Val Asp Phe Ser Leu
100 105 110
Pro Ala Glu Gly Val Ser His Glu Glu Leu Leu Ala Leu Thr Arg Asn
115 120 125
Val Ile Lys Tyr Ser Val Lys Thr Gly His Pro His Phe Val Asn Gln
130 135 140
Leu Phe Ser Ser Leu Asp Pro Tyr Gly Leu Leu Gly Gln Trp Leu Thr
145 150 155 160
Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser Pro Val Phe
165 170 175
Ser Leu Met Glu Glu Asp Val Leu Arg Glu Met Arg Arg Ile Ile Gly
180 185 190
Trp Lys Gly Gly Glu Gly Leu Phe Cys Pro Gly Gly Ser Met Ala Asn
195 200 205
Gly Tyr Ala Ile Asn Leu Ala Arg His His Arg Tyr Pro Asn Met Lys
210 215 220
Gln Thr Gly Leu Ser Gln Met Pro Arg Leu Val Ile Phe Thr Ser Glu
225 230 235 240
Asp Ala His Tyr Ser Val Lys Lys Leu Ala Ala Phe Leu Gly Ile Gly
245 250 255
Tyr Asp Asn Val Tyr Ser Val Lys Val Asp Ser Arg Gly Lys Met Leu
260 265 270
Val Ser Asp Leu Glu Ala Gln Ile Ala Arg Ala Thr Arg Glu Gly Ala
275 280 285
Val Pro Leu Met Val Ser Ser Thr Ala Gly Thr Thr Val Leu Gly Ala
290 295 300
Phe Asp Pro Leu Lys Asp Ile Ala Glu Val Cys Arg Lys His Arg Leu
305 310 315 320
Trp Phe His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Val Ser Arg
325 330 335
Thr Tyr Arg Arg Leu Leu Asp Gly Val Glu Leu Ala Asp Ser Ile Thr
340 345 350
Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln Gln Cys Ser Thr Leu
355 360 365
Leu Leu Arg His Glu Gly Leu Leu Gln Ser Ala His Gly Cys Gly Ala
370 375 380
Ser Tyr Leu Phe Gln Asn Asp Lys Phe Tyr Asp Ser Ser Tyr Asp Cys
385 390 395 400
Gly Asp Arg His Val Gln Cys Gly Arg Arg Ala Asp Val Val Lys Phe
405 410 415
Trp Tyr Met Trp Lys Ala Lys Gly Thr Arg Gly Leu Glu Glu His Val
420 425 430
Asp His Val Phe Ala Leu Ser Arg Tyr Phe Ala Asp Leu Val Arg Thr
435 440 445
Arg Asp Gly Trp His Leu Leu Ala Glu Pro Glu Cys Thr Asn Val Cys
450 455 460
Phe Arg Tyr Ile Pro Pro Ser Met Arg Asp Leu Ala Gly Arg Gln Leu
465 470 475 480
Asp Gln Ala Ile His Lys Val Ala Pro Met Ile Lys Glu Arg Met Val
485 490 495
Arg Ala Gly Thr Met Leu Met Thr Tyr Gln Pro Leu Arg Gly Thr Pro
500 505 510
Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly Leu Ser Glu Ile Asp
515 520 525
Met Gln Phe Phe Val Glu Glu Ile Glu Arg Leu Ala Ala Asp Leu
530 535 540
<210> 186
<211> 548
<212> PRT
<213> 切叶蚁(Acromyrmex echinatior)
<400> 186
Met Pro Ala Asn Glu Asp Thr Ser Asn Asp Glu Thr Ser Leu Arg Met
1 5 10 15
Lys His Glu Arg Ala Gly Pro Ser Met Glu Ser Ser Trp Arg Lys Met
20 25 30
Ser Glu Glu His Pro Arg Ala Asn Pro Ser Pro Ile Pro Asn Leu Pro
35 40 45
Gly Leu Val Asn Gly Ile Glu Glu Arg Gln Gly Ile Glu Lys Trp Asp
50 55 60
Phe Arg Ser Met Pro Arg Arg Asp Ala His Glu Lys Leu Phe Arg Asp
65 70 75 80
Phe Phe Glu Leu Val Leu Gln Arg Ala Val Phe Gln Pro Thr Ser Gly
85 90 95
Lys Asp Arg Val Val Glu Trp Val Asp Pro Tyr Asp Leu Arg Ser Val
100 105 110
Val Asp Leu Ser Leu Pro Ala Glu Gly Val Ser His Glu Lys Leu Leu
115 120 125
Thr Leu Thr Arg Asp Ile Ile Lys Tyr Ser Val Lys Thr Ser His Pro
130 135 140
His Phe Val Asn Gln Leu Phe Ser Ser Leu Asp Pro Tyr Gly Leu Leu
145 150 155 160
Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu
165 170 175
Val Ser Pro Val Phe Ser Leu Met Glu Glu Asp Val Leu Arg Glu Met
180 185 190
Arg Ala Ile Ile Ser Trp Gln Glu Gly Glu Gly Leu Phe Cys Pro Gly
195 200 205
Gly Ser Met Ala Asn Gly Tyr Ala Ile Asn Leu Ala Arg His His Arg
210 215 220
Tyr Pro Asn Leu Lys Gln Ser Gly Leu Ser Gln Met Pro Arg Leu Val
225 230 235 240
Val Phe Thr Ser Glu Asp Ala His Tyr Ser Val Lys Lys Leu Ala Ala
245 250 255
Phe Leu Gly Ile Gly Tyr Asp Asn Val Tyr Leu Val Lys Val Asp Ser
260 265 270
Arg Gly Lys Met Met Val Ser Asp Leu Glu Ala Gln Ile Ala Arg Ala
275 280 285
Val Glu Glu Gly Ala Ala Pro Leu Met Val Ser Ala Thr Ala Gly Thr
290 295 300
Thr Val Ile Gly Ala Phe Asp Pro Leu Arg Glu Ile Ala Glu Val Cys
305 310 315 320
Arg Lys His Glu Leu Trp Phe His Val Asp Ala Ala Trp Gly Gly Gly
325 330 335
Ala Leu Ile Ser Glu Thr Tyr Arg Gly Leu Leu Asp Gly Ile Gln Phe
340 345 350
Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln
355 360 365
Gln Cys Ser Thr Leu Leu Leu Arg His Glu Gly Leu Leu Gln Ala Ala
370 375 380
His Gly Cys Gly Ala Ser Tyr Leu Phe Gln Asn Asp Lys Phe Tyr Asp
385 390 395 400
Ala Ser Phe Asp Cys Gly Asp Arg His Val Gln Cys Gly Arg Arg Ala
405 410 415
Asp Val Val Lys Phe Trp Tyr Met Trp Lys Ala Lys Gly Thr Arg Gly
420 425 430
Leu Glu Ala His Val Asp Arg Leu Phe Ala Leu Ser Arg His Phe Thr
435 440 445
Asp Leu Ile Arg Thr Arg Asp Gly Trp His Leu Leu Val Glu Pro Glu
450 455 460
Cys Ile Asn Val Cys Phe Arg Tyr Ile Pro Pro Ser Lys Arg His Leu
465 470 475 480
Thr Gly Gln Glu Leu Glu Gln Ala Leu His Lys Ile Ala Pro Ile Ile
485 490 495
Lys Glu Arg Met Val Arg Ala Gly Thr Met Leu Ile Thr Tyr Gln Thr
500 505 510
Leu Arg Asn Met Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly
515 520 525
Leu Thr Glu Ala Asp Met Lys Phe Phe Val Glu Glu Ile Glu Arg Leu
530 535 540
Ala Met Asp Leu
545
<210> 187
<211> 544
<212> PRT
<213> 佛罗里达弓背蚁(Camponotus floridanus)
<400> 187
Met Pro Ala Asn Glu Asp Thr Ser Ala Asn Asp Phe Ala Phe Lys Arg
1 5 10 15
Ile Lys Pro Glu Arg Ala Asn Pro Ala Glu Ser Gln Pro Lys Met Ser
20 25 30
Arg Lys Asp Asp Ser Ser Thr Asn Leu Ile Leu Ser Asn Phe Thr Ala
35 40 45
Ser Val Glu Glu Arg Lys Glu Ala Glu Lys Gly Asp Phe Arg Ser Ile
50 55 60
Pro Cys Arg Ser Val His Glu Lys Leu Phe Arg Asp Phe Phe Glu Leu
65 70 75 80
Met Leu Gln Arg Ala Val Phe Gln Ser Thr Ser Gly Glu Glu Arg Val
85 90 95
Val Glu Trp Ile Asp Pro Asn Asn Leu Arg Ser Val Val Asp Leu Ser
100 105 110
Leu Pro Thr Glu Gly Val Ser His Glu Glu Leu Leu Thr Leu Thr Arg
115 120 125
Asp Ile Ile Lys Tyr Ser Val Lys Thr Gly His Pro His Phe Val Asn
130 135 140
Gln Leu Phe Ser Ser Leu Asp Pro Tyr Gly Leu Leu Gly Gln Trp Leu
145 150 155 160
Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser Pro Val
165 170 175
Phe Ser Leu Met Glu Glu Asp Val Leu Arg Glu Met Arg Ala Ile Ile
180 185 190
Gly Trp Gln Gly Gly Glu Gly Leu Phe Cys Pro Gly Gly Ser Met Ala
195 200 205
