CN107849521B - 用于由羧酸合成聚合物前体的过程和微生物 - Google Patents

用于由羧酸合成聚合物前体的过程和微生物 Download PDF

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CN107849521B
CN107849521B CN201680042118.8A CN201680042118A CN107849521B CN 107849521 B CN107849521 B CN 107849521B CN 201680042118 A CN201680042118 A CN 201680042118A CN 107849521 B CN107849521 B CN 107849521B
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A·亚库宁
A·库什努特诺瓦
J·陈洲
R·马哈德文
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Abstract

提供了一种由羧酸生物合成聚合物前体的方法，所述聚合物前体包括己二酸、1,6‑己二醇、6‑羟基己酸和6‑氨基己酸。提供了一种通过用酶处理由具有a,β‑烯酸酯还原酶活性的六碳二元羧酸生物合成己二酸的方法。描述了使用新型羧酸酯还原酶、醛还原酶和氨基转移酶将脂肪族和羟基羧酸生物催化转化为对应的醛、醇和胺。还提供了用于生物合成过程的遗传工程化微生物。

Description

用于由羧酸合成聚合物前体的过程和微生物
本申请包括序列表作为其说明书的一部分，所述序列表包括111个序列并且以电子形式与本申请一起提交，并且此序列表整体并入本申请。
技术领域
本公开涉及由羧酸酶促合成聚合物前体。
技术背景
一元羧酸和二元羧酸(诸如己二酸、富马酸、葡糖二酸和琥珀酸)、二胺(尸胺、腐胺、六亚甲基二胺)和二醇(丙二醇和丁二醇)是化学工业中用于缩聚反应的最常见的聚合物前体。聚合物材料在多种工业中应用广泛，但大多数聚合物材料仍然是由石油生产的。
己二酸是用于合成尼龙-6,6聚酰胺(约60亿美元的全球市场)的最重要的商业脂肪族二元羧酸。每年有260万公吨的己二酸是由石油来源的苯生产的，但已知的化学过程会产生有毒的副产物，诸如一氧化二氮(N2O)。
通常由石油制造的其他重要的聚合物前体包括1,4-丁二醇、1,6-己二醇、6-羟基己酸和6-氨基己酸。
由于人们越来越关注气候变化、能源安全和人类健康，因此需要可再生生物质代替石油来可持续生产商品化学品。
简述
一方面，提供了一种其中引入了外源核酸的宿主微生物，其中所述外源核酸编码具有α,β-烯酸酯还原酶活性的酶，其中所述酶包含SEQ ID NO:1的氨基酸序列或其活性片段或同源物。在一个实施方案中，所述酶包含SEQ ID NO:1的氨基酸(363)至(382)。在一个实施方案中，所述酶包含与SEQ ID NO:1的氨基酸序列至少80％、85％、90％、95％或100％相同的氨基酸序列。
另一个方面，提供了一种其中引入了外源核酸的宿主微生物，其中所述外源核酸编码具有α,β-烯酸酯还原酶活性的酶，其中所述酶包含SEQ ID NO:2的氨基酸序列或其活性片段或同源物。
在一个实施方案中，具有α,β-烯酸酯还原酶活性的酶来源于芽孢杆菌属(Bacillus)、梭菌属(Clostridium)或穆尔氏菌属(Moorella)的微生物。在一个实施方案中，所述微生物为凝结芽孢杆菌(Bacillus coagulans)。
在一个实施方案中，所述宿主微生物为大肠杆菌(Escherichia coli)。
还提供了用于生产己二酸的过程，所述过程包括在生产己二酸的条件下培养如上所述的宿主微生物足够长的时间以生产己二酸。
还提供了用于生产己二酸的过程，所述过程包括使用2-烯酸酯还原酶将6-碳不饱和二元羧酸酶促转化为己二酸。在一个实施方案中，所述酶包含SEQ ID NO:1或2的氨基酸序列，或者是包含SEQ ID NO:1或2的氨基酸序列的酶的片段或同源物。
在一个实施方案中，不饱和6-碳二元羧酸是粘康酸或2-己烯二酸。在一个实施方案中，6-碳不饱和二元羧酸由生物合成方法来生产。
在一个实施方案中，所述过程包括在生产己二酸的条件下培养如上所述的宿主微生物足够长的时间以生产己二酸。
在所述过程的一个实施方案中，培养基基本上是有氧的。
还提供了一种组合物，所述组合物包含如上所述的宿主微生物和粘康酸和/或2-己烯二酸。
另一个方面，提供了一种其中引入了外源核酸的宿主微生物，其中所述外源核酸编码具有羧酸酯还原酶活性的酶，其中所述酶包含SEQ ID NO:36的氨基酸序列或其活性片段或同源物。在一个实施方案中，所述酶包含与SEQ ID NO:36的氨基酸序列至少80％、85％、90％、95％或100％相同的氨基酸序列。在一个实施方案中，所述宿主微生物为大肠杆菌。
还提供了包含编码具有羧酸酯还原酶活性的酶的外源核酸的宿主微生物，其中所述酶包含SEQ ID NO:35、36、37、38或70中任一个的氨基酸序列或其活性片段或同源物。在一个实施方案中，宿主微生物还共表达编码磷酸泛酰巯基乙胺基转移酶的基因。
还提供了包含编码具有醛还原酶活性的酶的外源核酸的宿主微生物，其中所述酶包含NO:40-52的氨基酸序列，优选SEQ ID NO:47、43、50、45、46或52中的一个氨基酸序列，或其活性片段或同源物。
还提供了包含编码具有氨基转移酶活性的酶的外源核酸的宿主微生物，其中所述酶包含SEQ ID NO:53-65中的一个氨基酸序列，优选SEQ ID NO:53、59、55、58或60中的一个氨基酸序列，或其活性片段或同源物。
另一个方面，提供了用于生产短链醛的过程，所述过程包括使用羧酸酯还原酶将C5-C8饱和的二元羧酸的羧基酶促转化为醛。在一个实施方案中，羧酸酯还原酶包含SEQ IDNO:35、36、37、38或70中任一个的氨基酸序列或其活性片段或同源物。
所述过程还可包括使用醛还原酶将醛基酶促转化为羟基。在一个实施方案中，醛还原酶包含SEQ ID NO:40-52中的一个氨基酸序列，优选SEQ ID NO:47、43、50、45、46或52中的一个氨基酸序列，或其活性片段或同源物。
所述过程还可包括使用羧酸酯还原酶将第二羧酸基团酶促转化为醛，并且任选地使用醛还原酶将所述醛酶促转化为羟基。
在另一个实施方案中，所述过程还可包括使用氨基转移酶将醛酶促转化为胺。在一个实施方案中，氨基转移酶包含SEQ ID NO:53-65中的一个氨基酸序列，优选SEQ ID NO:53、59、55、58或60中的一个，或其活性片段或同源物。所述过程还可包括使用羧酸酯还原酶将第二羧酸基团酶促转化为醛，并且任选地使用氨基转移酶将所述醛酶促转化为胺。
在上述过程中，二元羧酸可为己二酸。
在一个实施方案中，醛为6-氧代己酸，并且酶转化的产物为6-羟基己酸。
在一个实施方案中，所述产物为1,6-己二醇。
在一个实施方案中，所述产物为6-氨基己酸。
在一个实施方案中，所述产物为六亚甲基二胺。
在一个实施方案中，所述二元羧酸是通过上述过程生产的己二酸。
还提供了用于生产6-羟基己酸的过程，所述过程包括在生产6-羟基己酸的条件下培养如上所述的宿主微生物足够长的时间以生产6-羟基己酸。
还提供了用于生产1,6-己二醇的过程，所述过程包括在生产1,6-己二醇的条件下培养如上所述的宿主微生物足够长的时间以生产1,6-己二醇。
还提供了用于生产6-氨基己酸的过程，所述过程包括在生产6-氨基己酸的条件下培养如上所述的宿主微生物足够长的时间以生产6-氨基己酸。
还提供了用于生产六亚甲基二胺的过程，所述过程包括在生产六亚甲基二胺的条件下培养如上所述的宿主微生物足够长的时间以生产六亚甲基二胺。
还提供了用于生产1,4-丁二醇的过程，所述过程包括在生产1,4-丁二醇的条件下培养宿主微生物足够长的时间以生产1,4-丁二醇，所述宿主微生物包含编码具有羧酸酯还原酶活性的酶的外源核酸，其中所述酶包含SEQ ID NO:36的氨基酸序列或其活性片段或同源物。在一个实施方案中，所述酶包含与SEQ ID NO:36的氨基酸序列至少80％、85％、90％、95％或100％相同的氨基酸序列。
附图简述
图1示出了使用2-烯酸酯还原酶(ERED)将不饱和六碳二元羧酸化学催化和生物催化氢化成己二酸。
图2示出了用于将羧酸生物转化为1,6-己二醇、6-羟基己酸和6-氨基己酸的酶和级联酶促反应。
图3示出了用于将4-羟基丁酸生物转化为1,4-丁二醇和4-氨基丁醇的酶和级联酶促反应。
图4使出了25种老黄酶(OYE；EC 1.6.99.1)的活性谱。OYE的酶活性的截断值为0.05U/mg蛋白质。在反应混合物(磷酸钾100mM和pH 7.0)中使用1mM底物、0.25mM NADPH和2.5μg蛋白质。2-己烯二酸的顺式异构体和反式异构体的混合物被用作底物。
图5示出了大肠杆菌中ERED表达的SDS-PAGE分析和蛋白质纯化。a)EREDCA、EREDCK和EREDBC。b)EREDMT、EREDCL和EREDCT。LC-MS分析显示，EREDCT的E泳道中75kDa的可溶性蛋白为假定蛋白ECD_02181(74.2kDA，基因库：ACT44004.1)、热休克蛋白90(71.4kDa，ACT42323.1)和分子伴侣DnaK(69.1kDa，ACT41916.1)(数据未示出)。M：标记，E：纯化的ERED，S：上清液(可溶性部分)，P：沉淀(不溶性部分)。
图6示出了表达重组ERED的大肠杆菌细胞将2-己烯二酸厌氧生物转化为己二酸的时间进程。2-己烯二酸的顺式异构体和反式异构体的混合物被用作底物，并且结果为来自至少两个独立测定的平均值。使用20mM 2-己烯二酸用于生物转化。
图7示出了表达重组a)EREDCL和b)EREDMT的大肠杆菌细胞将2-己烯二酸完全生物转化为己二酸。在厌氧条件下使用20mM 2-己烯二酸用于生物转化。
图8示出了表达重组a)EREDBC、b)EREDCA、c)EREDMT、d)EREDCK和e)EREDCL的大肠杆菌细胞将顺式,顺式-粘康酸24小时厌氧生物转化为己二酸。使用饱和顺式,顺式-粘康酸溶液(0.7mM)用于生物转化。底物和产物通过LC-MS进行检测。1：顺式,顺式-粘康酸，2：反式,反式-粘康酸，3：2-己烯二酸，4：己二酸，5：3-己烯二酸样化合物，m/z为143.0319。
图9示出了纯化的EREDBC和EREDCA的紫外可见吸收光谱。在空气(氧化，空气线)或3mM NADH(还原，NADH线)的存在下将纯化的蛋白保持在冰上。随后将6mM反式肉桂酸加入还原蛋白质中导致部分再氧化(在450nm处吸光度增加)。分光光度比色皿示出了在这些实验中使用的纯化EREDBC和EREDCA的棕色制剂。
图10示出了纯化EREDBC的体外生物化学表征。EREDBC的还原酶活性作为a)2-己烯二酸、b)2-环己烯-1-酮、c)3-甲基-2-己烯酮和d)反式肉桂酸浓度的函数。
图11示出了纯化EREDCA的体外生物化学表征。EREDCA的还原酶活性作为在NADH存在下的a)反式肉桂酸浓度或b)2-己烯二酸浓度的函数。
图12示出了纯化EREDBC和EREDCA的体外生物化学表征。还原酶活性作为在NADH存在下溶解在含有异丙醇的反应混合物中的2-己烯二酸的函数。
图13示出了纯化的a)EREDBC和b)EREDCA在21％的氧气存在下的体外氧气耐受性(残余活性)。
图14示出了研究的ERED的多序列比对。分别通过Cluster Omega和ESPript 3.0进行多序列比对和可视化。
图15示出了在厌氧条件下表达重组EREDBC(黑)和EREDCA(白)的大肠杆菌细胞将2-己烯二酸(15mM)全细胞生物转化为己二酸。细胞生长并且蛋白质厌氧表达。
图16示出了纯化的羧酸酯还原酶(CAR)的底物筛选：A–庚二酸、辛二酸、壬二酸(各5mM)；B-巴豆酸、丁酸、丙酸(各10mM)；C-苯甲酸(5mM)和肉桂酸(10mM)；D-己二酸和4-羟基丁酸(各5mM)。在NADPH消耗的氧化之后在340nm处通过分光光度法测量还原酶活性。
图17示出了A-在1mM NADH、1mM丙酮酸存在下的BSU31930用NH4Cl的饱和。B-在1mMNADPH、1mM酮戊二酸存在下的YOR375C。
图18示出了在50mM甲酸盐存在下PP33160双突变体(D222Q/H224N)用NADPH和NADH辅因子的饱和。
图19示出了与10mM己二酸或10mM 4-羟基丁酸(12h)的醛还原酶(AR)PP_3370转化偶联的体外CAR。反应由供应有50mM甲酸盐和5mM多磷酸盐的ATP和NADPH-再生酶供应。A–最终产品浓度的图形视图。B–转换效率的表格。
图20示出了表达CAR的大肠杆菌对10mM己二酸或10mM 4-羟基丁酸的体内转化。A–最终的1,6-己二醇和6-羟基己酸形成的时间进程。B–最后的1,4-丁二醇形成的时间过程C-转换效率的表格。
详述
如本文所用，“酶”包括由能够催化生物化学反应的细胞产生的蛋白质。此外，除非上下文另外规定，否则如本文所用，“酶”包括保留相关催化活性的蛋白质片段，并且可包括合成以保持相关的催化活性的人造酶。
一方面，本公开提供了用于由可再生原料生产聚合物前体的新型酶以及用于生产所述新型酶的微生物。
一方面，本公开提供了能够还原不饱和二元羧酸的C＝C键的新型酶以及用于生产所述新型酶的微生物。
一方面，本公开提供了用于将可由本文所述的不饱和二元羧酸生产的羟基羧酸转化为对应的醛、醇和胺的酶。
一方面，本公开提供了使用对α,β-烯酸酯具有氢化活性的生物催化剂来生产用于由不饱和六碳二元羧酸生物化学合成己二酸的细胞和生物体的过程。在一个实施方案中，如图1所示，己二酸可以使用2-烯酸酯还原酶由粘康酸或2-己烯二酸来生产。
在一个实施方案中，己二酸的生物化学合成在基本有氧的环境中执行。
另一个方面，本公开提供了将一元羧酸和二元羧酸酶促转化为对应的醛、醇和胺的过程。
一方面，本公开提供了短链饱和一元羧酸和二元羧酸(在一个实施方案中为C3至C8饱和一元羧酸和二元羧酸)酶促转化为对应的醛、醇和胺的过程。在一个实施方案中，所述羧酸为C5至C7二元羧酸。在一个实施方案中，所述羧酸为C6二元羧酸，在一个实施方案中为己二酸。如图2所示，己二酸的酶促转化产物可为1,6-己二醇、6-羟基己酸和6-氨基羧酸。
在另一个实施方案中，描述了可用末端羟基取代的一元羧酸的酶促转化，在一个实施方案中，所述羧酸为C3或C4一元酸，在一个实施方案中，如图3所示为4-羟基丁酸(4-hydroxybutyric acid)(4-羟基丁酸(4-hydroxybutanoic acid))酶促转化为对应的4-羟基丁醛，其可以如本文所述通过醛还原酶和氨基转移酶酶促转化为1,4-丁二醇或4-氨基丁醇。
羧酸酶促转化为对应的醛、醇和胺采用新型的羧酸酯还原酶(CAR)、醛还原酶(AR)和/或氨基转移酶(AT)。还提供了生产用于这些醛、醇和胺的生物化学合成的细胞和生物体的过程。
由于人们越来越关注气候变化、能源安全和人类健康，因此已经做了很大的努力来开发可再生生物质代替石油来可持续生产商品化学品。
使用可再生原料(诸如糖、淀粉、木质素、纤维素和半纤维素)生产聚合物前体可以帮助克服化石资源的消耗，而不改变当前聚合物合成的技术过程。使用工程化莽草酸酯[Curran K.A.,Leavitt J.M.,Karim A.S.,Alper H.S.Metabolic engineering ofmuconic acid production in Saccharomyces cerevisiae.Metab.Eng.15,55–66(2013)]或酮己二酸酯、逆转的β-氧化途径[YuJ L.,Xia X.X.,Zhong J.J.,Qian Z.G.Directbiosynthesis of adipic acid from a synthetic pathway in recombinantEscherichia coli.Biotechnol Bioeng,111,2580-2586(2014)]生产二元羧酸的工程化菌株的最近发展能够积累脂肪酸。
已经由葡萄糖通过顺序化学反应生产生物基己二酸，所述顺序化学反应即葡萄糖氧化为葡糖二酸并且葡糖二酸加氢脱氧为己二酸，所述顺序化学反应仅可以由化学催化剂催化(US20100317823A1)。己二酸的生物基生产也已经尝试使用代谢工程方法。具有工程化芳香族氨基酸生物合成途径的工业微生物(诸如大肠杆菌或酿酒酵母(Saccharomycescerevisiae))可以由葡萄糖生产顺式,顺式-粘康酸，然后所述顺式,顺式-粘康酸使用化学催化剂氢化为己二酸(参见例如Niu,W.,Draths,K.M.&Frost,J.W.Benzene-freesynthesis of adipic acid.Biotechnology Progress 18,201-211(2002))。然而，由2-己烯二酸或粘康酸生产己二酸的生物合成途径尚未被实验证明。
本公开提供了使用对α,β-烯酸酯具有氢化活性的生物催化剂由不饱和六碳二元羧酸生物化学合成己二酸。在一个实施方案中，己二酸使用2-烯酸酯还原酶(ERED；EC1.3.1.31)由粘康酸或2-己烯二酸来生产。然而，存在若干已知的从常见的可再生原料开始生产粘康酸和2-己烯二酸的途径，并且在一个实施方案中，一种或多种此类途径被适当地引入宿主微生物中。
本公开还提供了使用新型羧酸酯还原酶(EC 1.2.1.30)、醛还原酶(EC 1.1.1.-)和/或氨基转移酶(EC 2.6.1.48；EC 2.6.1.19)使用生物催化剂将羟基羧酸生物化学合成醛、醇和胺。在一个实施方案中，使用AR(EC 1.1.1.-)由6-羟基己醛生产1-6-己二醇。在一个实施方案中，使用AR(EC 1.1.1.-)由6-氧代己酸生产6-羟基己酸。在一个实施方案中，使用AT(EC 2.6.1.48；EC 2.6.1.19)由6-氧代己酸生产6-氨基己酸。
已经努力使用羧酸还原酶生产1,4-丁二醇，如Burke等2013WO2013184602A2中所述，其中鉴定了来自爱荷华诺卡氏菌(Nocardia iowensis)、耻垢分枝杆菌(Mycobacteriumsmegmatis)mc(2)155、鸟分枝杆菌(Mycobacterium avium)类结核(paratuberculosis)亚种K-10、海洋分枝杆菌(Mycobacterium marinum)M(指定为892)的CAR。
为了生产1,4-丁二醇，可以将羧基还原酶整合到产生4-羟基丁酸的宿主中。4羟基丁酸可以由2-酮戊二酸通过脱羧作用生成琥珀酸半醛然后还原来制备，或者由4-氨基丁酸和2-酮戊二酸通过转氨作用生成4-羟基丁酸和L-谷氨酸来制备[Yim等,Metabolicengineering of Escherichia coli for direct production of 1,4-butanediol.NatChem Biol.2011May 22；7(7):445-52.]。
与酶或(多)肽有关的表述“来源于”表示所述酶或多(肽)是从(微)生物中分离的，或者所述酶或(多)肽包括从(微)生物中分离或表征的酶或(多)肽的氨基酸序列的全部或生物学活性部分。
在某些实施方案中，如本文所提供的ERED来源于芽孢杆菌属、梭菌属或穆尔氏菌属的生物体。在一个实施方案中，ERED来源于丙酮丁醇梭菌(Clostridiumacetobutylicum)种。在一个实施方案中，ERED来源于芽孢杆菌属。在一个实施方案中，ERED来源于凝结芽孢杆菌种。
在一个实施方案中，ERED包含以下或者由以下组成：SEQ ID NO:1的氨基酸或其活性片段或同源物。
在另一个实施方案中，ERED包含以下或者由以下组成：SEQ ID NO:2的氨基酸或其活性片段或同源物。
一方面，所述酶与SEQ ID NO:1或SEQ ID NO:2的氨基酸序列至少80％、85％、90％、95％或100％相同。
在某些实施方案中，如本文所提供的CAR来源于分枝杆菌属(Mycobacterium)的生物体。在一个实施方案中，CAR来源于脓肿分枝杆菌(Mycobacterium abscessus)种。在一个实施方案中，CAR来源于类结核分枝杆菌(Mycobacterium paratuberculosis)种。在一个实施方案中，CAR来源于耻垢分枝杆菌种。
在一个实施方案中，所使用的CAR包含以下或者由以下组成：SEQ ID NO:35、36、37、38或70中的一个的氨基酸或其活性片段或同源物。
一方面，所述酶与SEQ ID NO:35、36、37、38或70中的一个氨基酸序列至少80％、85％、90％、95％或100％相同。
本发明人惊奇地发现来自脓肿分枝杆菌DSM444196的MAB_4714c相对于4-羟基丁酸和己二酸分别转化为4-羟基丁醛和6-氧代己酸具有高活性。
在一个实施方案中，所使用的CAR包含以下或者由以下组成：SEQ ID NO:36的氨基酸或其活性片段或同源物。
在一个实施方案中，所述酶与SEQ ID NO:36的氨基酸序列至少80％、85％、90％、95％或100％相同。
在一个实施方案中，如本文所述的CAR的活性片段或同源物包括磷酸泛酰巯基乙胺结合基序：
F-X(2)-L-G-G-D-S-X(2)-A
其中X(2)是任何两个氨基酸。
先前已经表明，CAR的翻译后磷酸泛酰巯基乙胺化是其活性所需要的[P.Venkitasubramanian等Reduction of Carboxylic Acids by Nocardia AldehydeOxidoreductase Requires a Phosphopantetheinylated Enzyme,January 5,2007TheJournal of Biological Chemistry.282,478-485.]。因此在一个实施方案中，如本文所述的CAR与编码磷酸泛酰巯基乙胺基转移酶的基因共表达。合适的磷酸泛酰巯基乙胺基转移酶基因将是本领域技术人员已知的。合适的磷酸泛酰巯基乙胺基转移酶基因包括例如来自枯草芽孢杆菌(Bacillus subtilis)和大肠杆菌K-12基因组DNA的BSU03570和entD。
在某些实施方案中，如本文所提供的AR来源于假单胞菌属(Pseudomonas)、红球菌属(Rhodococcus)、链霉菌属(Streptomyces)或栖热袍菌属(Thermotoga)的生物体。在一个实施方案中，AR来源于铜绿假单胞菌(Pseudomonas aeruginosa)种。在一个实施方案中，AR来源于恶臭假单胞菌(Pseudomonas putida)种。在一个实施方案中，AR来源于丁香假单胞菌(Pseudomonas syringae)种。在一个实施方案中，AR来源于乔斯特红球菌(Rhodococcusjostii)种。在一个实施方案中，AR来源于天蓝色链霉菌(Streptomyces coelicolor)种。在一个实施方案中，AR来源于海栖热袍菌(Thermotoga maritima.)种。
在一个实施方案中，AR包含以下或者由以下组成：SEQ ID NO:40-52中的一个的氨基酸或其活性片段或同源物。一方面，所述酶与SEQ ID NO:40-52中的一个氨基酸序列至少80％、85％、90％、95％或100％相同。
在一个实施方案中，AR包含以下或者由以下组成：SEQ ID NO:47、43、50、45、46或52中的一个的氨基酸或其活性片段或同源物。
一方面，所述酶与SEQ ID NO:47、43、50、45、46或52中的一个氨基酸序列至少80％、85％、90％、95％或100％相同。
在一个实施方案中，SEQ ID NO:43、45、47或50的氨基酸被用于将6-氧代己酸酶促转化为6-羟基己酸。
在某些实施方案中，如本文所提供的AT来源于埃希氏菌属(Escherichia)、中华根瘤菌属(Sinorhizobium)或链霉菌属的生物体。在一个实施方案中，所述AT来源于大肠杆菌种。在一个实施方案中，AT来源于苜蓿中华根瘤菌(Sinorhizobium meliloti)种。在一个实施方案中，AT来源于阿维链霉菌(Streptomyces avermitilis)种。在一个实施方案中，AT来源于苜蓿中华根瘤菌种。
在一个实施方案中，AT包含以下或者由以下组成：SEQ ID NO:53-65中的一个的氨基酸或其活性片段或同源物。一方面，所述酶与SEQ ID NO:53-65中的一个氨基酸序列至少80％、85％、90％、95％或100％相同。
在一个实施方案中，AT包含以下或者由以下组成：SEQ ID NO:53、59、55、58或60中的一个的氨基酸或其活性片段或同源物。
一方面，所述酶与SEQ ID NO:53、59、55、58或60中的一个氨基酸序列至少80％、85％、90％、95％或100％相同。
CAR腺苷酰化结构域使用ATP的2个磷酸根用于底物活化，形成酰基-AMP中间体转移到还原结构域，其中一个NADPH分子被氧化用于释放醛。将CAR与AR一起应用于将羧基完全还原为醇需要2个NADPH分子和1个ATP。己二酸完全生物转化为1,6-己二醇将总计耗费4个NADPH和2个ATP分子。辅因子依赖性酶(诸如氧化还原酶或转移酶)的体外应用需要用于再生消耗的辅因子以使系统成本有效的酶促系统。
在一个实施方案中，可以使用在ATP的再生中常采用的四种酶中的一种或多种：丙酮酸激酶(使用磷酸烯醇丙酮酸酯)、乙酸激酶(使用乙酰磷酸酯)、肌酸激酶(使用磷酸肌酸)和多磷酸激酶(使用多磷酸盐)。更多详情参见Andexer和Richter,Emerging Enzymesfor ATP Regeneration in Biocatalytic Process CHEMBIOCHEM 2015,16,380-386。大多数ATP再生酶只处理一步ADP磷酸化。在一个实施方案中可以使用多磷酸激酶，在一个实施方案中可以使用多磷酸激酶家族II；在一个实施方案中可以使用来自苜蓿中华根瘤菌的SMc02148和来自铜绿假单胞菌的PA3455用于ATP再生。
CAR、AR和氨基供体再生酶使用吡啶核苷酸作为辅因子。已经开发了用于再生NAD(P)H的若干种酶促方法，诸如用甲酸脱氢酶、葡萄糖脱氢酶、葡萄糖-6-磷酸脱氢酶和醇脱氢酶的还原。在一个实施方案中，宿主微生物可以被工程化以增加NADH和/或NADPH的辅因子库。在一个实施方案中，微生物可以被工程化以表达甲酸脱氢酶。在一个实施方案中，SEQID NO:108的甲酸脱氢酶或其活性片段或同源物。
在一个实施方案中，如果将大肠杆菌用作宿主生物体，则可以缺失葡萄糖磷酸异构酶(pgi)基因以将通量转向戊糖磷酸途径以增加NADPH库。其他策略可能涉及用来源于丙酮丁醇梭菌的外源NADPH依赖性甘油醛-3-磷酸脱氢酶将内源NADH-依赖性甘油醛-3-磷酸脱氢酶(GAPDH)转换到宿主大肠杆菌菌株。在另一种方法中，可以将NADH激酶(Pos5P)从酿酒酵母中引入宿主大肠杆菌菌株中。后者成功地用于增加通过NADPH依赖性途径产生的若干种产物[Lee,W.-H.,Kim,M.-D.,Jin,Y.-S.,&Seo,J.-H.(2013).Engineering of NADPHregenerators in Escherichia coli for enhanced biotransformation.AppliedMicrobiology and Biotechnology.97(7):2761-72]。如本领域技术人员将显而易见的，如果选择大肠杆菌作为宿主生物体，则可以通过限制竞争途径来增加NADH库，尽管编码NADH依赖性酶的基因的缺失包括但不限于：醇脱氢酶(adhE)、乳酸脱氢酶(ldhA)和丙酮酸-甲酸裂解酶(pflB)。
本文公开的AT消耗丙氨酸和/或谷氨酸作为需要再生的氨基供体。因此，在一个实施方案中，宿主微生物可以被工程化以增加丙氨酸和/或谷氨酸的生产。在一个实施方案中，这些宿主微生物可被工程化以表达谷氨酸脱氢酶或丙氨酸脱氢酶。发现两种酶适用于此目的–YOR375C，来自酿酒酵母的NADPH依赖性谷氨酸脱氢酶，和BSU31930，来自枯草芽孢杆菌的NADH依赖性丙氨酸脱氢酶。
在各个方面，每种酶的同源物是指与对应于所述酶的SEQ ID NO的氨基酸序列具有至少25％、至少50％、至少60％、至少70％、至少80％、至少85％、至少90％、至少95％或至少99％的同一性并且保留所述酶的活性的蛋白质。用于确定序列同一性的算法是公开可用的，并且包括例如通过美国国家生物技术信息中心(NCBI)可用的BLAST。本领域技术人员可以确定序列是否在表示同源性的程度上相似，并且因此确定相似或相同的功能。
本领域技术人员可以通过使用诸如位点特异性诱变的方法(Nucleic AcidRes.10,第6487页(1982)，Methods in Enzymol.100,第448页(1983)，Molecular Cloning第2版,Cold Spring Harbor Laboratory Press(1989)，PCR A Practical Approach IRL出版第200页(1991))适当地向由本文公开的核苷酸序列组成的酶的DNA中引入取代、缺失、插入和/或添加来获得编码每种酶的同源物的多核苷酸。编码每种酶的同源物的多核苷酸可以被引入宿主中并且在宿主中表达以获得同源物。
如本文所述的酶可以由非天然存在的微生物产生。
如本文所用，术语“微生物”旨在意指作为显微细胞存在的任何生物体，并且涵盖具有纤维尺寸的原核或真核细胞或生物体，并且包括所有物种的细菌、古细菌和真细菌以及真核微生物，诸如酵母菌和真菌。所述术语还包括可以培养用于生产生物化学物质的任何物种的细胞培养物。
如本文所用，当关于微生物使用时，术语“非天然存在的”是指具有至少一种遗传改变的微生物，所述遗传改变通常在所提及的物种的天然存在的菌株(包括所提及的物种的野生型菌株)中不存在。遗传改变包括例如引入编码代谢多肽的可表达核酸的修饰、其他核酸添加、核酸缺失和/或微生物遗传物质的其他功能性破坏。此类修饰包括例如所提及的物种的异源多肽、同源多肽或异源和同源多肽的编码区和其功能片段。另外的修饰包括例如，其中所述修饰改变基因或操纵子的表达的非编码调节区。
术语“内源的”是指源自宿主微生物的所提及的分子或活性。类似地，当关于编码核酸的表达使用时，所述术语是指包含在微生物内的编码核酸的表达。
如本文所用，术语“外源的”是指引入宿主微生物中的分子或活性。例如可以通过将编码核酸引入宿主遗传物质中，诸如通过整合到宿主染色体中或作为非染色体遗传物质(诸如质粒)，来引入分子。关于编码核酸的表达，所述术语是指将编码核酸以可表达的形式引入微生物中。当关于生物合成活性使用时，所述术语是指被引入参考宿主生物体的活性。来源可以是例如表达引入宿主微生物后的活性的编码核酸。
术语“异源的”是指来源于除所提及的物种之外的来源的分子或活性，而“同源的”是指来源于宿主微生物的分子或活性。因此，编码核酸的外源表达可以使用异源或同源编码核酸中的任一种或两者。
如本文所用，当关于微生物使用时，术语“分离的”是指当所提及的微生物在自然界中被发现时，基本上不含至少一种组分的生物体。所述术语包括当微生物在其自然环境中被发现时去除了一些或全部组分的所述微生物，并且包括基本上纯的微生物以及在非天然存在的培养基中培养的微生物。
如本文所用，术语“可操作地连接”是指一个或多个表达控制序列与待表达的多核苷酸中的编码区之间的连接，以此种方式使得在与表达控制序列相容的条件下实现表达。
在一个实施方案中，如本文所述的过程中使用的微生物是遗传修饰以便含有编码如本文所述的2-烯酸酯还原酶的核酸分子的微生物。
编码如本文所述的酶的核酸分子可以单独使用或作为载体的一部分使用。
在一个实施方案中，所述酶包含SEQ ID NO:1的氨基酸序列或其活性片段或同源物。在一个实施方案中，所述酶包含与SEQ ID NO:1的氨基酸序列至少80％、85％、90％、95％或100％相同的氨基酸序列。一方面，所述酶包含SEQ ID NO:1的氨基酸(363)至(382)。在一个实施方案中，所述酶包含SEQ ID NO:2的氨基酸序列或其活性片段或同源物。在一个实施方案中，所述酶包含与SEQ ID NO:2的氨基酸序列至少80％、85％、90％、95％或100％相同的氨基酸序列。另一个方面，所述酶包含SEQ ID NO:2的氨基酸(363)至(382)。
在一个实施方案中，所述酶包含4Fe-4S簇：
CX1SCX2EGCMGRX3QX4YSX5LX6C
其中
X1为任何氨基酸，优选疏水氨基酸，优选亮氨酸或异亮氨酸，优选亮氨酸；
X2为任何氨基酸，优选极性氨基酸，优选组氨酸或谷氨酸，优选组氨酸；
X3为任何氨基酸，优选疏水氨基酸，优选异亮氨酸或缬氨酸，优选异亮氨酸；
X4为任何氨基酸，优选谷氨酸或组氨酸，优选谷氨酸；
X5为任何氨基酸，优选极性氨基酸，优选丝氨酸或甲硫氨酸，优选丝氨酸；
X6为任何氨基酸，优选甘氨酸或天冬酰胺，优选甘氨酸。
在一个实施方案中，所述酶包含或基本上包含SEQ ID NO:1与SEQ ID NO:2之间(>80％、>85％、>90％、>95％或>99％)的共有序列。
在一个实施方案中，所述酶基本上是空气中稳定的，所述酶在一个实施方案中包含当在空气中储存3天时保留其活性的至少20％、至少25％或至少30％的酶。
核酸分子还可以包括与包含在核酸分子中的多核苷酸可操作连接的表达控制序列。这些表达控制序列可适合在细菌或真菌中确保可翻译RNA的转录和合成。表达是指异源DNA序列的转录，优选转录成可翻译mRNA。在细菌、酵母或真菌、蓝细菌和藻类中确保表达的调节元件是本领域技术人员众所周知的，并且涵盖启动子、增强子、终止信号、靶向信号等。与核酸分子联合使用的启动子在其来源和/或待表达的基因方面可以是同源的或异源的。合适的启动子是，例如，本身就是组成型表达的启动子。然而，也可以使用仅在由外部影响确定的时间点活化的启动子。可使用人造启动子和/或化学诱导型启动子。
引入微生物中的多核苷酸被表达以便导致产生具有如本文所述的酶活性的多肽。
不同表达系统的概述包含在以下中，例如，Bitter等(Methods in Enzymology153(1987),516-544)和Sawers等(Applied Microbiology and Biotechnology 46(1996),1-9)，Billman-Jacobe(Current Opinion in Biotechnology 7(1996),500-4)，Hockney(Trends in Biotechnology 12(1994),456-463)，Griffiths等,(Methods in MolecularBiology 75(1997),427-440)酵母表达系统的概述在以下中给出，例如，Hensing等(Antonie van Leuwenhoek 67(1995),261-279)，Bussineau等(Developments inBiological Standardization 83(1994),13-19)，Gellissen等(Antonie van Leuwenhoek62(1992),79-93，Fleer(Current Opinion in Biotechnology3(1992),486-496)，Vedvick(Current Opinion in Biotechnology 2(1991),742-745)以及Buckholz(Bio/Technology9(1991),1067-1072)。
表达载体已经在文献中被广泛描述。一般来讲，它们不仅包含选择标记基因以及在所选宿主中确保复制的复制起点，而且包含细菌或病毒启动子，并且在大多数情况下包含用于转录的终止信号。在启动子和终止信号之间，一般存在至少一个能够插入编码DNA序列的限制性位点或多接头。如果天然控制对应基因转录的DNA序列在选择的宿主生物体中是活性的，则它可以被用作启动子序列。然而，这个序列也可以交换为其他启动子序列。可以使用确保基因组成型表达的启动子以及允许有意控制基因表达的诱导型启动子。文献中详细描述了拥有这些特性的细菌和病毒启动子序列。在本领域技术人员已知的文献中描述了在微生物(包括大肠杆菌和酿酒酵母)中表达的调节序列。允许下游序列特别高表达的启动子为，例如，T7启动子(Studier等,Methods in Enzymology 185(1990),60-89)、lacUV5、trp、trp-lacUV5(DeBoer等,Rodriguez和Chamberlin编,Promoters,Structure andFunction；Praeger,New York,(1982),462-481；DeBoer等,Proc.Natl.Acad.Sci.USA(1983),21-25)、Ip1、rac(Boros等,Gene 42(1986),97-100)。
可以使用可提供比组成型启动子更高的多肽产量的诱导型启动子。合适地，在一个实施方案中，使用两个步骤的过程：首先在最佳条件下培养宿主细胞至相对高的细胞密度；然后诱导转录。
文献中也描述了用于转录的终止信号。
另外，可以通过分子生物学中众所周知的方法将不同的突变插入到多核苷酸中(参见例如，Sambrook和Russell(2001),Molecular Cloning:A Laboratory Manual,CSHPress,Cold Spring Harbor,N.Y.,USA)，从而能够合成具有修饰的生物学特性的多肽。在一个实施方案中，在此处的氨基酸序列修饰增加酶的催化活性或稳定性的位置处引入点突变。
另选地，也可以使用定向进化来改变酶的底物偏好。可以通过在蛋白质全长上使用随机诱变来完成特定活性的增强(Sheryl B.Rubin-Pitel等(2007),“Chapter 3:Directed Evolution Tools in Bioproduct and Bioprocess Development”inBioprocessing for Value-Added Products from Renewable Resources,S.-T.Yang,编,Elsevier,Amsterdam,The Netherlands.)。可以通过定点诱变来增加蛋白质溶解度，以在蛋白质表面上实现疏水性到亲水性的突变(Saul R.Trevino等Journal of MolecularBiology 366(2007),449-460)。
此外，可以用本领域众所周知的技术对基因进行密码子优化，以实现蛋白质的优化表达。
可以将对核酸序列的去除或包括靶向序列的适当修饰引入外源核酸序列中以赋予所需的特性。
如本领域技术人员将理解的，当两种或更多种外源编码核酸待共表达时，两种核酸都可以例如插入到一个表达载体中或插入分开的表达载体中。对于单载体表达，编码核酸可以与常见表达控制序列可操作地连接或与不同的表达控制序列(诸如一个诱导型启动子和一个组成型启动子)连接。
应理解，在一个实施方案中，产生途径中间体或产物的非天然存在的微生物可与表达下游或上游途径酶的另一种生物体(或其他生物体)组合使用以产生所需的产物。例如，工程化生物体可以用于生产和积累己二酸。然后己二酸可以用作另一种工程化生物体的底物，所述另一种工程化生物体表达一种或多种途径基因以将己二酸转化为1,6-己二醇、6-羟基己酸或6-氨基己酸。
宿主微生物可以选自并且非天然存在的微生物产生于，例如细菌、酵母、真菌或适用于发酵过程的多种其他微生物中的任一种。
宿主微生物可以选自并且非天然存在的微生物产生于，例如，细菌、酵母、真菌或可用作宿主生物体的多种其他微生物中的任一种。合适的宿主微生物可以选自细菌门，像变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinocacteria)、热袍菌门(Thermotogae)、拟杆菌属(Bacteroides)、蓝细菌门(Cyanobacteria)。古菌门广古菌门(Euryarchaeota)，真菌门子囊菌门(Ascomycota)和接合菌门(Zygomycota)，或真核生物门(Eukariota)像绿藻门(Chlorophyta)、甲藻门(Dinoflagellata)、硅藻门(Bacillariophyta)、眼点藻纲(Eustigmatophyceae)、定鞭藻门(Haptophyta)和不等鞭毛门(Heterokontophyta)。
合适的细菌和古菌物种包括：
大肠杆菌、枯草芽孢杆菌、地衣芽孢杆菌(Bacillus licheniformis)、蜡样芽孢杆菌(Bacillus cereus)、巨大芽胞杆菌(Bacillus megaterium)、短芽孢杆菌(Bacillusbrevis)、短小芽孢杆菌(Bacillus pumilus)、谷氨酸棒状杆菌(Corynebacteriumglutamicum)、运动发酵单胞菌(Zymomonas mobilis)、丙酮丁醇梭菌、丁酸杆菌(Clostridium butylicum)、克氏梭菌(Clostridium kluyveri)、产乙醇梭菌(Clostridiumautoethanogenum)、热醋穆尔氏菌(Moorella thermoacetica)、醋酸梭菌(Clostridiumaceticum)、拜氏梭菌(Clostridium beijerinckii)、杨氏梭菌(Clostridiumljungdahlii)、糖丁醇丙酮梭菌(Clostridium saccharoperbutylacetonicum)、产气荚膜梭菌(Clostridium perfringens)、艰难梭菌(Clostridium difficile)、肉毒梭菌(Clostridium botulinum)、酪丁酸梭菌(Clostridium tyrobutyricum)、破伤风形梭菌(Clostridium tetanomorphum)、破伤风梭菌(Clostridium tetani)、丙酸梭菌(Clostridium propionicum)、氨基丁酸梭菌(Clostridium aminobutyricum)、近端梭菌(Clostridium subterminale)、斯氏梭菌(Clostridium sticklandii)、富养罗尔斯顿氏菌(Ralstonia eutropha)、牛结核分枝杆菌(Mycobacterium bovis)、结核分枝杆菌(Mycobacterium tuberculosis)、牙龈卟啉单胞菌(Porphyromonas gingivalis)、荧光假单胞菌Pseudomonas fluorescens)、恶臭假单胞菌、铜绿假单胞菌、食羧假单胞菌(Pseudomonas carboxidovorans(Oligotropha carboxidovorans))、斯氏假单胞菌(Pseudomonas stutzeri)、肺炎克雷伯氏菌(Klebsiella pneumonia)、产酸克雷伯氏菌(Klebsiella oxytoca)、产琥珀酸厌氧螺菌(Anaerobiospirillum succiniciproducens)、产琥珀酸放线杆菌(Actinobacillus succinogenes)、产琥珀酸曼氏杆菌(Mannheimiasucciniciproducens)、菜豆根瘤菌(Rhizobium etli)、氧化葡糖杆菌(Gluconobacteroxydans)、乳酸乳球菌(Lactococcus lactis)、植物乳杆菌(Lactobacillus plantarum)、天蓝色链霉菌(Streptomyces coelicolor)、弗氏柠檬酸杆菌(Citrobacter freundii)、无丙二酸柠檬酸杆菌(Citrobacter amalonaticus)、醋酸钙不动杆菌(Acinetobactercalcoaceticus)、贝利不动杆菌(Acinetobacter baylyi)、海栖热袍菌、嗜盐杆菌(Halobacterium salinarum)、粘质沙雷菌(Serratia marcescens)、深红红螺菌(Rhodospirillum rubrum)、艾德尼拉菌属物种(Ideonella sp.)、荚膜红细菌(Rhodobacter capsulatus)、荚膜甲基球菌(Methylococcus capsulatus)、发孢甲基弯菌(Methylosinus trichosporium)、扭脱甲基杆菌(Methylobacterium extorquens)、甲基孢囊菌属GB25(Methylocystis GB25)、荚膜嗜甲基菌(Methylotrophus capsulatus)、甲基单胞菌属物种16a(Methylomonas sp.16a)、强烈火球菌(Pyrococcus furiosus)。
合适的真菌包括：酿酒酵母、粟酒裂殖酵母(Schizosaccharomyces pombe)、山楂覆膜孢酵母(Saccharomycopsis crataegensis)、乳酸克鲁维酵母(Kluyveromyceslactis)、马克斯克鲁维酵母(Kluyveromyces marxianus)、土曲霉(Aspergillusterreus)、黑曲霉(Aspergillus niger)、树干毕赤酵母(Pichia stipitis)、巴斯德毕赤酵母(Pichia pastoris)、少根根霉(Rhizopus arrhizus)、米根霉(Rhizopus oryzae)、解脂耶氏酵母(Yarrowia lipolytica)、东方伊萨酵母(Issatchenkia orientalis)、西方伊萨酵母(Issatchenkia occidentalis)、郎比可假丝酵母(Candida lambica)、山梨木糖假丝酵母(Candida sorboxylosa)、泽普兰假丝酵母(Candida zemplinina)、光滑假丝酵母(Candida geochares)、膜醭毕赤酵母(Pichia membranifaciens)、()、红茶接合酵母(Zygosaccharomyces kombuchaensis)、山梨糖同化假丝酵母(Candida sorbosivorans)、温德华假丝酵母(Candida vanderwaltii)、山梨糖假丝酵母(Candida sorbophila)、双孢接合酵母(Zygosaccharomyces bisporus)、缓慢接合酵母(Zygosaccharomyces lentus)、贝酵母(Saccharomyces bayanus)、巴德利酵母(Saccharomyces bulderi)、卡氏德巴利酵母(Debaryomyces castellii)、博伊丁假丝酵母(Candida boidinii)、埃切假丝酵母Candida etchellsii)、杰丁毕赤酵母(Pichia jadinii)、异常毕赤酵母(Pichiaanomala)、产黄青霉(Penicillium chrysogenum)、热带假丝酵母(Candida tropicalis)。
在一个实施方案中，宿主微生物是在有氧环境中可以表达如本文所述的酶的需氧生物体。
合适的蓝细菌包括：Acaryochloris marina MBIC11017、鱼腥藻物种(Anabaenasp.)PCC 7120、多变鱼腥藻(Anabaena variabilis)ATCC29413、四倍阿格门氏藻(Agmenellum quadruplicatum)、绿硫细菌(Chlorobium tepidum)TLS、蓝丝菌属物种(Cyanothece sp.)ATCC51142、无类囊体蓝藻(Gloeobacter violaceus)PCC 7421、铜绿微囊藻(Microcystis aeruginosa)NIES-843、点状念珠藻(Nostoc punctiforme)ATCC29133、海洋原绿球藻(Prochlorococcus marinus)MED4、海洋原绿球藻MIT9313、海洋原绿球藻SS120、海洋原绿球藻str.AS9601、海洋原绿球藻str.MIT 9211、海洋原绿球藻str.MIT9215、海洋原绿球藻str.MIT 9301、海洋原绿球藻str.MIT 9303、海洋原绿球藻str.MIT9312、海洋原绿球藻str.MIT 9515、海洋原绿球藻str.NATL1A、海洋原绿球藻str.NATL2A、沼泽红假单胞菌(Rhodopseudomonas palustris)CGA009、细长聚球藻(Synechococcus elongatus)PCC 6301、细长聚球藻PCC 7942、聚球藻属物种(Synechococcussp.)CC9311、聚球藻属物种CC9605、、聚球藻属物种CC9902、、聚球藻属物种JA-2-3B\'a(2-13)、聚球藻属物种JA-3-3Ab、聚球藻属物种PCC 7002、聚球藻属物种RCC307、聚球藻属物种WH 7803、聚球藻属物种WH8102、集胞藻属物种(Synechocystis sp.)PCC 6803、细长热聚球藻(Thermosynechococcus elongatus)BP-1、红海束毛藻(Trichodesmium erythraeum)IMS101。
合适的藻类包括：布朗葡萄藻(Botryococcus braunii)、莱茵衣藻(Chlamydomonas reinhardii)、小球藻属物种(Chlorella sp.)、寇氏隐甲藻(Crypthecodinium cohnii)、细柱藻属物种(Cylindrotheca sp.)、杜氏藻(Dunaliellaprimolecta)、等鞭金藻属物种(Isochrysis sp.)、盐生单肠藻(Monallanthus salina)、微球藻属物种(Nannochloris sp.)、微拟球藻属物种(Nannochloropsis sp.)、富油新绿藻(Neochloris oleoabundans)菱形藻属物种(Nitzschia sp.)、三角褐指藻(Phaeodactylumtricornutum)、裂殖壶菌属物种(Schizochytrium sp.)、干扁藻(Tetraselmis sueica)。
在一个实施方案中，宿主微生物是在有氧环境中可以表达具有α,β-酸酯还原酶活性的酶的需氧生物体。在有氧或微氧环境下此种酶的表达可获得改进的生长速率，导致改进的生产力。此外，这种酶可以用于由需要需氧生物合成途径的化合物来制备化学品。
大肠杆菌是特别有用的宿主生物体，因为它是适用于遗传工程良好表征的微生物。
用于构建并测试非天然存在的宿主的表达水平的方法可以通过重组和检测方法使用本领域众所周知的技术来执行。此类方法可以被发现描述于以下中，例如，Sambrook等,Molecular Cloning:A Laboratory Manual,第三版,Cold Spring HarborLaboratory,New York(2001)；以及Ausubel等,Current Protocols in MolecularBiology,John Wiley and Sons,Baltimore,Md.(1999)。用于构建并测试产生本文所述的一种或多种酶的大肠杆菌宿主的表达水平的具体方法在实施例中提供。
当提及包括在微生物中的多于一个的外源核酸时，应理解，这是指所提及的编码核酸或生物化学活性，而不是被引入宿主生物体中的分开的核酸数量。如本领域技术人员将理解的，此类外源核酸可在分开的核酸分子上、多顺反子的核酸分子上或其组合上被引入宿主生物体中。例如，当编码不同酶活性的两种或更多种外源核酸被引入宿主生物体中时，所述两种或更多种外源核酸可以例如在单一质粒上、在分开的质粒上作为单一核酸被引入，可以在单一位点或多个位点被整合到宿主染色体中，并且仍然被认为是两种或更多种外源核酸。
如对本技术领域技术人员将显而易见的，所描述的途径酶的核酸可以根椐所选择的宿主微生物被引入宿主生物体中。如对本技术领域技术人员将显而易见的，当宿主微生物表达途径基因时，可工程化所述微生物，使得编码酶的基因过表达并且/或者编码竞争途径的酶或蛋白质的基因可被缺失。
在一个实施方案中，方法用活细胞进行。在另一个实施方案中，所述过程在体外用裂解细胞或者用部分或完全纯化的酶或者用透化的细胞进行。在一个实施方案中，所述方法在体外进行并且所述酶被固定化。用于将酶固定在不同支撑体上的手段和方法是本领域技术人员众所周知的。此类体外过程可以类似于以下中描述的过程：(1)Dudley,Q.M.,Karim,A.S.,和Jewett,M.C.2014.Cell-Free Metabolic Engineering:Biomanufacturingbeyond the cell.Biotechnology Journal.10:69–82.，2)Zhang Y-HP*.2014.Productionof biofuels and biochemicals by in vitro synthetic biosystems:opportunitiesand challenges.Biotechnology Advances,Epub,DOI:10.1016/j.biotechadv.2014.10.009)
在一个实施方案中，所述方法用产生酶的宿主生物体在培养物中进行。生长培养基可以包括，例如，任何无机或有机碳源，其可以供应碳水化合物的来源或者宿主生物体可以天然使用或工程化使用的其他前体。在一个实施方案中，向宿主生物体提供糖的原料。此类来源包括例如糖，诸如葡萄糖、木糖、半乳糖、甘露糖、果糖和淀粉。葡萄糖可以从各种含碳水化合物的来源获得，所述来源包括常规的生物可再生来源，诸如玉米(玉蜀黍)、小麦、马铃薯、木薯和水稻以及另选的来源，诸如能源作物、植物生物质、农业废物、林业残留物、糖加工残留物和植物来源的家庭废物。
碳水化合物的来源包括可再生的原料和生物质，例如纤维素生物质、半纤维素生物质和木质素原料。此类生物质原料含有例如可以通过生物合成途径产生的碳水化合物底物、不饱和六碳二元羧酸、粘康酸和2-己烯二酸。另外，可以将酶引入能够使用其他底物(诸如甲醇、合成气、甘油、CO2)并产生粘康酸和2-己烯二酸的宿主微生物中。其他可再生原料和生物质将是本领域技术人员已知的。
可根据本发明使用的生物可再生原料来源包括任何包含碳水化合物来源的可再生有机物质。这些包括例如草、树(硬木和软木)、植物和作物残留物。其他来源可以包括例如有机废物材料(例如，废纸、绿色废物、城市废物等)。可使用本领域已知的方法从生物可再生材料中分离合适的碳水化合物，包括葡萄糖。参见例如，Centi和van Santen,Catalysis for Renewables,Wiley-VCH,Weinheim 2007；Kamm,Gruber和Kamm,Biorefineries-Industrial Processes and Products,Wiley-VCH,Weinheim 2006；Shang-Tian Yang,Bioprocessing for Value-Added Products from RenewableResources New Technologies and Applications,Elsevier B.V.2007；Furia,Starch inthe Food Industry,第8章,CRC Handbook of Food Additives第2版CRC出版,1973。还参见Kirk-Othmer Encyclopedia of Chemical Technology第5版,John Wiley and Sons2001中关于淀粉、糖和糖浆的章节。将淀粉转化为葡萄糖的过程也是本领域众所周知的，参见例如，Schenck,“Glucose and Glucose containing Syrups”in Ullmann'sEncyclopedia of Industrial Chemistry,Wiley-VCH 2009.。此外，将纤维素转化为葡萄糖的方法是在本领域已知的，参见例如，Centi和van Santen,Catalysis for Renewables,Wiley-VCH,Weinheim 2007；Kamm,Gruber和Kamm,Biorefineries-Industrial Processesand Products,Wiley-VCH,Weinheim2006；Shang-Tian Yang,Bioprocessing for Value-Added Products from Renewable Resources New Technologies and Applications,Elsevier B.V.2007。
在一个实施方案中，所提供的过程可在发酵罐中进行。
工程化生物体可以在多种反应器系统中培养，并且所述过程可以以不同的操作模式来进行。最常用的生物反应器是搅拌罐生物反应器或充气发酵罐。发酵罐配备有无菌气体供应，气泡分散的混合通过机械搅拌来实现，并且温度可使用使蒸汽或冷却水循环的夹套或螺旋管来维持。对于充气容器，可选择高高度/直径比(>3)以增加气泡与液相之间的接触时间。生物反应器的其他变型是气升式生物反应器，其中在没有机械搅拌的情况下实现混合，并且在固定生物催化剂时使用填充床或流化床生物反应器。
发酵可以以三种不同的模式进行：分批、补料分批和连续模式。标准分批式生物反应器被认为是“封闭的”系统。在分批模式下，所有的培养基组分都加入生物反应器中，同时确保无菌。一旦已经制备了培养基，则生物反应器接种适当的接种物，并且允许继续进行发酵直到结束而不对培养基做任何改变，即没有任何另外组分的补料。然而，可以加入诸如酸和/或碱的组分以维持pH，并且可以加入空气/氧气以维持溶解氧水平。在分批发酵中，生物质和产物浓度随时间变化，直到发酵完成。细胞经历经典的迟滞期、指数生长期、稳定期生长，接着是死亡期。
分批模式的变型是补料分批模式，其中将包括碳源的营养物随着过程进展加入发酵罐中。
除了分批或补料分批模式之外，还可以使用连续的发酵模式。与分批模式相比，连续系统被认为是“开放的”系统。在连续模式中，将确定成分生产培养基连续加入生物反应器中，并且以相同的速率去除等量的生物反应器内容物。连续操作可以在去除容器内容物(包括细胞)的恒化器中或者在使用灌注培养的生物反应器中进行，所述灌注培养允许将活细胞再循环回到生物反应器中，得到高细胞密度。
常用的发酵罐设计和不同的操作模式在文献中已经非常完善[BiochemicalEngineering Fundamentals,第2版J.E.Bailey and D.F.Ollis,McGraw Hill,New York,1986；Development of Sustainable Bioprocesses:Modeling and Assessment,E.Heinzle,A.P.Biwer和C.L.Cooney,John Wiley&Sons,Ltd.,2006；BioprocessEngineering:Basic Concepts,第2版,M.L.Shuler和F.Kargi,Prentice Hall,2001]。
可以采用分批、补料分批或连续发酵程序。
生物体可以在任何合适的生长培养基(诸如Luria-Bertani肉汤、Terrific肉汤或酵母提取物-蛋白胨-右旋糖(YPD)培养基)中生长。为了生产，根据宿主的选择，可以使用合成基本培养基，诸如M9基本培养基、酵母合成基本培养基、酵母氮基、BG-11或其变型。合适的基本培养基可含有至少一种碳源、至少一种氮源、盐、辅因子、缓冲剂以及生长和维持重组微生物所需的其他组分。碳源可以是前述碳源的一种或多种，氮源可以是铵盐或硝酸盐，其包括但不限于(NH4)2SO4、NH4Cl、(NH4)2HPO4、NH4OH、KNO3、NaNO3。培养基可增补有复合或有机氮源，诸如尿素、酵母提取物、酪蛋白氨基酸、蛋白胨、胰蛋白胨、大豆粉、玉米浸液或酪蛋白水解物。另外，可以向基本培养基供应痕量金属，其包括但不限于H3BO3、MnCl2、ZnSO4、Na2MoO4、CuSO4、Co(NO3)2、CuCl2、ZnCl2、CoCl2、FeCl3、KI。基本培养基可增补有维生素和/或非维生素化合物，其包括但不限于生物素、泛酸盐、叶酸、肌醇、烟酸、对氨基苯甲酸、吡哆醇、核黄素、硫胺素、氰钴胺素、柠檬酸、乙二胺四乙酸(EDTA)、柠檬酸铁铵。培养基可以由二氧化碳通过直接喷射或以NaHCO3或Na2CO3的形式供应。
在一个实施方案中，所述过程在基本有氧的条件下进行。
如本文所用，当关于培养或生长条件使用时，术语“基本有氧的”意指在一个实施方案中，氧的量等于或大于液体培养基中溶解氧的饱和度的约10％。在一个实施方案中，所述术语包括维持气氛大于约1％氧的液体或固体培养基的密封室。
在一个实施方案中，本文所述的过程可适当地在约4至约8之间的pH范围内执行。
尽管在一个实施方案中，对本文所述的方法执行的温度没有特别的限制，但一般来讲，本文所述的过程可在约20℃至约60℃之间的温度下执行。
本领域技术人员将能够容易地确定每单位量的底物所使用的酶的有效量，并且在一个实施方案中，所述有效量不受特别限制。本发明人已经发现每200μg底物1μg酶的比例有效，尽管酶与底物的比例更高(例如1:100)和更低(例如1:1000)可能是有效的，并且适当的比例可以可由本领域技术人员根据具体反应条件容易地确定。
可以使用本领域已知的方法来确定培养基中产物的量，所述方法诸如高效液相色谱(HPLC)、气相色谱(GC)、液相色谱-质谱(LC-MS)、气相色谱-质谱(GC-MS)。
测定本文所述的聚合物前体的生产的方法是本领域技术人员已知的，并且下面进一步举例说明。例如，可以通过诸如HPLC(高效液相色谱)、GC-MS(气相色谱-质谱)和LC-MS(液相色谱-质谱)的方法来分析产物、中间体和副产物的形成。也可以使用本领域众所周知的方法来测定来自外源DNA序列的单个酶活性。
在一些实施方案中，本文公开的过程还包括纯化所述过程的产物和本文所述方法的产物。此类纯化方法是本领域技术人员已知的，并且包括例如通过过滤、蒸馏或蒸发。从发酵肉汤中分离化合物取决于所需化合物的最终纯度。分离技术可包括：离心、微滤、超滤、纳滤、蒸发、结晶、蒸馏和离子交换。典型的下游加工操作将包括一系列过程，其包括使用离心或微滤分离细胞，使用超滤去除肉汤中的另外的固体，使用纳滤、离子交换或蒸发结晶从肉汤中去除盐，以及最后使用蒸馏纯化己二酸。
工业适用性
己二酸具有本领域技术人员已知的各种用途。所生产的己二酸大部分用作通过与六亚甲基二胺的形成6,6-尼龙的缩聚反应生产尼龙的单体。其他主要应用还涉及聚合物：己二酸是生产聚氨酯的单体，并且己二酸的酯用作增塑剂，油漆是在聚氯乙烯(PVC)中。
聚合物前体1,4-丁二醇、1,6-己二醇、6-羟基己酸和6-氨基己酸具有不同的工业用途。
1,4-丁二醇在工业上用作溶剂，并且用于制造一些塑料、弹性纤维和聚氨酯。1,4-丁二醇也用于合成γ-丁内酯(GBL)，并且在磷酸的存在和高温下，脱水为溶剂四氢呋喃。
1,6-己二醇广泛用于工业聚酯和聚氨酯生产。它也是丙烯酸树脂、粘合剂和染料的中间体。
氨基己酸是尼龙-6聚合过程中的中间体，其中尼龙-6由己内酰胺的开环水解形成。还发现了它在制药和施用中的用途。
本文提及的所有文件都以引用的方式并入，然而，应理解，以引用的方式全部或部分地并入的任何专利、公布、或其他公开材料是仅在所并入的材料不与本公开中所阐述的定义、陈述、或其他公开材料冲突的程度上并入本文的。因此，并且在必要的程度上，如本文所明确列出的公开取代以引用的方式并入本文的任何冲突的材料。
应理解，本领域技术人员将做出许多修改。因此，上面的描述和附图应该被认为是对本发明的说明，而不具有限制意义。还应理解，本发明旨在覆盖一般来讲符合本发明原理的、包括虽然不属于本发明所公开内容范畴但属于本发明所属领域的公知技术或常用的技术手段并可以应用于上文中阐述过的以及如以下所附权利要求范围示出的必要特征中的任何变型、用途或者变更。
上述本发明的实施方案仅旨在为示例性的。因此，本发明的范围旨在仅仅由所附权利要求书的范围来限定。
实施例
已经研究了两种黄素酶家族，即老黄酶(OYE；EC 1.6.99.1)和2-烯酸酯还原酶(ERED；EC 1.3.1.31)，用于烯烃的生物催化氢化，并对带有吸电子基团(诸如醛、酮和羧酸)的α,β-不饱和底物表现出广泛的底物特异性。分析了来自不同生物体的OYE和ERED的C＝C还原活性。
实施例1-蛋白质的克隆、表达和纯化
将编码选择的OYE和ERED蛋白质的基因从基因组DNA中进行PCR扩增，并通过如D.Bonsor,S.F.Butz,J.Solomons,S.Grant,I.J.S.Fairlamb,M.J.Fogg,G.Grogan,Organicand Biomolecular Chemistry 2006,4,1252-1260中所述的连接非依赖性方法克隆到p15Tv-Lic质粒中。
将重组ERED质粒转化到大肠杆菌BL21(DE3)ΔiscR菌株中用于过表达含铁硫的蛋白质(根据M.K.Akhtar,P.R.Jones,Applied Microbiology and Biotechnology 2008,78,853-862中所述的方法)。将大肠杆菌转化株在增补有100μg/mL氨苄青霉素的Terrific肉汤(TB)培养基(1L)中在37℃下有氧培养，直到光密度(OD 600nm)达到0.6-0.8。此时，用磁力搅拌棒将培养物转移到密闭的烧瓶中，并且在封闭的烧瓶中厌氧预培养30分钟后用0.4mMIPTG诱导蛋白质表达。另外，将培养物与作为最终电子受体的DMSO(50mM)一起在磁力搅拌板上在室温下孵育15-19小时。通过离心(9,000g)收获大肠杆菌细胞，并将细胞沉淀储存在液N2中用于蛋白纯化。对于全细胞生物转化细胞，要立即使用。在80％N2、10％H2和10％CO2的气氛下，在厌氧手套箱(Coy Laboratory Products,Grass Lake,MI,USA)中执行蛋白质纯化。所有的缓冲液在使用前都被脱气并用Ar喷射。将细胞沉淀重悬于裂解缓冲液(400mMNaCl、50mM HEPES、pH 7.5、5％甘油、5mM咪唑、0.05％吐温20、1mg/mL溶菌酶、3U/mL核酸酶、0.5mM EDTA和1mM DTT)中并在室温下孵育30分钟。通过离心澄清细胞裂解物，并将上清液与Ni亲和树脂(Qiagen,Valencia,CA,USA)一起在4℃下孵育1小时。然后将所述树脂用100mL洗涤缓冲液(400mM NaCl、50mM HEPES、pH 7.5、5％甘油、30mM咪唑、0.5mM EDTA和1mMDTT)洗涤，并用洗脱缓冲液(400mM NaCl、50mM HEPES、pH 7.5、5％甘油、250mM咪唑、0.5mMEDTA和1mM DTT)洗脱。纯化的ERED被冷冻并储存在液N2中。使用Bradford测定法测定蛋白质浓度，并在10％SDS-PAGE凝胶上评估蛋白质纯度。
将重组OYE质粒转化到大肠杆菌BL21-Gold(DE3)菌株(Stratagene,La Jolla,CA,USA)中。将大肠杆菌转化株在增补有100μg/mL氨苄青霉素的TB培养基中在37℃下培养，直到在600nm处的OD达到0.6-0.8。用0.4mM IPTG诱导蛋白质表达，并且大肠杆菌细胞在16℃下生长过夜。通过离心(9,000g)收获大肠杆菌细胞，并且将沉淀重悬于裂解缓冲液(400mMNaCl、50mM HEPES、pH 7.5、5％甘油和5mM咪唑)中，接着通过超声破碎细胞。通过离心澄清裂解物，并将上清液与Ni亲和树脂一起在4℃下孵育30分钟。然后将所述树脂用洗涤缓冲液(400mM NaCl、50mM HEPES、pH 7.5、5％甘油和30mM咪唑)洗涤，并用洗脱缓冲液(400mMNaCl、50mM HEPES、pH 7.5、5％甘油和250mM咪唑)洗脱。纯化的OYE被冷冻在液N2中并储存在-80℃下。
实施例2–比较实施例
本研究中的所有结果均来自至少两个独立的测定的平均值。对照实验平行执行以校正OYE的辅因子的底物非依赖性氧化。在厌氧反应中使用的所有缓冲液和试剂在使用之前用Ar严格喷射以除去痕量的氧。
OYE不含铁硫簇，但是它们可以以与ERED类似的方式通过NAD(P)H到FMN的氢负离子转移来催化活化烯烃的氢化。基于FMN氧化还原酶家族PF00724的Pfam结构域来克隆25个假定的OYE基因(表1)。将纯化的OYE针对丙烯醛、2-环己烯-1-酮、顺式,顺式-粘康酸、反式,反式-粘康酸和2-己烯二酸的氢化活性。在厌氧室中在室温下孵育20分钟后，通过分光光度法在96孔板中测量底物特异性。缓冲液和试剂也被脱气并用Ar喷射。反应混合物含有终体积为200μL的磷酸钾(50mM和pH 7.0)、NaCl(10mM)、NADPH(0.25mM)、各种底物(1mM)和蛋白质(2.5μg)。通过跟踪在340nm(ε340nm＝6,200/M·cm)处的吸光度降低来监测酶反应，作为辅因子NADPH向NADP+转化的量度。
表1.测试OYE蛋白质的生物催化氢化
Figure BDA0001550363180000341
Figure BDA0001550363180000351
OYE的酶活性的截断值为0.05U/mg蛋白质。ND＝未检测出。
将25种推定OYE在大肠杆菌中过表达并使用Ni-NTA树脂进行纯化。用典型的OYE底物(即丙烯醛和2-环己烯-1-酮)测试纯化蛋白的OYE活性，并且25种蛋白中有21种示出OYE活性(表1)。然而，没有纯化的OYE可以氢化2-己烯二酸或粘康酸。类似地，用三种过表达OYE的大肠杆菌菌株进行全细胞生物转化：来自大肠杆菌的EC5625(SEQ ID NO:10)、来自酿酒酵母的YST4373(SEQ ID NO:31)和来自枯草芽孢杆菌的BSU2381(SEQ ID NO:8)，显示没有将2-己烯二酸氢化为己二酸。这个结果与先前的报道一致，即OYE不能在没有另外的吸电子基团(诸如第二酸或酯、卤素或腈)的情况下容易地还原α,β-不饱和羧酸。
实施例3-酶筛选：ERED家族(EC 1.3.1.31)
本研究中的所有结果均来自至少两个独立的测定的平均值。对照实验平行执行以校正辅因子和ERED的底物非依赖性氧化。在厌氧反应中使用的所有缓冲液和试剂在使用之前用Ar严格喷射以除去痕量的氧。
检查了ERED的C＝C还原活性。在厌氧室中在30℃下孵育3-5分钟之后，在96孔板中通过分光光度法测量纯化ERED的酶活性，如M.Bühler,H.Simon,Hoppe-Seyler'sZeitschrift fur Physiologische Chemie 1982,363,609-625中所述。缓冲液和试剂被脱气并用Ar喷射。反应混合物含有终体积为200μL的磷酸钾(100mM和pH 7.0)、NADH(0.5mM)、指示底物(和蛋白质(0.1-50μg)。由于NADH氧化为NAD+，通过跟踪在340nm(ε340nm＝6,220/M·cm)处的吸光度降低来监测酶反应。
已知来自酪丁酸梭菌EREDCT，SEQ ID NO:5CAA71086.1)的ERED在体内对α,β-不饱和羧酸酯表现出广泛的底物特异性。(Buhler和Simon 1982)EREDCT示出与来自以下的ERED(图14)50％～82％的序列同一性：凝结芽孢杆菌36D1(EREDBC,SEQ ID NO:1)、丙酮丁醇梭菌(EREDCA,SEQ ID NO:2)、克氏梭菌(EREDCK,SEQ ID NO:3)、扬氏梭菌(EREDCL,SEQ IDNO:4)和热醋穆尔氏菌(EREDMT,SEQ ID NO:6)。使用大肠杆菌BL21(DE3)ΔiscR中表达的这些ERED的2-己烯二酸(20mM)的全细胞厌氧生物转化(图3)显示所有ERED(EREDCT除外)产生己二酸(图6)。EREDBC、EREDCA和EREDCK在3小时内催化将2-己烯二酸完全转化为己二酸，而EREDMT和EREDCL分别在6小时和48小时后示出2-己烯二酸的完全氢化(图7)。EREDCT的无活性可以归因于其在大肠杆菌中的低表达(图5b)，所述低表达可以使用在大肠杆菌中已建立的重组蛋白表达的密码子优化的方法来改进。虽然研究的EREDCT已知是氧敏感酶(Bühler,M.&Simon,H.On the kinetics and mechanism of enoate reductase.Hoppe-Seyler'sZeitschrift fur Physiologische Chemie 363,609-625(1982))，对于厌氧生长的EREDBC(SEQ ID NO:1)和EREDCA(SEQ ID NO:2)执行2-己烯二酸(15mM)的有氧生物转化。所有使用的缓冲液和溶剂均在有氧条件下制备并储存。在有氧孵育条件下，两种培养物在20分钟内催化将2-己烯二酸完全转化为己二酸(图15)。这个结果可以由用于生物转化的高细胞密度(平均每ml 100mg干细胞生物质)和在细胞内维持厌氧条件的大肠杆菌细胞中的大肠杆菌高呼吸速率来解释。
表2.测试ERED蛋白质的生物催化氢化
Figure BDA0001550363180000371
粘康酸(0.7mM)的顺式,顺式-异构体和反式,反式-异构体的全细胞生物转化显示EREDBC、EREDCA和EREDMT也将这些底物氢化为己二酸。孵育24小时后，在培养物中未检测到粘康酸或2-己烯二酸，并且获得了99％的己二酸产率(图8a、8b和8c)。EREDCK和EREDCL也表现出来自反式,反式-粘康酸的99％的己二酸产率，但是来自顺式,顺式异构体产量较低(<35％)(图8d和8e)。有趣的是，这些ERED用后一种底物产生了三种另外的产物，其被鉴定为反式,反式-粘康酸、2-己烯二酸和具有m/z 143.0319的3-己烯二酸样化合物(图8d和8e)。由顺式,顺式异构体形成反式,反式-粘康酸表明在EREDCK和EREDCL中存在顺式-反式异构酶活性。3-己烯二酸样化合物似乎是这些ERED的低效底物。这与来自大肠杆菌的突变型2,4-二烯酰基-CoA还原酶类似，其催化2,4-二烯酰基CoA氢化为3-烯酰基CoA(Tu,X.,Hubbard,P.A.,Kim,J.J.P.&Schulz,H.Two distinct proton donors at the activesite ofEscherichia coli 2,4-dienoyl-CoA reductase are responsible for theformation of different products.Biochemistry 47,1167-1175(2008).)。因此，结果表明所研究的ERED可以催化六碳二元羧酸的两个C＝C键的顺序氢化，但似乎具有不同的异构体偏好。
实施例4-EREDCA和EREDBC的生物化学表征
六种克隆ERED的厌氧过表达和亲和纯化产生大量的EREDBC、EREDCA和EREDCK的可溶性蛋白(>5mg/L)，而其他三种ERED示出较低的表达(图5)。纯化的EREDBC和EREDCA在溶液中表现出棕色以及在380nm处具有肩峰(shoulder)和在450nm处具有黄素样最大值的吸收光谱(图9)。纯化的EREDBC和EREDCA的棕色以及其吸收光谱中的380nm肩峰都表明存在功能性[4Fe-4S]簇。我们的光谱研究显示纯化的EREDBC和EREDCA可以被氧气(在空气存在下)完全氧化，用过量的NADH(3mM)还原，然后通过底物加入(即6mM反式肉桂酸)部分地再氧化。
对于体外生物化学表征，用2-己烯二酸、顺式-顺式-粘康酸、反式-反式粘康酸、反式肉桂酸、反式-2-甲基-2-丁烯酸、反式-2-丁烯酸、丙烯醛、2-环己烯-1-酮和3-甲基-2-环己烯酮来测量纯化的ERED的酶活性。纯化的EREDBC和EREDCA示出NADH依赖性还原酶活性，而其他纯化的ERED对于测试的底物无活性，可能是由于铁硫簇或黄素辅因子的丢失或者不同的底物偏好。[Feng,J.等Discovery and Characterization of BlsE,a Radical S-Adenosyl-L-methionine Decarboxylase Involved in the Blasticidin SBiosynthetic Pathway.PLoS ONE 8(2013).]。在图9中示出的EREDBC和EREDCA光谱中，相关的黄素和FeS簇辅因子的峰清晰可见，这可以帮助区分活性酶和无活性酶(同时纯化)。
厌氧条件下的酶学测定显示，纯化的EREDCA对反式肉桂酸(3.5U/mg蛋白质)具有高氢化活性，而对反式-2-丁烯酸(0.008U/mg蛋白质)具有低活性。反式肉桂酸的动力学常数kcat和Km)分别为5.2s-1和0.17mM，并且反式肉桂酸的比活性来自产芽孢梭菌(C.sporogenes)的肉桂酸还原酶(5U/mg蛋白质)的比活性相当。EREDCA通过反式肉桂酸在底物浓度高于0.5mM时以1.2mM的抑制常数(Ki)表现出底物抑制(图11a)。纯化的EREDCA针对2-己烯二酸(高达0.05U/mg蛋白质)也有活性，但是在4至28mM的浓度范围内，示出此底物没有饱和(图11b)。
纯化的EREDBC针对2-己烯二酸(0.09U/mg蛋白质)、2-环己烯-1-酮(0.08U/mg蛋白质)、3-甲基-2-环己烯酮(0.13U/mg蛋白质)和反式肉桂酸(0.39U/mg蛋白质)表现出显著的还原酶活性，并且针对反式-2-丁烯酸(巴豆酸，0.036U/mg蛋白质、和丙烯醛(0.037U/mg蛋白质)表现出可检测的活性。与EREDCA相比，EREDBC具有广泛的底物特异性(表3)。提出了生物相容性钯催化剂用于在包括肉桂酸变型和二元羧酸的生长培养基中的烯烃代谢物的非酶促氢化。EREDBC表现出与钯催化剂相当的底物光谱(Sirasani,G.,Tong,L.&Balskus,E.P.A biocompatible alkene hydrogenation merges organic synthesis withmicrobial metabolism.Angew.Chem.Int.Ed.(2014)，提供证据表明EREDBC可以在生物化学品的微生物生产中成为烯烃氢化的良好替代品。
表3.EREDBC针对不饱和羰基底物的体外C＝C还原酶活性。
Figure BDA0001550363180000391
[a]2-己烯二酸溶解在水性缓冲液(29mM)中或0.25mM异丙醇(35mM)中。
[b]用1N NaOH滴定的反式肉桂酸用于酶测定。
[c]ND：未检测出。
使用2-己烯二酸、2-环己烯-1-酮、3-甲基-2-环己烯酮和反式肉桂酸作为底物来执行纯化EREDBC的动力学研究(图10)。EREDBC示出2-己烯二酸、2-环己烯-1-酮和3-甲基-2-环己烯酮没有饱和，但是对于反式肉桂酸观察到饱和动力学(图10d)。反式肉桂酸的动力学常数(kcat和Km)分别为0.50s-1和0.51mM。EREDBC通过反式肉桂酸在底物浓度高于1.0mM时以1.4mM的抑制常数Ki表现出中等底物抑制。与反式肉桂酸相反，EREDBC示出2-己烯二酸溶解于水性缓冲溶液中的S形样曲线，但尽管其具有显著的体外活性(0.09U/mg蛋白质)(图10a)，但未观察到饱和，这可能是由于2-己烯二酸在水性溶液中溶解度有限。
为了增加溶解的底物浓度，将2-己烯二酸溶解在含有0.25mM(终浓度)异丙醇的反应混合物中，并且测定EREDBC和EREDCA的动力学常数。两种ERED酶均在2.5至35mM的浓度范围内示出S形动力学(图12)。与EREDCA相比，EREDBC对于2-己烯二酸表现出更高的周转率(kcat，1.86相对于0.138s-1)和相似的亲和性(Km，18.9相对于20.47mM)，导致与EREDCA相比EREDBC的催化效率高14.6倍(kcat/Km，0.984相对于0.0674s-1mM-1)。
实施例5–氧气耐受性
已知ERED是氧气敏感的酶，其由于在严格保守的基序C-2X-C-3X-C-11X-C中存在由4个半胱氨酸残基配位的氧气敏感的[4Fe-4S]铁-硫簇而被氧气快速灭活(图14)[Gall,M.等Enzymatic conversion of flavonoids using bacterial chalcone isomerase andenoate reductase.Angewandte Chemie International Edition 53,1439-1442(2014).]。然而，在本实验中，纯化EREDBC对氧气灭活表现出显著的抗性，而EREDCA仅在空气中孵育两天后被灭活(图13)。
将纯化ERED放置在密封有气密橡胶塞的5mL玻璃管中。厌氧管充满厌氧气体混合物(80％N2、10％H2和10％CO2)，而好氧管充满大气。将所述管在冰上保持一周，并且每天用注射器取出少量等份ERED(5ul)以测量反应混合物中的残余活性，所述反应混合物含有100mM磷酸钾缓冲液(pH 7.0)、0.5mM NADH、指示底物(用于EREDBC的0.25mM 2-己烯二酸和用于EREDCA的0.75mM肉桂酸)和蛋白质(1μg)，终体积200μL。
在4℃不存在氧气的情况下储存三天之后，EREDBC和EREDCA都分别保留了35％和19％的初始活性。在空气(21％氧气)的存在下，EREDBC示出显著的氧气耐受性(即在储存三天之后保留初始活性的33％之后没有氧气灭活)(图13a)。相反，EREDCA在储存一天之后表现出30％的残余活性，并且然后在储存两天之后被氧气完全灭活(图13b)。EREDBC和EREDCA的氧气耐受性可能与限制氧气进入其[4Fe-4S]相关。[Jervis,A.J.等The O2sensitivityof the transcription factor FNR is controlled by Ser24modulating the kineticsof[4Fe-4S]to[2Fe-2S]conversion.Proceedings of the National Academy ofSciences of the United States of America 106,4659-4664(2009).]。
材料-实施例6和7
己二醛(≥95％)购自Activate Scientific(UK)，6-羟基己酸购自Alfa Aesar(Ward Hill,Massachusetts,USA)，4-羟基丁酸购自Arking Pharma(Guelph,ON,Canada)，己二酸(≥99.5％)、丁醛(≥99％)和氨基己酸(≥99％)、1,6-己二醇(≥99％)购自igma(St.Louis,MO,USA)。
Figure BDA0001550363180000411
高保真DNA聚合酶和DpnI购自New England Biolabs(Whitby,Ontario,Canada)。用于连接非依赖性克隆的In-Fusion HD plus EcoDry从Clontech(Mountain View,CA,USA)获得。所有其他化学品购自Sigma。
体外实验的所有结果均来自至少三个独立的测定的平均值。对照实验平行执行以校正辅因子的底物非依赖性氧化。
HPLC和LC-MS产物分析-实施例6和7
使用配备有Aminex HPX-87H柱(300×7.8mm)(Bio-Rad Laboratories Inc.,Hercules,CA,USA)的Varian ProStar HPLC系统(Varian Medical Systems,Palo Alto,CA,USA)来分析1.6-己二醇、1,4-丁二醇、6-羟基己酸和己二酸标准物和反应样品。将样品用5mM H2SO4以0.6ml/min的流速、柱温50℃进行洗脱。基于210nm UV或RI处的峰面积通过线性回归来确定1,6-己二醇的浓度。
6-氨基己酸标准品和反应样品根据用于胺衍生化的柱前程序的Interchim网站方案(http://www.interchim.fr/ft/0/02727A.pdf)用邻苯二甲醛衍生化制备。在30℃衍生化2分钟之后，使用配备有荧光检测器(激发-340nm，发射-450nm)和Pursuit 5-C18(150x4.0mm)柱的Varian ProStar HPLC系统(Varian Medical Systems,Palo Alto,CA,USA)来分析样品。样品用甲醇/乙腈/水(45/45/10)(B)–40mM磷酸盐缓冲液pH 7.6(A)梯度洗脱。溶剂梯度为：0min，0％B；20min，60％B；22min，60％B；35min，0％B；接着用0％B平衡4分钟。通过基于峰面积的线性回归来确定底物和产物的浓度。
LC-MS平台由Dionex Ultimate 3000UHPLC系统和配备HESI源的Q-Exactive质谱仪组成(均来自Thermo Scientific)。使用Thermo XCalibur 2.2软件和Chromeleon 7.2软件执行系统控制和数据处理。通过液相色谱分离在配备有保护柱的Hypersil Gold C18柱(50mm x2.1mm，1.9μm粒径，Thermo Scientific)上进行。泵以200μL/min的流速运行。溶剂A是含有0.1％甲酸的水；溶剂B是乙腈。梯度为0min，0％B；检测4min，0％B；15min，100％B；24min，100％B；25min，0％B；30min，0％B；接着用0％B平衡5分钟。自动进样器温度维持在8℃，并且注射体积为10μL。数据采集以正离子模式完成，其中扫描范围m/z 100-300，在1Hz下分辨率70000，AGC目标3e6，并且最大注射时间200ms。1,6-己二醇的标准溶液(m/z119.1062)以及6-羟基己酸(m/z 133.0864)用于验证保留时间和m/z。
实施例6
6.1基因克隆、表达和蛋白质的纯化
将编码选择的CAR、AR、AT、谷氨酸脱氢酶YOR375C、丙氨酸脱氢酶BSU31930、多磷酸激酶PA3455和SMc02148蛋白质的基因从基因组DNA中进行PCR扩增并通过如先前所述的连接非依赖性方法克隆到具有N末端6His标签的p15Tv-Lic质粒中[Bonsor D.,Butz S.F.,Solomons J.,Grant S.,Fairlamb I.J.S.,Fogg M.J.,Grogan G.Ligation independentcloning(LIC)as a rapid route to families of recombinant biocatalysts fromsequenced prokaryotic genomes.Organic and Biomolecular Chemistry]。先前已经表明，CAR的翻译后磷酸泛酰巯基乙胺化是其活性所需要的[P.Venkitasubramanian等Reduction of Carboxylic Acids by Nocardia Aldehyde Oxidoreductase Requires aPhosphopantetheinylated Enzyme,January 5,2007The Journal of BiologicalChemistry.282,478-485.]。因此，从枯草芽孢杆菌和大肠杆菌K-12基因组DNA扩增编码磷酸泛酰巯基乙胺基转移酶BSU03570和entD的基因，并使用标准的连接依赖性方案将其克隆到无标签的pCDFDuet质粒中[Sambrook J,Russell D.Molecular Cloning-A LaboratoryManual.Cold Spring Harbor Laboratory Pres.(4th ed.)(2012)]。将甲酸脱氢酶基因P33160克隆到具有C末端标签的pET23b中。使用标准方案执行定点诱变[KunkelT.A..Rapid and efficient site-specific mutagenesis without phenotypicselection.Proc Natl Acad Sci U.S.A.82,488-492(1985)]。
将重组质粒转化到大肠杆菌BL21(DE3)菌株中用于过表达。将具有磷酸泛酰巯基乙胺基转移酶基因的质粒与含有CAR基因的质粒一起共转化。将大肠杆菌转化株在增补有100μg/mL氨苄青霉素(p15Tv-Lic，pET23b)和/或50μg/mL链霉素(pCDFDuet)的Terrific肉汤(TB)培养基(1L)中在37℃下有氧培养，直到培养物光密度(OD 600nm)达到0.8-1。此时，培养温度降至16℃，并且用0.4mM IPTG诱导蛋白质表达。过夜诱导之后，通过离心(4,500g)收获大肠杆菌细胞，并将细胞沉淀在蛋白质纯化之前储存在-20℃。将细胞沉淀重悬于溶解缓冲液(400mM NaCl、50mM HEPES、pH 7.5、5％甘油、5mM咪唑)中，并且在5秒脉冲中在冰上超声处理20分钟，接着冷却5秒。去除不溶性细胞材料(40,000g)，并将上清液与Ni亲和树脂(Qiagen,Valencia,CA,USA)一起在4℃下孵育1小时。然后将所述树脂用100mL洗涤缓冲液(400mM NaCl、50mM HEPES、pH 7.5、5％甘油和30mM咪唑)洗涤，并用洗脱缓冲液(400mMNaCl、50mM HEPES、pH 7.5、5％甘油和250mM咪唑)洗脱。纯化的蛋白质在液N2中冷冻成液滴并储存在-80℃。使用Bradford测定法测定蛋白质浓度，并使用13％SDS-PAGE凝胶评估蛋白质纯度。
6.2.酶筛选:CAR(1.2.1.30)
已经表明，爱荷华诺卡氏菌CAR具有广泛的底物范围，尽管其最大活性通常用芳族底物(诸如苯甲酸和肉桂酸)来观察。与爱荷华诺卡氏菌CAR(Uniprot编号：Q6RKB1)同源的CAR基因序列选自GenBank数据库。基于伴随磷酸泛酰巯基乙胺基结合基序(PF00530)的腺苷酸化(结合AMP的PF00501)和还原(结合NAD的PF07993)结构域的存在，克隆并在大肠杆菌中表达13个CAR基因(大部分来自分枝杆菌属)。(表4)。使用来自大肠杆菌(P19925)和枯草芽孢杆菌(P39135)的磷酸泛酰巯基乙胺基转移酶(PPT)作为CAR活化酶，其与来自分开质粒的选择的CAR基因一起共表达。使用1升大肠杆菌培养物并通过IPTG过夜诱导，使用亲和色谱法在Ni-NTA树脂上纯化12种可溶性CAR蛋白质。基于NADPH氧化测定，8种蛋白质对两种常见CAR底物表现出还原酶活性：苯甲酸和肉桂酸(图16-A)。纯化的酶针对脂肪族二元羧酸的筛选显示对较长链(C6)二元羧酸的显著还原酶活性，而对较短链二元酸(丙二酸、琥珀酸和戊二酸)没有发现可检测的活性(图16)。然而，发现具有类似链长度的一元羧酸C3-C4一元酸(例如4-羟基丁酸、巴豆酸、丁酸和丙酸)是这些酶的阳性底物(图16-B、D)。选择对己二酸和4-羟基丁酸具有最高还原酶活性的5种CAR酶(MSMEI_5586、MAB_4714c、MSMEG_5739、MAP_1040c和MSMEG_2956)SEQ ID NO:35-39，用于生物化学表征和生物转化反应。
表4.克隆、纯化并用4-羟基丁酸(5mM)和己二酸(5mM)作为底物测试还原酶活性的13种CAR。
Figure BDA0001550363180000451
Figure BDA0001550363180000461
*ND＝未检测到活性。NS＝未检测到可溶性蛋白。
6.3-酶筛选：AR和AT
克隆并在大肠杆菌中过表达来自不同细菌的28种醛还原酶。在NADPH和NADH二者的存在下，将使用亲和色谱法在Ni-NTA树脂上纯化的酶用戊二醛和丁醛筛选还原酶活性(表5)。使用丁醛作为商购不可获得的4-氧代-丁醛的底物替代物。分光光度法跟踪反应，辅因子在340nm处氧化。六种AR酶(RHA1_ro05279(SEQ ID NO:47)、PA1146(SEQ ID NO:43)、SCO0229(SEQ ID NO:50)、PP_3370(SEQ ID NO:45)、PSPTO_2097(SEQ ID NO:46)和TM_0111(SEQ ID NO:52)证明了针对这些底物的最高还原酶活性，并且它们被选择用于生物化学表征和生物转化。
纯化AR的酶活性在96孔板中通过分光光度法测量。反应混合物含有100mM HEPES缓冲液(pH 7.5)、0.4mM NADH/NADPH、10mM乙二醛或丁醛和蛋白质(10μg用于己二醛筛选，2μg用于丁醛筛选)，终体积200μL。用于乙二醛筛选的反应混合物含有1.8％DMSO。由于NAD(P)H氧化为NAD(P)+，通过跟踪在340nm(ε340nm＝6,220/M·cm)处的吸光度降低来监测酶反应。
将28种PLP依赖性氨基转移酶(来自I、II和III类)克隆并在大肠杆菌中过表达(表6)。将酶使用亲和色谱法在Ni-NTA树脂上进行纯化，并使用丙氨酸和谷氨酸作为氨基供体筛选酶对己二醛的还原酶活性。在过量的氯化铵存在下，使用与NADPH/NADH依赖性丙氨酸和谷氨酸脱氢酶的偶联反应，通过分光光度法测量酶活性。五种AT酶(GabT(SEQ ID NO:53)、SM2011_b20379(SEQ ID NO:59)、SAV2583(SEQ ID NO:55)、SM2011_c04388(SEQ IDNO:58)和SM2011_b20423(SEQ ID NO:60))表现出对己二醛/丁醛的最高活性，并且它们被用于生物化学表征。
在96孔板中通过分光光度法测量AT的酶活性，其与NAD(P)H依赖性丙氨酸和谷氨酸脱氢酶(YOR375C(SEQ ID NO:107)和BSU31930(SEQ ID NO:106))偶联。通过跟踪在340nm(ε340nm＝6,220/M·cm)处NAD(P)H至NAD(P)+的吸光度降低来监测反应。偶联的酶可以在过量的NH4Cl中胺化由AT产生的丙酮酸酯/酮戊二酸酯。反应混合物含有100mM HEPES缓冲液(pH 7.5)、0.5mM NADPH、0.5mM NADH、1mM丙氨酸、1mM谷氨酸、10mM乙二醛、5％乙醇、37.5mM NaCl、50mM氯化铵、50μM PLP、YOR375C和BSU31930(各2.5μg)和AT蛋白质(20μg)，终体积200μL。
表5.克隆、纯化并测试其己二醛(10mM)和丁醛(10mM)的还原的28种AR。
Figure BDA0001550363180000471
Figure BDA0001550363180000481
*ND＝未检测到活性。
表6.克隆、纯化并测试己二醛(10mM)的胺化的28种AT。
Figure BDA0001550363180000482
Figure BDA0001550363180000491
*ND＝未检测到活性。
6.4再生酶：NADPH、ATP、谷氨酸和丙氨酸。
CAR腺苷酰化结构域使用ATP的2个磷酸根用于底物活化，形成酰基-AMP中间体转移到还原结构域，其中一个NADPH分子被氧化用于释放醛。将CAR与AR一起应用于将羧基完全还原为醇需要2个NADPH分子和1个ATP。己二酸完全生物转化为1,6-己二醇将总计耗费4个NADPH和2个ATP分子。辅因子依赖性酶(诸如氧化还原酶或转移酶)的体外应用需要用于再生消耗的辅因子以使系统成本有效的酶促系统。在ATP的再生中存在四种常用的酶：丙酮酸激酶(使用磷酸烯醇丙酮酸酯)、乙酸激酶(使用乙酰磷酸酯)、肌酸激酶(使用磷酸肌酸)和多磷酸激酶(使用多磷酸盐)[Donk W.A.,Zhao H.Recent developments in pyridinenucleotide regeneration.Current Opinion in Biotechnology 14,421-426(2003)]。大多数ATP再生酶只处理一步ADP磷酸化。考虑到ATP再生的起始底物的价格和稳定性，选择多磷酸激酶。多磷酸激酶家族II包含能够以两个步骤再生AMP至ATP的酶。对于ATP再生，应用来自苜蓿中华根瘤菌的SMc02148(SEQ ID NO:105)和来自铜绿假单胞菌的PA3455(SEQ IDNO:104)(表7)[Nocek B.,Kochinyan S.,Proudfoot M.,Brown G.,Evdokimova E.,Osipiuk J.,Edwards A.M.,Savchenko A.,Joachimiak A.,Yakunin A.F.Polyphosphate-dependent synthesis of ATP and ADP by the family-2polyphosphate kinases in bacteria.Proc.Natl.Acad.Sci.U.S.A.388,17730-17735(2008)]。将酶进行克隆并在大肠杆菌中过表达。通过使用Ni-NTA树脂的亲和色谱法纯化的酶应用于CAR+AR/AT偶联反应。通过HPLC反相色谱证实了在多磷酸盐(10-14磷酸盐链长)的存在下酶对于ATP分子再生的分开步骤以及全AMP至ATP再生的活性。
筛选的AT酶消耗丙氨酸和/或谷氨酸作为还需要再生的氨基供体。发现两种酶适用于此目的–YOR375C(SEQ ID NO:107)，来自酿酒酵母的NADPH依赖性谷氨酸脱氢酶，和BSU31930(SEQ ID NO:106)，来自枯草芽孢杆菌的NADH依赖性丙氨酸脱氢酶(表7)。将酶进行克隆并在大肠杆菌中表达。用Ni-NTA树脂上的亲和色谱法纯化之后，测试酶的丙氨酸(图17A)和谷氨酸再生(图17B)。两种酶均用于初级AT筛选和AT生物化学表征作为偶联酶，使得在还原辅因子NADH和NADPH以及NH4Cl过量的存在下检测丙酮酸酯和酮戊二酸酯。
通过分光光度法在96孔板中测量甲酸脱氢酶P33160(D222Q/H224N)活性，通过在340nm(ε340nm＝6,220/M·cm)处NAD+/NADP+吸光度的增加来监测。反应混合物含有100mMTris-HCl缓冲液(pH 8.0)、50mM甲酸钠、1μg酶。
使用反相Pursuit5-C18柱HPLC在乙腈-具有8mM TBAH的50mM KH2PO4的线性梯度中测量多磷酸激酶活性。检测波长-245nm。反应混合物含有100mM Tris-HCl缓冲液(pH 8.0)、10mM MgCl2、0.25mM多磷酸盐、3mM ADP/AMP、0.5μg酶。
CAR、AR和氨基供体再生酶使用吡啶核苷酸作为辅因子。已经开发了用于再生NAD(P)H的若干种酶促方法，诸如用甲酸脱氢酶、葡萄糖脱氢酶、葡萄糖-6-磷酸脱氢酶和醇脱氢酶的还原[Woodyer R.D.,Johannes T.,Zhao H.Cofactor Regeneration forBiocatalytic Applications.In Enzyme Technology,(A.Pandey,C.Webb,C.S.Soccol,and C.Larroche,Eds.)第5章,Asiatech Publishers Inc.,New Delhi,India,83-101(2004)]。甲酸脱氢酶广泛应用于NADH的再生，并使用廉价的底物甲酸盐，释放出CO2，其可以很容易地从反应混合物中去除。NADPH依赖性甲酸脱氢酶比NADH依赖性酶要更不常见。基于Masaki Ihara的工作[Ihara M.,Okabe A.,Kawano Y.,和Urano M.Light DrivenCO2Fixation by using cyanobacterial photosystem I and NADPH-dependent formatedehydrogenase.Plos One.8,e71581(2013)]，在铜绿假单胞菌甲酸脱氢酶P33160的腺嘌呤核糖结合环中引入两个突变(D222Q和H224N)，改变事实铜绿假单胞菌甲酸脱氢酶P33160对NADH至NADPH的特异性(图18)。尽管NAD结合显著减少，但酶仍然能够再生，这使其适用于两种辅因子再生。
表7.纯化并测试体外偶联反应的辅因子再生酶
Figure BDA0001550363180000511
Figure BDA0001550363180000521
实施例7
7.1.体外生物转化
体外酶促生物转化在200ul反应混合物中在30℃、800rpm下执行12小时。
CAR+PP_3370反应混合物含有100mM HEPES(pH 7.5)、1mM NADPH、2mM ATP、10mMMgCl2、10mM己二酸或4-羟基丁酸、50mM甲酸钠、5mM多磷酸盐、80μg CAR、20μg PP_3370、10μg P33160(D222Q/H224N)、10μg PA3455和10μg SMc02148。
MAB_4714c+AT反应混合物含有100mM HEPES(pH 7.5)、0.5mM NADPH、1mM谷氨酸酯、2mM ATP、10mM MgCl2、20mM己二酸、50mM甲酸钠、60m M氯化铵、5mM多磷酸盐、80μg MAB_4714c、20μg gabT或SAV2583、10μg P33160(D222Q/H224N)、5μg YOR375C、10μg PA3455和10μg SMc02148。
12小时之后，将反应物离心(16,000g)，用10kDa过滤器过滤上清液，使用HPLC分析产物形成。
用所有的酶执行阴性对照实验，但不是CAR。
7.2.CAR的生物化学表征
将示出来自初始筛选的最高活性的酶(图16)在体外对于己二酸和羟基丁酸饱和进行表征(表8)。两种底物的最低Km属于MAB_4714c(分别为35.87±2.68mM和26.52±1.77mM)，也作为最高催化效率(89.55±3.07M-1·s-1和71±3M-1·s-1)。大部分CAR对苯甲酸的比活性与来自爱荷华诺卡氏菌的CAR(2U/mg)在相同范围内[Akhtsr K.M.,Turner N.J.,Jones R.P.Carboxylic acid reductase is a versatile enzyme for the conversionof fatty acids into fuels and chemical commodities.Proc Natl Acad Sci U SA.110,87–92(2013)]。考虑到己二酸的羧基的连续还原和作为中间体的1,6-己酸的形成，还表征了CAR对6-羟基己酸的活性。
表8.对ATP/NADPH依赖性己二酸和4-羟基丁酸还原的CAR动力学参数。
Figure BDA0001550363180000531
表9.对ATP/NADPH依赖性6-羟基己酸还原的CAR动力学参数。
Figure BDA0001550363180000532
7.3-AR/AT的生物化学表征
将AR RHA1_ro05279、PA1146、SCO0229、PP_3370对己二醛饱和进行表征，将PP_3370、PSPTO_2097、RHA1_ro05279、SCO0229、TM_0111、PA1146对丁醛饱和进行表征(表10)。关于CAR辅因子对NADPH而非NADH的偏爱，选择对NADPH更有活性的AR。对于两种底物，通过来自恶臭假单胞菌的AR PP_3370示出具有最小Km值的最大kcat/Km。观察到的催化效率被测定为己二醛(2.08±0.09)×104M-1·s-1和丁醛(5.58±0.14)×103M-1·s-1，Km值分别被评估为0.29±0.02和1.53±0.11mM。
将在己二醛下显示出最高活性的AT GabT、SM2011_b20379、SAV2583、SM2011_c04388、SM2011_b20423用于生物化学表征(表11)。最大催化效率(kcat/Km)和最低Km值被测定为大肠杆菌gabT转氨酶的(9.22±0.27)×102M-1·s-1和0.67mM。
表10.对己二醛和丁醛还原的AR动力学参数。
Figure BDA0001550363180000541
*反应物增补有1.8％DMSO以增加己二醛溶解度。
Figure BDA0001550363180000542
Figure BDA0001550363180000551
表11.对己二醛的AT动力学参数
Figure BDA0001550363180000552
7.4-在体外和体内将CAR与AR偶联以产生二醇
为了研究己二酸和4-羟基丁酸转化成对应的二醇的效率，将偶联的CAR与AR进行测试。将10mM的底物用于12小时转化。测试了对己二酸和4-羟基丁酸有活性的MSMEG_5739、MAB_4714c、MAP_1040c、MSMEI_5586和MSMEG_2956与PP_3370AR的偶联反应(图19)。ATP和NADPH再生酶提供还原辅因子的持续供应。由于PP_3370底物转化产率比任何CAR高最低2个数量级，因此其活性不应该是偶联反应的限制步骤。通过HPLC、RI-检测器、Aminex-87H柱分析产物形成。MAB_4714c表现出4-羟基丁酸至1,4-丁二醇的32.9±2.2％的最大转化，以及己二酸至1,6-己二醇的38.7±0.9％的转化。MMAB_4714c将3.2±0.2％的己二酸转化为6-羟基己酸(表12)。
接下来，使用表达重组CAR的大肠杆菌细胞进行一系列体内4-羟基丁酸和己二酸生物转化。选择用于体外转化的相同CAR(与BSU4205PPTase共表达)用于10mM底物的全细胞生物转化。大肠杆菌基本上拥有若干种具有不同底物偏好的醛还原酶。在不存在另外的AR表达的情况下，用内源性大肠杆菌AR成功实现了醛还原为醇的第二步骤。用于己二酸生物转化的PP_3370的另外的表达降低了转化率。表达MSMEG_5739的细胞催化24小时后4-羟基丁酸的最大转化率为86.1±5.2％，以及己二酸转化为1,6-己二醇的效率为53.8±1.4％，伴随最小的至6-羟基己酸的转化9.1％±0.7％(图20B)。特征在于在体内对所有底物的转化效率最高的MAB_4714c将63.4±23.8％的4-羟基丁酸和5.3±0.3％的己二酸转化为1,6-己二醇，尽管节省了53.8±1.4％的中间体6-羟基己酸(表13)。通过LC-MS分析证实了1,6-己二醇、1,4-丁二醇和6-羟基己酸的形成。
将表达具有BSU03570磷酸泛酰巯基乙胺基转移酶的重组CAR的大肠杆菌细胞在具有100μM氨苄青霉素、50μM链霉素且增补有2％甘油的LB培养基上在37℃下好氧生长，直到培养物光密度(OD600nm)达到0.6-0.8。此时，用0.4mM IPTG诱导蛋白质表达，并且加入最终浓度10mM的底物(己二酸或4-羟基丁酸)。在6小时、24小时和48小时取样，离心，通过10kDa离心过滤器过滤上清液。用HPLC或LC-MS来分析产物。使用携带空质粒的大肠杆菌细胞执行阴性对照实验。体内实验的所有结果均来自至少两个独立的测定的平均值。
表12.通过与醛还原酶PP3370偶联的CAR体外转化10mM己二酸或10mM 4-羟基丁酸(12小时)的效率。反应由供应有50mM甲酸盐和5mM多磷酸盐的ATP和NADPH-再生酶供应。
Figure BDA0001550363180000561
表13.表达CAR的大肠杆菌在体内转化10mM己二酸或10mM4-羟基丁酸(24小时)的效率。
Figure BDA0001550363180000571