Asn Gly Tyr Ala Ile Asn Leu Ala Arg His His Arg Tyr Pro Asn Leu
210 215 220
Lys Gln Ser Gly Leu Ser Gln Met Pro Arg Leu Val Val Phe Thr Ser
225 230 235 240
Glu Asp Ala His Tyr Ser Val Lys Lys Leu Ala Ala Phe Leu Gly Ile
245 250 255
Gly Tyr Asp Asn Val Tyr Leu Ile Lys Val Asp Ser Arg Gly Lys Met
260 265 270
Val Val Thr Asp Leu Glu Ala Gln Ile Val Arg Ala Ile Asn Glu Gly
275 280 285
Ala Val Pro Leu Met Val Ser Ala Thr Ala Gly Thr Thr Val Met Gly
290 295 300
Thr Phe Asp Pro Leu Lys Lys Ile Ala Glu Val Cys Arg Lys His Gly
305 310 315 320
Leu Trp Phe His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Val Ser
325 330 335
Arg Thr Tyr Arg Gly Leu Leu Asp Gly Ile Gln Leu Ala Asp Ser Ile
340 345 350
Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro Gln Gln Cys Ser Thr
355 360 365
Leu Leu Leu Arg His Glu Gly Leu Leu Gln Glu Ala His Gly Cys Gly
370 375 380
Ala Ser Tyr Leu Phe Gln Asn Asp Lys Phe Tyr Asp Ala Thr Phe Asp
385 390 395 400
Tyr Gly Asp Arg His Val Gln Cys Gly Arg Arg Ala Asp Val Val Lys
405 410 415
Phe Trp Tyr Met Trp Lys Ala Lys Gly Thr Arg Gly Leu Glu Ala His
420 425 430
Val Asp Cys Val Phe Ala Leu Ser Arg Tyr Phe Ala Asp Leu Ile Arg
435 440 445
Thr Arg Asp Gly Trp Arg Leu Leu Ala Glu Pro Glu Cys Thr Asn Val
450 455 460
Cys Phe Arg Tyr Ile Pro Leu Ser Lys Arg His Leu Thr Gly Arg Glu
465 470 475 480
Leu Asp Gln Ala Leu His Lys Ile Ala Pro Met Ile Lys Glu Arg Met
485 490 495
Met Arg Ala Gly Thr Met Leu Ile Thr Tyr Gln Thr Leu Arg Asp Met
500 505 510
Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser Gly Leu Thr Glu Ala
515 520 525
Asp Met Lys Phe Phe Val Glu Glu Ile Glu Arg Leu Ala Val Asp Leu
530 535 540
<210> 188
<211> 532
<212> PRT
<213> 人虱(Pediculus humanus)
<400> 188
Met Phe Pro Val Ile Ser Thr Gln Lys Gln Ser Phe Gly Ile Ile Asn
1 5 10 15
Leu Ser Leu Leu Glu Asp Asn Asn Phe Leu Ile Lys Thr Asn Ser Val
20 25 30
Val Asp Lys Ser Asp Lys Glu Asp Glu Lys Leu Asn Gln Lys His Trp
35 40 45
Ser Leu Pro Leu Glu Lys Tyr His Tyr Asp Phe Ile Val Lys Cys Val
50 55 60
Gly Ile Ile Met Lys Glu Ala Val Phe Asp Gly Thr Asn Arg Asn Ser
65 70 75 80
Lys Val Val Gln Trp Gln Asp Pro Glu Lys Leu Lys Lys Ser Phe Asp
85 90 95
Phe Ser Leu Asn Lys Tyr Ser Glu Thr Glu Gly Lys Leu Leu His Leu
100 105 110
Ile Ser Thr Ile Ile Lys Phe Ser Val Lys Thr Gly His Pro Tyr Phe
115 120 125
Val Asn Gln Leu Phe Ser Gly Val Asp Pro Tyr Gly Leu Ile Gly Gln
130 135 140
Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ser
145 150 155 160
Pro Val Phe Ser Ile Met Glu Glu Val Val Leu Glu Glu Met Arg Lys
165 170 175
Phe Ile Gly Phe Pro Asn Gly Lys Gly Asp Gly Thr Phe Cys Pro Gly
180 185 190
Gly Ser Ile Ser Asn Gly Phe Gly Ile Ser Cys Ala Arg Tyr His Leu
195 200 205
Phe Pro Gln Val Lys Lys Leu Gly Ile Tyr Gly Ile Gly Leu Arg Leu
210 215 220
Val Leu Phe Thr Ser Arg Asp Ala His Tyr Ser Ile Val Lys Leu Ala
225 230 235 240
Thr Phe Met Gly Leu Gly Ser Asp Asn Val Ile Ser Ile Lys Thr Asp
245 250 255
Glu Ser Gly Lys Met Lys Pro Glu Glu Leu Glu Lys Ala Ile Leu Lys
260 265 270
Val Leu Gln Glu Gly Gly Thr Pro Phe Met Val Ser Ala Thr Ser Gly
275 280 285
Thr Thr Val Leu Gly Ala Phe Asp Pro Leu Asp Ser Ile Ala Asp Ile
290 295 300
Cys Glu Lys Tyr Lys Leu Trp Phe His Val Asp Ala Ala Trp Gly Gly
305 310 315 320
Gly Cys Leu Met Ser Ser Ile His Lys Lys Lys Leu Gln Gly Ile His
325 330 335
Arg Thr Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Gly Val Pro
340 345 350
Gln Gln Cys Ser Ala Phe Leu Thr Lys His Lys Asn Leu Leu Lys Asn
355 360 365
Val His Cys Ala Lys Ala Thr Tyr Leu Phe Gln Lys Asp Lys Phe Tyr
370 375 380
Asp Val Lys Tyr Asp Thr Gly Asp Lys His Ile Gln Cys Gly Arg Arg
385 390 395 400
Ala Asp Val Leu Lys Phe Trp Phe Met Trp Lys Ala Lys Gly Ser Ser
405 410 415
Gly Phe Glu Lys His Ile Asn Lys Ile Phe Glu Thr Ala Leu Tyr Phe
420 425 430
Lys Lys Ser Ile Glu Asn Lys Pro Asp Phe Gln Leu Val Leu Ser Glu
435 440 445
Pro Glu Cys Thr Asn Ile Cys Phe Trp Tyr Ile Pro Pro Arg Leu Gln
450 455 460
Asn Ser Lys Tyr Asn Asn Asp Asp Leu Asn Lys Val Ala Pro Arg Met
465 470 475 480
Lys Glu Lys Met Met Lys Asp Gly Ser Met Met Ile Thr Tyr Gln Pro
485 490 495
Leu Arg His Leu Pro Asn Phe Phe Arg Leu Val Ile Val Asn Ser Gly
500 505 510
Leu Asp Thr His Asp Met Asp Arg Leu Ile Thr Ile Ile Gln Asn Ala
515 520 525
Gly Ala Ser Ile
530
<210> 189
<211> 549
<212> PRT
<213> 大头美切叶蚁(Atta cephalotes)
<400> 189
Met Pro Ala Asn Glu Asp Thr Ser Asn Asp Glu Thr Ser Leu Arg Met
1 5 10 15
Lys His Glu Arg Ala Asp Ser Ser Ala Glu Ser Ser Trp Arg Lys Met
20 25 30
Ser Glu Glu Arg Pro Arg Ala Asn Ser Ser Ser Leu Thr Leu Asn Leu
35 40 45
Pro Gly Leu Val Asn Gly Thr Glu Glu Arg Gln Arg Ala Glu Lys Arg
50 55 60
Asp Phe Arg Ser Met Pro Arg Arg Asp Ala His Glu Lys Leu Phe Arg
65 70 75 80
Asp Phe Phe Glu Leu Val Leu Gln Arg Ala Val Phe Gln Ser Thr Ser
85 90 95
Gly Lys Asp Arg Val Val Glu Trp Val Asp Pro Tyr Asp Leu Arg Ser
100 105 110
Val Val Asp Leu Ser Leu Pro Ala Glu Gly Val Ser His Glu Glu Leu
115 120 125
Leu Thr Leu Thr Arg Asp Ile Ile Lys Tyr Ser Val Lys Thr Gly His
130 135 140
Pro His Phe Val Asn Gln Leu Phe Ser Ser Leu Asp Pro Tyr Gly Leu
145 150 155 160
Leu Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val Tyr Thr Tyr
165 170 175
Glu Val Ser Pro Val Phe Ser Leu Met Glu Glu Asp Val Leu Arg Glu
180 185 190
Met Arg Ala Ile Ile Gly Trp Gln Gly Gly Glu Gly Leu Phe Cys Pro
195 200 205
Gly Gly Ser Met Ala Asn Gly Tyr Ala Ile Asn Leu Ala Arg His His
210 215 220
Arg Tyr Pro Asn Leu Lys Gln Ser Gly Leu Ser Gln Met Pro Arg Leu
225 230 235 240
Val Val Phe Thr Ser Glu Asp Ala His Tyr Ser Val Lys Lys Leu Ala
245 250 255
Ala Phe Leu Gly Ile Gly Tyr Asp Asn Val Tyr Leu Val Lys Val Asp
260 265 270
Ser Arg Gly Lys Met Met Val Ser Asp Leu Glu Thr Gln Ile Ala Gln
275 280 285
Ala Val Lys Glu Gly Ala Ala Pro Leu Met Val Ser Ala Thr Ala Gly
290 295 300
Thr Thr Val Ile Gly Ala Phe Asp Pro Leu Arg Glu Ile Ala Glu Val
305 310 315 320