7.5-用于在体外将己二酸转化为氨基己酸的CAR与AT的偶联。
筛选的CAR对胺化的底物没有表现出活性。MAB_4714c对己二酸的Km值为35.87±2.68mM(表8)。在最佳条件下，用于用己二醛表征AT的转化产率比用CAR高至少10倍以上。在此种情况下，大部分单醛将被氨基转移酶捕获，并且除了6-氨基己酸形式的催化转化之外。将MAB_4714c与GabT或SAV2583AT偶联，使用谷氨酸作为氨基供体。用ATP、NAD(P)H和谷氨酸再生酶及其底物(5mM多磷酸酯(链长10-14个磷酸根)、40mM甲酸、50mM NH4Cl)来供应反应物。将20mM的底物用于12小时转化。使用C18柱前O-邻苯二醛衍生化通过HPLC来分析产物形成。己二酸至6-氨基己酸的最大转化效率被估计为21.3±0.8％(表14)。
表14.在ATP、NAD(P)H和谷氨酸再生酶的存在下通过CAR与AT的偶联反应在体外(12小时)将20mM己二酸转化成6-氨基己酸的转化效率。
Figure BDA0001550363180000572
序列表
<110> 多伦多大学
<120> 用于由羧酸合成聚合物前体的过程和微生物
<130> 05014925-102USPR
<140> 05014925-102USPR
<141> 2015-04-23
<160> 111
<170> PatentIn 3.5 版
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<221> 4Fe-4S结构域
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Met Lys Tyr Lys Lys Leu Phe Glu Thr Val Lys Ile Arg Asn Val Glu
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Leu Lys Asn Arg Tyr Ala Met Ala Pro Met Gly Pro Leu Gly Leu Ala
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Asp Ala Glu Gly Gly Phe Asn Gln Arg Gly Ile Glu Tyr Tyr Thr Ala
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Arg Ala Arg Gly Gly Thr Ala Leu Ile Ile Thr Gly Val Thr Phe Val
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Asp Asn Glu Val Glu Glu His Gly Met Pro Asn Val Pro Cys Pro Thr
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His Asn Pro Val His Phe Val Arg Thr Ser Lys Glu Met Thr Glu Arg
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Ile His Ala Tyr Asp Ser Lys Ile Phe Leu Gln Met Ser Ala Gly Phe
100 105 110
Gly Arg Val Thr Ile Pro Thr Asn Leu Gly Glu Tyr Pro Pro Val Ala
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Pro Ser Pro Ile Pro His Arg Trp Leu Asp Lys Thr Cys Arg Glu Leu
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Thr Val Glu Glu Ile His Ser Ile Val Arg Lys Phe Gly Asp Gly Ala
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Phe Asn Ala Lys Arg Ala Gly Phe Asp Gly Val Gln Ile His Ala Val
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His Glu Gly Tyr Leu Leu Asp Gln Phe Ala Ile Ala Phe Phe Asn Lys
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Arg Thr Asp Ala Tyr Gly Gly Pro Leu Glu Asn Arg Leu Arg Phe Ala
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Arg Glu Ile Val Glu Glu Ile Lys Gln Arg Cys Gly Glu Asp Phe Pro
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Val Thr Leu Arg Phe Ser Pro Lys Ser Phe Ile Lys Asp Trp Arg Glu
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Gly Ala Leu Pro Gly Glu Glu Phe Glu Glu Lys Gly Arg Asp Leu Asp
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Glu Gly Ile Glu Ala Ala Lys Leu Leu Val Ser Tyr Gly Tyr Asp Ala
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Leu Asp Val Asp Val Gly Ser Tyr Asp Ser Trp Trp Trp Ser His Pro
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Pro Met Tyr Gln Lys Lys Gly Leu Tyr Ile Pro Tyr Ala Arg Leu Val
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Lys Glu Ala Val Asp Val Pro Val Leu Cys Ala Gly Arg Met Asp Asn
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Pro Asp Leu Ala Leu Ala Ala Leu Glu Asp Gly Ala Cys Asp Ile Ile
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Ser Leu Gly Arg Pro Leu Leu Ala Asp Pro Asp Tyr Val Asn Lys Leu
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Arg Ile Gly Gln Val Ala Asp Ile Arg Pro Cys Leu Ser Cys His Glu
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Glu Arg Ala Lys Gly Gly Thr Gly Leu Ile Ile Thr Gly Val Thr Phe
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Ala Arg Glu Ile Val Glu Glu Ile Lys Asn Arg Cys Gly Glu Asp Phe
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Pro Val Thr Leu Arg Tyr Ser Pro Lys Ser Phe Ile Lys Asp Leu Arg
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Asp Gly Ala Leu Pro Gly Glu Glu Phe Val Glu Lys Gly Arg Asp Leu
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Asp Glu Gly Val Glu Ala Ala Lys Leu Leu Val Ser Tyr Gly Tyr Asp
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Pro Pro Met Tyr Gln Glu Lys Gly Leu Tyr Arg Lys Tyr Ala Lys Leu
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Met Lys Asp Thr Val Asp Val Pro Val Ile Cys Ala Gly Arg Met Asp
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Asp Pro Asp Met Ala Leu Glu Ala Val Glu Asn Gly Thr Cys Asp Val
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Ile Ser Leu Gly Arg Pro Leu Leu Ala Asp Pro Asp Tyr Val Asn Lys
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Leu Arg Ser Asn Lys Cys Lys Ser Ile Arg Pro Cys Ile Ser Cys Gln
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Glu Gly Cys Met Gly Arg Val Gln His Tyr Ser Met Leu Asn Cys Ala
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Val Asn Pro Gln Ala Cys Lys Glu Arg Ala Asn Ser Leu Thr Pro Ile
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Ile Lys Ser Lys Lys Val Leu Ile Val Gly Gly Gly Val Ala Gly Cys
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Glu Ala Ala Arg Val Leu Ala Leu Arg Gly His Glu Pro Val Leu Tyr
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Glu Lys Ser Asn Arg Leu Gly Gly Asn Leu Ile Pro Gly Gly Ala Pro
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Ser Phe Lys Glu Asp Asp Ile Ala Leu Ala Asp Trp Tyr Thr Asn Thr
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Val Val Gly Gly Gly Leu Val Gly Cys Glu Leu Ala Leu Asp Leu Ala
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Glu Leu Pro Cys Asp Ser Val Ile Leu Ser Val Gly Tyr Lys Glu Glu
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Ala Phe Glu Val Ala Asn His Ile
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Val Lys Asn Lys Ile Ala Met Ala Pro Met Gly Ala Phe Gly Leu Val
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Gly Arg Ser Gly Ala Pro Gly Gly Leu Leu Thr Ser Gln Pro Val Ser
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Ala Ser Ala Val Pro Asn Tyr Trp Asp Pro Thr Val Thr Cys Arg Glu
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Ala Lys Ile Ala His Lys Ala Gly Phe Asp Gly Val Glu Ile His Ala
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Arg Arg Thr Asp Lys Tyr Gly Gly Asp Leu Arg Gly Arg Leu Gln Leu
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Pro Ile Glu Ile Val Glu Ala Ile Lys Thr Glu Val Gly Ser Asn Phe
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Pro Val Gly Leu Arg Tyr Ser Val Lys Ser Cys Ile Lys Asp Trp Gly
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Gln Gly Gly Leu Ala Glu Glu Asp Tyr Val Glu Lys Gly Arg Asp Leu
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Glu Glu Gly Leu Glu Ala Ala Lys Ile Leu Glu Ala Ala Gly Tyr Asp
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Ala Phe Asn Ala Asp Leu Gly Thr Tyr Asp Ala Trp Tyr Trp Ala His
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Pro Pro Leu Tyr Gln Lys Asp Gly Leu Tyr Leu Pro Tyr Thr Lys Glu
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Leu Lys Lys Val Val Lys Ile Pro Val Met Val Ala Gly Lys Met Gly
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Met Pro Asp Val Ala Glu Gly Ala Leu Glu Asp Glu Ala Ala Asp Met
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Val Thr Leu Gly Arg Pro Leu Leu Cys Asp Ala Tyr Trp Pro Lys Lys
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Val Phe Thr Gly Gln Ile Asp Arg Ile Arg Pro Cys Ile Gly Cys His
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Thr Gly Cys Met Gly Arg Gly Phe Glu Gly Arg Pro Leu Ser Cys Thr
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Val Asn Pro Ala Ala Gly Arg Glu Arg Tyr Tyr Glu Val Lys Pro Ala
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Ala Ala Pro Lys Lys Val Met Ile Val Gly Gly Gly Val Ala Gly Met
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Glu Ala Ala Arg Ile Thr Ala Met Arg Gly His Lys Val Ser Met Tyr
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Glu Gly Thr Lys Glu Leu Gly Gly Gln Val Ile Pro Ala Ser Val Pro
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Asp Phe Lys Ile Asp Asp Arg Arg Leu Leu Asp Trp Tyr Arg Asn Glu
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Met Lys Glu Leu Lys Val Lys Leu Val Leu Asp Thr Asn Val Thr Glu
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Ala Lys Glu Ile Lys Leu Asn Leu Pro Gly Ile Glu Lys Asp Lys Val
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Ala Thr Val Ile Glu Val Leu Lys Gly Ser Lys Gln Val Gly Glu Asn
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Val Leu Met Val Gly Gly Gly Leu Ala Gly Cys Glu Thr Ala Leu Tyr
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Leu Ala Lys Gln Gly Lys Lys Val Thr Ile Ile Glu Ala Arg Asp Thr
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Ile Leu Asn Ala Gly Lys Pro Val Pro His Met Asn Lys Ile Met Leu
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Ile Asp Leu Leu Lys Asn Ser Gly Val Asn Ile Ile Thr Glu Thr Ser
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Leu Leu Glu Val Thr Asp Arg Gly Ala Ile Leu Ile Asp Asn Lys Phe
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Lys Lys Gln Asn Ile Asp Ala Asp Thr Val Val Ile Ala Val Gly Phe
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Lys Ala Asp Arg Glu Leu Tyr Asn Lys Leu Arg Asp Lys Val Ala Asp
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Leu Tyr Leu Val Gly Asp Ala Asn Glu Ser Ala Asn Ile Met Asn Ala
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Ile Trp Ser Ala Asn Glu Ile Ala Leu Asn Cys
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Met Lys Asn Lys Ser Leu Phe Gln Pro Ile Lys Ile Gly Lys Met Glu
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Val Lys Asn Lys Ile Ser Met Ala Pro Met Gly Ala Phe Gly Leu Val
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Asp Asn Glu Gly Cys Tyr Asn Gln Arg Ala Ile Asp Tyr Tyr Val Glu
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Arg Ala Lys Gly Gly Thr Gly Leu Ile Ile Thr Ser Ile Thr Lys Val
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Glu Asn Glu Leu Asp Lys Val Val Pro Gly Ile Ile Pro Val Ile Ser
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Thr Asn Pro Gly Arg Phe Leu Met Thr Ser Ser Glu Met Thr Glu Arg
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Val His Ala Tyr Gly Ala Lys Ile Phe Leu Gln Leu Thr Met Gly Phe
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Gly Arg Ser Gly Ala Pro Gly Thr Leu Leu Thr Ser Gln Pro Val Ser
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Ala Ser Asp Ile Pro Asn Tyr Trp Asp Pro Thr Val Thr Cys Arg Ala
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Leu Thr Thr Ser Glu Val Glu Trp Ile Val Ser Lys Phe Ile Glu Gly
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Ala Asp Ile Ala Gln Lys Ala Asp Phe Asp Gly Val Glu Ile His Ala
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Val His Glu Gly Tyr Leu Leu Asp Gln Phe Thr Leu Ser Ile Phe Asn
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Arg Arg Thr Asp Lys Tyr Gly Gly Asp Leu Arg Gly Arg Leu Gln Leu
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Pro Ile Glu Ile Val Gln Gly Ile Lys Ala Arg Leu Gly Asp Asp Phe
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Pro Val Gly Leu Arg Tyr Ser Val Lys Ser Cys Ile Lys Asp Trp Arg
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Gln Gly Gly Leu Pro Asp Glu Asp Tyr Leu Glu Lys Gly Arg Asp Leu
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Lys Glu Gly Leu Asp Ala Ala Lys Ile Leu Glu Ala Ala Gly Tyr Asp
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Pro Pro Ile Tyr Gln Lys Asn Gly Leu Tyr Leu Pro Tyr Thr Lys Glu
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Leu Lys Lys Val Val Lys Ile Pro Val Ile Val Ala Gly Lys Leu Gly
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Ile Pro Glu Asp Ala Glu Lys Ala Leu Asp Asp Lys Ala Ala Asp Met
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Ile Gly Leu Gly Arg Pro Leu Leu Ala Asp Pro Tyr Trp Pro Lys Lys
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Val Leu Ser Asp Asn Ser Glu Arg Ile Arg Pro Cys Ile Gly Cys His
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Thr Gly Cys Leu Gly Arg Gly Phe Glu Gly Lys Pro Leu Ser Cys Thr
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Val Asn Pro Ala Val Gly Arg Glu Arg Tyr Tyr Glu Ile Lys Pro Thr
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Leu Thr Pro Lys Lys Val Met Val Val Gly Gly Gly Val Ala Gly Met
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Glu Ala Ala Arg Ile Ile Lys Met Arg Gly His Asp Val Thr Ile Tyr
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Val Leu Met Val Gly Gly Gly Leu Val Gly Cys Glu Ala Ala Leu Tyr
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Leu Ala Lys Gln Gly Lys Asn Val Thr Ile Val Glu Ala Lys Asp Thr
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Leu Leu Asn Ser Ser Lys Pro Ile Pro His Met Asn Lys Ile Met Leu
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Ile Asp Leu Leu Asn Met Tyr Asn Val Lys Ala Ile Thr Asn Asn Ser
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Leu Leu Glu Val Thr Asp Lys Gly Ala Val Leu Ile Asn Asn Lys Phe
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Lys Lys Gln Glu Val Ser Ala Asp Thr Val Thr Ile Ala Val Gly Phe
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Lys Ser Asp Arg Glu Leu Tyr Asn Lys Leu Asn Gly Asn Ile Ala Asp
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Leu Tyr Leu Ile Gly Asp Ala Tyr Gln Ser Ala Asn Ile Met Asn Ala
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Ile Trp Ser Ala Asn Glu Ile Gly Leu Asn Cys
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Met Lys Asn Lys Ser Leu Phe Glu Pro Ile Lys Ile Gly Lys Val Glu
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Val Lys Asn Lys Ile Ser Met Ala Pro Met Gly Ala Phe Gly Leu Val
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Asp Asn Glu Gly Cys Tyr Asn Gln Arg Ala Val Asp Tyr Tyr Val Glu
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Arg Ala Lys Gly Gly Thr Gly Leu Ile Ile Thr Ser Ile Thr Lys Val
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Glu Asn Glu Ile Asp Lys Val Val Pro Gly Val Ile Pro Ile Ile Ser
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Ile Asn Pro Gly Arg Phe Ile Met Thr Ser Ser Glu Met Thr Glu Arg
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Val His Ser Tyr Gly Ser Lys Ile Phe Leu Gln Leu Thr Met Gly Phe
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Gly Arg Ser Gly Ala Pro Gly Thr Leu Leu Thr Ser Gln Pro Val Ser
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Ala Ser Ser Val Pro Asn Tyr Trp Asp Pro Thr Val Thr Cys Arg Glu
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Leu Thr Thr Ser Glu Val Glu Trp Ile Val Ala Lys Phe Ile Gln Gly
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Ala Ala Ile Ala Gln Lys Ala Gly Phe Asp Gly Val Glu Ile His Ala
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Val His Glu Gly Tyr Leu Leu Asp Gln Phe Thr Leu Ser Ile Phe Asn
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Pro Ile Glu Ile Val Gln Gly Ile Lys Ala Gln Val Gly Ser Asp Phe
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Pro Pro Leu Tyr Gln Lys Asp Gly Leu Tyr Leu Pro Tyr Thr Lys Glu
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Leu Lys Lys Val Val Lys Ile Pro Val Ile Val Ala Gly Lys Leu Gly
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Val Pro Gln Glu Ala Glu Lys Ala Leu Asp Glu Gly Gly Ala Asp Met
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Ile Gly Leu Ala Arg Pro Leu Leu Ser Asp Ala Tyr Trp Pro Lys Lys
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Val Leu Ser Gly His Pro Glu Arg Ile Arg Pro Cys Ile Gly Cys His
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Val Gly Cys Leu Gly Arg Gly Phe Glu Gly Lys Pro Leu Ser Cys Ala
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Val Asn Pro Ala Ala Gly Arg Glu Arg Tyr Tyr Glu Ile Arg Pro Ala
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Ala Ile Pro Lys Lys Val Leu Ile Ala Gly Gly Gly Val Ala Gly Met
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Glu Ala Ala Arg Met Ala Val Leu Arg Gly His Lys Val Thr Leu Tyr
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Glu Ser Thr Asp Gln Leu Gly Gly Glu Ile Val Pro Gly Ser Val Pro
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Asp Phe Lys Ile Asp Asp Arg Arg Leu Leu Asp Trp Tyr Arg Asn Glu
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Lys Leu Val Gly Lys Glu Gln Pro Asp Val Val Ile Val Ala Thr Gly
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Ala Asn Asp Val Lys Ile Lys Leu Pro Gly Ile Glu Lys Asp Lys Val
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Ser Thr Ala Val Asp Ile Leu Asn Gly Ala Lys Lys Ser Gly Lys Asn
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Val Leu Ile Val Gly Gly Gly Leu Val Gly Cys Glu Thr Ala Leu Tyr
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Leu Ala Lys Ala Gly Lys Lys Val Ala Ile Val Glu Ala Lys Asp Lys
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Ile Leu Asp Ala Gly Lys Pro Ile Pro His Met Asn Lys Ile Met Leu
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Glu Asp Leu Ile Lys Lys Tyr Asn Ile Lys Val Ile Thr Gly Asn Ser
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Leu Leu Glu Val Thr Gly Asp Gly Ala Val Leu Ile Asp Asn Lys Phe
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Lys Gln Gln Glu Val Tyr Ala Asp Thr Ile Val Ile Ser Ile Gly Phe
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Lys Ser Asn Arg Lys Leu Tyr Asn Lys Leu His Gly Lys Val Thr Asp
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Leu Tyr Leu Ile Gly Asp Ala Tyr Gln Thr Ala Asn Ile Met Asp Ala
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Ile Trp Ser Gly Asn Glu Ile Gly Leu Asn Cys
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Met Val Val Ala Tyr Thr Arg Leu Phe Glu Pro Ile Lys Ile Gly Lys
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Val Glu Ile Lys Asn Lys Ile Ala Met Ala Pro Met Gly Val Leu Gly
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Leu Ala Thr Gln Asp Gly Cys Phe Ser Lys Arg Val Val Asp Tyr Tyr
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Val Glu Arg Ala Lys Gly Gly Thr Gly Leu Ile Ile Thr Ser Val Thr
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Lys Val Asp Asn Glu Ile Glu Arg Phe Lys Ala Gly Ala Val Pro Val
65 70 75 80
Ala Thr Ala Asn Pro Leu His Phe Ile Ala Thr Ala Gly Glu Leu Thr
85 90 95
Glu Arg Val His Ala Tyr Gly Thr Lys Ile Phe Leu Gln Leu Gly Met
100 105 110
Gly Phe Gly Arg Val Ala Ala Pro Ile Leu Leu Glu Ser His Pro Val
115 120 125
Ala Pro Ser Ala Leu Pro Asn Phe Trp Asp Pro Ser Ile Thr Cys Arg
130 135 140
Glu Leu Thr Thr Ala Glu Val Glu Thr Leu Val Gln Arg Ala Ser Glu
145 150 155 160
Ala Ala Glu Ile Ala Val Glu Ala Gly Phe Asp Gly Val Glu Ile His
165 170 175
Ala Met His Glu Gly Tyr Leu Leu Asp Gln Phe Thr Ile Ala Leu Phe
180 185 190
Asn Arg Arg Gly Asp Lys Tyr Gly Gly Ala Leu Glu Asp Arg Leu Thr
195 200 205
Phe Pro Ile Glu Ile Val Arg Ala Ile Lys Asp Arg Val Gly Lys Asp
210 215 220
Phe Pro Val Val Leu Arg Phe Ser Ile Lys Asn Tyr Ile Lys Asp Trp
225 230 235 240
Arg Gln Gly Gly Leu Pro Gly Glu Asn Phe Gln Glu Lys Gly Arg Asp
245 250 255
Val Glu Glu Ala Leu Ala Ala Ala Lys Ile Leu Glu Gly Ala Gly Tyr
260 265 270
Asp Gly Phe Asp Ala Asp Ala Gly Ser Tyr Asp Ala Trp Tyr Trp Ala
275 280 285
His Pro Pro Val Tyr Gln Lys His Gly Cys Tyr Leu Pro Leu Thr Gln
290 295 300
Arg Leu Lys Glu Val Val Lys Val Pro Val Ile Val Ala Gly Arg Leu
305 310 315 320
Glu Ile Pro Glu Leu Ala Glu Glu Ala Leu Val Lys Gly Gln Ala Asp
325 330 335
Met Ile Ala Ile Gly Arg Gly Leu Leu Thr Asp Pro Tyr Trp Val Asn
340 345 350
Lys Val Met Thr Gly Arg Ser Lys Asn Ile Arg Pro Cys Ile Gly Cys
355 360 365
His Asp Gly Cys Leu Gly Arg Gly Phe Leu Gly Arg Pro Leu Ser Cys
370 375 380
Thr Val Asn Pro Ala Cys Gly Arg Glu Glu Glu Tyr Ala Ile Asp Arg
385 390 395 400
Ala Pro Glu Ala Lys Gln Val Met Val Ile Gly Gly Gly Val Ala Gly
405 410 415
Met Glu Ala Ala Arg Val Ala Ala Leu Arg Gly His Arg Val Ser Leu
420 425 430
Tyr Glu Lys Ser Asp Arg Leu Gly Gly His Val Val Glu Ala Ala Val
435 440 445
Pro Asp Phe Lys Ala Asp Asp Gly Arg Leu Leu Glu Trp Tyr Lys Thr
450 455 460
Glu Leu Gly Glu Leu Gln Val Glu Ile His Leu Asn Gln Glu Val Thr
465 470 475 480
Pro Glu Phe Val Glu Glu Lys Asn Pro Asp Val Val Val Val Ala Thr
485 490 495
Gly Ser Thr Pro Ala Ile Pro Asp Ile Pro Gly Val Asn Lys Asp Lys
500 505 510
Val Thr Thr Val Ser Asp Leu Leu Leu Gly Lys Lys Gln Ala Gly Asp
515 520 525
Arg Val Val Ile Ile Gly Gly Gly Leu Ala Gly Cys Glu Thr Ala Leu
530 535 540
Trp Leu Ala Gln Gln Gly Lys Asp Val Thr Ile Ile Glu Ile Leu Asp
545 550 555 560
Asp Leu Met Arg Ala Gly Ile Pro Val Pro Tyr Met Asn Arg Met Met
565 570 575
Leu Leu Asp Leu Leu Lys Met Asn Gly Val Lys Trp Leu Thr Glu Thr
580 585 590
Ser Val Leu Glu Val Thr Asp Asp Gly Val Thr Leu Ile Gly Lys Thr
595 600 605
Tyr Gln Arg Ser Pro Leu Pro Ala Asp Thr Val Ile Leu Ala Val Gly
610 615 620
Phe Ala Ala Asp Gln Arg Leu Tyr Asn Ala Leu Arg Asp Lys Ile Pro
625 630 635 640
Asn Leu Tyr Leu Ile Gly Asp Ser Arg Glu Pro Arg Asn Ile Leu Ala
645 650 655
Ala Ile Trp Glu Gly Tyr Glu Val Gly Arg Gly Ile
660 665
<210> 7
<211> 338
<212> PRT
<213> 耐盐嗜碱芽孢杆菌C-125
<400> 7
Met Glu His Lys Leu Phe Ser Ser Tyr Val Val Lys Gly Val Thr Leu
1 5 10 15
Lys Asn Arg Ile Val Met Ser Pro Met Cys Met Tyr Ser Ser Asp Gln
20 25 30
Lys Asp Gly Lys Ile Arg Pro Phe His Ile Ser His Tyr Glu Ser Arg
35 40 45
Ala Ala Gly Gln Val Gly Leu Ile Ile Val Glu Ala Thr Ala Val Thr
50 55 60
Pro Gln Gly Arg Ile Ser Pro Tyr Asp Leu Gly Ile Trp Ser Asp Asp
65 70 75 80
His Ile Ser Gly Leu Thr Glu Thr Val Glu Arg Ile His Ala His Gly
85 90 95
Ala Lys Ala Ala Ile Gln Leu Ala His Ala Gly Arg Lys Ala Glu Leu
100 105 110
Asp Gly Pro Ile Ile Ala Pro Ser Ala Ile Ser Tyr Asp Lys Met Lys
115 120 125
Thr Pro Asp Ala Met Thr Glu Glu Gln Ile Ser Glu Thr Ile Glu Ala
130 135 140
Phe Lys Leu Gly Ala Leu Arg Ala Lys Lys Ala Gly Phe Asp Ile Ile
145 150 155 160
Glu Ile His Gly Ala His Gly Tyr Leu Ile Asn Glu Phe Leu Ser Pro
165 170 175
Leu Thr Asn Lys Arg Thr Asp Ala Tyr Gly Gly Ser Leu Glu Asn Arg
180 185 190
Tyr Arg Leu Leu Arg Glu Ile Ile Ser Glu Ile Gln Thr Val Trp Asp
195 200 205
Gly Pro Leu Phe Val Arg Ile Ser Ala Ala Glu Tyr Ala Glu Gly Gly
210 215 220
Asn Glu Leu Ser Asp Phe Ile Thr Leu Ala Lys Trp Met Lys Lys Gln
225 230 235 240
Gly Ile Asp Leu Ile Asp Cys Ser Ser Gly Ala Val Val Pro Ala Pro
245 250 255
Ile Pro Val Tyr Pro Gly Tyr Gln Val Pro Leu Ala Glu Ala Ile Arg
260 265 270
His Glu Ala Asn Ile Ala Thr Gly Ala Val Gly Leu Ile Thr Ser Gly
275 280 285
Ile Gln Ala Glu Glu Ile Leu Gln Asn Glu Arg Ala Asp Leu Ile Phe
290 295 300
Val Ala Arg Glu Leu Leu Arg Asn Pro Tyr Trp Pro Arg Glu Ala Ala
305 310 315 320
Leu Glu Leu Gly Thr Thr Ile Ser Gly Pro Ser Gln Tyr Asp Arg Ala
325 330 335
Trp Leu
<210> 8
<211> 338
<212> PRT
<213> 枯草芽孢杆菌亚种枯草菌株168
<400> 8
Met Ala Arg Lys Leu Phe Thr Pro Ile Thr Ile Lys Asp Met Thr Leu
1 5 10 15
Lys Asn Arg Ile Val Met Ser Pro Met Cys Met Tyr Ser Ser His Glu
20 25 30
Lys Asp Gly Lys Leu Thr Pro Phe His Met Ala His Tyr Ile Ser Arg
35 40 45
Ala Ile Gly Gln Val Gly Leu Ile Ile Val Glu Ala Ser Ala Val Asn
50 55 60
Pro Gln Gly Arg Ile Thr Asp Gln Asp Leu Gly Ile Trp Ser Asp Glu
65 70 75 80
His Ile Glu Gly Phe Ala Lys Leu Thr Glu Gln Val Lys Glu Gln Gly
85 90 95
Ser Lys Ile Gly Ile Gln Leu Ala His Ala Gly Arg Lys Ala Glu Leu
100 105 110
Glu Gly Asp Ile Phe Ala Pro Ser Ala Ile Ala Phe Asp Glu Gln Ser
115 120 125
Ala Thr Pro Val Glu Met Ser Ala Glu Lys Val Lys Glu Thr Val Gln
130 135 140
Glu Phe Lys Gln Ala Ala Ala Arg Ala Lys Glu Ala Gly Phe Asp Val
145 150 155 160
Ile Glu Ile His Ala Ala His Gly Tyr Leu Ile His Glu Phe Leu Ser
165 170 175
Pro Leu Ser Asn His Arg Thr Asp Glu Tyr Gly Gly Ser Pro Glu Asn
180 185 190
Arg Tyr Arg Phe Leu Arg Glu Ile Ile Asp Glu Val Lys Gln Val Trp
195 200 205
Asp Gly Pro Leu Phe Val Arg Val Ser Ala Ser Asp Tyr Thr Asp Lys
210 215 220
Gly Leu Asp Ile Ala Asp His Ile Gly Phe Ala Lys Trp Met Lys Glu
225 230 235 240
Gln Gly Val Asp Leu Ile Asp Cys Ser Ser Gly Ala Leu Val His Ala
245 250 255
Asp Ile Asn Val Phe Pro Gly Tyr Gln Val Ser Phe Ala Glu Lys Ile
260 265 270
Arg Glu Gln Ala Asp Met Ala Thr Gly Ala Val Gly Met Ile Thr Asp
275 280 285
Gly Ser Met Ala Glu Glu Ile Leu Gln Asn Gly Arg Ala Asp Leu Ile
290 295 300
Phe Ile Gly Arg Glu Leu Leu Arg Asp Pro Phe Phe Ala Arg Thr Ala
305 310 315 320
Ala Lys Gln Leu Asn Thr Glu Ile Pro Ala Pro Val Gln Tyr Glu Arg
325 330 335
Gly Trp
<210> 9
<211> 364
<212> PRT
<213> 紫色色杆菌ATCC 12472
<400> 9
Met Asn Ala Asp Lys Leu Leu Thr Pro Leu Thr Met Gly Ala Val Ala
1 5 10 15
Leu Ser Asn Arg Val Val Met Ala Pro Leu Thr Arg Leu Arg Asn Ile
20 25 30
Glu Pro Gly Asp Val Pro Gly Pro Leu Ala Lys Glu Tyr Tyr Arg Gln
35 40 45
Arg Ala Ser Ala Gly Leu Ile Val Ala Glu Gly Thr His Ile Ser Pro
50 55 60
Thr Ala Lys Gly Tyr Ala Gly Ala Pro Gly Ile Tyr Ser Glu Glu Gln
65 70 75 80
Val Arg Ala Trp Ser Glu Val Thr Gly Ala Val His Gln Asp Gly Gly
85 90 95
Lys Ile Ala Leu Gln Leu Trp His Thr Gly Arg Ile Ser His Arg Ser
100 105 110
Leu Gln Pro Asn Gly Asp Ala Pro Val Gly Pro Ser Ala Ile Gln Ala
115 120 125
Asp Ser Arg Thr Asn Ile Arg Ala Ala Asp Gly Ser Leu Val Arg Glu
130 135 140
Gln Cys Asp Thr Pro Arg Ala Leu Glu Ile Glu Glu Ile Glu Asp Ile
145 150 155 160
Ile Glu Asp Tyr Arg Arg Ala Ala Asp Asn Ala Arg Arg Ala Gly Phe
165 170 175
Asp Met Val Glu Ile His Gly Ala His Gly Tyr Leu Ile Asp Gln Phe
180 185 190
Leu Ser Pro Ala Ala Asn Val Arg Thr Asp Gln Tyr Gly Gly Ser Val
195 200 205
Glu Asn Arg Ala Arg Phe Leu Leu Glu Val Val Asp Ala Val Val Ala
210 215 220
Glu Trp Asp Ala Asp His Val Gly Ile Arg Ile Ser Pro Leu Gly Ile
225 230 235 240
Phe Asn Gly Val Ser Asn Thr Asp Gln Leu Asp Met Ala Leu Tyr Leu
245 250 255
Ala Glu Gln Leu Ala Lys Arg Lys Leu Ala Phe Leu His Ile Ser Glu
260 265 270
Pro Asp Trp Ala Gly Gly Pro Thr Leu Asp Asp Gly Phe Arg Ala Glu
275 280 285
Leu Arg Gln Arg Tyr Pro Gly Val Ile Ile Gly Ala Gly Gly Tyr Ser
290 295 300
Ala Glu Lys Ala Glu Thr Leu Leu Lys Lys Gly Phe Ile Asp Ala Ala
305 310 315 320
Ala Phe Gly Arg Ser Tyr Ile Ala Asn Pro Asp Leu Val Glu Arg Leu
325 330 335
Lys Gln Asn Ala Pro Leu Asn Pro Pro Lys Pro Asp Thr Phe Tyr Gly
340 345 350
Gly Gly Ala Glu Gly Tyr Thr Asp Tyr Pro Thr Leu
355 360
<210> 10
<211> 365
<212> PRT
<213> 大肠杆菌 K12
<400> 10
Met Ser Ser Glu Lys Leu Tyr Ser Pro Leu Lys Val Gly Ala Ile Thr
1 5 10 15
Ala Ala Asn Arg Ile Phe Met Ala Pro Leu Thr Arg Leu Arg Ser Ile
20 25 30
Glu Pro Gly Asp Ile Pro Thr Pro Leu Met Ala Glu Tyr Tyr Arg Gln
35 40 45
Arg Ala Ser Ala Gly Leu Ile Ile Ser Glu Ala Thr Gln Ile Ser Ala
50 55 60
Gln Ala Lys Gly Tyr Ala Gly Ala Pro Gly Ile His Ser Pro Glu Gln
65 70 75 80
Ile Ala Ala Trp Lys Lys Ile Thr Ala Gly Val His Ala Glu Asn Gly
85 90 95
His Met Ala Val Gln Leu Trp His Thr Gly Arg Ile Ser His Ala Ser
100 105 110
Leu Gln Pro Gly Gly Gln Ala Pro Val Ala Pro Ser Ala Leu Ser Ala
115 120 125
Gly Thr Arg Thr Ser Leu Arg Asp Glu Asn Gly Gln Ala Ile Arg Val
130 135 140
Glu Thr Ser Met Pro Arg Ala Leu Glu Leu Glu Glu Ile Pro Gly Ile
145 150 155 160
Val Asn Asp Phe Arg Gln Ala Ile Ala Asn Ala Arg Glu Ala Gly Phe
165 170 175
Asp Leu Val Glu Leu His Ser Ala His Gly Tyr Leu Leu His Gln Phe
180 185 190
Leu Ser Pro Ser Ser Asn His Arg Thr Asp Gln Tyr Gly Gly Ser Val
195 200 205
Glu Asn Arg Ala Arg Leu Val Leu Glu Val Val Asp Ala Gly Ile Glu
210 215 220
Glu Trp Gly Ala Asp Arg Ile Gly Ile Arg Val Ser Pro Ile Gly Thr
225 230 235 240
Phe Gln Asn Thr Asp Asn Gly Pro Asn Glu Glu Ala Asp Ala Leu Tyr
245 250 255
Leu Ile Glu Gln Leu Gly Lys Arg Gly Ile Ala Tyr Leu His Met Ser
260 265 270
Glu Pro Asp Trp Ala Gly Gly Glu Pro Tyr Thr Asp Ala Phe Arg Glu
275 280 285
Lys Val Arg Ala Arg Phe His Gly Pro Ile Ile Gly Ala Gly Ala Tyr
290 295 300
Thr Val Glu Lys Ala Glu Thr Leu Ile Gly Lys Gly Leu Ile Asp Ala
305 310 315 320
Val Ala Phe Gly Arg Asp Trp Ile Ala Asn Pro Asp Leu Val Ala Arg
325 330 335
Leu Gln Arg Lys Ala Glu Leu Asn Pro Gln Arg Ala Glu Ser Phe Tyr
340 345 350
Gly Gly Gly Ala Glu Gly Tyr Thr Asp Tyr Pro Thr Leu
355 360 365
<210> 11
<211> 338
<212> PRT
<213> 单核细胞增生李斯特菌 EGD-e
<400> 11
Met Ser Lys Leu Phe Ser Glu Tyr Lys Leu Lys Asp Val Thr Leu Lys
1 5 10 15
Asn Arg Ile Val Met Ser Pro Met Cys Met Tyr Ser Val Glu Asn Lys
20 25 30
Asp Gly Ile Ala Thr Asp Phe His Phe Ala His Tyr Val Ser Arg Ala
35 40 45
Ala Gly Gly Thr Gly Leu Val Ile Leu Glu Ala Thr Ala Val Gln Glu
50 55 60
Val Gly Arg Ile Ser Glu Phe Asp Leu Gly Leu Trp Asn Asp Glu Gln
65 70 75 80
Val Pro Ala Leu Lys Lys Leu Val Gly Gly Leu His Tyr His Gly Ala
85 90 95
Lys Ala Gly Ile Gln Leu Ala His Ala Gly Arg Lys Ala Val Leu Pro
100 105 110
Gly Glu Ile Val Ala Pro Ser Ala Ile Ala Phe Asp Glu Lys Ser Asp
115 120 125
Lys Pro Val Glu Leu Thr Lys Glu Ala Ile Lys Glu Val Val Ala Asp
130 135 140
Phe Lys Arg Ala Ala Tyr Arg Ala Lys Glu Ala Gly Phe Asp Val Ile
145 150 155 160
Glu Ile His Ala Ala His Gly Tyr Leu Ile His Gln Phe Leu Ser Pro
165 170 175
Ile Thr Asn Arg Arg Glu Asp Asn Tyr Gly Gly Pro Ala Gly Asn Arg
180 185 190
Tyr Lys Ile Leu Ser Asp Ile Ile Lys Ala Val Lys Glu Val Trp Asp
195 200 205
Gly Pro Ile Ile Val Arg Val Ser Ala Thr Asp Tyr Ala His Gly Gly
210 215 220
Leu Gln Leu Glu Asp His Ile Pro Phe Ala Lys Trp Met Lys Ala Asp
225 230 235 240
Gly Val Glu Leu Ile Asp Val Ser Thr Gly Gly Leu Val Asn Val Ala
245 250 255
Pro Pro Val Phe Pro Gly Tyr Gln Val Pro Phe Ala Asp Glu Ile Arg
260 265 270
Arg Gly Ala Gly Ile Ala Thr Gly Ala Leu Gly Leu Ile Thr Arg Gly
275 280 285
Glu Gln Ala Glu Glu Ile Leu Cys Asn Glu Arg Ala Asp Leu Ile Ile
290 295 300
Val Gly Arg Glu Leu Leu Arg Asn Pro Tyr Phe Ala Lys Asp Ala Ala
305 310 315 320
Lys Gln Leu Gly Glu Thr Ile Glu Ala Pro Lys Gln Tyr Ser Arg Ala
325 330 335
Trp Lys
<210> 12
<211> 354
<212> PRT
<213> 欧洲亚硝化单胞菌 ATCC 19718
<400> 12
Met Leu Phe Asn Pro Leu Gln Val Gly Ser Leu Thr Leu Pro Asn Arg
1 5 10 15
Ile Leu Leu Ala Pro Leu Thr Arg Ala Arg Ala Asp Ala Gly His Met
20 25 30
Pro Asn Ala Leu Met Ala Glu Tyr Tyr Ser Gln Arg Ala Thr Gly Gly
35 40 45
Leu Leu Ile Ser Glu Cys Thr Met Val Ala Pro Gly Thr Ser Ala Phe
50 55 60
Val Asn Glu Pro Gly Ile Tyr Asn Asp Ala Gln Ile Ala Ala Trp Arg
65 70 75 80
Gln Val Thr Asp Ala Val His Ala Lys Gly Gly Arg Ile Phe Met Gln
85 90 95
Ile Trp His Ala Gly Arg Ala Ala Tyr Pro Gly Ala Ala Asp Gly Ala
100 105 110
Pro Ile Val Ser Ser Ser Ala Thr Ala Ile Glu Gly Glu Ile His Thr
115 120 125
Pro Gln Gly Lys Val Pro His Ala Val Pro Arg Pro Leu Thr Val Asp
130 135 140
Glu Ile Pro Gly Ile Val Ala Ala Phe Ala Gln Gly Ala Arg Asn Ala
145 150 155 160
Ile Ala Ala Gly Phe Asp Gly Val Glu Val His Gly Ala Asn Gly Tyr
165 170 175
Leu Ile Asp Gln Phe Leu Arg Asp Thr Pro Asn Gln Arg Thr Asp Ala
180 185 190
Tyr Gly Gly Ser Leu Glu Asn Arg Ala Arg Leu Leu Phe Glu Val Leu
195 200 205
Thr Ala Val Thr Gln Ala Ile Gly Ser Glu Arg Val Gly Leu Arg Leu
210 215 220
Ser Pro Leu Asn Ser Phe Asn Ser Met Lys Asp Ser Asp Pro Leu Ala
225 230 235 240
Leu Ile Gly Phe Leu Ala Asp Arg Leu Asn Ala Phe Lys Leu Ala Tyr
245 250 255
Leu His Val Met Arg Ala Asp Phe Phe Gly Val Gln Lys Ala Asp Val
260 265 270
Met Pro Val Ala Arg Glu Lys Tyr Lys Gly Val Leu Val Gly Asn Met
275 280 285
Gly Tyr Ser Ala Asp Glu Ala Glu Ala Ala Ile Ala Glu Gly Arg Leu
290 295 300
Asp Ala Val Ala Phe Gly Thr Ala Phe Leu Ala Asn Pro Asp Leu Pro
305 310 315 320
Ala Arg Ile Arg Ala Lys Ala Pro Leu Asn Ala Pro Asp Ser Asn Thr
325 330 335
Phe Tyr Ala Gly Gly Ala Lys Gly Tyr Thr Asp Tyr Pro Thr Leu Gln
340 345 350
Leu Ala
<210> 13
<211> 369
<212> PRT
<213> 铜绿假单胞菌 PAO1
<400> 13
Met Ser Asn Leu Leu Leu Ser Pro Leu Ala Val Gly Asn Leu Ala Leu
1 5 10 15
Arg Asn Arg Ile Val Met Ala Pro Met Thr Arg Ser Arg Ala Gln Gln
20 25 30
Pro Gly Asp Val Pro Thr Ala Leu Asn Ala Leu Tyr Tyr Ala Gln Arg
35 40 45
Ala Gly Ala Gly Leu Ile Val Ser Glu Gly Thr Gln Ile Ser His Leu
50 55 60
Gly Gln Gly Tyr Ala Tyr Thr Pro Gly Ile Tyr Ser Glu Ala Gln Leu
65 70 75 80
Ala Gly Trp Arg Gln Val Thr Glu Ala Val His Ala Ala Gly Gly Leu
85 90 95
Ile Ala Ala Gln Leu Trp His Val Gly Arg Met Ser His Arg Ser Leu
100 105 110
Gln Ala Gly Gly Glu Ala Pro Ile Ala Pro Ser Pro Ile Gln Ala Lys
115 120 125
Ala Gln Val Phe Ile Ala Asp Gly Gln Gly Gly Gly Ser Met Ala Pro
130 135 140
Ala Asp Ala Pro Arg Glu Met Thr Leu Glu Asp Ile Arg Arg Val Arg
145 150 155 160
Asp Glu Phe Val Arg Ala Ala Arg Asn Ala Leu Asp Ala Gly Phe Asp
165 170 175
Leu Val Glu Leu His Gly Ala Asn Gly Tyr Leu Ile Asp Gln Phe Leu
180 185 190
Ala Ser Ala Ser Asn Arg Arg Ser Asp Ala Tyr Gly Gly Ser Leu Glu
195 200 205
Asn Arg Ala Arg Phe Leu Leu Glu Ile Val Asp Ala Leu Val Ala Ala
210 215 220
Val Gly Ala Glu Arg Val Gly Leu Arg Leu Ser Pro Trp Gly Thr Ile
225 230 235 240
Asn Asp Met His Asp Asp Glu Pro Glu Ala Met Thr Leu Tyr Leu Ala
245 250 255
Glu Ala Leu Gln Arg Arg Gly Ile Ala Tyr Leu His Leu Ala Glu Trp
260 265 270
Glu Trp Ser Gly Gly Pro Ala Tyr Pro Gln Gly Phe Arg Glu Arg Leu
275 280 285
Arg Glu Arg Phe Arg Ala Pro Leu Ile Val Cys Gly Asn Tyr Asp Ala
290 295 300
Glu Arg Ala Glu Ala Ile Leu Gln Ala Gly Leu Ala Asp Ala Val Ala
305 310 315 320
Ile Gly Arg Pro Phe Ile Ala Asn Pro Asp Leu Val Glu Arg Ile Arg
325 330 335
Leu Gly Ala Pro Leu Ala Glu Ala Asn Gln Ala Arg Phe Tyr Gly Gly
340 345 350
Asp Ala Ala Gly Tyr Thr Asp Tyr Pro Thr Leu Gly Gln Ser Ala Thr
355 360 365
Ala
<210> 14
<211> 350
<212> PRT
<213> 铜绿假单胞菌 PAO1
<400> 14
Met Ala Thr Leu Phe Asp Pro Ile Val Leu Gly Asp Leu Glu Leu Pro
1 5 10 15
Asn Arg Ile Val Met Ala Pro Leu Thr Arg Cys Arg Ala Asp Glu Gly
20 25 30
Arg Val Pro Asn Ala Leu Met Ala Glu Tyr Tyr Ala Gln Arg Ala Asp
35 40 45
Ala Gly Leu Ile Leu Ser Glu Ala Thr Ala Val Thr Pro Met Gly Val
50 55 60
Gly Tyr Pro Asp Thr Pro Gly Ile Trp Ser Asp Asp Gln Val Arg Gly
65 70 75 80
Trp Ser Asn Val Thr Lys Ala Val His Ala Ala Gly Gly Arg Ile Phe
85 90 95
Leu Gln Leu Trp His Val Gly Arg Ile Ser Asp Pro Leu Tyr Leu Asn
100 105 110
Gly Glu Leu Pro Val Ala Pro Ser Ala Ile Ala Ala Glu Gly His Val
115 120 125
Ser Leu Val Arg Pro Lys Arg Pro Tyr Val Thr Pro Arg Ala Leu Asp
130 135 140
Thr Glu Glu Ile Ala Asp Ile Val Glu Ala Tyr Arg Gln Gly Ala Glu
145 150 155 160
Arg Ala Lys Ala Ala Gly Phe Asp Gly Val Glu Ile His Gly Ala Asn
165 170 175
Gly Tyr Leu Leu Asp Gln Phe Leu Gln Asp Ser Thr Asn Lys Arg Thr
180 185 190
Asp Arg Tyr Gly Gly Ser Ile Glu Asn Arg Ala Arg Leu Leu Leu Glu
195 200 205
Val Thr Asp Ala Ala Ile Ser Val Trp Gly Ala Gln Arg Val Gly Val
210 215 220
His Leu Ala Pro Arg Ala Asp Ser His Asp Met Gly Asp Ser Asn Arg
225 230 235 240
Leu Glu Thr Phe Ser His Val Ala Arg Glu Leu Gly Lys Arg Gly Ile
245 250 255
Ala Phe Ile Cys Ala Arg Glu Ala Gln Ala Asp Asp Ser Ile Gly Val
260 265 270
Ala Leu Lys Lys Ala Phe Gly Gly Pro Tyr Ile Ala Asn Glu Gln Phe
275 280 285
Thr Leu Asp Ser Ala Asn Ala Ile Leu Ala Lys Gly Asp Ala Asp Ala
290 295 300
Val Ala Phe Gly Val Pro Phe Ile Ala Asn Pro Asp Leu Val Glu Arg
305 310 315 320
Leu Arg Gln Gly Ala Glu Leu Asn Pro Pro Arg Pro Glu Thr Phe Tyr
325 330 335
Thr Gly Gly Thr Glu Gly Tyr Leu Asp Tyr Pro Thr Leu Ala
340 345 350
<210> 15
<211> 349
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 15
Met Thr Thr Leu Phe Asp Pro Ile Lys Leu Gly Asp Leu Gln Leu Pro
1 5 10 15
Asn Arg Ile Ile Met Ala Pro Leu Thr Arg Cys Arg Ala Asp Glu Gly
20 25 30
Arg Val Pro Asn Ala Leu Met Ala Glu Tyr Tyr Val Gln Arg Ala Ser
35 40 45
Ala Gly Leu Ile Leu Ser Glu Ala Thr Ser Val Ser Pro Met Gly Val
50 55 60
Gly Tyr Pro Asp Thr Pro Gly Ile Trp Asn Asp Glu Gln Val Arg Gly
65 70 75 80
Trp Asn Asn Val Thr Lys Ala Val His Ala Ala Gly Gly Arg Ile Phe
85 90 95
Leu Gln Leu Trp His Val Gly Arg Ile Ser His Pro Ser Tyr Leu Asn
100 105 110
Gly Glu Leu Pro Val Ala Pro Ser Ala Ile Gln Pro Lys Gly His Val
115 120 125
Ser Leu Val Arg Pro Leu Ser Asp Tyr Pro Thr Pro Arg Ala Leu Glu
130 135 140
Thr Glu Glu Ile Asn Asp Ile Val Glu Ala Tyr Arg Ser Gly Ala Glu
145 150 155 160
Asn Ala Lys Ala Ala Gly Phe Asp Gly Val Glu Ile His Gly Ala Asn
165 170 175
Gly Tyr Leu Leu Asp Gln Phe Leu Gln Ser Ser Thr Asn Gln Arg Thr
180 185 190
Asp Arg Tyr Gly Gly Ser Leu Glu Asn Arg Ala Arg Leu Leu Leu Glu
195 200 205
Val Thr Asp Ala Ala Ile Glu Val Trp Gly Ala Gln Arg Val Gly Val
210 215 220
His Leu Ala Pro Arg Ala Asp Ala His Asp Met Gly Asp Ala Asp Arg
225 230 235 240
Ala Glu Thr Phe Thr Tyr Val Ala Arg Glu Leu Gly Lys Arg Gly Ile
245 250 255
Ala Phe Ile Cys Ser Arg Glu Arg Glu Ala Asp Asp Ser Ile Gly Pro
260 265 270
Leu Ile Lys Glu Ala Phe Gly Gly Pro Tyr Ile Val Asn Glu Arg Phe
275 280 285
Asp Lys Ala Ser Ala Asn Ala Ala Leu Ala Ser Gly Lys Ala Asp Ala
290 295 300
Val Ala Phe Gly Val Pro Phe Ile Ala Asn Pro Asp Leu Pro Ala Arg
305 310 315 320
Leu Ala Ala Asp Ala Pro Leu Asn Glu Ala His Pro Glu Thr Phe Tyr
325 330 335
Gly Lys Gly Pro Val Gly Tyr Ile Asp Tyr Pro Arg Leu
340 345
<210> 16
<211> 363
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 16
Met Ser Ala Leu Phe Glu Pro Tyr Thr Leu Lys Asp Val Thr Leu Arg
1 5 10 15
Asn Arg Ile Ala Ile Pro Pro Met Cys Gln Tyr Met Ala Glu Asp Gly
20 25 30
Met Ile Asn Asp Trp His His Val His Leu Ala Gly Leu Ala Arg Gly
35 40 45
Gly Ala Gly Leu Leu Val Val Glu Ala Thr Ala Val Ala Pro Glu Gly
50 55 60
Arg Ile Thr Pro Gly Cys Ala Gly Ile Trp Ser Asp Ala His Ala Gln
65 70 75 80
Ala Phe Val Pro Val Val Gln Ala Ile Lys Ala Ala Gly Ser Val Pro
85 90 95
Gly Ile Gln Ile Ala His Ala Gly Arg Lys Ala Ser Ala Asn Arg Pro
100 105 110
Trp Glu Gly Asp Asp His Ile Ala Ala Asp Asp Ala Arg Gly Trp Glu
115 120 125
Thr Ile Ala Pro Ser Ala Ile Ala Phe Gly Ala His Leu Pro Lys Val
130 135 140
Pro Arg Glu Met Thr Leu Asp Asp Ile Ala Arg Val Lys Gln Asp Phe
145 150 155 160
Val Asp Ala Ala Arg Arg Ala Arg Asp Ala Gly Phe Glu Trp Ile Glu
165 170 175
Leu His Phe Ala His Gly Tyr Leu Gly Gln Ser Phe Phe Ser Glu His
180 185 190
Ser Asn Lys Arg Thr Asp Ala Tyr Gly Gly Ser Phe Asp Asn Arg Ser
195 200 205
Arg Phe Leu Leu Glu Thr Leu Ala Ala Val Arg Glu Val Trp Pro Glu
210 215 220
Asn Leu Pro Leu Thr Ala Arg Phe Gly Val Leu Glu Tyr Asp Gly Arg
225 230 235 240
Asp Glu Gln Thr Leu Glu Glu Ser Ile Glu Leu Ala Arg Arg Phe Lys
245 250 255
Ala Gly Gly Leu Asp Leu Leu Ser Val Ser Val Gly Phe Thr Ile Pro
260 265 270
Asp Thr Asn Ile Pro Trp Gly Pro Ala Phe Met Gly Pro Ile Ala Glu
275 280 285
Arg Val Arg Arg Glu Ala Lys Leu Pro Val Thr Ser Ala Trp Gly Phe
290 295 300
Gly Thr Pro Gln Leu Ala Glu Ala Ala Leu Gln Ala Asn Gln Leu Asp
305 310 315 320
Leu Val Ser Val Gly Arg Ala His Leu Ala Asp Pro His Trp Ala Tyr
325 330 335
Phe Ala Ala Lys Glu Leu Gly Val Glu Lys Ala Ser Trp Thr Leu Pro
340 345 350
Ala Pro Tyr Ala His Trp Leu Glu Arg Tyr Arg
355 360
<210> 17
<211> 368
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 17
Met Ser Leu Leu Leu Glu Pro Tyr Thr Leu Arg Gln Leu Thr Leu Pro
1 5 10 15
Asn Arg Ile Ala Val Ser Pro Met Cys Gln Tyr Ser Ala Ile Asp Gly
20 25 30
Leu Ala Asn Asp Trp His Leu Val His Leu Gly Cys Arg Ala Val Gly
35 40 45
Gly Ala Gly Leu Val Ile Ser Glu Ala Val Ala Val Thr Ala Asp Gly
50 55 60
Arg Ile Thr Ala Glu Asp Leu Gly Leu Trp Asn Asp Asp Gln Ile Val
65 70 75 80
Pro Leu Gln Arg Ile Thr Arg Phe Ile Thr Ala Gln Gly Ala Val Pro
85 90 95
Gly Ile Gln Leu Ala His Ala Gly Arg Lys Ala Ser Thr Tyr Arg Pro
100 105 110
Trp Leu Gly Lys Gln Gly Ser Val Lys Leu Glu Asp Gly Gly Trp Gln
115 120 125
Pro Val Gly Pro Ser Lys Ile Ala Phe Asp Pro Gln His Thr Pro Pro
130 135 140
Arg Glu Leu Ser Lys Asp Glu Ile Gln Asp Val Ile Asp Ala Phe Val
145 150 155 160
Ala Ser Thr Glu Arg Ala Leu Lys Ala Gly Phe Lys Val Val Glu Ile
165 170 175
His Ala Ala His Gly Tyr Leu Leu His Gln Phe Leu Ser Pro Leu Ser
180 185 190
Asn Gln Arg Arg Asp Glu Tyr Gly Ser Cys Phe Glu Asn Arg Ile Arg
195 200 205
Leu Thr Leu Gln Val Val Glu Ala Val Arg Lys Val Trp Pro Ala Glu
210 215 220
Leu Pro Leu Phe Val Arg Val Ser Ala Thr Asp Trp Val Glu Asp Gly
225 230 235 240
Trp Asn Pro Asp Glu Thr Val Glu Leu Ala Arg Arg Leu Arg Val Leu
245 250 255
Gly Val Asp Leu Ile Asp Val Ser Ser Gly Gly Thr Ser Val Asn Ala
260 265 270
Glu Ile Pro Thr Gly Pro Gly Tyr Gln Thr Arg Phe Ala Glu Arg Val
275 280 285
Arg Lys Glu Ala Glu Ile Ala Thr Gly Thr Val Gly Met Ile Thr Glu
290 295 300
Pro Ala Gln Ala Glu His Ile Leu Arg Thr Gly Gln Ala Asp Val Ile
305 310 315 320
Phe Leu Ala Arg Glu Leu Leu Arg Asp Pro Tyr Trp Pro Leu His Ala
325 330 335
Asp Asp Asp Leu Gly Gly Asn Lys Ala Thr Trp Pro Ala Gln Tyr Gln
340 345 350
Arg Ala Thr Ser Arg Ala Asn Pro Ile His Glu Ser Asp Leu Arg Asp
355 360 365
<210> 18
<211> 367
<212> PRT
<213> 丁香假单胞菌番茄致病变种菌株 DC3000
<400> 18
Met Ser Ala Ala Ala Leu Phe Thr Pro Phe Arg Leu Gly Gly Leu Glu
1 5 10 15
Leu Ser Ser Arg Val Val Met Ala Pro Met Thr Arg Ser Phe Ser Pro
20 25 30
Gly His Val Pro Asn Ser Lys Val Ile Glu Tyr Tyr Arg Arg Arg Ala
35 40 45
Ala Ala Gly Val Gly Leu Ile Ile Thr Glu Gly Thr Thr Val Asn His
50 55 60
Pro Ala Ser Asn Gly Tyr Pro Asn Val Pro Gln Phe Phe Gly Asp Ala
65 70 75 80
Pro Leu Ala Gly Trp Lys Lys Val Val Asp Ala Val His Ala Glu Gly
85 90 95
Gly Lys Ile Ala Pro Gln Leu Trp His Val Gly Ala Val Arg Arg Leu
100 105 110
Gly Thr Glu Pro Asp Gly Ser Ile Pro Ala Tyr Gly Pro Met Glu Lys
115 120 125
Val Lys Asp Gly Gln Val Leu Val His Gly Met Ser Lys Gln Asp Ile
130 135 140
Asp Glu Val Val Ala Ala Phe Ala Gln Ala Ala Ala Asp Ala Lys Ala
145 150 155 160
Ile Gly Met Asp Gly Val Glu Ile His Gly Ala His Gly Tyr Leu Val
165 170 175
Asp Gln Phe Phe Trp Glu Gly Ser Asn Gln Arg Ser Asp Glu Tyr Gly
180 185 190
Gly Ser Leu Ala Asn Arg Ser Arg Phe Ala Ile Glu Leu Ile Lys Ala
195 200 205
Val Arg Ala Ala Val Gly Pro Asp Tyr Pro Ile Ile Phe Arg Phe Ser
210 215 220
Gln Trp Lys Gln Gln Asp Tyr Ser Ala Arg Leu Val Gln Thr Pro Glu
225 230 235 240
Ala Leu Gly Glu Phe Val Gln Pro Leu Ser Asp Ala Gly Val Asp Ile
245 250 255
Phe His Cys Ser Thr Arg Arg Phe Trp Glu Pro Glu Phe Glu Gly Ser
260 265 270
Glu Leu Asn Leu Ala Gly Trp Thr Arg Lys Leu Thr Gly Lys Pro Thr
275 280 285
Ile Thr Val Gly Ser Val Gly Leu Asp Gly Glu Phe Leu Gln Phe Met
290 295 300
Val Asn Thr Glu Lys Val Ala Gln Pro Ala Ser Leu Glu Asn Leu Leu
305 310 315 320
Glu Arg Leu Gly Lys Asp Glu Phe Asp Leu Val Ala Val Gly Arg Ala
325 330 335
Leu Leu Val Asp Pro Asp Trp Ala Val Lys Val Arg Glu Gly Arg Glu
340 345 350
Gln Asp Ile Leu Pro Phe Ser Arg Glu Ala Leu Thr Thr Leu Val
355 360 365
<210> 19
<211> 363
<212> PRT
<213> 丁香假单胞菌番茄致病变种菌株 DC3000
<400> 19
Met Ser Ala Leu Phe Glu Pro Phe Lys Leu Lys Asp Val Thr Leu Arg
1 5 10 15
Asn Arg Ile Ala Ile Pro Pro Met Cys Gln Tyr Ser Ala Ile Asp Gly
20 25 30
Val Val Asn Asp Trp His His Val His Leu Ala Ser Met Ala Arg Gly
35 40 45
Gly Ala Gly Leu Leu Val Val Glu Ala Thr Ala Val Ala Pro Glu Gly
50 55 60
Arg Ile Thr Pro Gly Cys Thr Gly Ile Trp Asn Asp Glu Gln Ala Gln
65 70 75 80
Ala Phe Val Pro Ile Val Lys Ala Ile Lys Asp Ala Gly Cys Val Pro
85 90 95
Gly Ile Gln Ile Ala His Ala Gly Arg Lys Ala Ser Ala Asn Arg Pro
100 105 110
Trp Glu Gly Asp Asp His Ile Val Ala Ser Asp Ser Arg Gly Trp Asp
115 120 125
Thr Ile Ala Pro Ser Ala Ile Ala Phe Gly Ala Asn Leu Pro Lys Val
130 135 140
Pro Arg Ala Met Thr Leu Asp Asp Ile Ala Arg Val Arg Gln Asn Phe
145 150 155 160
Val Asp Ala Ala Arg Arg Ala Arg Asp Ala Gly Phe Glu Trp Ile Glu
165 170 175
Leu His Phe Ala His Gly Tyr Leu Gly Gln Ser Phe Phe Ser Glu His
180 185 190
Ser Asn Gln Arg Thr Asp Glu Tyr Gly Gly Ser Phe Asp Asn Arg Ser
195 200 205
Arg Phe Leu Leu Glu Thr Leu Ala Ala Val Arg Glu Val Trp Pro Glu
210 215 220
Asn Leu Pro Leu Thr Ala Arg Phe Gly Val Leu Glu Tyr Asp Gly Arg
225 230 235 240
Asp Glu Gln Thr Leu Thr Glu Ser Ile Glu Leu Ala Arg Arg Phe Lys
245 250 255
Ala Gly Gly Leu Asp Leu Leu Ser Val Ser Val Gly Phe Ser Thr Pro
260 265 270
Asp Ala Asn Ile Pro Trp Gly Pro Ala Phe Met Gly Pro Ile Ala Glu
275 280 285
Arg Val Arg Arg Glu Ala Ala Ile Pro Val Thr Ser Ala Trp Gly Phe
290 295 300
Gly Glu Pro Lys Leu Ala Glu Glu Ala Val Lys Ser Gly Gln Leu Asp
305 310 315 320
Leu Val Ser Ile Gly Arg Ala His Leu Ala Asp Pro His Trp Ala Tyr
325 330 335
Phe Ala Ala Lys Glu Leu Gly Val Glu Lys Ser Ala Trp Thr Leu Pro
340 345 350
Ala Pro Tyr Ala His Trp Leu Glu Arg Tyr Arg
355 360
<210> 20
<211> 349
<212> PRT
<213> 丁香假单胞菌番茄致病变种菌株 DC3000
<400> 20
Met Thr Thr Ile Phe Asp Pro Ile Thr Leu Gly Asp Leu Gln Leu Pro
1 5 10 15
Asn Arg Ile Ile Leu Ala Pro Leu Thr Arg Cys Arg Ala Asp Glu Gly
20 25 30
Arg Val Pro Asn Ala Leu Met Ser Glu Tyr Tyr Val Gln Arg Ala Ser
35 40 45
Ala Gly Leu Ile Leu Thr Glu Ala Thr Ser Val Thr Pro Met Gly Val
50 55 60
Gly Tyr Pro Asp Thr Pro Gly Ile Trp Ser Asn Asp Gln Val Arg Gly
65 70 75 80
Trp Ser Asn Ile Thr Gln Ala Val His Asn Ala Gly Gly Arg Ile Val
85 90 95
Leu Gln Leu Trp His Val Gly Arg Ile Ser His Pro Ser Tyr Leu Asn
100 105 110
Gly Glu Thr Pro Val Ala Pro Ser Ala Ile Ala Ala Gln Gly His Val
115 120 125
Ser Leu Met Arg Pro Ile Thr Pro Leu Pro Thr Pro Arg Ala Leu Glu
130 135 140
Leu Ala Glu Ile Gly Asp Ile Val Glu Ala Tyr Arg Val Gly Ala Glu
145 150 155 160
Asn Ala Lys Ala Ala Gly Phe Asp Gly Val Glu Val His Gly Ala Asn
165 170 175
Gly Tyr Leu Leu Glu Gln Phe Leu Leu Thr Gly Ser Asn Gln Arg Thr
180 185 190
Asp Gln Tyr Gly Gly Ser Leu Glu Asn Arg Ala Arg Leu Leu Leu Glu
195 200 205
Val Thr Asp Ala Val Ile Asp Val Trp Gly Ala Gly Arg Val Gly Val
210 215 220
His Leu Ser Pro Arg Phe Asp Met His Asn Met Ser Asp Glu Asn Arg
225 230 235 240
Ala Glu Thr Phe Ser Tyr Val Ala Lys Glu Leu Gly Lys Arg Gly Ile
245 250 255
Ala Phe Ile Cys Ala Arg Glu Arg Asp Ala Glu Asp Ser Leu Gly Pro
260 265 270
Gln Leu Lys Lys Asp Phe Gly Gly Val Tyr Ile Ala Asn Glu Lys Phe
275 280 285
Thr Lys Asp Ser Ala Asn Ala Trp Leu Ala Glu Gly Lys Ala Asp Ala
290 295 300
Phe Ala Phe Gly Val Pro Tyr Ile Ala Asn Pro Asp Leu Pro Glu Arg
305 310 315 320
Leu Ala Ser Asp Ala Pro Leu Asn Glu Ala His Pro Glu Thr Phe Tyr
325 330 335
Gly Lys Gly Pro Val Gly Tyr Ile Asp Tyr Pro Arg Leu
340 345
<210> 21
<211> 367
<212> PRT
<213> 丁香假单胞菌菜豆致病变种 1448A
<400> 21
Met Ser Val Glu Ala Leu Phe Thr Pro Phe Arg Leu Gly Ser Leu Glu
1 5 10 15
Leu Ser Ser Arg Val Val Met Ala Pro Met Thr Arg Ser Phe Ser Pro
20 25 30
Gly His Val Pro Asn Ser Lys Val Ile Glu Tyr Tyr Arg Arg Arg Ala
35 40 45