Cys Lys Lys His Gly Leu Trp Phe His Val Asp Ala Ala Trp Gly Gly
325 330 335
Gly Ala Leu Val Ser Gly Thr Tyr Arg Asp Leu Leu Asp Gly Ile Gln
340 345 350
Phe Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Ala Ala Pro
355 360 365
Gln Gln Cys Ser Thr Leu Leu Leu Arg His Glu Gly Leu Leu Gln Ala
370 375 380
Ala His Gly Cys Gly Ala Ser Tyr Leu Phe Gln Asn Asp Lys Phe Tyr
385 390 395 400
Asp Ala Ser Phe Asp Cys Gly Asp Arg His Val Gln Cys Gly Arg Arg
405 410 415
Ala Asp Val Ile Lys Phe Trp Tyr Met Trp Lys Ala Lys Gly Met Arg
420 425 430
Gly Phe Glu Ala His Val Asp His Leu Phe Ala Leu Ser Arg His Phe
435 440 445
Thr Asp Leu Ile Arg Ile Arg Asp Gly Trp His Leu Leu Val Glu Pro
450 455 460
Glu Cys Ile Asn Val Cys Phe Arg Tyr Ile Pro Pro Ser Lys Arg His
465 470 475 480
Leu Ala Gly Gln Glu Leu Glu Gln Ala Leu His Lys Ile Ala Pro Ile
485 490 495
Ile Lys Glu Arg Met Val Arg Ala Gly Thr Met Leu Ile Thr Tyr Gln
500 505 510
Thr Leu Arg Asn Met Pro Asn Phe Phe Arg Leu Val Leu Gln Asn Ser
515 520 525
Gly Leu Thr Glu Val Asp Met Lys Phe Phe Val Glu Glu Ile Glu Arg
530 535 540
Leu Ala Met Asp Leu
545
<210> 190
<211> 497
<212> PRT
<213> 蚤状溞(Daphnia pulex)
<400> 190
Met Glu Asn Pro Ala Leu Ile Gly Trp Phe Gln Thr Glu Pro Ser Gln
1 5 10 15
Glu Gln His Glu Gln Phe Met Arg Lys Val Met Asp Ile Val Leu Lys
20 25 30
Glu Ala Val Phe Glu Gly Thr Ser Arg Asn Asn Leu Val Ile Glu Trp
35 40 45
Met Glu Pro Glu Ser Leu Leu Thr Leu Leu Gly Lys Glu Leu Pro Lys
50 55 60
Asn Pro Gln Ser Asp Glu Thr Leu Val Glu Leu Ile Lys Asn Val Val
65 70 75 80
Arg Tyr Ser Val Lys Thr Gly His Pro His Phe Ile Asn Gln Leu Phe
85 90 95
Ser Ser Leu Asp Pro Tyr Gly Leu Val Ala Gln Trp Val Thr Asp Ser
100 105 110
Leu Asn Pro Ser Val Tyr Thr Tyr Glu Val Ala Pro Val Phe Thr Leu
115 120 125
Leu Glu His Gln Ile Leu Arg Glu Met Arg Arg Trp Val Gly Phe Pro
130 135 140
Asp Gly Ser Gly Asp Gly Val Phe Cys Pro Gly Gly Ser Met Ala Asn
145 150 155 160
Ile Tyr Gly Ile Gln Cys Ala Arg His Arg Ala Met Pro Ser Leu Lys
165 170 175
Glu Thr Gly Thr Phe Gly Ser Pro Arg Leu Val Val Leu Thr Ser Lys
180 185 190
Asp Ala His Tyr Ser Val Lys Lys Ala Cys Phe Leu Leu Gly Ile Gly
195 200 205
Val Ser Asn Leu Tyr Leu Val Asp Val Asp Thr Ser Gly Arg Met Asp
210 215 220
Leu Val His Leu Arg Gln Glu Val Gln Arg Ala Leu Asn Glu Asn Ala
225 230 235 240
Arg Pro Phe Met Val Ser Ala Thr Ala Gly Thr Thr Val Leu Gly Ala
245 250 255
Thr Asp Pro Leu Asp Gly Ile Ala Asp Ile Cys Gln Glu Phe Gly Met
260 265 270
Trp Met His Val Asp Ala Ala Trp Gly Gly Gly Ala Leu Met Ser Thr
275 280 285
Lys His Arg His Ile Leu Lys Gly Ile Glu Arg Ala Asp Ser Val Thr
290 295 300
Trp Asn Pro His Lys Leu Leu Gly Val Pro Gln Gln Cys Ser Thr Phe
305 310 315 320
Leu Thr Arg His Ala Asp Leu Leu Leu Glu Ala Asn Ser Ala Ser Ala
325 330 335
Ser Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Pro Lys Trp Asp Val
340 345 350
Gly Asp Lys Tyr Leu Gln Cys Gly Arg Arg Ala Asp Val Leu Lys Phe
355 360 365
Trp Leu Met Trp Gln Ala Lys Gly Ser Leu Gly Leu Glu Lys His Val
370 375 380
Asp Thr Leu Phe Glu Asn Val Ala Tyr Phe Thr Ser Phe Ile Arg Asn
385 390 395 400
Arg Lys Gly Phe Gln Leu Val Leu Glu Glu Pro Pro Phe Val Asn Val
405 410 415
Cys Phe Trp Tyr Ile Pro Pro Ser Leu Gln Gly Ala Gln His Asp Glu
420 425 430
Asp Tyr Glu Glu Lys Leu His Lys Ile Ala Pro Lys Ile Lys Glu Arg
435 440 445
Met Ile Lys Lys Gly Ser Met Met Ile Thr Tyr Gln Pro Leu Arg Asn
450 455 460
Leu Pro Asn Phe Phe Arg Leu Val Leu Gln Ser Ser Ala Val Thr Ile
465 470 475 480
Asp Asp Met Glu Phe Phe Ala Glu Glu Ile Glu Arg Leu Gly Cys Asp
485 490 495
Leu
<210> 191
<211> 581
<212> PRT
<213> 霸王莲花青螺(Lottia gigantea)
<400> 191
Met Glu Thr Lys Arg Phe Asp Asn Met Ser Ile Asn Asp Glu Ala Thr
1 5 10 15
Lys Glu Gln Phe Asn Glu Leu Lys Gln Arg Leu Leu Glu Gly Lys Ser
20 25 30
Leu Lys Ser Lys Gln Val Ser Lys Val Met Ile Asp Val Asp Ser Glu
35 40 45
Ser Glu Asp Ser Ala Leu Ser Leu Gln Asp Ser Ser Ala Asn Thr Asp
50 55 60
Glu Ser Asp Pro Asp Glu Ser Asn Val Glu Thr Cys Thr Thr Gln Leu
65 70 75 80
Gln His Ser Gln Arg Val Asn Ser Arg Arg Lys Ser Thr Lys Ala Lys
85 90 95
Phe Leu Thr Ile Pro Gln Val His His Asp Asp Phe Leu Ser Asp Thr
100 105 110
Phe Asp Leu Ile Met Lys Glu Ile Val Gln Lys Ala Gly Asp Arg Ser
115 120 125
Gln Lys Val Val Glu Trp Lys Ala Pro Glu Glu Leu Arg Glu Leu Met
130 135 140
Asp Leu Asp Pro Thr Ala Val Gly Glu Thr His Glu Lys Leu Leu Met
145 150 155 160
Arg Leu Gln Asp Ile Ile Lys Tyr Ser Val Lys Thr Gly His Pro Arg
165 170 175
Phe Val Asn Gln Leu Phe Ser Ser Leu Asp Pro Tyr Gly Leu Ala Gly
180 185 190
Gln Ile Val Thr Asp Ala Leu Asn Thr Ser Gln Tyr Thr Tyr Glu Thr
195 200 205
Ala Pro Val Phe Thr Leu Met Glu Glu Thr Val Leu Lys Thr Met Arg
210 215 220
Thr Cys Ile Gly Tyr Thr Glu Gly Asn Gly Ile Phe Cys Pro Gly Gly
225 230 235 240
Ser Leu Ser Asn Ile Met Ala Val Asn Cys Ala Arg His Phe Met Phe
245 250 255
Pro Gln Thr Lys Lys Thr Gly Met Phe Gly Leu Pro Pro Leu Val Ile
260 265 270
Tyr Thr Ser Glu Leu Ala His Tyr Ser Ile Lys Lys Ala Gly Tyr Leu
275 280 285
Leu Gly Phe Gly Asp Asp Asn Val Lys Leu Val Lys Thr Asp Glu Leu
290 295 300
Gly Lys Ile Ile Pro Glu Asp Leu Glu Arg Gln Ile Gln Glu Thr Ile
305 310 315 320
Asp Glu Gly Cys Thr Pro Phe Met Ile Val Ala Thr Ala Gly Thr Thr
325 330 335
Val Phe Gly Ala Phe Asp Pro Ile Asp Lys Met Ala Asp Val Ala Gln
340 345 350
Lys Tyr Gly Leu Trp Tyr His Val Asp Gly Ala Trp Gly Gly Gly Val
355 360 365
Leu Met Ser Lys Lys His Arg Asp Met Met Lys Gly Val Asp Arg Ala
370 375 380
Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu Gly Val Pro Gln Gln
385 390 395 400
Cys Ser Cys Phe Leu Thr Lys His Val Asn Ile Leu Gln Gln Cys His
405 410 415
Arg Ala Asp Ala Lys Tyr Leu Phe Gln Lys Asp Lys Phe Tyr Asp Thr
420 425 430
Ser Tyr Asp Thr Gly Asp Lys Thr Phe Gln Cys Gly Arg Lys Val Asp
435 440 445
Val Leu Lys Phe Trp Leu Met Trp Lys Ala Lys Gly Ser Glu Gly Phe
450 455 460
Glu Gln His Ile Asp Lys Leu Phe Asp Asn Thr Arg Tyr Ile Val Glu
465 470 475 480
Lys Leu Lys Gln Arg Glu Gly Phe Arg Met Val Val Asn Glu Pro Asp
485 490 495
Cys Thr Asn Val Cys Phe Trp Tyr Val Pro Pro Ser Leu Arg Asn Met
500 505 510
Pro Glu Asp Glu Glu Phe Trp Asn Arg Leu His Thr Val Ala Pro Lys
515 520 525
Val Lys Glu Gly Met Ile Arg Asp Gly Thr Met Met Ile Thr Tyr Gln
530 535 540
Pro Gln Lys Asp Leu Val Asn Phe Phe Arg Leu Val Leu Gln Asn Ser
545 550 555 560
Ala Thr Thr Tyr Glu Asp Met Asp Phe Phe Ile Asp Glu Ile Glu Arg
565 570 575
Leu Gly Lys Phe Leu
580
<210> 192
<211> 538
<212> PRT
<213> 佛罗里达文昌鱼(Branchiostoma floridae)
<400> 192
Met Ser Asn Ile Leu Thr Ser Leu Glu Asp Ser Ser Phe Ser Cys Ser
1 5 10 15
Cys Ala Lys Glu Met Ile Asn Gly Gly Ile Asp Gly His Leu Asn Asp
20 25 30
Cys Ser Val Arg Asn Leu Lys Ser Ser Leu Asn Asp Asp Thr Ala Asn
35 40 45
Val Lys Ala Pro Lys Lys Asp Pro Leu Pro Lys Asp Glu Glu Asp Phe
50 55 60
Leu Lys Ala Val Phe Gln Val Ile Leu Glu Asp Gly Val Arg Lys Gly
65 70 75 80
Arg Asp Ile Thr Gln Lys Val Val Asp Phe His Gln Pro Asp Glu Leu
85 90 95
Arg Thr Leu Leu Asp Leu Glu Ile Arg Asp Thr Pro Glu Asp His Gln
100 105 110
Ala Leu Ile Lys His Met Lys Asp Thr Val Lys Tyr Ser Val Arg Thr
115 120 125
Ser His Pro Arg Phe Phe Asn Gln Leu Phe Ser Gly Gln Asn Thr Tyr
130 135 140
Ala Leu Ala Gly Gln Trp Leu Thr Glu Thr Leu Asn Thr Ser Gln Tyr
145 150 155 160
Thr Phe Glu Val Ala Pro Val Phe Thr Ile Met Glu Asn Val Val Leu
165 170 175
His Lys Met Arg Asp Ile Val Gly Tyr Ser Gly Gly Asp Gly Ile Phe
180 185 190
Cys Pro Gly Gly Ser Ile Ser Asn Leu Tyr Ala Leu Asn Val Ala Arg
195 200 205
Tyr Arg Tyr Met Pro Asp Ile Lys Lys Thr Gly Leu Phe Gly Leu Pro
210 215 220
Arg Leu Val Val Phe Thr Ser Lys Gln Ser His Tyr Ser Ile Lys Lys
225 230 235 240
Ala Ala Ser Val Leu Gly Ile Gly Thr Asn Asn Val Val Leu Val Asp
245 250 255
Cys Asp Glu Arg Gly Lys Met Ile Ala Ser Asp Leu Glu Ala Gln Ile
260 265 270
Leu Arg Val Lys Ala Glu Gly Ala Val Pro Phe Phe Val Asn Cys Thr
275 280 285
Ser Gly Thr Thr Val Leu Gly Ala Tyr Asp Pro Leu Asp Glu Val Ser
290 295 300
Asp Ile Cys Glu Lys His Gly Leu Trp Met His Val Asp Ala Ala Trp
305 310 315 320
Gly Gly Gly Val Met Met Ser Pro Lys Tyr Arg Ala Ser Arg Met Arg
325 330 335
Gly Val Glu Arg Ser Asp Ser Ile Thr Trp Asn Pro His Lys Met Met
340 345 350
Gly Ala Gly Gln Gln Cys Ser Ala Phe Leu Leu Lys His Glu Asn Leu
355 360 365
Leu Gln His Cys His Glu Ala Lys Ala Lys Tyr Leu Phe Gln Gln Asp
370 375 380
Lys Phe Tyr Asp Val Ser Tyr Asp Thr Gly Asp Lys Ser Ile Gln Cys
385 390 395 400
Gly Arg Lys Val Asp Val Phe Lys Leu Trp Leu Met Trp Lys Ala Lys
405 410 415
Gly Ser Gln Gly Phe His Gln Asp Met Asp Ala Ile Phe Asp Lys Thr
420 425 430
Arg Tyr Leu Val Glu Lys Val Lys Ala Arg Glu Gly Phe Lys Met Val
435 440 445
Leu Asp Glu Pro Glu Cys Ser Asn Val Cys Phe Trp Tyr Ile Pro Pro
450 455 460
Ser Leu Arg Gly Lys Glu Asp Glu Ala Asp Tyr Lys Asp Lys Leu His
465 470 475 480
Gln Val Ala Pro Arg Ile Lys Glu Arg Met Val Leu Ser Gly Thr Met
485 490 495
Leu Val Gly Tyr Gln Pro Leu Gly Asn Lys Pro Asn Phe Phe Arg Gln
500 505 510
Val Phe Ser Ser Pro Ser Val Thr Glu Glu Asp Leu Asp Phe Leu Leu
515 520 525
Asp Glu Ile Glu Arg Leu Gly Glu Asp Leu
530 535
<210> 193
<211> 335
<212> PRT
<213> 油菜花露尾甲(Meligethes aeneus)
<400> 193
Lys Glu Met Arg Gln Ile Val Gly Phe Lys Asn Gly Asp Gly Asp Gly
1 5 10 15
Ile Phe Cys Pro Gly Gly Ser Met Ala Asn Gly Tyr Ala Ile Ser Cys
20 25 30
Ala Arg Tyr Lys Phe Met Pro Glu Ile Lys Gln Lys Gly Leu His Ala
35 40 45
Leu Pro Arg Leu Val Leu Phe Thr Ser Arg Asp Ala His Tyr Ser Ile
50 55 60
Lys Lys Leu Ser Ser Phe Leu Gly Ile Gly Thr Asp Asn Val Tyr Ala
65 70 75 80
Ile Asn Thr Asp Glu Lys Gly Lys Met Asp Met Lys His Leu Glu Glu
85 90 95
Glu Val Glu Arg Ser Ile Lys Glu Gly Gly Ala Pro Phe Met Val Ser
100 105 110
Ala Thr Ser Gly Thr Thr Val Ile Gly Ala Phe Asp Pro Leu Glu Lys
115 120 125
Ile His Glu Ile Cys Gln Lys Tyr Gly Met Trp Met His Val Asp Ala
130 135 140
Ala Trp Gly Gly Gly Ala Leu Met Ser Lys Lys His Arg Tyr Leu Leu
145 150 155 160
Lys Gly Ile Glu Lys Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu
165 170 175
Leu Thr Ala Pro Gln Gln Cys Ser Thr Leu Leu Leu Lys Gln Asp Gly
180 185 190
Ile Leu Ser Ala Met Asn Ser Ala Asn Ala Thr Tyr Leu Phe Gln Lys
195 200 205
Asp Lys Phe Tyr Asp Thr Lys Tyr Asp Ile Gly Asp Lys His Ile Gln
210 215 220
Cys Gly Arg Arg Pro Asp Val Ile Lys Phe Trp Phe Met Trp Lys Ala
225 230 235 240
Lys Gly Thr Ser Gly Phe Glu Gln His Ile Asp Lys Val Phe Glu Asn
245 250 255
Ala Lys Phe Phe Thr Asp Thr Ile Arg Glu Arg Glu Gly Phe Glu Met
260 265 270
Val Ile Pro Glu Pro Glu Cys Thr Asn Ile Cys Phe Trp Tyr Val Pro
275 280 285
Pro Ser Leu Arg Asn Arg Lys Ser Asp Pro Asp Tyr Gln Asp Lys Leu
290 295 300
His Lys Val Ala Pro Lys Ile Lys Glu Lys Met Met Arg Glu Gly Thr
305 310 315 320
Met Met Val Thr Tyr Gln Pro Leu Arg Glu Thr Pro Asn Phe Phe
325 330 335
<210> 194
<211> 401
<212> PRT
<213> 红火蚁(Solenopsis invicta)
<400> 194
Ser Thr Ser Gly Glu Asp Arg Val Val Glu Trp Val Asp Pro Tyr Glu
1 5 10 15
Leu Arg Ser Val Val Asp Leu Ser Leu Pro Ala Glu Gly Val Ser His
20 25 30
Glu Glu Leu Leu Lys Leu Thr Arg Asp Val Ile Lys Tyr Ser Val Lys
35 40 45
Thr Gly His Pro His Phe Val Asn Gln Leu Phe Ser Ser Leu Asp Pro
50 55 60
Tyr Gly Leu Leu Gly Gln Trp Leu Thr Asp Ala Leu Asn Pro Ser Val
65 70 75 80
Tyr Thr Tyr Glu Val Ser Pro Val Phe Ser Leu Met Glu Glu Asp Val
85 90 95
Leu Arg Glu Met Arg Ser Ile Ile Gly Trp His Asn Gly Glu Gly Leu
100 105 110
Phe Cys Pro Gly Gly Ser Met Ala Asn Gly Tyr Ala Ile Asn Leu Ala
115 120 125
Arg His His Arg Tyr Pro His Leu Lys Gln Thr Gly Leu Ser Gln Met
130 135 140
Pro Arg Leu Val Val Phe Thr Ser Glu Asp Ala His Tyr Ser Val Lys
145 150 155 160
Lys Leu Ala Ala Phe Leu Gly Ile Gly Tyr Asp Asn Val Tyr Leu Ala
165 170 175
Lys Val Asp Ser Arg Gly Lys Met Val Val Ser Asp Leu Glu Ala Gln
180 185 190
Ile Ala Arg Ala Ile Glu Glu Gly Ala Ala Pro Leu Met Val Ser Ala
195 200 205