Ala Ala Gly Val Gly Leu Ile Val Thr Glu Gly Thr Thr Val Asn His
50 55 60
Lys Ala Ser Asn Gly Tyr Pro Asn Val Pro Gln Phe Phe Gly Asp Ala
65 70 75 80
Pro Leu Ala Gly Trp Arg Lys Val Val Glu Ala Val His Ala Glu Gly
85 90 95
Gly Lys Ile Val Pro Gln Leu Trp His Val Gly Ala Val Arg Arg Pro
100 105 110
Gly Thr Glu Pro Asp Gly Ser Val Pro Ala Tyr Gly Pro Met Glu Lys
115 120 125
Val Lys Asp Gly Gln Val Leu Val His Gly Met Ser Lys Gln Asp Ile
130 135 140
Asp Asp Val Val Ala Ala Phe Ala Gln Ala Ala Val Asp Ala Lys Ala
145 150 155 160
Ile Gly Met Asp Gly Val Glu Ile His Gly Ala His Gly Tyr Leu Ile
165 170 175
Asp Gln Phe Phe Trp Glu Gly Ser Asn Gln Arg Ser Asp Glu Tyr Gly
180 185 190
Gly Ser Leu Ala Lys Arg Ser Arg Phe Ala Ile Glu Leu Ile Lys Ala
195 200 205
Val Arg Val Ala Val Gly Ala Asp Tyr Pro Ile Ile Phe Arg Phe Ser
210 215 220
Gln Trp Lys Gln Gln Asp Tyr Thr Ala Arg Leu Val Gln Thr Pro Glu
225 230 235 240
Ala Leu Gly Glu Phe Leu Gln Pro Leu Ala Asp Ala Gly Val Asp Ile
245 250 255
Phe His Cys Ser Thr Arg Arg Phe Trp Glu Pro Glu Phe Glu Gly Ser
260 265 270
Asp Leu Asn Leu Ala Gly Trp Thr Arg Lys Leu Thr Gly Lys Pro Thr
275 280 285
Ile Thr Val Gly Ser Val Gly Leu Asp Gly Glu Phe Leu Gln Phe Met
290 295 300
Val Asn Thr Asp Lys Val Ala Gln Pro Ala Ser Leu Glu Asn Leu Leu
305 310 315 320
Lys Arg Leu Gly Asn Asp Glu Phe Asp Leu Val Ala Val Gly Arg Ala
325 330 335
Leu Leu Val Asp Pro Asp Trp Ala Leu Lys Val Arg Glu Gly Arg Glu
340 345 350
Gln Asp Ile Leu Pro Phe Ser Arg Glu Ala Leu Thr Thr Leu Val
355 360 365
<210> 22
<211> 368
<212> PRT
<213> 丁香假单胞菌菜豆 1448A
<400> 22
Met Ser Leu Leu Leu Glu Pro Tyr Thr Leu Arg Gln Leu Thr Leu Arg
1 5 10 15
Asn Arg Ile Ala Val Ser Pro Met Cys Gln Tyr Ser Ser Val Asp Gly
20 25 30
Leu Ala Asn Asp Trp His Leu Val His Leu Gly Ser Arg Ala Val Gly
35 40 45
Gly Ala Gly Leu Val Ile Thr Glu Ala Met Ala Val Thr Pro Asp Gly
50 55 60
Arg Ile Thr Pro Glu Asp Leu Gly Leu Trp Asn Asp Glu Gln Ile Glu
65 70 75 80
Pro Leu Gln Arg Ile Thr Arg Phe Ile Asn Ala Gln Gly Ala Val Ala
85 90 95
Gly Ile Gln Leu Ala His Ala Gly Arg Lys Ala Ser Thr Trp Arg Pro
100 105 110
Trp Leu Gly Lys His Gly Ser Ile Ser Leu Asn Glu Gly Gly Trp Thr
115 120 125
Pro Val Gly Pro Ser Ala Ile Ala Phe Asp Pro Gln His Thr Ala Pro
130 135 140
Val Gln Leu Ser Glu Thr Gln Ile Gln Glu Leu Ile Lys Ala Phe Val
145 150 155 160
Asp Ser Ala Arg Arg Ala Leu Thr Ala Gly Phe Lys Val Val Glu Ile
165 170 175
His Ala Ala His Gly Tyr Leu Leu His Gln Phe Leu Ser Pro Leu Ser
180 185 190
Asn Gln Arg Thr Asp Gln Tyr Gly Gly Ser Phe Glu Asn Arg Ile Arg
195 200 205
Leu Thr Leu Gln Val Thr Glu Ala Val Arg Ala Val Trp Pro Glu Glu
210 215 220
Leu Pro Leu Phe Val Arg Val Ser Ala Thr Asp Trp Val Glu Asp Gly
225 230 235 240
Trp Asn Ala Asp Glu Thr Val Glu Leu Ala Arg Arg Leu Lys Ala Leu
245 250 255
Gly Thr Asp Leu Ile Asp Val Ser Ser Gly Gly Thr Ser Ala Asn Ala
260 265 270
Glu Ile Pro Val Gly Pro Gly Tyr Gln Thr Arg Phe Ala Glu Arg Val
275 280 285
Arg Lys Glu Ser Glu Ile Ala Thr Gly Thr Val Gly Met Ile Thr Asp
290 295 300
Pro Ala Gln Ala Glu His Ile Leu Arg Thr Gly Gln Ala Asp Ile Ile
305 310 315 320
Leu Leu Ala Arg Glu Leu Leu Arg Asp Pro Tyr Trp Pro Leu Arg Ala
325 330 335
Asp Glu Asp Leu Gly Gly Arg Leu Ala Thr Trp Pro Ala Gln Tyr Gln
340 345 350
Arg Ala Thr His Arg Asp Gln Pro Ile His Glu Ser Asp Leu Arg Asp
355 360 365
<210> 23
<211> 349
<212> PRT
<213> 丁香假单胞菌菜豆 1448A
<400> 23
Met Thr Thr Ile Phe Asp Pro Ile Thr Ile Gly Asp Leu Gln Leu Pro
1 5 10 15
Asn Arg Ile Ile Met Ala Pro Leu Thr Arg Cys Arg Ala Asp Glu Gly
20 25 30
Arg Val Pro Asn Ala Leu Met Ser Glu Tyr Tyr Val Gln Arg Ala Ser
35 40 45
Ala Gly Leu Ile Leu Thr Glu Ala Thr Ser Val Thr Pro Met Gly Val
50 55 60
Gly Tyr Pro Asp Thr Pro Gly Ile Trp Ser Asn Asp Gln Val Arg Gly
65 70 75 80
Trp Ser Asn Ile Thr Lys Ala Val His Thr Ala Gly Gly Arg Ile Val
85 90 95
Leu Gln Leu Trp His Val Gly Arg Ile Ser His Pro Ala Tyr Leu Asn
100 105 110
Gly Glu Thr Pro Val Ala Pro Ser Ala Ile Ala Ala Glu Gly His Val
115 120 125
Ser Leu Met Arg Pro Ile Thr Pro Leu Pro Thr Pro Arg Ala Leu Glu
130 135 140
Leu Ala Glu Ile Gly Asp Ile Val Glu Ala Tyr Arg Val Gly Ala Glu
145 150 155 160
Asn Ala Lys Ala Ala Gly Phe Asp Gly Val Glu Val His Gly Ala Asn
165 170 175
Gly Tyr Leu Leu Glu Gln Phe Leu Leu Thr Gly Ser Asn Gln Arg Thr
180 185 190
Asp Glu Tyr Gly Gly Ser Val Glu Asn Arg Ala Arg Leu Leu Leu Glu
195 200 205
Val Thr Asp Ala Val Ile Glu Val Trp Gly Ala Gly Arg Val Gly Val
210 215 220
His Leu Ser Pro Arg Phe Asp Met His Asp Met Ser Asp Ala Asn Arg
225 230 235 240
Thr Glu Thr Phe Ser Tyr Val Ala Lys Glu Leu Gly Lys Arg Gly Ile
245 250 255
Ala Phe Ile Cys Ala Arg Glu Arg Asp Ala Glu Asp Ser Leu Gly Pro
260 265 270
Gln Leu Lys Lys Asp Phe Gly Gly Val Tyr Ile Ala Asn Glu Lys Phe
275 280 285
Thr Lys Asp Thr Ala Asn Thr Trp Leu Ala Glu Gly Lys Ala Asp Ala
290 295 300
Ile Ala Phe Gly Val Pro Tyr Ile Ala Asn Pro Asp Leu Pro Glu Arg
305 310 315 320
Leu Ala Ser Asp Ala Pro Leu Asn Glu Ala His Pro Glu Thr Phe Tyr
325 330 335
Gly Lys Gly Pro Val Gly Tyr Ile Asp Tyr Pro Arg Leu
340 345
<210> 24
<211> 370
<212> PRT
<213> 红球菌属物种 RHA1
<400> 24
Met Gln His Thr Ser Thr Thr Ala Ser Thr Leu Phe Ser Pro Leu Lys
1 5 10 15
Leu Gly Ser Thr Glu Leu Ala Asn Arg Val Ala Leu Ala Pro Met Thr
20 25 30
Arg Val Ser Gly Asp Glu Asp Gly Ser Ala Asn Glu Ser Ile Ala Ser
35 40 45
Tyr Tyr Glu Val Phe Ala Lys Gly Gly Phe Gly Leu Leu Ile Thr Glu
50 55 60
Gly Ile Tyr Pro Asp Thr Lys Tyr Ser Gln Gly Tyr Leu Asn Gln Pro
65 70 75 80
Gly Leu Ala Thr Ala Gln His Ala Asp Ser Trp Lys Pro Val Val Glu
85 90 95
Arg Val His Thr Ala Gly Ala Arg Ile Phe Ala Gln Leu Met His Ala
100 105 110
Gly Ala Gln Ser Gln Gly Asn Arg Tyr Val Thr Glu Thr Ile Ala Pro
115 120 125
Ser Thr Thr Pro Pro Leu Gly Glu Gln Leu Gly Phe Tyr Gly Gly Thr
130 135 140
Gly Pro Tyr Ala Thr Pro Arg Ala Leu Ser Leu Arg Asp Ile Asp Asp
145 150 155 160
Val Cys Ala Gly Phe Ala Gln Ala Ala Gln His Ala Val Ala Ala Gly
165 170 175
Phe Asp Gly Val Glu Ile His Gly Ala Asn Gly Tyr Leu Leu Asp Gln
180 185 190
Phe Leu Thr Asp Tyr Met Asn Gln Arg Asp Asp Glu Tyr Gly Gly Ser
195 200 205
Leu Ala Asn Arg Leu Lys Ile Tyr Arg Glu Val Ile Ala Ala Val Arg
210 215 220
Gln Val Val Gly Ser Lys Val Thr Val Gly Val Arg Ile Ser Gln Ser
225 230 235 240
Lys Val Ser Asp Tyr Asn His Arg Trp Ala Gly Gly Glu Ser Asp Ala
245 250 255
Ala Glu Ile Phe Gln Gly Leu Ala Glu Thr Gly Val Asp Tyr Ile His
260 265 270
Thr Thr Glu Phe Asp Ala Thr Ala Pro Ala Phe Ala Asp Ser Gly Thr
275 280 285
Thr Leu Ala Ala Phe Ala Thr Lys Tyr Ser Gly Leu Pro Val Ile Ala
290 295 300
Asn Gly Gln Leu Gly Asp Pro Ala Ala Ala Asp Ala Leu Ile Glu Asn
305 310 315 320
Gly Ser Ala Gln Val Ile Ser Leu Gly Lys Pro Ala Leu Ala Asn Arg
325 330 335
Asp Trp Pro Gln Arg Ala Leu Asp Ala Lys Pro Ile Gln Ser Asp Val
340 345 350
Asp Pro Ser Val Phe Ser Pro Met Ala Thr Val Lys Ser Trp Glu Leu
355 360 365
Ser Ala
370
<210> 25
<211> 408
<212> PRT
<213> 红球菌属物种 RHA1
<400> 25
Met Ser Arg Gln Ala Lys Trp Glu Glu Leu Ser Ser Pro Ile Glu Met
1 5 10 15
Arg Gly Val Lys Phe Lys Asn Arg Met Met Arg Ser Gly Met Tyr Glu
20 25 30
Gly Met Ala Thr Glu Arg Gly Glu Val Thr Pro Glu Leu Glu Thr Trp
35 40 45
Ile Ala Arg Gln Ala Ala Gly Gly Ala Ala Cys Ile Phe Pro Gly Tyr
50 55 60
Thr Asn Val Cys Leu Pro Arg Cys Met Pro Tyr Gln Thr Gly Ser Tyr
65 70 75 80
Ser Asp Ser His Ile Pro Gly Leu Thr Lys Leu Ala Glu Ile Ile His
85 90 95
Ser Tyr Gly Ala Val Ala Gly Leu Gln Leu Ala Ala Ala Gly Arg Gln
100 105 110
Ala Asn Pro Asn Leu Phe Phe Ser Asp Ala Glu Ala Val Met Gly Pro
115 120 125
Ser Ala Met Gly Pro Ser Pro Leu Tyr Gln Ala Ala Cys Arg Glu Met
130 135 140
Thr Glu Ala Glu Ile Trp Gln Ala Val Glu Asp Gln Ala Ala Ala Ala
145 150 155 160
Ala Arg Ala Lys Glu Ala Gly Phe Asp Ile Val Leu Pro Asn Ala Ala
165 170 175
His Gly Tyr Met Leu Ala Gln Phe Leu Ser Pro His Thr Asn Lys Arg
180 185 190
Thr Asp Ala Trp Gly Gly Thr Pro Glu Lys Arg Phe Lys Phe Phe Glu
195 200 205
Glu Thr Phe Arg Ala Val Arg Glu Ala Val Gly Pro Glu Met Leu Val
210 215 220
Trp Thr Lys Ile Ser Val Asp Glu Pro Trp Glu Asp Gly Ile Asn Leu
225 230 235 240
Asp Glu Ala Arg Ile Phe Ile Pro Glu Val Ala Lys Leu Val Asp Ala
245 250 255
Ile Glu Ile Ser Gly Gly Thr Ile Val Asp Asn Val Phe Met Met Thr
260 265 270
Arg Gly Glu Ile Pro Ile Gly Thr Leu Arg Lys Gly Ile Gly Gly Pro
275 280 285
Val Glu Gly Ile Ser Ala Glu Leu Val Glu Gly Leu Tyr Gly Ile Arg
290 295 300
Asp Ser Val Lys Phe Glu Glu Thr Tyr Trp Tyr Asp His Ala Leu Ala
305 310 315 320
Leu Lys Pro Leu Cys Gly Asp Thr Pro Leu Ile Leu Thr Gly Gly His
325 330 335
Lys Tyr Pro Ala Thr Met Asn Gln Ile Ile Lys Glu Gly Thr Ala Asp
340 345 350
Met Leu Ser Leu Ala Arg Val Leu Ala Ala Glu Pro Asn Phe Pro Asn
355 360 365
Glu Val Val Ala Gly Arg Glu Asp Pro Gly Lys Cys Ser Asn Cys Asn
370 375 380
Lys Cys Leu Val Glu Val Val Leu Gly Asn Lys Leu Arg Cys Tyr Leu
385 390 395 400
Pro Asp Pro Gly Tyr Thr Tyr Ala
405
<210> 26
<211> 375
<212> PRT
<213> 金黄色葡萄球菌亚种金色 Mu50
<400> 26
Met Lys Ser Lys Tyr Glu Pro Leu Phe Asp Lys Val Glu Leu Pro Asn
1 5 10 15
Gly Val Glu Leu Arg Asn Arg Phe Val Leu Ala Pro Leu Thr His Ile
20 25 30
Ser Ser Asn Asp Asp Gly Thr Ile Ser Asp Val Glu Leu Pro Tyr Ile
35 40 45
Glu Lys Arg Ser Gln Asp Val Gly Ile Thr Ile Asn Ala Ala Ser Asn
50 55 60
Val Ser Asp Val Gly Lys Ala Phe Pro Gly Gln Pro Ser Ile Ala His
65 70 75 80
Asp Ser Asp Ile Glu Gly Leu Lys Arg Leu Ala Thr Ala Met Lys Lys
85 90 95
Asn Gly Ala Lys Ala Leu Val Gln Ile His His Gly Gly Ala Gln Ala
100 105 110
Leu Pro Glu Leu Ile Pro Asp Gly Asp Val Val Ala Pro Ser Pro Ile
115 120 125
Ser Leu Lys Ser Phe Gly Gln Lys Gln Glu His Ser Ala Arg Glu Met
130 135 140
Thr Asn Glu Glu Ile Glu Gln Ala Ile Lys Asp Phe Gly Glu Ala Thr
145 150 155 160
Arg Arg Ala Ile Glu Ala Gly Phe Asp Gly Val Glu Ile His Gly Ala
165 170 175
Asn His Tyr Leu Ile His Gln Phe Val Ser Pro Tyr Tyr Asn Arg Arg
180 185 190
Asn Asp Val Trp Ala Asn Gln Tyr Lys Phe Pro Val Ala Val Ile Glu
195 200 205
Glu Val Leu Lys Ala Lys Glu Ala Tyr Gly Asn Lys Asp Phe Ile Val
210 215 220
Gly Tyr Arg Leu Ser Pro Glu Glu Ala Glu Ser Pro Gly Ile Thr Met
225 230 235 240
Glu Ile Thr Glu Glu Leu Val Asn Lys Ile Ser His Met Pro Ile Asp
245 250 255
Tyr Ile His Val Ser Met Met Asp Thr His Ala Thr Thr Arg Glu Gly
260 265 270
Lys Tyr Ala Gly Gln Glu Arg Leu Pro Leu Ile His Lys Trp Ile Asn
275 280 285
Gly Arg Met Pro Leu Ile Gly Ile Gly Ser Ile Phe Thr Ala Asp Glu
290 295 300
Ala Leu Asp Ala Val Glu Asn Val Gly Val Asp Leu Val Ala Ile Gly
305 310 315 320
Arg Glu Leu Leu Leu Asp Tyr Gln Phe Val Glu Lys Ile Lys Asp Gly
325 330 335
Arg Glu Asp Glu Ile Ile Asn Tyr Phe Asp Pro Glu Arg Glu Asp Asn
340 345 350
His His Leu Thr Pro Asn Leu Trp His Gln Phe Asn Glu Gly Phe Tyr
355 360 365
Pro Leu Pro Arg Lys Asp Lys
370 375
<210> 27
<211> 359
<212> PRT
<213> 阿维链霉菌 MA-4680
<400> 27
Met Ser Ala Leu Phe Glu Pro Tyr Ser Leu Arg Asp Leu Thr Ile Pro
1 5 10 15
Asn Arg Val Trp Met Ala Pro Met Cys Gln Tyr Ser Ala Ala Pro Gln
20 25 30
Gly Pro Ala Ala Gly Ala Pro His Asp Trp His Phe Ala His Tyr Ala
35 40 45
Ala Arg Ala Thr Gly Gly Thr Gly Leu Ile Val Val Glu Ala Thr Ala
50 55 60
Val Ser Pro Glu Gly Arg Ile Ser Pro Tyr Asp Leu Gly Ile Trp Asn
65 70 75 80
Asp Thr Gln Val Glu Ala Phe Arg Arg Ile Thr Arg Phe Leu Ala Ala
85 90 95
Gln Gly Thr Val Pro Ala Ile Gln Leu Ala His Gly Gly Arg Lys Ala
100 105 110
Ser Thr Asp Arg Pro Trp Lys Gly Gly Ala Pro Leu Ala Ala Asp Glu
115 120 125
His Gly Trp Gln Pro Leu Gly Pro Ser Pro Val Ala Phe Asp Glu Arg
130 135 140
His Pro Val Pro Thr Glu Leu Ser Val Ala Glu Ile Arg Gln Ile Val
145 150 155 160
Gly Gln Phe Ala Asp Ala Ala Arg Arg Ala Leu Ala Ala Gly Phe Glu
165 170 175
Ile Ala Glu Ile His Gly Ala His Gly Tyr Leu Ile Asn Glu Phe Leu
180 185 190
Ser Pro His Ser Asn Arg Arg Thr Asp Glu Tyr Gly Gly Ser Tyr Glu
195 200 205
Asn Arg Val Arg Phe Ala Leu Glu Val Val Asp Ala Val Arg Glu Val
210 215 220
Trp Pro Ala Gly Lys Pro Leu Phe Phe Arg Val Ser Ala Thr Asp Trp
225 230 235 240
Leu Asp Glu Gly Gly Trp Thr Ala Asp Asp Thr Val Arg Phe Ala Ala
245 250 255
Glu Leu Gln Ala His Gly Val Asp Leu Leu Asp Val Ser Thr Gly Gly
260 265 270
Asn Ala Ser Gly Val Arg Ile Pro Thr Gly Pro Gly Tyr Gln Val Pro
275 280 285
Phe Ala Ala Arg Val Lys Ala Gly Thr Ser Leu Pro Val Ala Ala Val
290 295 300
Gly Leu Ile Thr Asp Ala Glu Gln Ala Glu Lys Ile Leu Ala Asn Gly
305 310 315 320
Glu Ala Asp Ala Ile Leu Leu Gly Arg Glu Leu Leu Arg Asn Pro Ser
325 330 335
Phe Ala Arg Leu Ala Ala Arg Glu Leu Gly Gly Asp Val His Val Pro
340 345 350
Ala Gln Tyr His Arg Ser Val
355
<210> 28
<211> 363
<212> PRT
<213> 苜蓿中华根瘤菌 1021
<400> 28
Met Glu Ile Pro Met Asn Pro Met Phe Thr Ser Val Arg Val Gly Arg
1 5 10 15
Tyr Met Leu Pro Asn Arg Leu Val Met Ala Pro Met Thr Arg Ser Arg
20 25 30
Ala Ala Phe Asp Gly Thr Pro Gly Glu Leu Ala Thr Glu Tyr Tyr Val
35 40 45
Gln Arg Ala Arg Leu Gly Leu Ile Val Thr Glu Gly Thr Gln Pro Ser
50 55 60
Asp Asp Gly Gln Gly Tyr Leu Thr Thr Pro Gly Ile Tyr Thr Pro Ala
65 70 75 80
His Ile Ala Gly Trp Arg Lys Ile Thr Ser Ala Val His Asp Lys Gly
85 90 95
Gly His Ile Phe Ile Gln Leu Met His Ala Gly Arg Met Ser His Pro
100 105 110
Asp Asn Thr Pro His His Arg Gln Gly Val Ala Pro Ser Ala Ile Ala
115 120 125
Pro Gly Ala Gly Met Phe Thr Ala Thr Gly Met Gln Asp Ile Pro Thr
130 135 140
Pro Arg Ala Leu Thr Thr Glu Glu Val Arg Arg Thr Val Ala Glu Phe
145 150 155 160
Ser His Ala Ala Arg Ser Ala Ile Glu Ala Gly Ala Asp Gly Ile Glu
165 170 175
Ile His Gly Ala Asn Ala Tyr Leu Val Gln Gln Phe Phe Ala Pro Ser
180 185 190
Ala Asn Thr Arg Thr Asp Glu Tyr Gly Gly Ser Ile Glu Asn Arg Ala
195 200 205
Arg Phe Ala Ile Glu Val Ala Thr Ala Ile Ala Glu Glu Val Gly Ala
210 215 220
Asp Arg Thr Ala Ile Arg Leu Ser Pro Gly Thr Ala Leu Trp Gly Ile
225 230 235 240
Asp Glu Gly Ala Glu Gly Pro Asp Leu Tyr Arg Tyr Leu Val Ala Glu
245 250 255
Leu Asp Lys Leu Gly Leu Ala Tyr Val His Ile Leu His Gln Ser Asn
260 265 270
Glu Pro Leu Leu Ala Asp Ile Arg Lys Leu Trp Arg Gln Pro Leu Ile
275 280 285
Leu Asn Arg Pro Gly Arg Pro Arg Gly Gln Ile Gly Ala Asp Val Ala
290 295 300
Ser Gly Leu Ala Asp Leu Glu Ala Phe Gly Gln Met Val Leu Ala Asn
305 310 315 320
Pro Asp Phe Val Ala Arg Leu Lys Ala Asp Ala Ala Met Asn Glu Ser
325 330 335
Asp Pro Lys Thr Phe Tyr Gly Gly Ala Ala Lys Gly Tyr Ile Asp Tyr
340 345 350
Pro Val Leu Ser Ala Arg Thr Asp Thr Ser Ser
355 360
<210> 29
<211> 378
<212> PRT
<213> 奥奈达希瓦氏菌 MR-1
<400> 29
Met Ser Val Phe Thr Ala Tyr Glu Ser Gly Ala Leu Thr Leu Gln Asn
1 5 10 15
Arg Ile Val Met Ala Pro Met Thr Arg Ala Arg Thr Thr Gln Pro Gly
20 25 30
Asn Ile Pro Asn Asp Leu Met Val Gln Tyr Tyr Ala Gln Arg Ser Ser
35 40 45
Ala Gly Leu Ile Ile Thr Glu Ala Thr Gln Ile Ser Asn Asp Ser Gln
50 55 60
Gly Tyr Ser Phe Thr Pro Gly Val Tyr Thr Glu Ala Gln Ile Asp Gly
65 70 75 80
Trp Lys Lys Val Thr Ala Ala Val His Glu Ala Gly Gly Lys Ile Phe
85 90 95
Asn Gln Ile Trp His Val Gly Arg Val Ser His Pro Ile Phe Gln Gln
100 105 110
Gly Asn Ala Pro Ile Ala Pro Ser Ala Ile Ala Pro Val Gly Thr Lys
115 120 125
Val Trp Ile Val Asp Glu Ala His Pro Glu Gly Gln Met Val Asp Cys
130 135 140
Pro Glu Pro Arg Glu Met Thr Gln Ala Asp Ile Asp Arg Val Val Ala
145 150 155 160
Asp Phe Ala Lys Ala Gly Ala Asn Ala Ile Ala Ala Gly Phe Asp Gly
165 170 175
Ile Glu Ile His Gly Gly Asn Gly Tyr Leu Ile Asp Gln Phe Leu Arg
180 185 190
Thr Asn Ser Asn His Arg Thr Asp Ala Tyr Gly Gly Ser Pro Glu Lys
195 200 205
Arg Ile Arg Phe Leu Leu Glu Val Val Glu Ala Val Ser Ala Gln Ile
210 215 220
Gly Ala Asp Lys Val Gly Val Arg Leu Ala Pro Tyr Val Thr Phe Lys
225 230 235 240
Asp Met Ala Cys Pro Glu Ile Val Glu Thr Ile Leu Leu Ala Ala Lys
245 250 255
Gln Leu Ser Ala Phe Gly Val Ala Tyr Leu His Leu Ser Glu Ala Asp
260 265 270
Trp Asp Asp Ala Pro Gln Ile Pro Glu Ser Phe Arg Ile Glu Leu Arg
275 280 285
Asn Val Phe Lys Gly Ser Ile Ile Val Ala Gly Arg Tyr Asp Val Glu
290 295 300
Arg Ala Asn Asp Val Ile Glu Lys Gly Tyr Ala Asp Leu Val Ala Phe
305 310 315 320
Gly Arg Ala Phe Ile Ala Asn Pro Asp Leu Pro Tyr Arg Leu Ala Asn
325 330 335
Gln Leu Pro Leu Ser Pro Phe Asp Lys Gly Pro Leu Phe Gly Gly Ser
340 345 350
Ala Ala Gly Tyr Thr Asp Tyr Pro Ser Tyr Gln Ala Ala Leu Arg Ala
355 360 365
Val Ile Arg Ser Ser Asp Asp Glu Val Ala
370 375
<210> 30
<211> 365
<212> PRT
<213> 野油菜黄单胞菌野油菜致病变种菌株 ATCC 33913
<400> 30
Met Ser Gln Leu Phe Thr Pro Ile Asn Phe Gly Pro Leu Ala Leu Ala
1 5 10 15
Asn Arg Ile Val Ile Ala Pro Met Cys Gln Tyr Ser Ala Gln Asp Gly
20 25 30
Arg Ala Thr Asp Trp His Arg Ile His Leu Gly Thr Leu Ser Gln Ser
35 40 45
Gly Ala Ala Leu Leu Ile Leu Glu Ala Ala Ala Val Gln Pro Gln Gly
50 55 60
Arg Ile Ser Tyr Ala Asp Leu Gly Leu Tyr Asp Asp Ala Thr Glu Gln
65 70 75 80
Ala Leu Gly Glu Val Leu Ala Ser Val Arg Arg Trp Ser Pro Met Pro
85 90 95
Ile Gly Ile Gln Leu Ala His Ala Gly Arg Lys Ala Ser Thr Asp Leu
100 105 110
Pro Trp Lys Gly Gly Glu Ala Ile Ala Pro Asp His Ala Asn Gly Trp
115 120 125
Gln Thr Val Ser Ala Ser Ala Leu Pro Phe Arg Asp Gly Gln Pro Leu
130 135 140
Pro Gln Ala Leu Asp Ala Ala Gly Ile Asp Ala Val Val Glu Ala Phe
145 150 155 160
Val Thr Ala Ala Lys Arg Ala Glu Arg Leu Glu Leu Gln Leu Ile Glu
165 170 175
Leu His Ala Ala His Gly Tyr Leu Met His Gln Phe Leu Ser Pro Leu
180 185 190
Ser Asn Gln Arg Ser Asp Ala Tyr Gly Gly Ser Leu Asp Asn Arg Met
195 200 205
Arg Leu Thr Leu Arg Ile Phe Asp Ala Val Arg Ala Ala Val Ser Asp
210 215 220
Gln Met Ala Ile Gly Val Arg Ile Ser Ala Thr Asp Trp Val Asp Gly
225 230 235 240
Gly Trp Asp Leu Pro Gln Ser Ile Ala Leu Ser Lys Ala Leu Asp Ala
245 250 255
Arg Gly Ala His Tyr Ile His Val Ser Ser Gly Gly Leu Asp Pro Arg
260 265 270
Gln Gln Ile Pro Val Gln Ala Ser Tyr Gln Ile Pro Phe Ala Glu Ala
275 280 285
Ile Arg Ala Gln Val Asp Thr Pro Val Ile Gly Val Gly Leu Ile Thr
290 295 300
Glu Pro Gln Gln Ala Glu Ala Ile Leu Arg Asp Gly Gln Ala Asp Ala
305 310 315 320
Ile Ala Leu Ala Arg Gly Ile Leu Tyr Asp Pro Arg Trp Pro Trp His
325 330 335
Ala Ala Ala Ala Leu Gly Ala Ser Val Thr Pro Ala Pro Gln Tyr Gln
340 345 350
Arg Cys Glu Pro Arg Glu Ala Arg Gly Val Phe Ala Ser
355 360 365
<210> 31
<211> 400
<212> PRT
<213> 酿酒酵母
<400> 31
Met Pro Phe Val Lys Asp Phe Lys Pro Gln Ala Leu Gly Asp Thr Asn
1 5 10 15
Leu Phe Lys Pro Ile Lys Ile Gly Asn Asn Glu Leu Leu His Arg Ala
20 25 30
Val Ile Pro Pro Leu Thr Arg Met Arg Ala Gln His Pro Gly Asn Ile
35 40 45
Pro Asn Arg Asp Trp Ala Val Glu Tyr Tyr Ala Gln Arg Ala Gln Arg
50 55 60
Pro Gly Thr Leu Ile Ile Thr Glu Gly Thr Phe Pro Ser Pro Gln Ser
65 70 75 80
Gly Gly Tyr Asp Asn Ala Pro Gly Ile Trp Ser Glu Glu Gln Ile Lys
85 90 95
Glu Trp Thr Lys Ile Phe Lys Ala Ile His Glu Asn Lys Ser Phe Ala
100 105 110
Trp Val Gln Leu Trp Val Leu Gly Trp Ala Ala Phe Pro Asp Thr Leu
115 120 125
Ala Arg Asp Gly Leu Arg Tyr Asp Ser Ala Ser Asp Asn Val Tyr Met
130 135 140
Asn Ala Glu Gln Glu Glu Lys Ala Lys Lys Ala Asn Asn Pro Gln His
145 150 155 160
Ser Ile Thr Lys Asp Glu Ile Lys Gln Tyr Val Lys Glu Tyr Val Gln
165 170 175
Ala Ala Lys Asn Ser Ile Ala Ala Gly Ala Asp Gly Val Glu Ile His
180 185 190
Ser Ala Asn Gly Tyr Leu Leu Asn Gln Phe Leu Asp Pro His Ser Asn
195 200 205
Asn Arg Thr Asp Glu Tyr Gly Gly Ser Ile Glu Asn Arg Ala Arg Phe
210 215 220
Thr Leu Glu Val Val Asp Ala Val Val Asp Ala Ile Gly Pro Glu Lys
225 230 235 240
Val Gly Leu Arg Leu Ser Pro Tyr Gly Val Phe Asn Ser Met Ser Gly
245 250 255
Gly Ala Glu Thr Gly Ile Val Ala Gln Tyr Ala Tyr Val Leu Gly Glu
260 265 270
Leu Glu Arg Arg Ala Lys Ala Gly Lys Arg Leu Ala Phe Val His Leu
275 280 285
Val Glu Pro Arg Val Thr Asn Pro Phe Leu Thr Glu Gly Glu Gly Glu
290 295 300
Tyr Asn Gly Gly Ser Asn Lys Phe Ala Tyr Ser Ile Trp Lys Gly Pro
305 310 315 320
Ile Ile Arg Ala Gly Asn Phe Ala Leu His Pro Glu Val Val Arg Glu
325 330 335
Glu Val Lys Asp Pro Arg Thr Leu Ile Gly Tyr Gly Arg Phe Phe Ile
340 345 350
Ser Asn Pro Asp Leu Val Asp Arg Leu Glu Lys Gly Leu Pro Leu Asn
355 360 365
Lys Tyr Asp Arg Asp Thr Phe Tyr Lys Met Ser Ala Glu Gly Tyr Ile
370 375 380
Asp Tyr Pro Thr Tyr Glu Glu Ala Leu Lys Leu Gly Trp Asp Lys Asn
385 390 395 400
<210> 32
<211> 660
<212> PRT
<213> 大肠杆菌 BL21-DE3
<400> 32
Met Lys Thr Val Val Phe Ala Tyr His Asp Met Gly Cys Leu Gly Ile
1 5 10 15
Glu Ala Leu Leu Ala Ala Gly Tyr Glu Ile Ser Ala Ile Phe Thr His
20 25 30
Thr Asp Asn Pro Gly Glu Lys Ala Phe Tyr Gly Ser Val Ala Arg Leu
35 40 45
Ala Ala Glu Arg Gly Ile Pro Val Tyr Ala Pro Asp Asn Val Asn His
50 55 60
Pro Leu Trp Val Glu Arg Ile Ala Gln Leu Ser Pro Asp Val Ile Phe
65 70 75 80
Ser Phe Tyr Tyr Arg His Leu Ile Tyr Asp Glu Ile Leu Gln Leu Ala
85 90 95
Pro Ala Gly Ala Phe Asn Leu His Gly Ser Leu Leu Pro Lys Tyr Arg
100 105 110
Gly Arg Ala Pro Leu Asn Trp Val Leu Val Asn Gly Glu Thr Glu Thr
115 120 125
Gly Val Thr Leu His Arg Met Val Lys Arg Ala Asp Ala Gly Ala Ile
130 135 140
Val Ala Gln Leu Arg Ile Ala Ile Ala Pro Asp Asp Ile Ala Ile Thr
145 150 155 160
Leu His His Lys Leu Cys His Ala Ala Arg Gln Leu Leu Glu Gln Thr
165 170 175
Leu Pro Ala Ile Lys His Gly Asn Ile Leu Glu Ile Ala Gln Arg Glu
180 185 190
Asn Glu Ala Thr Cys Phe Gly Arg Arg Thr Pro Asp Asp Ser Phe Leu
195 200 205
Glu Trp His Lys Pro Ala Ser Val Leu His Asn Met Val Arg Ala Val
210 215 220
Ala Asp Pro Trp Pro Gly Ala Phe Ser Tyr Val Gly Asn Gln Lys Phe
225 230 235 240
Thr Val Trp Ser Ser Arg Val His Pro His Ala Ser Lys Ala Gln Pro
245 250 255
Gly Ser Val Ile Ser Val Ala Pro Leu Leu Ile Ala Cys Gly Asp Gly
260 265 270
Ala Leu Glu Ile Val Thr Gly Gln Ala Gly Asp Gly Ile Thr Met Gln
275 280 285
Gly Ser Gln Leu Ala Gln Thr Leu Gly Leu Val Gln Gly Ser Arg Leu
290 295 300
Asn Ser Gln Pro Ala Cys Thr Ala Arg Arg Arg Thr Arg Val Leu Ile
305 310 315 320
Leu Gly Val Asn Gly Phe Ile Gly Asn His Leu Thr Glu Arg Leu Leu
325 330 335
Arg Glu Asp His Tyr Glu Val Tyr Gly Leu Asp Ile Gly Ser Asp Ala
340 345 350
Ile Ser Arg Phe Leu Asn His Pro His Phe His Phe Val Glu Gly Asp
355 360 365
Ile Ser Ile His Ser Glu Trp Ile Glu Tyr His Val Lys Lys Cys Asp
370 375 380
Val Val Leu Pro Leu Val Ala Ile Ala Thr Pro Ile Glu Tyr Thr Arg
385 390 395 400
Asn Pro Leu Arg Val Phe Glu Leu Asp Phe Glu Glu Asn Leu Arg Ile
405 410 415
Ile Arg Tyr Cys Val Lys Tyr Arg Lys Arg Ile Ile Phe Pro Ser Thr
420 425 430
Ser Glu Val Tyr Gly Met Cys Ser Asp Lys Tyr Phe Asp Glu Asp His
435 440 445
Ser Asn Leu Ile Val Gly Pro Val Asn Lys Pro Arg Trp Ile Tyr Ser
450 455 460
Val Ser Lys Gln Leu Leu Asp Arg Val Ile Trp Ala Tyr Gly Glu Lys
465 470 475 480
Glu Gly Leu Gln Phe Thr Leu Phe Arg Pro Phe Asn Trp Met Gly Pro
485 490 495
Arg Leu Asp Asn Leu Asn Ala Ala Arg Ile Gly Ser Ser Arg Ala Ile
500 505 510
Thr Gln Leu Ile Leu Asn Leu Val Glu Gly Ser Pro Ile Lys Leu Ile
515 520 525
Asp Gly Gly Lys Gln Lys Arg Cys Phe Thr Asp Ile Arg Asp Gly Ile
530 535 540
Glu Ala Leu Tyr Arg Ile Ile Glu Asn Ala Gly Asn Arg Cys Asp Gly
545 550 555 560
Glu Ile Ile Asn Ile Gly Asn Pro Glu Asn Glu Ala Ser Ile Glu Glu
565 570 575
Leu Gly Glu Met Leu Leu Ala Ser Phe Glu Lys His Pro Leu Arg His
580 585 590
His Phe Pro Pro Phe Ala Gly Phe Arg Val Val Glu Ser Ser Ser Tyr
595 600 605
Tyr Gly Lys Gly Tyr Gln Asp Val Glu His Arg Lys Pro Ser Ile Arg
610 615 620
Asn Ala His Arg Cys Leu Asp Trp Glu Pro Lys Ile Asp Met Gln Glu
625 630 635 640
Thr Ile Asp Glu Thr Leu Asp Phe Phe Leu Arg Thr Val Asp Leu Thr
645 650 655
Asp Lys Pro Ser
660
<210> 33
<211> 624
<212> PRT
<213> 大肠杆菌 BL21-DE3
<400> 33
Met Lys Gly Gln Glu Thr Arg Gly Phe Gln Ser Glu Val Lys Gln Leu
1 5 10 15
Leu His Leu Met Ile His Ser Leu Tyr Ser Asn Lys Glu Ile Phe Leu
20 25 30
Arg Glu Leu Ile Ser Asn Ala Ser Asp Ala Ala Asp Lys Leu Arg Phe
35 40 45
Arg Ala Leu Ser Asn Pro Asp Leu Tyr Glu Gly Asp Gly Glu Leu Arg
50 55 60
Val Arg Val Ser Phe Asp Lys Asp Lys Arg Thr Leu Thr Ile Ser Asp
65 70 75 80
Asn Gly Val Gly Met Thr Arg Asp Glu Val Ile Asp His Leu Gly Thr
85 90 95
Ile Ala Lys Ser Gly Thr Lys Ser Phe Leu Glu Ser Leu Gly Ser Asp
100 105 110
Gln Ala Lys Asp Ser Gln Leu Ile Gly Gln Phe Gly Val Gly Phe Tyr
115 120 125
Ser Ala Phe Ile Val Ala Asp Lys Val Thr Val Arg Thr Arg Ala Ala
130 135 140
Gly Glu Lys Pro Glu Asn Gly Val Phe Trp Glu Ser Ala Gly Glu Gly
145 150 155 160
Glu Tyr Thr Val Ala Asp Ile Thr Lys Glu Asp Arg Gly Thr Glu Ile
165 170 175
Thr Leu His Leu Arg Glu Gly Glu Asp Glu Phe Leu Asp Asp Trp Arg
180 185 190
Val Arg Ser Ile Ile Ser Lys Tyr Ser Asp His Ile Ala Leu Pro Val
195 200 205
Glu Ile Glu Lys Arg Glu Glu Lys Asp Gly Glu Thr Val Ile Ser Trp
210 215 220
Glu Lys Ile Asn Lys Ala Gln Ala Leu Trp Thr Arg Asn Lys Ser Glu
225 230 235 240
Ile Thr Asp Glu Glu Tyr Lys Glu Phe Tyr Lys His Ile Ala His Asp
245 250 255
Phe Asn Asp Pro Leu Thr Trp Ser His Asn Arg Val Glu Gly Lys Gln
260 265 270
Glu Tyr Thr Ser Leu Leu Tyr Ile Pro Ser Gln Ala Pro Trp Asp Met
275 280 285
Trp Asn Arg Asp His Lys His Gly Leu Lys Leu Tyr Val Gln Arg Val
290 295 300
Phe Ile Met Asp Asp Ala Glu Gln Phe Met Pro Asn Tyr Leu Arg Phe
305 310 315 320
Val Arg Gly Leu Ile Asp Ser Ser Asp Leu Pro Leu Asn Val Ser Arg
325 330 335
Glu Ile Leu Gln Asp Ser Thr Val Thr Arg Asn Leu Arg Asn Ala Leu
340 345 350
Thr Lys Arg Val Leu Gln Met Leu Glu Lys Leu Ala Lys Asp Asp Ala
355 360 365
Glu Lys Tyr Gln Thr Phe Trp Gln Gln Phe Gly Leu Val Leu Lys Glu
370 375 380
Gly Pro Ala Glu Asp Phe Ala Asn Gln Glu Ala Ile Ala Lys Leu Leu
385 390 395 400
Arg Phe Ala Ser Thr His Thr Asp Ser Ser Ala Gln Thr Val Ser Leu
405 410 415
Glu Asp Tyr Val Ser Arg Met Lys Glu Gly Gln Glu Lys Ile Tyr Tyr
420 425 430
Ile Thr Ala Asp Ser Tyr Ala Ala Ala Lys Ser Ser Pro His Leu Glu
435 440 445
Leu Leu Arg Lys Lys Gly Ile Glu Val Leu Leu Leu Ser Asp Arg Ile
450 455 460
Asp Glu Trp Met Met Asn Tyr Leu Thr Glu Phe Asp Gly Lys Pro Phe
465 470 475 480
Gln Ser Val Ser Lys Val Asp Glu Ser Leu Glu Lys Leu Ala Asp Glu
485 490 495
Val Asp Glu Ser Ala Lys Glu Ala Glu Lys Ala Leu Thr Pro Phe Ile
500 505 510
Asp Arg Val Lys Ala Leu Leu Gly Glu Arg Val Lys Asp Val Arg Leu
515 520 525
Thr His Arg Leu Thr Asp Thr Pro Ala Ile Val Ser Thr Asp Ala Asp
530 535 540
Glu Met Ser Thr Gln Met Ala Lys Leu Phe Ala Ala Ala Gly Gln Lys
545 550 555 560
Val Pro Glu Val Lys Tyr Ile Phe Glu Leu Asn Pro Asp His Val Leu
565 570 575
Val Lys Arg Ala Ala Asp Thr Glu Asp Glu Ala Lys Phe Ser Glu Trp
580 585 590
Val Glu Leu Leu Leu Asp Gln Ala Leu Leu Ala Glu Arg Gly Thr Leu
595 600 605
Glu Asp Pro Asn Leu Phe Ile Arg Arg Met Asn Gln Leu Leu Val Ser
610 615 620
<210> 34
<211> 638
<212> PRT
<213> 大肠杆菌 BL21-DE3
<400> 34
Met Gly Lys Ile Ile Gly Ile Asp Leu Gly Thr Thr Asn Ser Cys Val
1 5 10 15
Ala Ile Met Asp Gly Thr Thr Pro Arg Val Leu Glu Asn Ala Glu Gly
20 25 30
Asp Arg Thr Thr Pro Ser Ile Ile Ala Tyr Thr Gln Asp Gly Glu Thr
35 40 45
Leu Val Gly Gln Pro Ala Lys Arg Gln Ala Val Thr Asn Pro Gln Asn
50 55 60
Thr Leu Phe Ala Ile Lys Arg Leu Ile Gly Arg Arg Phe Gln Asp Glu
65 70 75 80
Glu Val Gln Arg Asp Val Ser Ile Met Pro Phe Lys Ile Ile Ala Ala
85 90 95
Asp Asn Gly Asp Ala Trp Val Glu Val Lys Gly Gln Lys Met Ala Pro
100 105 110
Pro Gln Ile Ser Ala Glu Val Leu Lys Lys Met Lys Lys Thr Ala Glu
115 120 125
Asp Tyr Leu Gly Glu Pro Val Thr Glu Ala Val Ile Thr Val Pro Ala
130 135 140
Tyr Phe Asn Asp Ala Gln Arg Gln Ala Thr Lys Asp Ala Gly Arg Ile
145 150 155 160
Ala Gly Leu Glu Val Lys Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala
165 170 175
Leu Ala Tyr Gly Leu Asp Lys Gly Thr Gly Asn Arg Thr Ile Ala Val
180 185 190
Tyr Asp Leu Gly Gly Gly Thr Phe Asp Ile Ser Ile Ile Glu Ile Asp
195 200 205
Glu Val Asp Gly Glu Lys Thr Phe Glu Val Leu Ala Thr Asn Gly Asp
210 215 220
Thr His Leu Gly Gly Glu Asp Phe Asp Ser Arg Leu Ile Asn Tyr Leu
225 230 235 240
Val Glu Glu Phe Lys Lys Asp Gln Gly Ile Asp Leu Arg Asn Asp Pro
245 250 255
Leu Ala Met Gln Arg Leu Lys Glu Ala Ala Glu Lys Ala Lys Ile Glu
260 265 270
Leu Ser Ser Ala Gln Gln Thr Asp Val Asn Leu Pro Tyr Ile Thr Ala
275 280 285
Asp Ala Thr Gly Pro Lys His Met Asn Ile Lys Val Thr Arg Ala Lys
290 295 300
Leu Glu Ser Leu Val Glu Asp Leu Val Asn Arg Ser Ile Glu Pro Leu
305 310 315 320
Lys Val Ala Leu Gln Asp Ala Gly Leu Ser Val Ser Asp Ile Asp Asp
325 330 335
Val Ile Leu Val Gly Gly Gln Thr Arg Met Pro Met Val Gln Lys Lys
340 345 350
Val Ala Glu Phe Phe Gly Lys Glu Pro Arg Lys Asp Val Asn Pro Asp
355 360 365
Glu Ala Val Ala Ile Gly Ala Ala Val Gln Gly Gly Val Leu Thr Gly
370 375 380
Asp Val Lys Asp Val Leu Leu Leu Asp Val Thr Pro Leu Ser Leu Gly
385 390 395 400
Ile Glu Thr Met Gly Gly Val Met Thr Thr Leu Ile Ala Lys Asn Thr
405 410 415
Thr Ile Pro Thr Lys His Ser Gln Val Phe Ser Thr Ala Glu Asp Asn
420 425 430
Gln Ser Ala Val Thr Ile His Val Leu Gln Gly Glu Arg Lys Arg Ala
435 440 445
Ala Asp Asn Lys Ser Leu Gly Gln Phe Asn Leu Asp Gly Ile Asn Pro
450 455 460
Ala Pro Arg Gly Met Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala
465 470 475 480
Asp Gly Ile Leu His Val Ser Ala Lys Asp Lys Asn Ser Gly Lys Glu
485 490 495
Gln Lys Ile Thr Ile Lys Ala Ser Ser Gly Leu Asn Glu Asp Glu Ile
500 505 510
Gln Lys Met Val Arg Asp Ala Glu Ala Asn Ala Glu Ala Asp Arg Lys
515 520 525
Phe Glu Glu Leu Val Gln Thr Arg Asn Gln Gly Asp His Leu Leu His
530 535 540
Ser Thr Arg Lys Gln Val Glu Glu Ala Gly Asp Lys Leu Pro Ala Asp
545 550 555 560
Asp Lys Thr Ala Ile Glu Ser Ala Leu Thr Ala Leu Glu Thr Ala Leu
565 570 575
Lys Gly Glu Asp Lys Ala Ala Ile Glu Ala Lys Met Gln Glu Leu Ala
580 585 590
Gln Val Ser Gln Lys Leu Met Glu Ile Ala Gln Gln Gln His Ala Gln
595 600 605
Gln Gln Thr Ala Gly Ala Asp Ala Ser Ala Asn Asn Ala Lys Asp Asp
610 615 620
Asp Val Val Asp Ala Glu Phe Glu Glu Val Lys Asp Lys Lys
625 630 635
<210> 35
<211> 1193
<212> PRT
<213> 耻垢分枝杆菌 ATCC 700084
<400> 35
Met His Gln Leu Thr Val Thr Gly Met Asn Ile Cys Glu Val Gln Arg
1 5 10 15
Leu Phe Pro Arg Met Thr Ser Asp Val His Asp Ala Thr Asp Gly Val
20 25 30
Thr Glu Thr Ala Leu Asp Asp Glu Gln Ser Thr Arg Arg Ile Ala Glu
35 40 45
Leu Tyr Ala Thr Asp Pro Glu Phe Ala Ala Ala Ala Pro Leu Pro Ala
50 55 60
Val Val Asp Ala Ala His Lys Pro Gly Leu Arg Leu Ala Glu Ile Leu
65 70 75 80
Gln Thr Leu Phe Thr Gly Tyr Gly Asp Arg Pro Ala Leu Gly Tyr Arg
85 90 95
Ala Arg Glu Leu Ala Thr Asp Glu Gly Gly Arg Thr Val Thr Arg Leu
100 105 110
Leu Pro Arg Phe Asp Thr Leu Thr Tyr Ala Gln Val Trp Ser Arg Val
115 120 125
Gln Ala Val Ala Ala Ala Leu Arg His Asn Phe Ala Gln Pro Ile Tyr
130 135 140
Pro Gly Asp Ala Val Ala Thr Ile Gly Phe Ala Ser Pro Asp Tyr Leu
145 150 155 160
Thr Leu Asp Leu Val Cys Ala Tyr Leu Gly Leu Val Ser Val Pro Leu
165 170 175
Gln His Asn Ala Pro Val Ser Arg Leu Ala Pro Ile Leu Ala Glu Val
180 185 190
Glu Pro Arg Ile Leu Thr Val Ser Ala Glu Tyr Leu Asp Leu Ala Val
195 200 205
Glu Ser Val Arg Asp Val Asn Ser Val Ser Gln Leu Val Val Phe Asp
210 215 220
His His Pro Glu Val Asp Asp His Arg Asp Ala Leu Ala Arg Ala Arg
225 230 235 240
Glu Gln Leu Ala Gly Lys Gly Ile Ala Val Thr Thr Leu Asp Ala Ile
245 250 255
Ala Asp Glu Gly Ala Gly Leu Pro Ala Glu Pro Ile Tyr Thr Ala Asp
260 265 270
His Asp Gln Arg Leu Ala Met Ile Leu Tyr Thr Ser Gly Ser Thr Gly
275 280 285
Ala Pro Lys Gly Ala Met Tyr Thr Glu Ala Met Val Ala Arg Leu Trp
290 295 300
Thr Met Ser Phe Ile Thr Gly Asp Pro Thr Pro Val Ile Asn Val Asn
305 310 315 320
Phe Met Pro Leu Asn His Leu Gly Gly Arg Ile Pro Ile Ser Thr Ala
325 330 335
Val Gln Asn Gly Gly Thr Ser Tyr Phe Val Pro Glu Ser Asp Met Ser
340 345 350
Thr Leu Phe Glu Asp Leu Ala Leu Val Arg Pro Thr Glu Leu Gly Leu
355 360 365
Val Pro Arg Val Ala Asp Met Leu Tyr Gln His His Leu Ala Thr Val
370 375 380
Asp Arg Leu Val Thr Gln Gly Ala Asp Glu Leu Thr Ala Glu Lys Gln
385 390 395 400
Ala Gly Ala Glu Leu Arg Glu Gln Val Leu Gly Gly Arg Val Ile Thr
405 410 415
Gly Phe Val Ser Thr Ala Pro Leu Ala Ala Glu Met Arg Ala Phe Leu
420 425 430
Asp Ile Thr Leu Gly Ala His Ile Val Asp Gly Tyr Gly Leu Thr Glu
435 440 445
Thr Gly Ala Val Thr Arg Asp Gly Val Ile Val Arg Pro Pro Val Ile
450 455 460
Asp Tyr Lys Leu Ile Asp Val Pro Glu Leu Gly Tyr Phe Ser Thr Asp
465 470 475 480
Lys Pro Tyr Pro Arg Gly Glu Leu Leu Val Arg Ser Gln Thr Leu Thr
485 490 495
Pro Gly Tyr Tyr Lys Arg Pro Glu Val Thr Ala Ser Val Phe Asp Arg
500 505 510
Asp Gly Tyr Tyr His Thr Gly Asp Val Met Ala Glu Thr Ala Pro Asp
515 520 525
His Leu Val Tyr Val Asp Arg Arg Asn Asn Val Leu Lys Leu Ala Gln
530 535 540
Gly Glu Phe Val Ala Val Ala Asn Leu Glu Ala Val Phe Ser Gly Ala
545 550 555 560
Ala Leu Val Arg Gln Ile Phe Val Tyr Gly Asn Ser Glu Arg Ser Phe
565 570 575
Leu Leu Ala Val Val Val Pro Thr Pro Glu Ala Leu Glu Gln Tyr Asp
580 585 590
Pro Ala Ala Leu Lys Ala Ala Leu Ala Asp Ser Leu Gln Arg Thr Ala
595 600 605
Arg Asp Ala Glu Leu Gln Ser Tyr Glu Val Pro Ala Asp Phe Ile Val
610 615 620
Glu Thr Glu Pro Phe Ser Ala Ala Asn Gly Leu Leu Ser Gly Val Gly
625 630 635 640
Lys Leu Leu Arg Pro Asn Leu Lys Asp Arg Tyr Gly Gln Arg Leu Glu
645 650 655
Gln Met Tyr Ala Asp Ile Ala Ala Thr Gln Ala Asn Gln Leu Arg Glu
660 665 670
Leu Arg Arg Ala Ala Ala Thr Gln Pro Val Ile Asp Thr Leu Thr Gln
675 680 685
Ala Ala Ala Thr Ile Leu Gly Thr Gly Ser Glu Val Ala Ser Asp Ala
690 695 700
His Phe Thr Asp Leu Gly Gly Asp Ser Leu Ser Ala Leu Thr Leu Ser
705 710 715 720
Asn Leu Leu Ser Asp Phe Phe Gly Phe Glu Val Pro Val Gly Thr Ile
725 730 735
Val Asn Pro Ala Thr Asn Leu Ala Gln Leu Ala Gln His Ile Glu Ala
740 745 750
Gln Arg Thr Ala Gly Asp Arg Arg Pro Ser Phe Thr Thr Val His Gly
755 760 765
Ala Asp Ala Thr Glu Ile Arg Ala Ser Glu Leu Thr Leu Asp Lys Phe
770 775 780
Ile Asp Ala Glu Thr Leu Arg Ala Ala Pro Gly Leu Pro Lys Val Thr
785 790 795 800
Thr Glu Pro Arg Thr Val Leu Leu Ser Gly Ala Asn Gly Trp Leu Gly
805 810 815
Arg Phe Leu Thr Leu Gln Trp Leu Glu Arg Leu Ala Pro Val Gly Gly
820 825 830
Thr Leu Ile Thr Ile Val Arg Gly Arg Asp Asp Ala Ala Ala Arg Ala
835 840 845
Arg Leu Thr Gln Ala Tyr Asp Thr Asp Pro Glu Leu Ser Arg Arg Phe
850 855 860
Ala Glu Leu Ala Asp Arg His Leu Arg Val Val Ala Gly Asp Ile Gly
865 870 875 880
Asp Pro Asn Leu Gly Leu Thr Pro Glu Ile Trp His Arg Leu Ala Ala
885 890 895
Glu Val Asp Leu Val Val His Pro Ala Ala Leu Val Asn His Val Leu
900 905 910
Pro Tyr Arg Gln Leu Phe Gly Pro Asn Val Val Gly Thr Ala Glu Val
915 920 925
Ile Lys Leu Ala Leu Thr Glu Arg Ile Lys Pro Val Thr Tyr Leu Ser
930 935 940
Thr Val Ser Val Ala Met Gly Ile Pro Asp Phe Glu Glu Asp Gly Asp
945 950 955 960
Ile Arg Thr Val Ser Pro Val Arg Pro Leu Asp Gly Gly Tyr Ala Asn
965 970 975
Gly Tyr Gly Asn Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala
980 985 990
His Asp Leu Cys Gly Leu Pro Val Ala Thr Phe Arg Ser Asp Met Ile
995 1000 1005
Leu Ala His Pro Arg Tyr Arg Gly Gln Val Asn Val Pro Asp Met
1010 1015 1020
Phe Thr Arg Leu Leu Leu Ser Leu Leu Ile Thr Gly Val Ala Pro
1025 1030 1035
Arg Ser Phe Tyr Ile Gly Asp Gly Glu Arg Pro Arg Ala His Tyr
1040 1045 1050
Pro Gly Leu Thr Val Asp Phe Val Ala Glu Ala Val Thr Thr Leu
1055 1060 1065
Gly Ala Gln Gln Arg Glu Gly Tyr Val Ser Tyr Asp Val Met Asn
1070 1075 1080
Pro His Asp Asp Gly Ile Ser Leu Asp Val Phe Val Asp Trp Leu
1085 1090 1095
Ile Arg Ala Gly His Pro Ile Asp Arg Val Asp Asp Tyr Asp Asp
1100 1105 1110
Trp Val Arg Arg Phe Glu Thr Ala Leu Thr Ala Leu Pro Glu Lys
1115 1120 1125
Arg Arg Ala Gln Thr Val Leu Pro Leu Leu His Ala Phe Arg Ala
1130 1135 1140
Pro Gln Ala Pro Leu Arg Gly Ala Pro Glu Pro Thr Glu Val Phe
1145 1150 1155
His Ala Ala Val Arg Thr Ala Lys Val Gly Pro Gly Asp Ile Pro
1160 1165 1170
His Leu Asp Glu Ala Leu Ile Asp Lys Tyr Ile Arg Asp Leu Arg
1175 1180 1185
Glu Phe Gly Leu Ile
1190
<210> 36
<211> 1178
<212> PRT
<213> 脓肿分枝杆菌 DSM44196
<400> 36
Met Thr Glu Thr Ile Ser Thr Ala Ala Val Pro Thr Thr Asp Leu Glu
1 5 10 15
Glu Gln Val Lys Arg Arg Ile Glu Gln Val Val Ser Asn Asp Pro Gln
20 25 30
Leu Ala Ala Leu Leu Pro Glu Asp Ser Val Thr Glu Ala Val Asn Glu
35 40 45
Pro Asp Leu Pro Leu Val Glu Val Ile Arg Arg Leu Leu Glu Gly Tyr
50 55 60
Gly Asp Arg Pro Ala Leu Gly Gln Arg Ala Phe Glu Phe Val Thr Gly
65 70 75 80
Asp Asp Gly Ala Thr Val Ile Ala Leu Lys Pro Glu Tyr Thr Thr Val
85 90 95
Ser Tyr Arg Glu Leu Trp Glu Arg Ala Glu Ala Ile Ala Ala Ala Trp
100 105 110
His Glu Gln Gly Ile Arg Asp Gly Asp Phe Val Ala Gln Leu Gly Phe
115 120 125
Thr Ser Thr Asp Phe Ala Ser Leu Asp Val Ala Gly Leu Arg Leu Gly
130 135 140
Thr Val Ser Val Pro Leu Gln Thr Gly Ala Ser Leu Gln Gln Arg Asn
145 150 155 160
Ala Ile Leu Glu Glu Thr Arg Pro Ala Val Phe Ala Ala Ser Ile Glu
165 170 175
Tyr Leu Asp Ala Ala Val Asp Ser Val Leu Ala Thr Pro Ser Val Arg
180 185 190
Leu Leu Ser Val Phe Asp Tyr His Ala Glu Val Asp Ser Gln Arg Glu
195 200 205
Ala Leu Glu Ala Val Arg Ala Arg Leu Glu Ser Ala Gly Arg Thr Ile
210 215 220
Val Val Glu Ala Leu Ala Glu Ala Leu Ala Arg Gly Arg Asp Leu Pro
225 230 235 240
Ala Ala Pro Leu Pro Ser Ala Asp Pro Asp Ala Leu Arg Leu Leu Ile
245 250 255
Tyr Thr Ser Gly Ser Thr Gly Thr Pro Lys Gly Ala Met Tyr Pro Gln
260 265 270
Trp Leu Val Ala Asn Leu Trp Gln Lys Lys Trp Leu Thr Asp Asp Val
275 280 285
Ile Pro Ser Ile Gly Val Asn Phe Met Pro Met Ser His Leu Ala Gly
290 295 300
Arg Leu Thr Leu Met Gly Thr Leu Ser Gly Gly Gly Thr Ala Tyr Tyr
305 310 315 320
Ile Ala Ser Ser Asp Leu Ser Thr Phe Phe Glu Asp Ile Ala Leu Ile
325 330 335
Arg Pro Ser Glu Val Leu Phe Val Pro Arg Val Val Glu Met Val Phe
340 345 350
Gln Arg Phe Gln Ala Glu Leu Asp Arg Ser Leu Ala Pro Gly Glu Ser
355 360 365
Asn Ser Glu Ile Ala Glu Arg Ile Lys Val Arg Ile Arg Glu Gln Asp
370 375 380
Phe Gly Gly Arg Val Leu Ser Ala Gly Ser Gly Ser Ala Pro Leu Ser
385 390 395 400
Pro Glu Met Thr Glu Phe Met Glu Ser Leu Leu Gln Val Pro Leu Arg
405 410 415
Asp Gly Tyr Gly Ser Thr Glu Ala Gly Gly Val Trp Arg Asp Gly Val
420 425 430
Leu Gln Arg Pro Pro Val Thr Asp Tyr Lys Leu Val Asp Val Pro Glu
435 440 445
Leu Gly Tyr Phe Thr Thr Asp Ser Pro His Pro Arg Gly Glu Leu Arg
450 455 460
Leu Lys Ser Glu Thr Met Phe Pro Gly Tyr Tyr Lys Arg Pro Glu Thr
465 470 475 480
Thr Ala Asp Val Phe Asp Asp Glu Gly Tyr Tyr Lys Thr Gly Asp Val
485 490 495
Val Ala Glu Leu Gly Pro Asp His Leu Lys Tyr Leu Asp Arg Val Lys
500 505 510
Asn Val Leu Lys Leu Ala Gln Gly Glu Phe Val Ala Val Ser Lys Leu
515 520 525
Glu Ala Ala Tyr Thr Gly Ser Pro Leu Val Arg Gln Ile Phe Val Tyr
530 535 540
Gly Asn Ser Glu Arg Ser Phe Leu Leu Ala Val Val Val Pro Thr Pro
545 550 555 560
Glu Val Leu Glu Arg Tyr Ala Asp Ser Pro Asp Ala Leu Lys Pro Leu
565 570 575
Ile Gln Asp Ser Leu Gln Gln Val Ala Lys Asp Ala Glu Leu Gln Ser
580 585 590
Tyr Glu Ile Pro Arg Asp Phe Ile Val Glu Thr Val Pro Phe Thr Val
595 600 605
Glu Ser Gly Leu Leu Ser Asp Ala Arg Lys Leu Leu Arg Pro Lys Leu
610 615 620
Lys Asp His Tyr Gly Glu Arg Leu Glu Ala Leu Tyr Ala Glu Leu Ala
625 630 635 640
Glu Ser Gln Asn Glu Arg Leu Arg Gln Leu Ala Arg Glu Ala Ala Thr
645 650 655
Arg Pro Val Leu Glu Thr Val Thr Asp Ala Ala Ala Ala Leu Leu Gly
660 665 670
Ala Ser Ser Ser Asp Leu Ala Pro Asp Val Arg Phe Ile Asp Leu Gly
675 680 685
Gly Asp Ser Leu Ser Ala Leu Ser Tyr Ser Glu Leu Leu Arg Asp Ile
690 695 700
Phe Glu Val Asp Val Pro Val Gly Val Ile Asn Ser Val Ala Asn Asp
705 710 715 720
Leu Ala Ala Ile Ala Arg His Ile Glu Ala Gln Arg Thr Gly Ala Ala
725 730 735
Thr Gln Pro Thr Phe Ala Ser Val His Gly Lys Asp Ala Thr Val Ile
740 745 750
Thr Ala Gly Glu Leu Thr Leu Asp Lys Phe Leu Asp Glu Ser Leu Leu
755 760 765
Lys Ala Ala Lys Asp Val Gln Pro Ala Thr Ala Asp Val Lys Thr Val
770 775 780
Leu Val Thr Gly Gly Asn Gly Trp Leu Gly Arg Trp Leu Val Leu Asp
785 790 795 800
Trp Leu Glu Arg Leu Ala Pro Asn Gly Gly Lys Val Tyr Ala Leu Ile
805 810 815
Arg Gly Ala Asp Ala Glu Ala Ala Arg Ala Arg Leu Asp Ala Val Tyr
820 825 830
Glu Ser Gly Asp Pro Lys Leu Ser Ala His Tyr Arg Gln Leu Ala Gln
835 840 845
Gln Ser Leu Glu Val Ile Ala Gly Asp Phe Gly Asp Gln Asp Leu Gly
850 855 860
Leu Ser Gln Glu Val Trp Gln Lys Leu Ala Lys Asp Val Asp Leu Ile
865 870 875 880
Val His Ser Gly Ala Leu Val Asn His Val Leu Pro Tyr Ser Gln Leu
885 890 895
Phe Gly Pro Asn Val Ala Gly Thr Ala Glu Ile Ile Lys Leu Ala Ile
900 905 910
Ser Glu Arg Leu Lys Pro Val Thr Tyr Leu Ser Thr Val Gly Ile Ala
915 920 925
Asp Gln Ile Pro Val Thr Glu Phe Glu Glu Asp Ser Asp Val Arg Val
930 935 940
Met Ser Ala Glu Arg Gln Ile Asn Asp Gly Tyr Ala Asn Gly Tyr Gly
945 950 955 960
Asn Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His Asp Leu
965 970 975
Ala Gly Leu Pro Val Arg Val Phe Arg Ser Asp Met Ile Leu Ala His
980 985 990
Ser Asp Tyr His Gly Gln Leu Asn Val Thr Asp Val Phe Thr Arg Ser
995 1000 1005
Ile Gln Ser Leu Leu Leu Thr Gly Val Ala Pro Ala Ser Phe Tyr
1010 1015 1020
Glu Leu Asp Ala Asp Gly Asn Arg Gln Arg Ala His Tyr Asp Gly
1025 1030 1035
Val Pro Gly Asp Phe Thr Ala Ala Ser Ile Thr Ala Ile Gly Gly
1040 1045 1050
Val Asn Val Val Asp Gly Tyr Arg Ser Phe Asp Val Phe Asn Pro
1055 1060 1065
His His Asp Gly Val Ser Met Asp Thr Phe Val Asp Trp Leu Ile
1070 1075 1080
Asp Ala Gly Tyr Lys Ile Ala Arg Ile Asp Asp Tyr Asp Gln Trp
1085 1090 1095
Leu Ala Arg Phe Glu Leu Ala Leu Lys Gly Leu Pro Glu Gln Gln
1100 1105 1110
Arg Gln Gln Ser Val Leu Pro Leu Leu Lys Met Tyr Glu Lys Pro
1115 1120 1125
Gln Pro Ala Ile Asp Gly Ser Ala Leu Pro Thr Ala Glu Phe Ser
1130 1135 1140
Arg Ala Val His Glu Ala Lys Val Gly Asp Ser Gly Glu Ile Pro
1145 1150 1155
His Val Thr Lys Glu Leu Ile Leu Lys Tyr Ala Ser Asp Ile Gln
1160 1165 1170
Leu Leu Gly Leu Val
1175
<210> 37
<211> 1173
<212> PRT
<213> 耻垢分枝杆菌 ATCC 700084
<400> 37
Met Thr Ser Asp Val His Asp Ala Thr Asp Gly Val Thr Glu Thr Ala
1 5 10 15
Leu Asp Asp Glu Gln Ser Thr Arg Arg Ile Ala Glu Leu Tyr Ala Thr
20 25 30
Asp Pro Glu Phe Ala Ala Ala Ala Pro Leu Pro Ala Val Val Asp Ala
35 40 45
Ala His Lys Pro Gly Leu Arg Leu Ala Glu Ile Leu Gln Thr Leu Phe
50 55 60
Thr Gly Tyr Gly Asp Arg Pro Ala Leu Gly Tyr Arg Ala Arg Glu Leu
65 70 75 80
Ala Thr Asp Glu Gly Gly Arg Thr Val Thr Arg Leu Leu Pro Arg Phe
85 90 95
Asp Thr Leu Thr Tyr Ala Gln Val Trp Ser Arg Val Gln Ala Val Ala
100 105 110
Ala Ala Leu Arg His Asn Phe Ala Gln Pro Ile Tyr Pro Gly Asp Ala
115 120 125
Val Ala Thr Ile Gly Phe Ala Ser Pro Asp Tyr Leu Thr Leu Asp Leu
130 135 140
Val Cys Ala Tyr Leu Gly Leu Val Ser Val Pro Leu Gln His Asn Ala
145 150 155 160
Pro Val Ser Arg Leu Ala Pro Ile Leu Ala Glu Val Glu Pro Arg Ile
165 170 175
Leu Thr Val Ser Ala Glu Tyr Leu Asp Leu Ala Val Glu Ser Val Arg
180 185 190
Asp Val Asn Ser Val Ser Gln Leu Val Val Phe Asp His His Pro Glu
195 200 205
Val Asp Asp His Arg Asp Ala Leu Ala Arg Ala Arg Glu Gln Leu Ala
210 215 220
Gly Lys Gly Ile Ala Val Thr Thr Leu Asp Ala Ile Ala Asp Glu Gly
225 230 235 240
Ala Gly Leu Pro Ala Glu Pro Ile Tyr Thr Ala Asp His Asp Gln Arg
245 250 255
Leu Ala Met Ile Leu Tyr Thr Ser Gly Ser Thr Gly Ala Pro Lys Gly
260 265 270
Ala Met Tyr Thr Glu Ala Met Val Ala Arg Leu Trp Thr Met Ser Phe
275 280 285
Ile Thr Gly Asp Pro Thr Pro Val Ile Asn Val Asn Phe Met Pro Leu
290 295 300
Asn His Leu Gly Gly Arg Ile Pro Ile Ser Thr Ala Val Gln Asn Gly
305 310 315 320
Gly Thr Ser Tyr Phe Val Pro Glu Ser Asp Met Ser Thr Leu Phe Glu
325 330 335
Asp Leu Ala Leu Val Arg Pro Thr Glu Leu Gly Leu Val Pro Arg Val
340 345 350
Ala Asp Met Leu Tyr Gln His His Leu Ala Thr Val Asp Arg Leu Val
355 360 365
Thr Gln Gly Ala Asp Glu Leu Thr Ala Glu Lys Gln Ala Gly Ala Glu
370 375 380
Leu Arg Glu Gln Val Leu Gly Gly Arg Val Ile Thr Gly Phe Val Ser
385 390 395 400
Thr Ala Pro Leu Ala Ala Glu Met Arg Ala Phe Leu Asp Ile Thr Leu
405 410 415
Gly Ala His Ile Val Asp Gly Tyr Gly Leu Thr Glu Thr Gly Ala Val
420 425 430
Thr Arg Asp Gly Val Ile Val Arg Pro Pro Val Ile Asp Tyr Lys Leu
435 440 445
Ile Asp Val Pro Glu Leu Gly Tyr Phe Ser Thr Asp Lys Pro Tyr Pro
450 455 460
Arg Gly Glu Leu Leu Val Arg Ser Gln Thr Leu Thr Pro Gly Tyr Tyr
465 470 475 480
Lys Arg Pro Glu Val Thr Ala Ser Val Phe Asp Arg Asp Gly Tyr Tyr
485 490 495
His Thr Gly Asp Val Met Ala Glu Thr Ala Pro Asp His Leu Val Tyr
500 505 510
Val Asp Arg Arg Asn Asn Val Leu Lys Leu Ala Gln Gly Glu Phe Val
515 520 525
Ala Val Ala Asn Leu Glu Ala Val Phe Ser Gly Ala Ala Leu Val Arg
530 535 540
Gln Ile Phe Val Tyr Gly Asn Ser Glu Arg Ser Phe Leu Leu Ala Val
545 550 555 560
Val Val Pro Thr Pro Glu Ala Leu Glu Gln Tyr Asp Pro Ala Ala Leu
565 570 575
Lys Ala Ala Leu Ala Asp Ser Leu Gln Arg Thr Ala Arg Asp Ala Glu
580 585 590
Leu Gln Ser Tyr Glu Val Pro Ala Asp Phe Ile Val Glu Thr Glu Pro
595 600 605
Phe Ser Ala Ala Asn Gly Leu Leu Ser Gly Val Gly Lys Leu Leu Arg
610 615 620
Pro Asn Leu Lys Asp Arg Tyr Gly Gln Arg Leu Glu Gln Met Tyr Ala
625 630 635 640
Asp Ile Ala Ala Thr Gln Ala Asn Gln Leu Arg Glu Leu Arg Arg Ala
645 650 655
Ala Ala Thr Gln Pro Val Ile Asp Thr Leu Thr Gln Ala Ala Ala Thr
660 665 670
Ile Leu Gly Thr Gly Ser Glu Val Ala Ser Asp Ala His Phe Thr Asp
675 680 685
Leu Gly Gly Asp Ser Leu Ser Ala Leu Thr Leu Ser Asn Leu Leu Ser
690 695 700
Asp Phe Phe Gly Phe Glu Val Pro Val Gly Thr Ile Val Asn Pro Ala
705 710 715 720
Thr Asn Leu Ala Gln Leu Ala Gln His Ile Glu Ala Gln Arg Thr Ala
725 730 735
Gly Asp Arg Arg Pro Ser Phe Thr Thr Val His Gly Ala Asp Ala Thr
740 745 750
Glu Ile Arg Ala Ser Glu Leu Thr Leu Asp Lys Phe Ile Asp Ala Glu
755 760 765
Thr Leu Arg Ala Ala Pro Gly Leu Pro Lys Val Thr Thr Glu Pro Arg
770 775 780
Thr Val Leu Leu Ser Gly Ala Asn Gly Trp Leu Gly Arg Phe Leu Thr
785 790 795 800
Leu Gln Trp Leu Glu Arg Leu Ala Pro Val Gly Gly Thr Leu Ile Thr
805 810 815
Ile Val Arg Gly Arg Asp Asp Ala Ala Ala Arg Ala Arg Leu Thr Gln
820 825 830
Ala Tyr Asp Thr Asp Pro Glu Leu Ser Arg Arg Phe Ala Glu Leu Ala
835 840 845
Asp Arg His Leu Arg Val Val Ala Gly Asp Ile Gly Asp Pro Asn Leu
850 855 860
Gly Leu Thr Pro Glu Ile Trp His Arg Leu Ala Ala Glu Val Asp Leu
865 870 875 880
Val Val His Pro Ala Ala Leu Val Asn His Val Leu Pro Tyr Arg Gln
885 890 895
Leu Phe Gly Pro Asn Val Val Gly Thr Ala Glu Val Ile Lys Leu Ala
900 905 910
Leu Thr Glu Arg Ile Lys Pro Val Thr Tyr Leu Ser Thr Val Ser Val
915 920 925
Ala Met Gly Ile Pro Asp Phe Glu Glu Asp Gly Asp Ile Arg Thr Val
930 935 940
Ser Pro Val Arg Pro Leu Asp Gly Gly Tyr Ala Asn Gly Tyr Gly Asn
945 950 955 960
Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His Asp Leu Cys
965 970 975
Gly Leu Pro Val Ala Thr Phe Arg Ser Asp Met Ile Leu Ala His Pro
980 985 990
Arg Tyr Arg Gly Gln Val Asn Val Pro Asp Met Phe Thr Arg Leu Leu
995 1000 1005
Leu Ser Leu Leu Ile Thr Gly Val Ala Pro Arg Ser Phe Tyr Ile
1010 1015 1020
Gly Asp Gly Glu Arg Pro Arg Ala His Tyr Pro Gly Leu Thr Val
1025 1030 1035
Asp Phe Val Ala Glu Ala Val Thr Thr Leu Gly Ala Gln Gln Arg