Thr Ala Gly Thr Thr Val Ile Gly Ala Phe Asp Pro Leu Lys Asp Ile
210 215 220
Ala Glu Val Cys Lys Lys Tyr Gly Leu Trp Phe His Val Asp Ala Ala
225 230 235 240
Trp Gly Gly Gly Ala Leu Val Ser Ala Ala Tyr Arg Gly Leu Leu Asp
245 250 255
Gly Leu His Leu Ala Asp Ser Val Thr Trp Asn Pro His Lys Leu Leu
260 265 270
Ala Ala Pro Gln Gln Cys Ser Thr Leu Leu Leu Arg His Lys Gly Leu
275 280 285
Leu Gln Ala Ala His Gly Cys Gly Ala Ser Tyr Leu Phe Gln Asn Asp
290 295 300
Lys Phe Tyr Asp Ala Ser Phe Asp Cys Gly Asp Arg His Val Gln Cys
305 310 315 320
Gly Arg Arg Ala Asp Val Val Lys Phe Trp Tyr Met Trp Lys Ala Lys
325 330 335
Gly Thr Arg Gly Leu Glu Ala His Val Asp Arg Val Phe Ala Leu Ser
340 345 350
Arg Tyr Phe Ala Asp Leu Ile Arg Ala Arg Glu Gly Trp His Leu Leu
355 360 365
Val Glu Pro Glu Cys Thr Asn Val Cys Phe Arg Tyr Val Pro Pro Ser
370 375 380
Lys Arg His Leu Ala Gly Gln Glu Leu Asp Gln Val Leu His Lys Val
385 390 395 400
Ser
<210> 195
<211> 555
<212> PRT
<213> 太平洋牡蛎(Crassostrea gigas)
<400> 195
Met Ala Ser Cys Met Thr Glu Ser Phe Lys Val His Ser Cys Ala His
1 5 10 15
Lys Arg Ile His Asp Lys Asn Asp Phe Glu Val Glu Pro Thr Glu Lys
20 25 30
Gln Arg Ile Thr Met Leu Ser Asp Lys Pro Val Arg Lys Ser Asn Lys
35 40 45
Glu Asn Ser Met Cys Asn Val Thr Ile Lys Thr Lys Glu Ser His Ser
50 55 60
Glu Asn His Lys Lys His Glu Lys Lys Ala Pro Lys Gln Glu Lys Ala
65 70 75 80
Lys His Glu Phe Leu Asp Lys Leu Tyr Glu Met Met Ile Lys Asp Gly
85 90 95
Phe Met Lys Ala Arg Asp Arg Asn Glu Lys Val Val Glu Phe Ser Tyr
100 105 110
Pro Glu Glu Leu Lys Gln Lys Ile Asp Phe Asp Leu Gly Ser Lys Thr
115 120 125
Ser Asp Glu Lys Ile Leu Ser Leu Cys Gln Asp Ile Ile Lys Tyr Ser
130 135 140
Val Lys Val Ala His Pro Arg Phe Phe Asn Gln Leu Tyr Gly Gly Leu
145 150 155 160
Asp Glu Tyr Ser Leu Gly Gly Cys Trp Leu Thr Glu Thr Met Asn Ala
165 170 175
Ser Leu Tyr Thr Tyr Glu Val Ser Pro Val Phe Ser Leu Met Glu Arg
180 185 190
Val Val Ile Asp Lys Met Leu Gly Lys Ile Gly Phe Glu Asp Gly Asp
195 200 205
Ala Met Phe Cys Pro Gly Gly Ser Ile Ser Asn Met Tyr Ala Leu Asn
210 215 220
Ile Ala Arg Tyr Phe Lys Tyr Pro Glu Val Lys Lys Lys Gly Ile Lys
225 230 235 240
Gly Ile Pro Asp Ile Cys Ala Phe Thr Ser Glu Lys Cys His Tyr Ser
245 250 255
Ile Gly Lys Gly Val Ala Phe Met Gly Met Gly Leu Asp Asn Leu Ile
260 265 270
Asn Val Lys Thr Asp Ala Asn Gly Lys Met Ile Pro Glu Asp Leu Glu
275 280 285
Lys Lys Ile Leu Glu Ala Lys Ala Glu Gly Lys Thr Pro Tyr Phe Val
290 295 300
Asn Ala Thr Ala Gly Thr Thr Val Phe Gly Ala Phe Asp Pro Ile Asp
305 310 315 320
Glu Ile Ala Asp Ile Cys Gln Lys Tyr Asn Leu Trp Met His Val Asp
325 330 335
Gly Ala Trp Gly Gly Gly Ala Leu Leu Ser Lys Thr Tyr Ser Pro Leu
340 345 350
Leu Lys Gly Val Glu Arg Ala Asp Ser Met Thr Trp Asn Pro His Lys
355 360 365
Leu Met Gly Val Pro Gln Gln Cys Ser Leu Val Phe Thr Lys His Lys
370 375 380
Gly Leu Leu Glu Gln Cys His Ser Ala Asn Ala Ser Tyr Leu Phe Gln
385 390 395 400
Gln Asp Lys Phe Tyr Asp Val Ser Tyr Asp Thr Gly Asp Lys Ser Ile
405 410 415
Gln Cys Gly Arg Lys Asn Asp Val Leu Lys Leu Trp Ile Met Trp Lys
420 425 430
Asn Lys Gly Asp Glu Gly Phe Glu Arg Asp Ile Asp Asn Gln Phe Glu
435 440 445
Cys Ala Lys Tyr Leu Ala Gln Leu Val Gln Glu Arg Glu Gly Phe Glu
450 455 460
Leu Met Leu Glu Pro Gln Cys Thr Asn Val Cys Phe Tyr Tyr Ile Pro
465 470 475 480
Lys Arg Leu Arg Gly Leu Glu Arg Thr Pro Glu Trp Trp Asn Glu Ile
485 490 495
Ser Lys Val Gly Pro Lys Val Lys Glu Gly Met Met Lys Ala Gly Ser
500 505 510
Met Met Val Gly Tyr Gln Pro Asp Gly Asp Phe Val Asn Phe Phe Arg
515 520 525
Met Ile Ile Ser Asn Leu Asp Thr Val Lys Ser Asp Met Asp Phe Val
530 535 540
Val Asp Glu Ile Asp Arg Leu Gly Lys Asp Leu
545 550 555
<210> 196
<211> 1020
<212> DNA
<213> 近平滑假丝酵母(Candida parapsilosis)
<400> 196
atggttaact acggcttcgt tggtttgggt cagatgggtc aacacatggc tagacacatc 60
tacaaccaat tggaagtcga cgacaagttg tacgtttacg acaccgttcc aactgccgtt 120
gaccaattcg tttcaaacgt tacccagcaa aacgctacca acaaggagaa gttggttacc 180
ttgccagact tgaagtcctt cgttaccggt gttgacggtc aattggactt catcgtcacc 240
atggttccag agggtaagca cgtcaagggt gttgttgagg acatcgttac ctctttcgaa 300
caaaacggtt actccccaga ctacaacact accatcatcg actcctccac catcgacatc 360
ccaacctcca gacaggttca cgaatacgtt aaggaaacct tgcctcaatt cgacttcatc 420
gacgcacctg tttccggtgg tgttgctggt gccagaaagg gcaccttgtc cttcatgttg 480
tccagagaaa cccaccaaga cgtctctcca gccttgacta ccttgttgaa caagatgggt 540
aagaacatct tcccatgtgg tgctacccac ggcaccggtt tggctgctaa gttgtccaac 600
aactacttgt tggctgtcac caacatcgca gttgcagact ccttccaatt ggcaaaggca 660
ttcggtttga acttgcaaaa ctacgctaag ttggttgcag tttccaccgg caagtcttgg 720
gcttctgttg acaactgccc aatccctggt gtttacccag aaaacaactt gcctgcagac 780
gttggttacc aaggtggttt catcaccaag ttgaccagaa aggacgttgt tttggctacc 840
gactgcgcca aggaccaagg tagattcttg ttcttgggtg acgcaggtag atactggtac 900
gacaaggcgt gtgaaagaga agacatcgcc aacagagact tggcagtttt gtacgaatgg 960
ttgggtgact tgaagcaaga agcagacggc accgttgtcg acaccaaggt ttccaagtaa 1020
<210> 197
<211> 339
<212> PRT
<213> 近平滑假丝酵母(Candida parapsilosis)
<400> 197
Met Val Asn Tyr Gly Phe Val Gly Leu Gly Gln Met Gly Gln His Met
1 5 10 15
Ala Arg His Ile Tyr Asn Gln Leu Glu Val Asp Asp Lys Leu Tyr Val
20 25 30
Tyr Asp Thr Val Pro Thr Ala Val Asp Gln Phe Val Ser Asn Val Thr
35 40 45
Gln Gln Asn Ala Thr Asn Lys Glu Lys Leu Val Thr Leu Pro Asp Leu
50 55 60
Lys Ser Phe Val Thr Gly Val Asp Gly Gln Leu Asp Phe Ile Val Thr
65 70 75 80
Met Val Pro Glu Gly Lys His Val Lys Gly Val Val Glu Asp Ile Val
85 90 95
Thr Ser Phe Glu Gln Asn Gly Tyr Ser Pro Asp Tyr Asn Thr Thr Ile
100 105 110
Ile Asp Ser Ser Thr Ile Asp Ile Pro Thr Ser Arg Gln Val His Glu
115 120 125
Tyr Val Lys Glu Thr Leu Pro Gln Phe Asp Phe Ile Asp Ala Pro Val
130 135 140
Ser Gly Gly Val Ala Gly Ala Arg Lys Gly Thr Leu Ser Phe Met Leu
145 150 155 160
Ser Arg Glu Thr His Gln Asp Val Ser Pro Ala Leu Thr Thr Leu Leu
165 170 175
Asn Lys Met Gly Lys Asn Ile Phe Pro Cys Gly Ala Thr His Gly Thr
180 185 190