1040 1045 1050
Glu Gly Tyr Val Ser Tyr Asp Val Met Asn Pro His Asp Asp Gly
1055 1060 1065
Ile Ser Leu Asp Val Phe Val Asp Trp Leu Ile Arg Ala Gly His
1070 1075 1080
Pro Ile Asp Arg Val Asp Asp Tyr Asp Asp Trp Val Arg Arg Phe
1085 1090 1095
Glu Thr Ala Leu Thr Ala Leu Pro Glu Lys Arg Arg Ala Gln Thr
1100 1105 1110
Val Leu Pro Leu Leu His Ala Phe Arg Ala Pro Gln Ala Pro Leu
1115 1120 1125
Arg Gly Ala Pro Glu Pro Thr Glu Val Phe His Ala Ala Val Arg
1130 1135 1140
Thr Ala Lys Val Gly Pro Gly Asp Ile Pro His Leu Asp Glu Ala
1145 1150 1155
Leu Ile Asp Lys Tyr Ile Arg Asp Leu Arg Glu Phe Gly Leu Ile
1160 1165 1170
<210> 38
<211> 1173
<212> PRT
<213> 类结核分枝杆菌 ATCC BAA-968
<400> 38
Met Ser Thr Ala Thr His Asp Glu Arg Leu Asp Arg Arg Val His Glu
1 5 10 15
Leu Ile Ala Thr Asp Pro Gln Phe Ala Ala Ala Gln Pro Asp Pro Ala
20 25 30
Ile Thr Ala Ala Leu Glu Gln Pro Gly Leu Arg Leu Pro Gln Ile Ile
35 40 45
Arg Thr Val Leu Asp Gly Tyr Ala Asp Arg Pro Ala Leu Gly Gln Arg
50 55 60
Val Val Glu Phe Val Thr Asp Ala Lys Thr Gly Arg Thr Ser Ala Gln
65 70 75 80
Leu Leu Pro Arg Phe Glu Thr Ile Thr Tyr Ser Glu Val Ala Gln Arg
85 90 95
Val Ser Ala Leu Gly Arg Ala Leu Ser Asp Asp Ala Val His Pro Gly
100 105 110
Asp Arg Val Cys Val Leu Gly Phe Asn Ser Val Asp Tyr Ala Thr Ile
115 120 125
Asp Met Ala Leu Gly Ala Ile Gly Ala Val Ser Val Pro Leu Gln Thr
130 135 140
Ser Ala Ala Ile Ser Ser Leu Gln Pro Ile Val Ala Glu Thr Glu Pro
145 150 155 160
Thr Leu Ile Ala Ser Ser Val Asn Gln Leu Ser Asp Ala Val Gln Leu
165 170 175
Ile Thr Gly Ala Glu Gln Ala Pro Thr Arg Leu Val Val Phe Asp Tyr
180 185 190
His Pro Gln Val Asp Asp Gln Arg Glu Ala Val Gln Asp Ala Ala Ala
195 200 205
Arg Leu Ser Ser Thr Gly Val Ala Val Gln Thr Leu Ala Glu Leu Leu
210 215 220
Glu Arg Gly Lys Asp Leu Pro Ala Val Ala Glu Pro Pro Ala Asp Glu
225 230 235 240
Asp Ser Leu Ala Leu Leu Ile Tyr Thr Ser Gly Ser Thr Gly Ala Pro
245 250 255
Lys Gly Ala Met Tyr Pro Gln Ser Asn Val Gly Lys Met Trp Arg Arg
260 265 270
Gly Ser Lys Asn Trp Phe Gly Glu Ser Ala Ala Ser Ile Thr Leu Asn
275 280 285
Phe Met Pro Met Ser His Val Met Gly Arg Ser Ile Leu Tyr Gly Thr
290 295 300
Leu Gly Asn Gly Gly Thr Ala Tyr Phe Ala Ala Arg Ser Asp Leu Ser
305 310 315 320
Thr Leu Leu Glu Asp Leu Glu Leu Val Arg Pro Thr Glu Leu Asn Phe
325 330 335
Val Pro Arg Ile Trp Glu Thr Leu Tyr Gly Glu Phe Gln Arg Gln Val
340 345 350
Glu Arg Arg Leu Ser Glu Ala Gly Asp Ala Gly Glu Arg Arg Ala Val
355 360 365
Glu Ala Glu Val Leu Ala Glu Gln Arg Gln Tyr Leu Leu Gly Gly Arg
370 375 380
Phe Thr Phe Ala Met Thr Gly Ser Ala Pro Ile Ser Pro Glu Leu Arg
385 390 395 400
Asn Trp Val Glu Ser Leu Leu Glu Met His Leu Met Asp Gly Tyr Gly
405 410 415
Ser Thr Glu Ala Gly Met Val Leu Phe Asp Gly Glu Ile Gln Arg Pro
420 425 430
Pro Val Ile Asp Tyr Lys Leu Val Asp Val Pro Asp Leu Gly Tyr Phe
435 440 445
Ser Thr Asp Arg Pro His Pro Arg Gly Glu Leu Leu Leu Arg Thr Glu
450 455 460
Asn Met Phe Pro Gly Tyr Tyr Lys Arg Ala Glu Thr Thr Ala Gly Val
465 470 475 480
Phe Asp Glu Asp Gly Tyr Tyr Arg Thr Gly Asp Val Phe Ala Glu Ile
485 490 495
Ala Pro Asp Arg Leu Val Tyr Val Asp Arg Arg Asn Asn Val Leu Lys
500 505 510
Leu Ala Gln Gly Glu Phe Val Thr Leu Ala Lys Leu Glu Ala Val Phe
515 520 525
Gly Asn Ser Pro Leu Ile Arg Gln Ile Tyr Val Tyr Gly Asn Ser Ala
530 535 540
Gln Pro Tyr Leu Leu Ala Val Val Val Pro Thr Glu Glu Ala Leu Ala
545 550 555 560
Ser Gly Asp Pro Glu Thr Leu Lys Pro Lys Ile Ala Asp Ser Leu Gln
565 570 575
Gln Val Ala Lys Glu Ala Gly Leu Gln Ser Tyr Glu Val Pro Arg Asp
580 585 590
Phe Ile Ile Glu Thr Thr Pro Phe Ser Leu Glu Asn Gly Leu Leu Thr
595 600 605
Gly Ile Arg Lys Leu Ala Trp Pro Lys Leu Lys Gln His Tyr Gly Glu
610 615 620
Arg Leu Glu Gln Met Tyr Ala Asp Leu Ala Ala Gly Gln Ala Asn Glu
625 630 635 640
Leu Ala Glu Leu Arg Arg Asn Gly Ala Gln Ala Pro Val Leu Gln Thr
645 650 655
Val Ser Arg Ala Ala Gly Ala Met Leu Gly Ser Ala Ala Ser Asp Leu
660 665 670
Ser Pro Asp Ala His Phe Thr Asp Leu Gly Gly Asp Ser Leu Ser Ala
675 680 685
Leu Thr Phe Gly Asn Leu Leu Arg Glu Ile Phe Asp Val Asp Val Pro
690 695 700
Val Gly Val Ile Val Ser Pro Ala Asn Asp Leu Ala Ala Ile Ala Ser
705 710 715 720
Tyr Ile Glu Ala Glu Arg Gln Gly Ser Lys Arg Pro Thr Phe Ala Ser
725 730 735
Val His Gly Arg Asp Ala Thr Val Val Arg Ala Ala Asp Leu Thr Leu
740 745 750
Asp Lys Phe Leu Asp Ala Glu Thr Leu Ala Ala Ala Pro Asn Leu Pro
755 760 765
Lys Pro Ala Thr Glu Val Arg Thr Val Leu Leu Thr Gly Ala Thr Gly
770 775 780
Phe Leu Gly Arg Tyr Leu Ala Leu Glu Trp Leu Glu Arg Met Asp Met
785 790 795 800
Val Asp Gly Lys Val Ile Ala Leu Val Arg Ala Arg Ser Asp Glu Glu
805 810 815
Ala Arg Ala Arg Leu Asp Lys Thr Phe Asp Ser Gly Asp Pro Lys Leu
820 825 830
Leu Ala His Tyr Gln Gln Leu Ala Ala Asp His Leu Glu Val Ile Ala
835 840 845
Gly Asp Lys Gly Glu Ala Asn Leu Gly Leu Gly Gln Asp Val Trp Gln
850 855 860
Arg Leu Ala Asp Thr Val Asp Val Ile Val Asp Pro Ala Ala Leu Val
865 870 875 880
Asn His Val Leu Pro Tyr Ser Glu Leu Phe Gly Pro Asn Ala Leu Gly
885 890 895
Thr Ala Glu Leu Ile Arg Leu Ala Leu Thr Ser Lys Gln Lys Pro Tyr
900 905 910
Thr Tyr Val Ser Thr Ile Gly Val Gly Asp Gln Ile Glu Pro Gly Lys
915 920 925
Phe Val Glu Asn Ala Asp Ile Arg Gln Met Ser Ala Thr Arg Ala Ile
930 935 940
Asn Asp Ser Tyr Ala Asn Gly Tyr Gly Asn Ser Lys Trp Ala Gly Glu
945 950 955 960
Val Leu Leu Arg Glu Ala His Asp Leu Cys Gly Leu Pro Val Ala Val
965 970 975
Phe Arg Cys Asp Met Ile Leu Ala Asp Thr Thr Tyr Ala Gly Gln Leu
980 985 990
Asn Leu Pro Asp Met Phe Thr Arg Leu Met Leu Ser Leu Val Ala Thr
995 1000 1005
Gly Ile Ala Pro Gly Ser Phe Tyr Glu Leu Asp Ala Asp Gly Asn
1010 1015 1020
Arg Gln Arg Ala His Tyr Asp Gly Leu Pro Val Glu Phe Ile Ala
1025 1030 1035
Ala Ala Ile Ser Thr Leu Gly Ser Gln Ile Thr Asp Ser Asp Thr
1040 1045 1050
Gly Phe Gln Thr Tyr His Val Met Asn Pro Tyr Asp Asp Gly Val
1055 1060 1065
Gly Leu Asp Glu Tyr Val Asp Trp Leu Val Asp Ala Gly Tyr Ser
1070 1075 1080
Ile Glu Arg Ile Ala Asp Tyr Ser Glu Trp Leu Arg Arg Phe Glu
1085 1090 1095
Thr Ser Leu Arg Ala Leu Pro Asp Arg Gln Arg Gln Tyr Ser Leu
1100 1105 1110
Leu Pro Leu Leu His Asn Tyr Arg Thr Pro Glu Lys Pro Ile Asn
1115 1120 1125
Gly Ser Ile Ala Pro Thr Asp Val Phe Arg Ala Ala Val Gln Glu
1130 1135 1140
Ala Lys Ile Gly Pro Asp Lys Asp Ile Pro His Val Ser Pro Pro
1145 1150 1155
Val Ile Val Lys Tyr Ile Thr Asp Leu Gln Leu Leu Gly Leu Leu
1160 1165 1170
<210> 39
<211> 1184
<212> PRT
<213> 脓肿分枝杆菌 DSM44196
<400> 39
Met Thr Val Thr Asn Glu Thr Asn Pro Gln Gln Glu Gln Leu Ser Arg
1 5 10 15
Arg Ile Glu Ser Leu Arg Glu Ser Asp Pro Gln Phe Arg Ala Ala Gln
20 25 30
Pro Asp Pro Ala Val Ala Glu Gln Val Leu Arg Pro Gly Leu His Leu
35 40 45
Ser Glu Ala Ile Ala Ala Leu Met Thr Gly Tyr Ala Glu Arg Pro Ala
50 55 60
Leu Gly Glu Arg Ala Arg Glu Leu Val Thr Asp Gln Asp Gly Arg Thr
65 70 75 80
Thr Leu Arg Leu Leu Pro Arg Phe Asp Thr Thr Thr Tyr Gly Glu Leu
85 90 95
Trp Ser Arg Thr Thr Ser Val Ala Ala Ala Trp His His Asp Ala Ala
100 105 110
His Pro Val Lys Ala Gly Asp Leu Val Ala Thr Leu Gly Phe Thr Ser
115 120 125
Ile Asp Tyr Thr Val Leu Asp Leu Ala Ile Met Ile Leu Gly Gly Val
130 135 140
Ala Val Pro Leu Gln Thr Ser Ala Pro Ala Ser Gln Trp Thr Thr Ile
145 150 155 160
Leu Ala Glu Ala Glu Pro Asn Thr Leu Ala Val Ser Ile Glu Leu Ile
165 170 175
Gly Ala Ala Met Glu Ser Val Arg Ala Thr Pro Ser Ile Lys Gln Val
180 185 190
Val Val Phe Asp Tyr Thr Pro Glu Val Asp Asp Gln Arg Glu Ala Phe
195 200 205
Glu Ala Ala Ser Thr Gln Leu Ala Gly Thr Gly Ile Ala Ile Glu Thr
210 215 220
Leu Asp Ala Val Ile Ala Arg Gly Ala Ala Leu Pro Ala Ala Pro Leu
225 230 235 240
Tyr Ala Pro Ser Ala Gly Asp Asp Pro Leu Ala Leu Leu Ile Tyr Thr
245 250 255
Ser Gly Ser Thr Gly Ala Pro Lys Gly Ala Met His Ser Glu Asn Ile
260 265 270
Val Arg Arg Trp Trp Ile Arg Glu Asp Val Met Ala Gly Thr Glu Asn
275 280 285
Leu Pro Met Ile Gly Leu Asn Phe Met Pro Met Ser His Ile Met Gly
290 295 300
Arg Gly Thr Leu Thr Ser Thr Leu Ser Thr Gly Gly Thr Gly Tyr Phe
305 310 315 320
Ala Ala Ser Ser Asp Met Ser Thr Leu Phe Glu Asp Met Glu Leu Ile
325 330 335
Arg Pro Thr Ala Leu Ala Leu Val Pro Arg Val Cys Asp Met Val Phe
340 345 350
Gln Arg Phe Gln Thr Glu Val Asp Arg Arg Leu Ala Ser Ser Asp Thr
355 360 365
Ala Ser Ala Glu Ala Val Ala Ala Glu Val Lys Ala Asp Ile Arg Asp
370 375 380
Asn Leu Phe Gly Gly Arg Val Ser Ala Val Met Val Gly Ser Ala Pro
385 390 395 400
Leu Ser Glu Glu Leu Gly Glu Phe Ile Glu Ser Cys Phe Glu Leu Asn
405 410 415
Leu Thr Asp Gly Tyr Gly Ser Thr Glu Ala Gly Met Val Phe Arg Asp
420 425 430
Gly Ile Val Gln Arg Pro Pro Val Ile Asp Tyr Lys Leu Val Asp Val
435 440 445
Pro Glu Leu Gly Tyr Phe Ser Thr Asp Lys Pro His Pro Arg Gly Glu
450 455 460
Leu Leu Leu Lys Thr Asp Gly Met Phe Leu Gly Tyr Tyr Lys Arg Pro
465 470 475 480
Glu Val Thr Ala Gly Val Phe Asp Ala Asp Gly Phe Tyr Met Thr Gly
485 490 495
Asp Ile Val Ala Glu Leu Ala His Asp Asn Ile Glu Ile Ile Asp Arg
500 505 510
Arg Asn Asn Val Leu Lys Leu Ser Gln Gly Glu Phe Val Ala Val Ala
515 520 525
Thr Leu Glu Ala Glu Tyr Ala Asn Ser Pro Val Val His Gln Ile Tyr
530 535 540
Val Tyr Gly Ser Ser Glu Arg Ser Tyr Leu Leu Ala Val Val Val Pro
545 550 555 560
Thr Pro Glu Ala Val Ala Ala Ala Lys Gly Asp Ala Ala Ala Leu Lys
565 570 575
Thr Thr Ile Ala Asp Ser Leu Gln Asp Ile Ala Lys Glu Ile Gln Leu
580 585 590
Gln Ser Tyr Glu Val Pro Arg Asp Phe Ile Ile Glu Pro Gln Pro Phe
595 600 605
Thr Gln Gly Asn Gly Leu Leu Thr Gly Ile Ala Lys Leu Ala Arg Pro
610 615 620
Asn Leu Lys Ala His Tyr Gly Pro Arg Leu Glu Gln Met Tyr Ala Glu
625 630 635 640
Ile Ala Glu Gln Gln Ala Ala Glu Leu Arg Ala Leu His Gly Val Asp
645 650 655
Pro Asp Lys Pro Ala Leu Glu Thr Val Leu Lys Ala Ala Gln Ala Leu
660 665 670
Leu Gly Val Ser Ser Ala Glu Leu Ala Ala Asp Ala His Phe Thr Asp
675 680 685
Leu Gly Gly Asp Ser Leu Ser Ala Leu Ser Phe Ser Asp Leu Leu Arg
690 695 700
Asp Ile Phe Ala Val Glu Val Pro Val Gly Val Ile Val Ser Ala Ala
705 710 715 720
Asn Asp Leu Ser Gly Val Ala Lys Phe Val Asp Glu Gln Arg Tyr Ser
725 730 735
Gly Gly Thr Arg Pro Thr Ala Glu Thr Val His Gly Ala Gly His Thr
740 745 750
Glu Ile Arg Ala Ala Asp Leu Thr Leu Asp Lys Phe Ile Asp Glu Ala
755 760 765
Thr Leu His Ala Ala Pro Ser Leu Pro Lys Ala Val Gly Ile Pro His
770 775 780
Thr Val Leu Leu Thr Gly Ser Asn Gly Tyr Leu Gly His Tyr Leu Ala
785 790 795 800
Leu Glu Trp Leu Glu Arg Leu Asp Lys Thr Glu Gly Lys Leu Ile Ala
805 810 815
Ile Val Arg Gly Lys Asn Ala Glu Ala Ala Tyr Arg Arg Leu Glu Glu
820 825 830
Ala Phe Asp Thr Gly Asp Thr Gln Leu Leu Ala His Phe Arg Ser Leu
835 840 845
Ala Asp Lys His Leu Glu Val Leu Ala Gly Asp Ile Gly Asp Pro Asn
850 855 860
Leu Gly Leu Asp Ala Asp Thr Trp Gln Arg Leu Ala Asp Thr Val Asp
865 870 875 880
Val Ile Val His Pro Ala Ala Leu Val Asn His Val Leu Pro Tyr Ser
885 890 895
Gln Leu Phe Gly Pro Asn Val Val Gly Thr Ala Glu Ile Ile Lys Leu
900 905 910
Ala Ile Thr Thr Lys Ile Lys Pro Val Thr Tyr Leu Ser Thr Val Ala
915 920 925
Val Ala Ala Tyr Val Asp Pro Thr Thr Phe Asp Glu Glu Ser Asp Ile
930 935 940
Arg Leu Ile Ser Ala Val Arg Pro Val Asp Glu Leu Tyr Ala Asn Gly
945 950 955 960
Tyr Gly Asn Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His
965 970 975
Asp Leu Cys Gly Leu Pro Val Ala Val Phe Arg Ser Asp Met Ile Leu
980 985 990
Ala His Ser Arg Tyr Thr Gly Gln Leu Asn Val Pro Asp Gln Phe Thr
995 1000 1005
Arg Leu Ile Leu Ser Leu Ile Ala Thr Gly Ile Ala Pro Gly Ser
1010 1015 1020
Phe Tyr Gln Ala His Ala Thr Gly Glu Arg Pro Leu Ala His Tyr
1025 1030 1035
Asp Gly Leu Pro Gly Asp Phe Thr Ala Glu Ala Ile Thr Thr Leu
1040 1045 1050
Gly Thr Gln Val Val Asp Ser Tyr Glu Thr Tyr Asp Cys Val Asn
1055 1060 1065
Pro His Ala Asp Gly Val Ser Leu Asp Asn Phe Val Asp Trp Leu
1070 1075 1080
Ile Glu Ala Gly Tyr Pro Ile Ala Arg Ile Asp Asn Tyr Thr Glu
1085 1090 1095
Trp Phe Thr Arg Phe Asp Thr Ala Ile Arg Ser Leu Pro Glu Lys
1100 1105 1110
Gln Lys Gln His Ser Leu Leu Pro Leu Leu His Ala Phe Glu Gln
1115 1120 1125
Pro Ser Ala Ala Glu Asn His Gly Val Val Pro Ala Lys Arg Phe
1130 1135 1140
Gln His Ala Val Gln Ala Ala Gly Ile Gly Pro Ala Gly Gln Asp
1145 1150 1155
Gly Thr Thr Asp Ile Pro His Leu Ser Arg Arg Leu Ile Val Lys
1160 1165 1170
Tyr Ala Lys Asp Leu Glu Gln Leu Gly Leu Leu
1175 1180
<210> 40
<211> 351
<212> PRT
<213> 根瘤农杆菌 ATCC 33970
<400> 40
Met Gln Pro Phe Val Tyr Met Ala Ala Pro Ala Arg Ile Val Phe Ser
1 5 10 15
Ala Gly Ser Ser Ala Asp Val Ala Glu Glu Ile Arg Arg Leu Gly Leu
20 25 30
Ser Arg Ala Leu Val Leu Ser Thr Pro Gln Gln Lys Gly Asp Ala Glu
35 40 45
Ala Leu Ala Ser Arg Leu Gly Arg Leu Ala Ala Gly Val Phe Ser Glu
50 55 60
Ala Ala Met His Thr Pro Val Glu Val Thr Lys Thr Ala Val Glu Ala
65 70 75 80
Tyr Arg Ala Ala Gly Ala Asp Cys Val Val Ser Leu Gly Gly Gly Ser
85 90 95
Thr Thr Gly Leu Gly Lys Ala Ile Ala Leu Arg Thr Asp Ala Ala Gln
100 105 110
Ile Val Ile Pro Thr Thr Tyr Ala Gly Ser Glu Val Thr Pro Ile Leu
115 120 125
Gly Gln Thr Glu Asn Gly Val Lys Thr Thr Met Arg Gly Pro Glu Ile
130 135 140
Leu Pro Glu Val Val Ile Tyr Asp Ala Glu Leu Thr Leu Gly Leu Pro
145 150 155 160
Val Ala Ile Ser Met Thr Ser Gly Leu Asn Ala Met Ala His Ala Ala
165 170 175
Glu Ala Leu Tyr Ala Arg Asp Arg Asn Pro Ile Ala Ser Met Met Ala
180 185 190
Val Glu Gly Leu Arg Ala Met Ile Glu Ala Leu Pro Val Val Arg Gln
195 200 205
Ala Pro His Asp Ile Gly Ala Arg Glu Thr Ala Leu Tyr Gly Ala Trp
210 215 220
Leu Cys Gly Thr Val Leu Gly Ala Val Gly Met Ser Leu His His Lys
225 230 235 240
Leu Cys His Thr Leu Gly Gly Ser Leu Asp Leu Pro His Ala Glu Thr
245 250 255
His Ala Val Leu Leu Pro His Thr Ile Ala Tyr Val Glu Glu Ala Ala
260 265 270
Pro Asn Leu Leu Ala Pro Leu Ala Ala Leu Val Gly Gly Arg Ala Gly
275 280 285
Ala Gly Leu Phe Asp Phe Ala Ala Arg Leu Gly Ala Pro Ser Ser Leu
290 295 300
Ala Ala Leu Gly Val Gly Ala Asp Asp Leu Asp Pro Met Ala Glu Leu
305 310 315 320
Ala Thr Ala Asn Pro Tyr Trp Cys Pro Arg Pro Ile Glu Lys Thr Ala
325 330 335
Ile Arg Asp Leu Leu Gln Arg Ala Phe Glu Gly Ala Arg Pro Ala
340 345 350
<210> 41
<211> 246
<212> PRT
<213> 耐盐嗜碱芽孢杆菌ATCC BAA-125
<400> 41
Met Leu Gln Gly Lys Thr Ala Ile Val Thr Gly Ala Ser Arg Gly Ile
1 5 10 15
Gly Arg Ala Thr Ala Met Glu Leu Ala Arg His Gly Ala Asn Val Val
20 25 30
Val Asn Tyr Ala Gly Asn Lys Glu Lys Ala Glu Lys Val Val Ala Glu
35 40 45
Ile Lys Glu Leu Gly Val Glu Ala Ile Ala Ile Gln Ala Asp Val Ala
50 55 60
Asp Ser Glu Ser Val Gln Ala Met Val Lys Glu Thr Ile Asp Thr Phe
65 70 75 80
Gly Ala Val Asp Ile Leu Val Asn Asn Ala Gly Ile Thr Arg Asp Asn
85 90 95
Leu Phe Met Arg Met Lys Glu Glu Asp Trp Asp Ala Val Ile Asp Thr
100 105 110
Asn Leu Lys Gly Val Phe His Cys Ser Lys Ala Val Thr Arg Pro Met
115 120 125
Met Lys Gln Arg Phe Gly Arg Ile Ile Asn Val Ser Ser Val Val Gly
130 135 140
Ala Ile Gly Asn Ala Gly Gln Ala Asn Tyr Val Ala Ala Lys Ala Gly
145 150 155 160
Val Ile Gly Leu Thr Lys Thr Leu Ala Arg Glu Leu Ala Asn Arg Asn
165 170 175
Ile Thr Val Asn Ala Val Ala Pro Gly Phe Ile Glu Thr Asp Met Thr
180 185 190
Gly Glu Leu Pro Glu Asp Val Lys Ala Gln Met Leu Gly Gln Ile Pro
195 200 205
Leu Ala Arg Leu Gly Gln Pro Glu Glu Val Ala Lys Ala Val Arg Phe
210 215 220
Leu Ala Ser Asp Asp Ala Ser Tyr Leu Thr Gly Gln Thr Ile His Val
225 230 235 240
Asn Gly Gly Met Val Met
245
<210> 42
<211> 250
<212> PRT
<213> 紫色色杆菌ATCC 12472
<400> 42
Met Arg Arg Leu Glu Gly Lys Gln Ala Leu Ile Thr Gly Gly Thr Ser
1 5 10 15
Gly Ile Gly Leu Glu Thr Ala Lys Gln Phe Leu Ala Glu Gly Ala Lys
20 25 30
Val Ala Ile Thr Gly Arg Ser Glu Ala Gly Leu Ala Gln Ala Asn Glu
35 40 45
Val Leu Gly Gly Arg Ala Leu Leu Leu Lys Ser Asp Ala Gly Ser Leu
50 55 60
Asp Asp Gln Arg Ala Leu Pro Ala Gln Leu Arg Ser His Gly Trp Ser
65 70 75 80
Arg Leu Asp Ala Ala Tyr Leu Asn Ala Gly Asp Val Thr His Leu Pro
85 90 95
Leu Ala Asp Trp Ser Glu Ala Asp Phe Asp Arg Val Met Ala Val Asn
100 105 110
Leu Lys Gly Pro Phe Phe Leu Leu Gln Ala Leu Ser Pro Leu Leu Ala
115 120 125
Asn Pro Ser Ser Val Ile Leu Cys Gly Ser Val Gly Ala Arg Ile Gly
130 135 140
Val Pro Ser Ser Ser Ala Tyr Ala Ala Ser Lys Ala Gly Leu Leu Ser
145 150 155 160
Leu Ala Arg Thr Leu Ser Ala Glu Trp Leu Asp Arg Gly Ile Arg Val
165 170 175
Asn Gly Leu Ser Pro Gly Pro Thr His Thr Pro Ala Phe Gly Lys Leu
180 185 190
Gly Met Pro Glu Asp Glu Leu Pro Ala Leu Arg Asp Arg Ile Arg Lys
195 200 205
Leu Val Pro Leu Gly Arg Leu Gly Arg Pro Glu Glu Leu Ala Lys Ala
210 215 220
Ala Val Phe Leu Ala Ser Asp Glu Ser Ser Tyr Met Leu Gly Ser Glu
225 230 235 240
Leu Leu Val Asp Gly Gly Val Gly Asn Val
245 250
<210> 43
<211> 393
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 43
Met Ser Asp Leu His Tyr Trp Asn Tyr Pro Thr Asp Ile Leu Cys Gly
1 5 10 15
Val Gly Ala Leu Glu Gln Leu Pro Arg Arg Cys Ala Leu Ala Gly Ala
20 25 30
Arg Arg Pro Leu Leu Val Thr Asp Pro Gly Met Arg Ala Leu Glu Pro
35 40 45
Leu Arg Leu Val Arg Glu Cys Leu Glu Arg Ala Gly Ile Asp His Asp
50 55 60
Leu Phe His Glu Leu Ser Ser Asn Pro Ser Leu Ala Glu Val Gln Gly
65 70 75 80
Gly Ala Arg Arg Phe Glu Ala Gly Gly His Asp Ala Leu Ile Ala Leu
85 90 95
Gly Gly Gly Ser Ala Leu Asp Ala Ala Lys Gly Ile Ala Leu Leu Ser
100 105 110
Arg Asp Pro His Gly Leu Glu Arg Phe Glu Trp Thr Gln Thr Leu Arg
115 120 125
Ser Tyr Pro Thr Leu Ala Asp Tyr Pro Pro Leu Gly Leu Pro Pro Leu
130 135 140
Leu Ala Leu Pro Thr Thr Ala Gly Thr Gly Ser Glu Leu Gly Arg Glu
145 150 155 160
Ala Val Leu Thr Asp Thr Gln Leu Gly Ile Lys Arg Val Val Gly His
165 170 175
Arg Glu Leu Leu Ala Ala Cys Val Phe Leu Asp Pro Arg Leu Thr Arg
180 185 190
Gly Leu Pro Pro Ala Leu Ser Ala Ala Thr Gly Met Asp Ala Leu Thr
195 200 205
His His Leu Glu Ala Leu Phe Ser Pro Leu Tyr His Pro Met Ser Ala
210 215 220
Gly Ile Ala Leu Glu Gly Val Arg Leu Val Arg Gln His Leu Glu Asn
225 230 235 240
Ala Val Arg Asp Gly Asn Asp Leu Ala Ala Arg Glu Gly Met Leu Val
245 250 255
Ala Ser Ala Ser Ala Ala Val Ala Phe Gln Lys Gly Leu Gly Gly Val
260 265 270
His Ala Leu Ala His Pro Leu Gly Ala Arg His His Leu His His Gly
275 280 285
Leu Leu Asn Ala Val Leu Leu Pro Tyr Val Leu Leu Ala Asn Arg Pro
290 295 300
Ala Ile Glu Ala Asp Ala Ala Arg Leu Ala Arg Tyr Leu Glu Leu Asp
305 310 315 320
Glu Ala Ser Phe Asp Gly Leu Leu Ala Trp Ile Leu Glu Leu Arg Ala
325 330 335
Arg Ile Gly Ile Pro Ala Asn Leu Ala Ala Leu Gly Leu Asp Gly Glu
340 345 350
Asp Ala Gln Trp Val Gly Glu Gln Ala Leu Ala Asp Leu Ser Ser Ser
355 360 365
Ala Thr Asn Ala Leu Pro Leu Asp Ala Arg Asp Tyr Ala Arg Ile Tyr
370 375 380
Arg Gln Ala Val Ala Gly Thr Leu Ala
385 390
<210> 44
<211> 257
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 44
Met Val Asp His Ser Ile Lys Gly Lys Thr Val Leu Ile Thr Gly Gly
1 5 10 15
Ala Lys Asn Leu Gly Gly Leu Ile Ala Arg Asp Leu Ala Ala His Gly
20 25 30
Ala Lys Ala Ile Thr Ile His Tyr Asn Ser Ala Ala Ser Lys Ala Asp
35 40 45
Ala Asp Ala Thr Val Ala Ala Leu Gln Ala Ala Gly Ala Lys Ala Val
50 55 60
Ala Leu Gln Gly Asp Leu Thr Ser Ala Ala Ala Met Glu Lys Leu Phe
65 70 75 80
Ala Asp Ala Ile Ala Ala Val Gly Lys Pro Asp Ile Ala Ile Asn Thr
85 90 95
Val Gly Lys Val Leu Lys Lys Pro Ile Thr Glu Ile Ser Glu Thr Glu
100 105 110
Tyr Asp Glu Met Ser Ala Val Asn Ser Lys Ser Ala Phe Phe Phe Leu
115 120 125
Arg Glu Ala Gly Lys His Val Asn Asp Asn Gly Lys Ile Cys Thr Leu
130 135 140
Val Thr Ser Leu Leu Gly Ala Tyr Thr Pro Tyr Tyr Ala Ala Tyr Ala
145 150 155 160
Gly Thr Lys Ala Pro Val Glu His Phe Thr Arg Ala Ala Ser Lys Glu
165 170 175
Phe Gly Ala Arg Gly Ile Ser Val Thr Ala Val Gly Pro Gly Pro Met
180 185 190
Asp Thr Pro Phe Phe Tyr Pro Ala Glu Gly Ala Asp Ala Val Asp Tyr
195 200 205
Leu Lys Asn Ala Ala Ala Leu Ser Pro Phe Ser Lys Thr Gly Leu Thr
210 215 220
Asp Ile Asp Asp Val Val Pro Phe Ile Arg His Leu Val Ser Glu Gly
225 230 235 240
Trp Trp Ile Thr Gly Gln Thr Ile Leu Ile Asn Gly Gly Phe Ser Thr
245 250 255
Lys
<210> 45
<211> 331
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 45
Met Gln Tyr Val Lys Leu Gly Ser Thr Gly Leu Asp Ile Ser Arg Leu
1 5 10 15
Cys Leu Gly Cys Met Thr Phe Gly Glu Pro Asp Ala Gly Thr His Pro
20 25 30
Trp Thr Leu Gly Glu Asp Ala Ser Arg Pro Ile Ile Arg His Ala Val
35 40 45
Glu Gln Gly Ile Asn Phe Phe Asp Thr Ala Asn Ser Tyr Ser Ala Gly
50 55 60
Thr Ser Glu Ile Ile Leu Gly Lys Leu Leu Arg Glu Phe Thr Arg Arg
65 70 75 80
Glu Glu Thr Val Ile Ala Thr Lys Val Phe Phe Pro Ala Asn Met Trp
85 90 95
Glu Gly Ala Ser Arg Pro Asn Glu Gln Gly Leu Ser Arg Lys Ala Ile
100 105 110
Met Ala Asn Ile Asp Ala Ser Leu Ser Arg Leu Gly Thr Asp Tyr Val
115 120 125
Asp Leu Tyr Gln Ile His Arg Trp Asp Tyr His Thr Pro Ile Glu Glu
130 135 140
Thr Met Glu Ala Leu His Asp Val Val Lys Ala Gly Lys Ala Arg Tyr
145 150 155 160
Ile Gly Ala Ser Ser Met Tyr Ala Trp Gln Phe Ala Lys Ala Gln Gln
165 170 175
Val Ala Ala Asn His Gly Trp Ser Arg Phe Val Ser Met Gln Asn Tyr
180 185 190
Leu Asn Leu Ile Tyr Arg Glu Glu Glu Arg Glu Met Ile Pro Leu Cys
195 200 205
Leu Asp Gln Gly Val Gly Leu Met Pro Trp Ser Pro Met Ala Arg Gly
210 215 220
Arg Leu Thr Arg Pro His Gly Gln Gln Thr Gln Arg Thr Arg Thr Asp
225 230 235 240
Val Ser Gly Gln Ser Phe Tyr Glu Lys Thr Glu Val Glu Asp Gly Arg
245 250 255
Val Ile Asp Val Val Glu Gln Ile Ala Ser Glu Arg Gly Val Pro Met
260 265 270
Ala Gln Val Ala Leu Ala Trp Val Leu Gly Arg Ala Gly Val Ser Ala
275 280 285
Pro Ile Val Gly Ala Ser Lys Pro Ala His Leu Asp Asp Ala Leu Gly
290 295 300
Ala Leu Ser Leu Gln Leu Ser Glu Asp Glu Val Ala Arg Leu Gln Ala
305 310 315 320
Pro Tyr Val Pro His Ala Val Thr Gly Phe Lys
325 330
<210> 46
<211> 249
<212> PRT
<213> 丁香假单胞菌番茄致病变种 DC3000
<400> 46
Met Ile Glu His Leu Lys Leu Ala Gly Lys Val Ala Leu Val Gln Gly
1 5 10 15
Gly Ser Arg Gly Ile Gly Ala Ala Ile Val Asn Arg Leu Ala Lys Glu
20 25 30
Gly Ala Ala Val Ala Phe Thr Tyr Ile Asn Ser Glu Val Asn Ala Leu
35 40 45
Glu Ile Gln Asp Ser Ile Asn Ala Asn Gly Gly Arg Ala Leu Ala Ile
50 55 60
Arg Ala Asp Ser Ala Asp Glu Lys Ala Ile Arg Gln Ala Val Gln Thr
65 70 75 80
Thr Ala Glu Thr Leu Gly Arg Leu Asp Ile Leu Val Asn Asn Ala Gly
85 90 95
Val Leu Ala Ile Ala Pro Leu Asn Glu Phe Ser Met Gln Asp Phe Asp
100 105 110
Lys Thr Leu Ala Ile Asn Val Arg Ser Val Phe Ile Ala Ser Gln Glu
115 120 125
Ala Ala Arg His Met Glu Glu Gly Gly Arg Ile Ile Asn Ile Gly Ser
130 135 140
Thr Asn Ala Glu Arg Met Pro Phe Ala Gly Gly Ala Thr Tyr Ala Met
145 150 155 160
Ser Lys Ser Ala Leu Ile Gly Leu Thr Lys Gly Met Ala Arg Asp Leu
165 170 175
Gly Pro Gln Gly Ile Thr Val Asn Asn Val Gln Pro Gly Pro Val Asp
180 185 190
Thr Asp Met Asn Pro Ala Gln Gly Glu Phe Ala Glu Ser Leu Lys Ala
195 200 205
Leu Met Ala Leu Pro Arg Tyr Gly Lys Ser Glu Glu Ile Ala Ser Phe
210 215 220
Val Ala Tyr Leu Ala Gly Pro Glu Ala Gly Tyr Ile Thr Gly Ala Ser
225 230 235 240
Leu Thr Ile Asp Gly Gly Phe Ser Ala
245
<210> 47
<211> 259
<212> PRT
<213> 乔斯特红球菌菌株 RHA1
<400> 47
Met Ser Ile Lys Gly Lys Val Ala Leu Val Thr Gly Ala Gly Gln Gly
1 5 10 15
Ile Gly Arg Ala Ile Ala Leu Arg Leu Ala Ser Asp Gly Ala Asp Ile
20 25 30
Ser Leu Val Asp Val Asn Gly Asp Arg Leu Asp Ala Val Ala Asp Glu
35 40 45
Val Arg Ala Val Gly Val Lys Ala Ile Ser Leu Val Ala Asp Val Thr
50 55 60
Asp Arg Asp Gln Val Gln Ser Ala Val Asp Arg Thr Glu Arg Glu Leu
65 70 75 80
Gly Gly Phe Asp Ile Ile Val Asn Asn Ala Gly Ile Ala Gln Val Asn
85 90 95