Gly Leu Ala Ala Lys Leu Ser Asn Asn Tyr Leu Leu Ala Val Thr Asn
195 200 205
Ile Ala Val Ala Asp Ser Phe Gln Leu Ala Lys Ala Phe Gly Leu Asn
210 215 220
Leu Gln Asn Tyr Ala Lys Leu Val Ala Val Ser Thr Gly Lys Ser Trp
225 230 235 240
Ala Ser Val Asp Asn Cys Pro Ile Pro Gly Val Tyr Pro Glu Asn Asn
245 250 255
Leu Pro Ala Asp Val Gly Tyr Gln Gly Gly Phe Ile Thr Lys Leu Thr
260 265 270
Arg Lys Asp Val Val Leu Ala Thr Asp Cys Ala Lys Asp Gln Gly Arg
275 280 285
Phe Leu Phe Leu Gly Asp Ala Gly Arg Tyr Trp Tyr Asp Lys Ala Cys
290 295 300
Glu Arg Glu Asp Ile Ala Asn Arg Asp Leu Ala Val Leu Tyr Glu Trp
305 310 315 320
Leu Gly Asp Leu Lys Gln Glu Ala Asp Gly Thr Val Val Asp Thr Lys
325 330 335
Val Ser Lys
<210> 198
<211> 996
<212> DNA
<213> 汉逊德巴利酵母(Debaryomyces hansenii)
<400> 198
atgaccaact acggcttcat cggtttgggt gaaatgggtc aacacatggg tagacacatc 60
tacaacaagt tggagccaga cgacaagttg tacgtttacg acttggaacc agcaaacacc 120
aagaagttcg tcgacaccgt taccgaagca tcccctgcaa acaagaagtt ggtcgttcca 180
ttggaatcca tctccgactt cgttcacaag gtcgactccc aattggagtt catcatgacc 240
atggttccag aaggtaagca cgtcaagtcc gttgtcgaag agttggtcca gaactacaag 300
aagtgtgcta ccgacatctc cggtgtctcc actaccttct tggactcctc caccgtcgac 360
atcccaacct ccatcgaagt ccacaagtac gtcaagcaac aaatcccaga cttcgacttc 420
atcgacaccc cagtttccgg tggtgttgcc ggtgccagaa agggcacctt gtccttcatg 480
ttgtccagag agacccacga agacatgtcc ccatccttga cctccttgtt gtccaagatg 540
ggcaccaaca tcttcccatg tggtaagaac cacggcaccg gtttggcagc aaagttgtcc 600
aacaactact tgttggctgt caccaacttg gcggtcgcag actccttcca attggcaaag 660
tccttcggtt tggacatgaa gaactacgct aagttggtct ccgtttccac cggtaagtcc 720
tgggcttccg ttgacaactg tccaatcaaa ggcgcttacc caaaggaaaa cgacttgcca 780
gcagacagag actgccacgg tggtttcatc accaagttgg ctagaaagga cttggtcttg 840
gctacccaat ccgcaaagtc caacaacaga ttcttgttct tgggtgaagt cggtaagaag 900
tggtacgaca aggcttgtga aagagaagac ttggcaacca aggacttgtc cgtcttgtac 960
gaatggttgg aagaattgtc cgaaaaggac aagtaa 996
<210> 199
<211> 331
<212> PRT
<213> 汉逊德巴利酵母(Debaryomyces hansenii)
<400> 199
Met Thr Asn Tyr Gly Phe Ile Gly Leu Gly Glu Met Gly Gln His Met
1 5 10 15
Gly Arg His Ile Tyr Asn Lys Leu Glu Pro Asp Asp Lys Leu Tyr Val
20 25 30
Tyr Asp Leu Glu Pro Ala Asn Thr Lys Lys Phe Val Asp Thr Val Thr
35 40 45
Glu Ala Ser Pro Ala Asn Lys Lys Leu Val Val Pro Leu Glu Ser Ile
50 55 60
Ser Asp Phe Val His Lys Val Asp Ser Gln Leu Glu Phe Ile Met Thr
65 70 75 80
Met Val Pro Glu Gly Lys His Val Lys Ser Val Val Glu Glu Leu Val
85 90 95
Gln Asn Tyr Lys Lys Cys Ala Thr Asp Ile Ser Gly Val Ser Thr Thr
100 105 110
Phe Leu Asp Ser Ser Thr Val Asp Ile Pro Thr Ser Ile Glu Val His
115 120 125
Lys Tyr Val Lys Gln Gln Ile Pro Asp Phe Asp Phe Ile Asp Thr Pro
130 135 140
Val Ser Gly Gly Val Ala Gly Ala Arg Lys Gly Thr Leu Ser Phe Met
145 150 155 160
Leu Ser Arg Glu Thr His Glu Asp Met Ser Pro Ser Leu Thr Ser Leu
165 170 175
Leu Ser Lys Met Gly Thr Asn Ile Phe Pro Cys Gly Lys Asn His Gly
180 185 190
Thr Gly Leu Ala Ala Lys Leu Ser Asn Asn Tyr Leu Leu Ala Val Thr
195 200 205
Asn Leu Ala Val Ala Asp Ser Phe Gln Leu Ala Lys Ser Phe Gly Leu
210 215 220
Asp Met Lys Asn Tyr Ala Lys Leu Val Ser Val Ser Thr Gly Lys Ser
225 230 235 240
Trp Ala Ser Val Asp Asn Cys Pro Ile Lys Gly Ala Tyr Pro Lys Glu
245 250 255
Asn Asp Leu Pro Ala Asp Arg Asp Cys His Gly Gly Phe Ile Thr Lys
260 265 270
Leu Ala Arg Lys Asp Leu Val Leu Ala Thr Gln Ser Ala Lys Ser Asn
275 280 285
Asn Arg Phe Leu Phe Leu Gly Glu Val Gly Lys Lys Trp Tyr Asp Lys
290 295 300
Ala Cys Glu Arg Glu Asp Leu Ala Thr Lys Asp Leu Ser Val Leu Tyr
305 310 315 320
Glu Trp Leu Glu Glu Leu Ser Glu Lys Asp Lys
325 330
<210> 200
<211> 1017
<212> DNA
<213> 季也蒙酵母(Meyerozyma guilliermondii)
<400> 200
atggcaaact acggtttcat cggtttgggt caaatgggcc aacacatggc gagacacatc 60
tacaaccaat tggaaccaaa cgactccttg tacgtccacg acgtctccag agacgcaacc 120
gagaccttcg ttaacaccgt cacctccgct tccccagaca gaaaggactg tttgaaggca 180
ttgtacaaca tctccgactt cgttaccggt gttaacgctc aattggacta catcatcacc 240
atggttcctg aaggtagaca cgttaagggt gttgttcaac agttgatcga aacctaccaa 300
caaggtgcac catccactac caagactacc atcttggact catctaccat cgacatccca 360
acctcaatcg aagttcacaa ctacgtcaag cagcaaatcc cagaattcga cttcatcgac 420
accccagtca gtggtggcgt tgctggtgcc agaaagggca ccttgtcctt catgttgtcc 480
agaccaaccg acgaatccat ctctccttct ttgaacacct tgttgaagaa gatgggtaag 540
aacatcttcc catgtggtgc aaaccacggt gcaggtttag ccgctaagtt gtccaacaac 600
tacttgttgg cagttaccaa cttggcggtt gccgactcct tcagattggc tcactccttc 660
ggtttggact tggtcaagta ctccaagttg gttgctgttt ccaccggcaa gtgttgggct 720
gcagttgaca actgcccaat cccaggtgtc tacccagccg agtacagatt gccagtcgac 780
gacggttaca acggtggttt catcaccaag ttgaccaaga aggacttggt tttggctacc 840
gacgctgcta agttcaacga ccgtttcttg ttcatgggtg acgtttccag acactggtac 900
gaaaaggctt gtgaaagaga agacatcgct tccagagact tggcagtttt gtacgaatgg 960
ttgggtgaca tggaacaaca agcagacggc accatcgtcg acaagaagga cgtctaa 1017
<210> 201
<211> 338
<212> PRT
<213> 季也蒙酵母(Meyerozyma guilliermondii)
<400> 201
Met Ala Asn Tyr Gly Phe Ile Gly Leu Gly Gln Met Gly Gln His Met
1 5 10 15
Ala Arg His Ile Tyr Asn Gln Leu Glu Pro Asn Asp Ser Leu Tyr Val
20 25 30
His Asp Val Ser Arg Asp Ala Thr Glu Thr Phe Val Asn Thr Val Thr
35 40 45
Ser Ala Ser Pro Asp Arg Lys Asp Cys Leu Lys Ala Leu Tyr Asn Ile
50 55 60
Ser Asp Phe Val Thr Gly Val Asn Ala Gln Leu Asp Tyr Ile Ile Thr
65 70 75 80
Met Val Pro Glu Gly Arg His Val Lys Gly Val Val Gln Gln Leu Ile
85 90 95
Glu Thr Tyr Gln Gln Gly Ala Pro Ser Thr Thr Lys Thr Thr Ile Leu
100 105 110
Asp Ser Ser Thr Ile Asp Ile Pro Thr Ser Ile Glu Val His Asn Tyr
115 120 125
Val Lys Gln Gln Ile Pro Glu Phe Asp Phe Ile Asp Thr Pro Val Ser
130 135 140
Gly Gly Val Ala Gly Ala Arg Lys Gly Thr Leu Ser Phe Met Leu Ser
145 150 155 160
Arg Pro Thr Asp Glu Ser Ile Ser Pro Ser Leu Asn Thr Leu Leu Lys
165 170 175
Lys Met Gly Lys Asn Ile Phe Pro Cys Gly Ala Asn His Gly Ala Gly
180 185 190
Leu Ala Ala Lys Leu Ser Asn Asn Tyr Leu Leu Ala Val Thr Asn Leu
195 200 205
Ala Val Ala Asp Ser Phe Arg Leu Ala His Ser Phe Gly Leu Asp Leu
210 215 220
Val Lys Tyr Ser Lys Leu Val Ala Val Ser Thr Gly Lys Cys Trp Ala
225 230 235 240
Ala Val Asp Asn Cys Pro Ile Pro Gly Val Tyr Pro Ala Glu Tyr Arg
245 250 255
Leu Pro Val Asp Asp Gly Tyr Asn Gly Gly Phe Ile Thr Lys Leu Thr
260 265 270
Lys Lys Asp Leu Val Leu Ala Thr Asp Ala Ala Lys Phe Asn Asp Arg
275 280 285
Phe Leu Phe Met Gly Asp Val Ser Arg His Trp Tyr Glu Lys Ala Cys
290 295 300
Glu Arg Glu Asp Ile Ala Ser Arg Asp Leu Ala Val Leu Tyr Glu Trp
305 310 315 320
Leu Gly Asp Met Glu Gln Gln Ala Asp Gly Thr Ile Val Asp Lys Lys
325 330 335
Asp Val
<210> 202
<211> 1023
<212> DNA
<213> 葡萄牙棒孢酵母(Clavispora lusitaniae)
<400> 202
atgaagaact tcggcttcat cggtttgggt caaatgggtc agcacatggc tagacacatc 60
tacaaccaat tggaacctca agacaccttg tacgtttacg acgcagtccc atccgcaacc 120
gacgcgttcg tcgaaaagaa ctccacccca gaaaaggctt ctcaattggt cccattgaag 180
tccttgtcct ccttcgttac cgacgttgac tcccaattgg acttcatcat caccatggtc 240
ccagaaggta agcacgtcaa ggctgtcatc caagacttgg tttcctccta ccaaaagcaa 300
ccatccttgc aaccaatgac cttcttggac tcctccacca tcgacatctc cacctccaga 360
gaagttcacg aattcgttaa gtccaccatc ccatccttcg acttcatcga caccccagtc 420
tctggtggcg ttgctggcgc tagaaaggcc tccttgtcct tcatgttgtc ccgtgaaacc 480
atcgaagacg tctcccctgc tttgacctcc ttgttgaaca agatgggtaa gaacatcttc 540
gcatgcggtc cttcccacgg ttccggcttg gcagcaaagt tggcaaacaa ctacttgttg 600
gctgtcacca acttggctgt tgcagactcc ttccaattgg caaacacctt caacttgaac 660
ttgcaacaat acgctaagtt ggttgctgtt tccaccggta agtcctgggc atccgttgac 720
aactgtccaa tcgctggtgc ttacccaaag gagtacaact tgccagcaga ctccggttac 780
gaaggtggtt tcatcaccaa gttgaccaag aaggacttgg ttttggcgac cgactgtgct 840
gcggaataca acagattctt gttcttgggc gacatctcca gaaagtggta cttgaaggca 900
tgtgagagag aagacttggg ttccagagac ttgggcgttt tgttcgaatg gttgggtcaa 960
atcgaagaaa gaaacggtga agttgttgac accaagacct ccgaaccatt gaagatcaac 1020
taa 1023
<210> 203
<211> 340
<212> PRT
<213> 葡萄牙棒孢酵母(Clavispora lusitaniae)
<400> 203
Met Lys Asn Phe Gly Phe Ile Gly Leu Gly Gln Met Gly Gln His Met
1 5 10 15
Ala Arg His Ile Tyr Asn Gln Leu Glu Pro Gln Asp Thr Leu Tyr Val
20 25 30
Tyr Asp Ala Val Pro Ser Ala Thr Asp Ala Phe Val Glu Lys Asn Ser
35 40 45
Thr Pro Glu Lys Ala Ser Gln Leu Val Pro Leu Lys Ser Leu Ser Ser
50 55 60
Phe Val Thr Asp Val Asp Ser Gln Leu Asp Phe Ile Ile Thr Met Val
65 70 75 80
Pro Glu Gly Lys His Val Lys Ala Val Ile Gln Asp Leu Val Ser Ser
85 90 95
Tyr Gln Lys Gln Pro Ser Leu Gln Pro Met Thr Phe Leu Asp Ser Ser
100 105 110
Thr Ile Asp Ile Ser Thr Ser Arg Glu Val His Glu Phe Val Lys Ser
115 120 125
Thr Ile Pro Ser Phe Asp Phe Ile Asp Thr Pro Val Ser Gly Gly Val
130 135 140
Ala Gly Ala Arg Lys Ala Ser Leu Ser Phe Met Leu Ser Arg Glu Thr
145 150 155 160
Ile Glu Asp Val Ser Pro Ala Leu Thr Ser Leu Leu Asn Lys Met Gly
165 170 175
Lys Asn Ile Phe Ala Cys Gly Pro Ser His Gly Ser Gly Leu Ala Ala
180 185 190
Lys Leu Ala Asn Asn Tyr Leu Leu Ala Val Thr Asn Leu Ala Val Ala
195 200 205
Asp Ser Phe Gln Leu Ala Asn Thr Phe Asn Leu Asn Leu Gln Gln Tyr
210 215 220
Ala Lys Leu Val Ala Val Ser Thr Gly Lys Ser Trp Ala Ser Val Asp
225 230 235 240
Asn Cys Pro Ile Ala Gly Ala Tyr Pro Lys Glu Tyr Asn Leu Pro Ala
245 250 255
Asp Ser Gly Tyr Glu Gly Gly Phe Ile Thr Lys Leu Thr Lys Lys Asp
260 265 270
Leu Val Leu Ala Thr Asp Cys Ala Ala Glu Tyr Asn Arg Phe Leu Phe
275 280 285
Leu Gly Asp Ile Ser Arg Lys Trp Tyr Leu Lys Ala Cys Glu Arg Glu
290 295 300
Asp Leu Gly Ser Arg Asp Leu Gly Val Leu Phe Glu Trp Leu Gly Gln
305 310 315 320
Ile Glu Glu Arg Asn Gly Glu Val Val Asp Thr Lys Thr Ser Glu Pro
325 330 335
Leu Lys Ile Asn
340
<210> 204
<211> 1035
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 204
atgagtaagc caaaagtctt gttgattgga ttcggtggtg ttggaaccat tgtatcttac 60
actttggagc accttggccg tgccgaggtc tctgcagttt cacgcccaga gacccatgac 120
tctatagtaa atggatttcg tatcgagtcc attgactatg gtatcgttga gaactatgtt 180
ccgaccaacg tttatgtcac agcaaaagaa gcttacaaac aacaaggccc atttgattat 240
atcattatca ccacaaagaa tatccctgat attgcaccag ttgttgatat gattgatggg 300
tgctacaacg agaaatccgt tattgtgttg attcaaaatg gcattgggat tgaaattcca 360
atttatagga gatatccaaa cgcaattata ttaagtgggg tcacgttgat tggcacaacg 420
ttgtacgaag ctacagtcaa gcatgtcgca agggatgata tcaagtttgg gccttttatc 480
aactataact tggataaaca gctgcagatt aacaagtgta aggaatttat tgaactttat 540
gaaaacgaca aaaacttggt tgaatatgag gaagatgtca agtttaccag atggaggaaa 600
ctcgtctaca atgcctgtat caatacaacc tgtgcgctag ctaatctaga tgcaggtaga 660
gtgcagatat ttggcggatt cgagaccctc gtcaaacctg ccatgttgga agtcattgcg 720
gttgctaaaa gtgagggtgt tgaattacca gcaaaggaag tgatggatac catgtgcaat 780
atgggcaaag atgtctacta tccaccttcc atgttgattg atgttcggaa cggcacgtac 840
ctggaacata ttgtcatcat tggcaatgtt gtcaaatatg ggtcccgtaa tggtgttcca 900
attccaacat tgacggtgtt gaacaacttg ttgaagctcg tccaaatgag aacaatggaa 960
gccaataaga ggtttgtctt gccagagaag aggccacttc cagaggaaaa ctaccagatt 1020
gaatacctct attga 1035
<210> 205
<211> 344
<212> PRT
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 205
Met Ser Lys Pro Lys Val Leu Leu Ile Gly Phe Gly Gly Val Gly Thr
1 5 10 15
Ile Val Ser Tyr Thr Leu Glu His Leu Gly Arg Ala Glu Val Ser Ala
20 25 30
Val Ser Arg Pro Glu Thr His Asp Ser Ile Val Asn Gly Phe Arg Ile
35 40 45
Glu Ser Ile Asp Tyr Gly Ile Val Glu Asn Tyr Val Pro Thr Asn Val
50 55 60
Tyr Val Thr Ala Lys Glu Ala Tyr Lys Gln Gln Gly Pro Phe Asp Tyr
65 70 75 80
Ile Ile Ile Thr Thr Lys Asn Ile Pro Asp Ile Ala Pro Val Val Asp
85 90 95
Met Ile Asp Gly Cys Tyr Asn Glu Lys Ser Val Ile Val Leu Ile Gln
100 105 110
Asn Gly Ile Gly Ile Glu Ile Pro Ile Tyr Arg Arg Tyr Pro Asn Ala
115 120 125
Ile Ile Leu Ser Gly Val Thr Leu Ile Gly Thr Thr Leu Tyr Glu Ala
130 135 140
Thr Val Lys His Val Ala Arg Asp Asp Ile Lys Phe Gly Pro Phe Ile
145 150 155 160
Asn Tyr Asn Leu Asp Lys Gln Leu Gln Ile Asn Lys Cys Lys Glu Phe
165 170 175
Ile Glu Leu Tyr Glu Asn Asp Lys Asn Leu Val Glu Tyr Glu Glu Asp
180 185 190
Val Lys Phe Thr Arg Trp Arg Lys Leu Val Tyr Asn Ala Cys Ile Asn
195 200 205
Thr Thr Cys Ala Leu Ala Asn Leu Asp Ala Gly Arg Val Gln Ile Phe
210 215 220
Gly Gly Phe Glu Thr Leu Val Lys Pro Ala Met Leu Glu Val Ile Ala
225 230 235 240
Val Ala Lys Ser Glu Gly Val Glu Leu Pro Ala Lys Glu Val Met Asp
245 250 255
Thr Met Cys Asn Met Gly Lys Asp Val Tyr Tyr Pro Pro Ser Met Leu
260 265 270
Ile Asp Val Arg Asn Gly Thr Tyr Leu Glu His Ile Val Ile Ile Gly
275 280 285
Asn Val Val Lys Tyr Gly Ser Arg Asn Gly Val Pro Ile Pro Thr Leu
290 295 300
Thr Val Leu Asn Asn Leu Leu Lys Leu Val Gln Met Arg Thr Met Glu
305 310 315 320
Ala Asn Lys Arg Phe Val Leu Pro Glu Lys Arg Pro Leu Pro Glu Glu
325 330 335
Asn Tyr Gln Ile Glu Tyr Leu Tyr
340
<210> 206
<211> 1020
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 206
caacttggtt caaaccaaga aagccgtcgt gtactttgcg tattttcaac ttcttccctt 60
caactccaac cttataaccc aatccgacag ttagcataca gtagaaaatc cataagttaa 120
ttgcaaaaac cacataattc ccaatgactg gcacaatttg catgaacggc gcaaaaccaa 180
cacctatcct tcttgacttg tggacaactt ctccggcatc ggctccagtt tctggtttag 240
gcgacacttt caaggtaccc ttgttgacta ggttctcagc aacatactcc tcctcgataa 300
ctttgaccgt ctttgtgcca aacacattgt tcaatagttt ggacgctcgc ataccgaaga 360
cagactcgtc catccaatag catgtactcc gaatgctctc aatcaacggt atcaaatcgc 420
ttggtgtctt tggaggcggt gttctaggtt tcatcttcgc cttcccgttt cctgtttctt 480
ggacttcaaa atacgggtca tatgcatctt cattggtgaa cctgaacagt ttgggccgag 540
tagtgccaaa caccttcaac agtataatct tcactccttg gccgattaga aacatgctta 600
tatgtttgcc ggtattctta aagtgtatac tggagccctt ttaatcacat tttttttact 660
tgtcagtctc catacggagt ttaatgtcct tatatcgatc ttcatcagct ccaacgggac 720
aggaatcaat aaccttcctt gcctgtccca aaaagaatga attttttttc aaaagcttta 780
cgatgcatac cacaaaagga agattattcc cacatgttcc agaagtgtgc ggagatacaa 840
agggttcatg aaaacgtgaa tcttctaaaa acttagcaca acaataaaaa tctacaatgt 900
tacagtaagt attattttct ttttgtcgac acactccaac ggttagattt ccaagtattc 960
aatccaatgt attacttgtc agacagccat ccactcccat cttagaacat cacttccgaa 1020
<210> 207
<211> 20
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 207
gatatgggcg gtagagaaga 20
<210> 208
<211> 20
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 208
gctccttcaa aggcaacaca 20
<210> 209
<211> 53
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 209
gtaaaacgac ggccagtgag ttcgttaacg gtagagtctc tccttctggt tcg 53
<210> 210
<211> 66
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 210
aaaaataaac tagtaaaata aattaattaa catgcggccg caagtgtgag agaaggagaa 60
agaccg 66
<210> 211
<211> 66
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 211
ctggagaata gatcttcaac gcgtttaaac tgtgcggccg cctgaagtgc cattcccatc 60
acatgg 66
<210> 212
<211> 51
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 212
gaccatgatt acgccaagct ccgcggtagc ggggaagata ttttttgaat a 51
<210> 213
<211> 27
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 213
ccacaagcca caatgatgat aaatggg 27
<210> 214
<211> 24
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 214
gttgtagagt gtatgcggca atgg 24
<210> 215
<211> 26
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 215
gttgaacaac ttgttgaagc tcgtcc 26
<210> 216
<211> 20
<212> DNA
<213> 东方伊萨酵母(Issatchenkia orientalis)
<400> 216
tcggcacggc caaggtgctc 20

Claims (28)

1.一种重组酵母细胞,包含(1)能够产生3-HP的主动3-HP途径和(2)对编码丙酮酸还原酶的内源基因的破坏,其中:
(a)该丙酮酸还原酶与SEQ ID NO:205具有至少97%序列一致性;或
(b)编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少97%序列一致性;
其中所述丙酮酸还原酶源自东方伊萨酵母(Issatchenkia orientalis)。
2.如权利要求1所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205具有至少98%序列一致性。
3.如权利要求1所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205具有至少99%序列一致性。
4.如权利要求1所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205具有100%序列一致性。
5.如权利要求1-4中任一项的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少98%序列一致性。
6.如权利要求1-4中任一项的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少99%序列一致性。
7.如权利要求1-4中任一项的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有100%序列一致性。
8.如权利要求1-7中任一项所述的重组细胞,其中该丙酮酸还原酶与SEQ ID NO:205相差不超过五个氨基酸。
9.如权利要求1-7中任一项所述的重组细胞,其中该细胞包含对编码丙酮酸还原酶的内源基因的破坏,该丙酮酸还原酶由SEQ ID NO:205组成。
10.如权利要求1-9中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少97%序列一致性。
11.如权利要求1-9中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少98%序列一致性。
12.如权利要求1-9中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有至少99%序列一致性。
13.如权利要求1-9中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列与SEQ ID NO:204具有100%序列一致性。
14.如权利要求1-7中任一项所述的重组细胞,其中编码该丙酮酸还原酶的内源基因的编码序列由SEQ ID NO:204组成。
15.如权利要求1-14中任一项所述的重组细胞,其中当在相同条件下培养时,与缺乏对编码该丙酮酸还原酶的内源基因的破坏的亲本菌株相比,细胞产生的D-乳酸盐/D-乳酸酯(D-lactate)减少至少50%。
16.如权利要求1-15中任一项所述的重组细胞,其中当在相同条件下培养时,与缺乏对编码该丙酮酸还原酶的内源基因的破坏的亲本菌株相比,该细胞产生的丙酮酸还原酶减少至少50%。
17.如权利要求1-16中任一项所述的重组细胞,其中使编码该丙酮酸还原酶的内源基因失活。
18.如权利要求1-17中任一项所述的重组细胞,其中该细胞包含选自以下各项的一种或多种异源多核苷酸:
编码丙酮酸脱氢酶(PDH)的异源多核苷酸;
编码乙酰辅酶A羧化酶(ACC)的异源多核苷酸;
编码丙二酰辅酶A还原酶的异源多核苷酸;和
编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。
19.如权利要求1-17中任一项所述的重组细胞,其中该细胞包含选自以下各项的一种或多种异源多核苷酸:
编码PEP羧化酶(PPC)的异源多核苷酸;
编码丙酮酸羧化酶(PYC)的异源多核苷酸;
编码天冬氨酸转氨酶(AAT)的异源多核苷酸;
编码天冬氨酸1-脱羧酶(ADC)的异源多核苷酸;
编码β-丙氨酸转氨酶(BAAT)或氨基丁酸转氨酶(gabT)的异源多核苷酸;和
编码3-HP脱氢酶(3-HPDH)的异源多核苷酸。
20.如权利要求1-19中任一项所述的重组细胞,其中该细胞属于选自以下各项的属:伊萨酵母属、假丝酵母属、克鲁维酵母属、毕赤酵母属、裂殖酵母属、有孢圆酵母属、接合酵母属和酵母属。
21.如权利要求20所述的重组细胞,其中该细胞选自东方伊萨酵母、郎比可假丝酵母以及布拉迪酵母(S.bulderi)。
22.如权利要求1-21中任一项所述的重组细胞,其中该细胞是CB1酵母细胞。
23.如权利要求1-22中任一项所述的重组细胞,其中该酵母细胞不能发酵戊糖。
24.一种生产3-HP的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如权利要求1-23中任一项所述的重组酵母细胞,以生产3-HP;并且
(b)回收该3-HP。
25.一种生产丙烯酸或其盐的方法,该方法包括:
(a)在适合的条件下,在可发酵的培养基中培养如权利要求1-23中任一项所述的重组细胞,以生产3-HP;
(b)回收该3-HP;
(c)在适合的条件下,将该3-HP脱水,以生产丙烯酸或其盐;并且
(d)回收该丙烯酸或其盐。
26.如权利要求24或25所述的方法,其中该可发酵的培养基包括少于1%的戊糖。
27.如权利要求24-26中任一项所述的方法,其中该重组细胞是CNB1酵母细胞。
28.一种用于获得如权利要求1-23中任一项所述的重组宿主细胞的方法,该方法包括:
(a)培养亲本菌株;
(b)(i)用一个或多个3-HP途径基因转化该亲本菌株以在(a)的亲本菌株中提供主动3-HP途径;
(b)(ii)破坏在(a)的亲本菌株中编码丙酮酸还原酶的内源基因;并且
(c)分离生成自(b)(i)和(b)(ii)的突变菌株。
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