Pro Ile Ala Asp Val Thr Pro Glu Glu Val Ser Arg Ile Leu Ala Val
100 105 110
Asn Val Glu Gly Val Leu Trp Gly Ile Gln Val Gly Ala Ala Lys Phe
115 120 125
Arg Glu Arg Gly His Gly Gly Lys Ile Ile Asn Ala Ser Ser Ile Ala
130 135 140
Gly His Glu Gly Phe Pro Met Leu Gly Val Tyr Ser Ala Thr Lys Phe
145 150 155 160
Ala Val Arg Gly Leu Thr Gln Ala Ala Ala Lys Glu Tyr Ala Ala Asp
165 170 175
Gly Ile Thr Val Asn Ala Tyr Cys Pro Gly Val Val Gly Thr Asp Met
180 185 190
Trp Val Thr Ile Asp Glu Arg Phe Ala Ala Leu Thr Gly Ala Pro Lys
195 200 205
Gly Ala Thr Phe Glu Lys Phe Val Gly Gly Ile Ala Leu Gly Arg Ala
210 215 220
Gln Thr Pro Glu Asp Val Ala Ala Tyr Val Ser Tyr Leu Ala Gly Pro
225 230 235 240
Asp Ser Asp Tyr Met Thr Gly Gln Ser Gly Leu Ile Asp Gly Gly Leu
245 250 255
Val Tyr Arg
<210> 48
<211> 391
<212> PRT
<213> 乔斯特红球菌 RHA1
<400> 48
Met Pro Glu Ile Val Lys Phe Glu Met Pro Arg Val Glu Thr Val Val
1 5 10 15
Gln Gly Val Gly Ala Val Arg Glu Ile Gly Ala Glu Leu Asp Arg Arg
20 25 30
Asp Leu Lys Arg Val Phe Leu Val Thr Gly His Ser Val Gly Ser Ser
35 40 45
Glu Ala Phe Ala Ala Leu Ala Ser Asp Leu Gly Asp Arg Val Val Gly
50 55 60
Ile Phe Asp Gln Val Lys Ala His Asn Pro Val Glu Leu Ile Val Glu
65 70 75 80
Leu Ile Thr Ala Ala Lys Ser Ala Gln Ala Glu Ala Phe Val Ala Val
85 90 95
Gly Gly Gly Ser Pro Val Asp Ala Ala Lys Leu Ala Ala Ile Gly Leu
100 105 110
Cys Glu Gly Ser Glu Ser Val Glu Asp Leu Ala Arg Asn Tyr Leu Val
115 120 125
Phe Glu Tyr Pro Asn Thr Ile His Gln Lys Pro Leu Thr Gly Thr Pro
130 135 140
Met Pro Val Phe Ala Val Pro Thr Thr Leu Ser Ala Ala Glu Trp Asp
145 150 155 160
Gly Phe Ala Gly Ser Val Asp His Thr Arg Asp Thr Lys Asp Leu Thr
165 170 175
Val Tyr Leu Glu Ala Thr Pro Gln Val Val Phe Leu Asp Pro Glu Phe
180 185 190
Cys Ala His Thr Pro Arg Asp Leu Trp Ala Thr Thr Gly Val Arg Ala
195 200 205
Leu Asp His Ala Val Glu Thr Ala Tyr Ala Lys Asn Ala His Pro Phe
210 215 220
Thr Thr Ala Leu Ala Asn Gly Ala Leu Thr Met Leu Ala Glu Asn Leu
225 230 235 240
Pro Arg Ser Val Lys Asp Pro His Asp Tyr Asp Ala Ala Leu Lys Cys
245 250 255
Leu Glu Ala Ala Trp Met Ser Ile Ile Gly Val His Asn Val Ser Leu
260 265 270
Gly Leu Ser His Ala Ile Gly His Gln Leu Gly Ala Val Gly Ile Pro
275 280 285
His Gly Val Thr Ser Cys Ile Met Leu Pro His Val Met Arg Phe Leu
290 295 300
Glu Pro Val Thr Ser Ala Glu Gln Ala Arg Met Ala Gln Ser Leu Ala
305 310 315 320
Gln Val Gln Gly Asp Gly Glu Asp Leu Pro Ala Ala Asp Arg Leu Glu
325 330 335
Arg Ile Leu Asp Glu Leu Gly Val Pro Arg Arg Val Ser Glu Phe Gly
340 345 350
Val Gly Arg Asp Lys Met Asp Gly Val Ala Arg Ala Thr Leu Gly Asp
355 360 365
Ile Val Val Arg Glu Ser Pro Arg Pro Val Asp Glu Thr Val Val Tyr
370 375 380
Gly Leu Leu Glu Thr Val Trp
385 390
<210> 49
<211> 245
<212> PRT
<213> 沼泽红假单胞菌 ATCC BAA98
<400> 49
Met Thr Pro Arg Asn Ala Thr Val Ala Val Ile Gly Ala Gly Asp Tyr
1 5 10 15
Ile Gly Ala Glu Ile Ala Lys Lys Phe Ala Ala Glu Gly Phe Thr Val
20 25 30
Phe Ala Gly Arg Arg Asn Gly Glu Lys Leu Ala Pro Leu Val Ala Glu
35 40 45
Ile Glu Ala Ala Gly Gly Arg Ile Val Ala Arg Ser Leu Asp Ala Arg
50 55 60
Asn Glu Asp Glu Val Thr Ala Phe Leu Asn Ala Ala Asp Ala His Ala
65 70 75 80
Pro Leu Glu Val Thr Ile Phe Asn Val Gly Ala Asn Val Asn Phe Pro
85 90 95
Ile Leu Glu Thr Thr Asp Arg Val Phe Arg Lys Val Trp Glu Met Ala
100 105 110
Cys Trp Ala Gly Phe Val Ser Gly Arg Glu Ser Ala Arg Leu Met Leu
115 120 125
Ala His Gly Gln Gly Lys Ile Phe Phe Thr Gly Ala Thr Ala Ser Leu
130 135 140
Arg Gly Gly Ser Gly Phe Ala Ala Phe Ala Ser Ala Lys Phe Gly Leu
145 150 155 160
Arg Ala Val Ala Gln Ser Met Ala Arg Glu Leu Met Pro Lys Asn Ile
165 170 175
His Val Ala His Leu Ile Ile Asp Ser Gly Val Asp Thr Ala Trp Val
180 185 190
Arg Glu Arg Arg Glu Gln Met Phe Gly Lys Asp Ala Leu Ala Asn Pro
195 200 205
Asp Leu Leu Met Pro Pro Ala Ala Val Ala Gly Ala Tyr Trp Gln Leu
210 215 220
Tyr Gln Gln Pro Lys Ser Ala Trp Thr Phe Glu Met Glu Ile Arg Pro
225 230 235 240
Tyr Gly Glu Lys Trp
245
<210> 50
<211> 282
<212> PRT
<213> 天蓝色链霉菌 ATCC BAA471
<400> 50
Met Gln Arg Gln Asn Trp Leu Ile Thr Gly Val Ser Thr Gly Leu Gly
1 5 10 15
Arg Ala Phe Ala Gln Ala Ala Leu Ser Ala Gly His Thr Val Val Gly
20 25 30
Thr Val Arg Ser Glu Asn Asp Val Arg Ala Phe Glu Glu Leu Gly Pro
35 40 45
Gly His Ala His Gly Arg Ile Leu Asp Val Thr Asp Gly Gly Ala Val
50 55 60
Asn Gly Val Val Ala Glu Val Glu Arg Ser Val Gly Pro Leu Asp Val
65 70 75 80
Val Val Ala Asn Ala Gly Tyr Gly Leu Glu Gly Thr Phe Glu Glu Thr
85 90 95
Pro Leu Ala Glu Val Arg Arg Gln Phe Asp Val Asn Val Phe Gly Ala
100 105 110
Met Ala Thr Leu Gln Ala Ala Leu Pro His Met Arg Ala Arg Arg Arg
115 120 125
Gly His Leu Met Ala Val Thr Ser Met Gly Gly Leu Met Ala Val Pro
130 135 140
Gly Met Ser Ala Tyr Cys Gly Ser Lys Phe Ala Leu Glu Gly Ile Leu
145 150 155 160
Glu Ala Leu Gly Lys Glu Val Ala Gln Phe Gly Ile His Val Thr Ala
165 170 175
Ile Glu Pro Gly Ser Phe Arg Thr Asp Trp Ala Gly Arg Ser Met Thr
180 185 190
Arg Ala Ala Arg Thr Val Ala Asp Tyr Asp Asp Leu Phe Ala Pro Ile
195 200 205
Arg Glu Ala Arg Gln Lys Ala Ser Gly Asn Gln Leu Gly Asn Pro Ala
210 215 220
Lys Ala Gly Glu Ala Val Val His Ile Thr Ser Val Asp Gln Pro Pro
225 230 235 240
Ala His Leu Val Leu Gly Ser Asp Ala Leu Arg Leu Val Thr Ala Ala
245 250 255
Arg Thr Ala Val Asp Glu Asp Ile Arg Ala Trp Glu Ala Leu Ser Arg
260 265 270
Thr Thr Asp Phe Pro Gln Gly Ala Gln Leu
275 280
<210> 51
<211> 252
<212> PRT
<213> 福氏志贺氏菌 Shi06HN006
<400> 51
Met His Tyr Gln Pro Lys Gln Asp Leu Leu Asn Asp Arg Ile Ile Leu
1 5 10 15
Val Thr Gly Ala Ser Asp Gly Ile Gly Arg Glu Ala Ala Met Thr Tyr
20 25 30
Ala Arg Tyr Gly Ala Thr Val Ile Leu Leu Gly Arg Asn Glu Glu Lys
35 40 45
Leu Arg Gln Val Ala Ser His Ile Asn Glu Glu Thr Gly Arg Gln Pro
50 55 60
Gln Trp Phe Ile Leu Asp Leu Leu Thr Cys Thr Ser Glu Asn Cys Gln
65 70 75 80
Gln Leu Ala Gln Arg Ile Val Val Asn Tyr Pro Arg Leu Asp Gly Val
85 90 95
Leu His Asn Ala Gly Leu Leu Gly Asp Val Cys Pro Met Ser Glu Gln
100 105 110
Asn Pro Gln Val Trp Gln Asp Val Met Gln Ile Asn Val Asn Ala Thr
115 120 125
Phe Met Leu Thr Gln Ala Leu Leu Pro Leu Leu Leu Lys Ser Asp Ala
130 135 140
Gly Ser Leu Val Phe Thr Ser Ser Ser Val Gly Arg Gln Gly Arg Ala
145 150 155 160
Asn Trp Gly Ala Tyr Ala Ala Ser Lys Phe Ala Thr Glu Gly Met Met
165 170 175
Gln Val Leu Ala Asp Glu Tyr Gln Gln Arg Leu Arg Val Asn Cys Ile
180 185 190
Asn Pro Gly Gly Thr Arg Thr Ala Met Arg Ala Ser Ala Phe Pro Thr
195 200 205
Glu Asp Pro Gln Lys Leu Lys Thr Pro Ala Asp Ile Met Pro Leu Tyr
210 215 220
Leu Trp Leu Met Gly Asp Asp Ser Arg Arg Lys Thr Gly Met Thr Phe
225 230 235 240
Asp Ala Gln Pro Gly Arg Lys Pro Gly Ile Ser Gln
245 250
<210> 52
<211> 387
<212> PRT
<213> 海栖热孢菌 ATCC 43589
<400> 52
Met Phe Lys Ile Ser Phe Tyr Leu Pro Thr Glu Ile Ile Phe Arg Val
1 5 10 15
Gly Ala Val Asp Glu Leu Glu Glu Arg Ala Lys Lys Leu Gly Lys Lys
20 25 30
Ala Leu Ile Val Thr Gly Arg Ser Ser Thr Lys Lys Thr Gly Leu Leu
35 40 45
Gln Arg Val Val Asp Leu Leu Lys Lys Ala Gly Val Glu Ser Phe Val
50 55 60
Phe Asp Lys Ile Val Pro Asn Pro Ile Ser Asp His Val Asp Glu Ala
65 70 75 80
Ala Glu Ile Val Arg Lys Glu Lys Ile Asp Phe Ile Ile Gly Leu Gly
85 90 95
Gly Gly Ser Pro Ile Asp Ser Ala Lys Ala Ile Ser Ile Thr Ala Pro
100 105 110
Asn Glu Gly Lys Phe Trp Asp Tyr Val Pro Val Gly Gly Gly Lys Ile
115 120 125
Pro Glu Lys Ser Ile Pro Val Val Ala Ile Pro Thr Thr His Gly Thr
130 135 140
Gly Thr Glu Ala Asp Pro Phe Ala Val Ile Thr Asn Pro Gln Thr Lys
145 150 155 160
Glu Lys Val Gly Ile Gly Tyr Arg Asn Thr Phe Pro Val Leu Ser Leu
165 170 175
Val Asp Pro Glu Val Met Lys Thr Leu Pro Lys Asp Gln Thr Ala Tyr
180 185 190
Thr Ser Met Asp Ala Phe Tyr His Ala Ile Glu Ala Phe Leu Asn Val
195 200 205
Asn Ala Asn Pro Tyr Ser Asp Val Leu Ala Leu Asp Ala Met Lys Arg
210 215 220
Ile Val Thr Tyr Leu Pro Val Ala Tyr Glu Asn Gly Glu Asp Met Glu
225 230 235 240
Ala Arg Thr Asn Leu Ala Trp Ala Ser Thr Glu Ala Gly Ile Thr Glu
245 250 255
Thr Leu Thr Gly Val Ile Ala Asn His Ala Leu Glu His Gly Leu Ser
260 265 270
Gly Phe Tyr Pro Glu Ile Thr His Gly Leu Gly Leu Cys Ile Thr Gly
275 280 285
Pro Tyr Leu Phe Glu Tyr Ile Phe Asp His Ala Tyr Glu Arg Leu Ala
290 295 300
Ile Val Gly Arg Glu Val Phe Gly Val Tyr Glu Thr Asp Asp Arg Lys
305 310 315 320
Ala Gly Arg Leu Ala Ile Lys Lys Leu Arg Asp Phe Gln Glu Met Phe
325 330 335
Gly Leu Asn Lys Arg Leu Ser Glu Leu Gly Val Lys Lys Glu Asp Ile
340 345 350
Pro Lys Met Ala Glu Thr Gly Tyr Arg Ile Leu Asn Gly Val Val Val
355 360 365
Val Thr Pro Gly Asn Leu Thr Ala Lys Asp Met Glu Glu Ile Phe Asn
370 375 380
Arg Cys Tyr
385
<210> 53
<211> 426
<212> PRT
<213> 大肠杆菌 K12
<400> 53
Met Asn Ser Asn Lys Glu Leu Met Gln Arg Arg Ser Gln Ala Ile Pro
1 5 10 15
Arg Gly Val Gly Gln Ile His Pro Ile Phe Ala Asp Arg Ala Glu Asn
20 25 30
Cys Arg Val Trp Asp Val Glu Gly Arg Glu Tyr Leu Asp Phe Ala Gly
35 40 45
Gly Ile Ala Val Leu Asn Thr Gly His Leu His Pro Lys Val Val Ala
50 55 60
Ala Val Glu Ala Gln Leu Lys Lys Leu Ser His Thr Cys Phe Gln Val
65 70 75 80
Leu Ala Tyr Glu Pro Tyr Leu Glu Leu Cys Glu Ile Met Asn Gln Lys
85 90 95
Val Pro Gly Asp Phe Ala Lys Lys Thr Leu Leu Val Thr Thr Gly Ser
100 105 110
Glu Ala Val Glu Asn Ala Val Lys Ile Ala Arg Ala Ala Thr Lys Arg
115 120 125
Ser Gly Thr Ile Ala Phe Ser Gly Ala Tyr His Gly Arg Thr His Tyr
130 135 140
Thr Leu Ala Leu Thr Gly Lys Val Asn Pro Tyr Ser Ala Gly Met Gly
145 150 155 160
Leu Met Pro Gly His Val Tyr Arg Ala Leu Tyr Pro Cys Pro Leu His
165 170 175
Gly Ile Ser Glu Asp Asp Ala Ile Ala Ser Ile His Arg Ile Phe Lys
180 185 190
Asn Asp Ala Ala Pro Glu Asp Ile Ala Ala Ile Val Ile Glu Pro Val
195 200 205
Gln Gly Glu Gly Gly Phe Tyr Ala Ser Ser Pro Ala Phe Met Gln Arg
210 215 220
Leu Arg Ala Leu Cys Asp Glu His Gly Ile Met Leu Ile Ala Asp Glu
225 230 235 240
Val Gln Ser Gly Ala Gly Arg Thr Gly Thr Leu Phe Ala Met Glu Gln
245 250 255
Met Gly Val Ala Pro Asp Leu Thr Thr Phe Ala Lys Ser Ile Ala Gly
260 265 270
Gly Phe Pro Leu Ala Gly Val Thr Gly Arg Ala Glu Val Met Asp Ala
275 280 285
Val Ala Pro Gly Gly Leu Gly Gly Thr Tyr Ala Gly Asn Pro Ile Ala
290 295 300
Cys Val Ala Ala Leu Glu Val Leu Lys Val Phe Glu Gln Glu Asn Leu
305 310 315 320
Leu Gln Lys Ala Asn Asp Leu Gly Gln Lys Leu Lys Asp Gly Leu Leu
325 330 335
Ala Ile Ala Glu Lys His Pro Glu Ile Gly Asp Val Arg Gly Leu Gly
340 345 350
Ala Met Ile Ala Ile Glu Leu Phe Glu Asp Gly Asp His Asn Lys Pro
355 360 365
Asp Ala Lys Leu Thr Ala Glu Ile Val Ala Arg Ala Arg Asp Lys Gly
370 375 380
Leu Ile Leu Leu Ser Cys Gly Pro Tyr Tyr Asn Val Leu Arg Ile Leu
385 390 395 400
Val Pro Leu Thr Ile Glu Asp Ala Gln Ile Arg Gln Gly Leu Glu Ile
405 410 415
Ile Ser Gln Cys Phe Asp Glu Ala Lys Gln
420 425
<210> 54
<211> 463
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 54
Met Asp Lys Gln Lys Gln Asp Leu Ile Ser Gln Gln Asp Arg Gly Ala
1 5 10 15
Val Leu His Pro Phe Thr Gln Leu Lys Asp Phe Ala Ser Gly Lys Ala
20 25 30
Gly Asp Pro Thr Ile Ile Glu Thr Ala Lys Gly Ile Thr Ile Thr Asp
35 40 45
Ala Thr Gly Arg Glu Tyr Ile Asp Gly Phe Ala Gly Leu Tyr Cys Met
50 55 60
Asn Val Gly Tyr Gly Arg Thr Glu Val Ala Glu Ala Ile Ser Lys Gln
65 70 75 80
Ala His Lys Leu Ala Tyr Tyr His Thr Tyr Ala Ala His Thr Thr Glu
85 90 95
Glu Leu Ala Arg Leu Ser Asp Arg Leu Val Lys Met Ala Pro Gly Lys
100 105 110
Met Ser Lys Val Phe Tyr Gly Met Ser Gly Ser Asp Ala Asn Glu Thr
115 120 125
Gln Ala Lys Leu Val Trp Tyr Tyr Asn Asn Leu Arg Gly Lys Pro Asn
130 135 140
Lys Lys Lys Ile Ile Ala Arg Glu Arg Gly Tyr His Gly Cys Ser Val
145 150 155 160
Val Ser Gly Ser Met Thr Gly Met Ser Phe Tyr His Asp His Met Asp
165 170 175
Leu Pro Ile Ala Gly Ile Leu Arg Thr Gly Ala Pro His Tyr Tyr Trp
180 185 190
Gly Ala Glu Pro Gly Glu Thr Glu Leu Glu Phe Ser Ala Arg Arg Ala
195 200 205
Arg Glu Leu Glu Glu Leu Ile Leu Arg Glu Gly Pro Asp Thr Ile Gly
210 215 220
Ala Phe Ile Ala Glu Pro Val Leu Gly Thr Gly Gly Ile Thr Pro Pro
225 230 235 240
Pro Glu Gly Tyr Trp Ala Ala Val Gln Pro Val Leu Lys Lys Tyr Asp
245 250 255
Ile Leu Leu Ile Ala Asp Glu Val Ile Thr Gly Phe Gly Arg Thr Gly
260 265 270
Ala Met Phe Gly Cys Asp Lys Tyr Gly Ile Glu Pro Asp Leu Ile Thr
275 280 285
Val Ala Lys Gly Leu Thr Ser Ala Tyr Ala Pro Leu Ser Ala Ser Ile
290 295 300
Val Gly Glu Lys Val Tyr Lys Val Met Glu Glu Gly Ala Asp Arg Val
305 310 315 320
Gly Ala Phe Ser His Gly Tyr Thr Tyr Ser Gly His Pro Ile Gly Val
325 330 335
Ala Ala Ala Asn Ala Val Leu Asp Ile Val Glu Gly Glu Asn Ile Pro
340 345 350
Ala His Val Asp Lys Val Gly Gln Tyr Phe Gln Lys Ser Met Gln Glu
355 360 365
Thr Phe Ser Ser Leu Pro Ile Val Gly Glu Val Arg Gly Val Gly Met
370 375 380
Met Ala Ala Ile Glu Phe Val Ala Asp Pro Ala Thr Lys Lys Arg Phe
385 390 395 400
Asp Pro Ala Leu Lys Val Gly Ala Arg Ile Ser Lys Ala Ala Arg Asp
405 410 415
Arg Asn Leu Ile Ala Arg Ala Met Pro His Gly Glu Ile Leu Gly Phe
420 425 430
Ala Pro Pro Leu Val Thr Thr Glu Gln Glu Val Asp Arg Ile Ile Ser
435 440 445
Ile Ala His Glu Ala Val Leu Ser Val Leu Asp Glu Leu Lys Val
450 455 460
<210> 55
<211> 444
<212> PRT
<213> 阿维链霉菌 ATCC 31267
<400> 55
Met Ser Ala Leu Pro Gln Glu Arg Arg Val Val Thr Ala Ile Pro Gly
1 5 10 15
Pro Lys Ser Gln Glu Leu Gln Ala Arg Arg Thr Ala Val Val Ala Ala
20 25 30
Gly Val Gly Ser Val Leu Pro Val Phe Thr Thr Arg Ala Gly Gly Gly
35 40 45
Ile Ile Glu Asp Val Asp Gly Asn Arg Leu Ile Asp Phe Gly Ser Gly
50 55 60
Ile Ala Val Thr Ser Val Gly Ala Ser Ala Glu Ala Val Val Arg Arg
65 70 75 80
Ala Ser Ala Gln Leu Gln Asp Phe Thr His Thr Cys Phe Met Val Thr
85 90 95
Pro Tyr Glu Gly Tyr Val Ala Val Ala Glu Ala Leu Ala Glu Leu Thr
100 105 110
Pro Gly Asp His Ala Lys Lys Ser Ala Leu Phe Asn Ser Gly Ala Glu
115 120 125
Ala Val Glu Asn Ala Val Lys Ile Ala Arg Ala Tyr Thr Lys Arg Gln
130 135 140
Ala Val Val Val Phe Asp His Gly Tyr His Gly Arg Thr Asn Leu Thr
145 150 155 160
Met Ala Leu Thr Ala Lys Asn Met Pro Tyr Lys Asn Gly Phe Gly Pro
165 170 175
Phe Ala Pro Glu Val Tyr Arg Val Pro Val Ala Tyr Gly Tyr Arg Trp
180 185 190
Leu Thr Gly Pro Glu Asn Ala Gly Ala Glu Ala Ser Ala Gln Ala Ile
195 200 205
Asp Met Ile Asn Lys Gln Ile Gly Ala Asp Asn Val Ala Ala Ile Ile
210 215 220
Ile Glu Pro Val Leu Gly Glu Gly Gly Phe Ile Glu Pro Ala Lys Gly
225 230 235 240
Phe Leu Pro Ala Ile Ser Lys Phe Ala Lys Asp Asn Gly Ile Val Phe
245 250 255
Val Ala Asp Glu Ile Gln Ser Gly Phe Cys Arg Thr Gly Gln Trp Phe
260 265 270
Ala Cys Glu Asp Glu Gly Ile Val Pro Asp Leu Ile Thr Thr Ala Lys
275 280 285
Gly Ile Ala Gly Gly Leu Pro Leu Ala Ala Val Thr Gly Arg Ala Glu
290 295 300
Ile Met Asp Ala Ala His Ala Gly Gly Leu Gly Gly Thr Tyr Gly Gly
305 310 315 320
Asn Pro Val Ala Cys Ala Gly Ala Leu Gly Ala Ile Glu Thr Met Lys
325 330 335
Glu Leu Asp Leu Asn Ala Lys Ala Lys Asn Ile Glu Ala Val Met Lys
340 345 350
Ala Arg Leu Gly Ala Met Ala Glu Lys Phe Asp Ile Ile Gly Asp Val
355 360 365
Arg Gly Arg Gly Gly Met Ile Ala Ile Glu Leu Val Lys Asp Arg Asp
370 375 380
Thr Lys Glu Pro Asn Pro Glu Ala Ala Gly Ala Leu Ala Lys Ala Cys
385 390 395 400
His Gln Glu Gly Leu Leu Val Leu Thr Cys Gly Thr Tyr Gly Asn Val
405 410 415
Leu Arg Phe Leu Pro Pro Leu Val Ile Gly Glu Asp Leu Leu Asn Glu
420 425 430
Gly Leu Asp Ile Ile Glu Gln Ala Phe Ser Arg Ile
435 440
<210> 56
<211> 444
<212> PRT
<213> 天蓝色链霉菌 ATCC BAA-471
<400> 56
Met Thr Ala Leu Pro Gln Glu Arg Arg Val Val Thr Ala Ile Pro Gly
1 5 10 15
Pro Lys Ser Gln Glu Leu Gln Ala Arg Arg Thr Ala Val Val Ala Gln
20 25 30
Gly Val Gly Ser Thr Leu Pro Val Phe Val Ala Arg Ala Gly Gly Gly
35 40 45
Val Ile Glu Asp Val Asp Gly Asn Arg Leu Ile Asp Phe Gly Ser Gly
50 55 60
Ile Ala Val Thr Ser Val Gly Ala Ser Ala Glu Ala Val Val Arg Lys
65 70 75 80
Ala Ser Ala Gln Leu Ala Asp Phe Thr His Thr Cys Phe Met Val Thr
85 90 95
Pro Tyr Glu Gly Tyr Val Ala Val Ala Glu Ala Leu Ala Glu Leu Thr
100 105 110
Pro Gly Asp His Ala Lys Lys Ser Ala Leu Phe Asn Ser Gly Ala Glu
115 120 125
Ala Val Glu Asn Ala Val Lys Ile Ala Arg Ala Tyr Thr Lys Arg Gln
130 135 140
Ala Val Val Val Phe Asp His Gly Tyr His Gly Arg Thr Asn Leu Thr
145 150 155 160
Met Ala Leu Thr Ala Lys Asn Met Pro Tyr Lys His Gly Phe Gly Pro
165 170 175
Phe Ala Pro Glu Val Tyr Arg Val Pro Val Ala Tyr Gly Tyr Arg Trp
180 185 190
Pro Thr Gly Ala Glu Asn Ala Gly Pro Glu Ala Ala Ala Gln Ala Ile
195 200 205
Asp Gln Ile Ser Lys Gln Val Gly Ala Glu Asn Val Ala Ala Val Ile
210 215 220
Ile Glu Pro Val Leu Gly Glu Gly Gly Phe Ile Glu Pro Ala Lys Gly
225 230 235 240
Phe Leu Pro Ala Ile Arg Glu Phe Ala Ser Asp Asn Gly Ile Val Phe
245 250 255
Val Ala Asp Glu Ile Gln Ser Gly Phe Cys Arg Thr Gly Gln Trp Phe
260 265 270
Ala Cys Glu Asp Glu Gly Ile Val Pro Asp Leu Ile Thr Thr Ala Lys
275 280 285
Gly Ile Ala Gly Gly Leu Pro Leu Ser Ala Val Thr Gly Arg Ala Glu
290 295 300
Ile Met Asp Ala Ala His Ser Gly Gly Leu Gly Gly Thr Tyr Gly Gly
305 310 315 320
Asn Pro Val Ala Cys Ala Gly Ala Leu Gly Ala Ile Glu Thr Met Lys
325 330 335
Glu Leu Asp Leu Asn Ala Arg Ala Lys Asp Ile Glu Ala Val Met Lys
340 345 350
Ser Arg Leu Ala Ala Met Ala Glu Lys Phe Asp Val Ile Gly Asp Val
355 360 365
Arg Gly Arg Gly Ala Met Ile Ala Ile Glu Leu Val Lys Asp Arg Glu
370 375 380
Thr Lys Glu Pro Asn Pro Ala Ala Thr Ala Ala Leu Ala Lys Ala Cys
385 390 395 400
His Gln Glu Gly Leu Leu Val Leu Thr Cys Gly Thr Tyr Gly Asn Val
405 410 415
Leu Arg Phe Leu Pro Pro Leu Val Ile Gly Glu Asp Leu Leu Asn Glu
420 425 430
Gly Leu Asp Ile Ile Glu Gln Ala Phe Thr Arg Val
435 440
<210> 57
<211> 442
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 57
Met Ser Asn Arg Leu Asn Thr Thr Pro Asn Asp Leu Ser Ala Phe Trp
1 5 10 15
Met Pro Phe Thr Ala Asn Arg Gln Phe Lys Lys Glu Pro Arg Leu Phe
20 25 30
Val Gly Ala Lys Asp Met Tyr Tyr Thr Thr His Asp Gly Arg Thr Val
35 40 45
Leu Asp Gly Thr Ala Gly Leu Trp Cys Val Asn Ala Gly His Cys Arg
50 55 60
Pro Lys Ile Thr Glu Ala Ile Arg Glu Gln Ala Gly Glu Leu Asp Tyr
65 70 75 80
Ala Pro Ala Phe Gln Leu Gly His Pro Lys Ala Phe Glu Leu Ala Asn
85 90 95
Arg Leu Val Asp Ile Ala Pro Glu Gly Met Asn His Val Leu Tyr Thr
100 105 110
Asn Ser Gly Ser Glu Ser Val Asp Thr Ala Leu Lys Val Ala Leu Ala
115 120 125
Tyr His Arg Ala Lys Gly Asn Gly Ser Arg Phe Arg Leu Ile Gly Arg
130 135 140
Glu Arg Gly Tyr His Gly Val Asn Phe Gly Gly Ile Ser Val Gly Gly
145 150 155 160
Ile Val Ala Asn Arg Lys Met Phe Gly Thr Leu Leu Thr Gly Val Asp
165 170 175
His Leu Pro His Thr His Leu Pro Ala Lys Asn Ala Phe Ser Arg Gly
180 185 190
Glu Pro Glu His Gly Ala Asp Leu Ala Ala Glu Leu Glu Arg Ile Val
195 200 205
Thr Leu His Asp Ala Ser Thr Val Ala Ala Val Ile Val Glu Pro Val
210 215 220
Ala Gly Ser Thr Gly Val Leu Ile Pro Pro Lys Gly Tyr Leu Gln Lys
225 230 235 240
Leu Arg Glu Ile Cys Thr Lys His Gly Ile Leu Leu Ile Phe Asp Glu
245 250 255
Val Ile Thr Gly Tyr Gly Arg Leu Gly Thr Pro Phe Ala Ala Gln Tyr
260 265 270
Phe Asp Val Lys Pro Asp Ile Ile Thr Thr Ala Lys Gly Leu Thr Asn
275 280 285
Gly Val Ile Pro Met Gly Ala Val Phe Val Thr Ser Glu Ile His Asp
290 295 300
Ala Phe Met Thr Gly Pro Glu His Leu Ile Glu Phe Phe His Gly Tyr
305 310 315 320
Thr Tyr Ser Gly Asn Pro Ile Ala Ser Ala Ala Ala Leu Gly Thr Leu
325 330 335
Asp Thr Tyr Arg Glu Glu Gly Leu Leu Thr Arg Ala Ala Glu Leu Ala
340 345 350
Pro Tyr Trp Glu Glu Ala Leu His Ser Leu Lys Asp Cys Pro His Val
355 360 365
Ile Asp Ile Arg Asn Ile Gly Leu Ile Gly Ala Val Glu Leu Glu Pro
370 375 380
Ile Ala Gly Glu Pro Thr Lys Arg Ala Phe Ser Ala Phe Leu Lys Ala
385 390 395 400
Tyr Glu Lys Gly Leu Leu Ile Arg Thr Thr Gly Asp Ile Ile Ala Leu
405 410 415
Ser Pro Pro Leu Ile Ile Glu Lys Arg Gln Ile Asp Glu Leu Phe Asp
420 425 430
Lys Leu Arg Asp Val Leu Lys Asn Asn Ile
435 440
<210> 58
<211> 445
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 58
Met Asn Ala His Asn Lys Pro Asn Ala Pro Val Leu Asp Ser Tyr Trp
1 5 10 15
Met Pro Phe Thr Ala Asn Arg Gln Phe Lys Ala Ala Pro Arg Leu Leu
20 25 30
Ala Ala Ala Glu Gly Met His Tyr Thr Ser Val Asp Gly Arg Thr Val
35 40 45
Leu Asp Gly Thr Ala Gly Leu Trp Cys Val Asn Ala Gly His Gly Arg
50 55 60
Arg Gln Ile Ala Ala Ala Val Glu Arg Gln Leu Ser Thr Met Asp Phe
65 70 75 80
Ala Pro Ser Phe Gln Met Gly His Pro Ile Ala Phe Asp Phe Ala Glu
85 90 95
Arg Leu Ala Glu Ile Ala Pro Gly Pro Ala Gly Ala Lys Leu Asp Arg
100 105 110
Val Phe Phe Thr Gly Ser Gly Ser Glu Ser Val Asp Thr Ala Leu Lys
115 120 125
Met Ala Leu Ala Tyr Gln Arg Ser Ile Gly Gln Gly Thr Arg Thr Arg
130 135 140
Leu Ile Gly Arg Glu Arg Gly Tyr His Gly Val Gly Phe Gly Gly Ile
145 150 155 160
Ser Val Gly Gly Ile Val Asn Asn Arg Arg Val Phe Pro Gln Leu Pro
165 170 175
Gly Ser Asp His Leu Arg His Thr His Asp Pro Ala Lys Asn Ala Phe
180 185 190
Val Lys Gly Gln Pro Glu His Gly Ala Glu Leu Ala Asp Asp Leu Glu
195 200 205
Arg Leu Val Ala Leu His Gly Ala Glu Thr Ile Ala Ala Cys Ile Val
210 215 220
Glu Pro Val Ala Gly Ser Thr Gly Val Leu Ile Pro Pro Lys Gly Tyr
225 230 235 240
Leu Glu Arg Leu Arg Ala Ile Cys Asp Lys His Gly Ile Leu Leu Ile
245 250 255
Phe Asp Glu Val Ile Thr Gly Phe Gly Arg Leu Gly Ala Ala Phe Ala
260 265 270
Thr Asp Tyr Phe Gly Val Thr Pro Asp Ile Val Thr Thr Ala Lys Gly
275 280 285
Leu Thr Asn Gly Ala Ile Pro Met Gly Ala Val Phe Ala Ser Arg Lys
290 295 300
Val His Asp Ala Leu Met His Gly Pro Glu Gly Gln Ile Glu Leu Phe
305 310 315 320
His Gly Tyr Thr Tyr Ser Gly His Pro Ala Ala Cys Ala Ala Gly Ile
325 330 335
Ala Thr Leu Asp Ile Tyr Arg Asp Glu Gly Leu Met Thr Arg Ala Ala
340 345 350
Glu Ile Glu Gly Asp Trp His Glu Ala Met His Ser Met Lys Gly Leu
355 360 365
Pro His Val Ile Asp Ile Arg Thr Ile Gly Leu Ile Ala Gly Ile Glu
370 375 380
Leu Gln Ser Arg Asp Gly Ala Pro Gly Ala Arg Ala Tyr Asp Val Phe
385 390 395 400
Val Asp Cys Phe Glu Arg Gly Leu Leu Ile Arg Val Thr Gly Asp Ile
405 410 415
Ile Ala Phe Ser Pro Pro Leu Ile Ala Glu Lys Gln His Phe Gly Glu
420 425 430
Ile Val Ser Ile Leu Ala Asp Ala Leu Lys Arg Val Lys
435 440 445
<210> 59
<211> 461
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 59
Met Asn Asp Arg Pro Asn Ser Leu His Ala Leu Asp Lys Gln Ser Leu
1 5 10 15
Val His Pro Tyr Thr Asn Leu Ala Val His Gln Glu Thr Gly Pro His
20 25 30
Val Ile Thr Gly Gly Asp Gly Ile Tyr Val Val Asp Asp Glu Gly Lys
35 40 45
Arg Tyr Ile Glu Gly Leu Ala Gly Leu Phe Cys Ala Gly Leu Gly Phe
50 55 60
Ser Glu Gln Arg Leu Val Glu Ala Ala Thr Arg Gln Leu Lys Thr Met
65 70 75 80
Pro Phe Tyr His Ser Phe Ala His Lys Ser Thr Glu Pro Gly Ile Arg
85 90 95
Leu Ala Glu Lys Leu Leu Ser Ile Ala Pro Val Pro Met Ser Lys Val
100 105 110
Phe Phe Ala Gly Ser Gly Ser Glu Ala Asn Asp Thr Ala Ile Lys Leu
115 120 125
Ile Trp Tyr Tyr Asn Asn Ala Leu Gly Arg Pro Glu Lys Lys Lys Ile
130 135 140
Ile Ser Arg Arg Lys Ala Tyr His Gly Val Thr Val Ala Thr Ala Ser
145 150 155 160
Leu Thr Gly Leu Pro Phe Asn His Arg Asp Phe Asp Leu Pro Ile Ala
165 170 175
Asn Ile Leu His Thr Asp Cys Pro His Tyr Trp Arg Phe Ala Glu Thr
180 185 190
Gly Glu Thr Glu Glu Asp Phe Ala Thr Arg Met Ala Asn Asn Leu Glu
195 200 205
Ala Ile Ile Leu Glu Glu Gly Pro Glu Thr Ile Ala Ala Phe Phe Ala
210 215 220
Glu Pro Val Met Val Ser Gly Gly Val Ile Thr Pro Pro Lys Thr Tyr
225 230 235 240
Phe Glu Lys Val Gln Ala Val Leu Arg Lys Tyr Asp Ile Leu Leu Ile
245 250 255
Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Met Phe Gly
260 265 270
Ser Glu Thr Tyr Gly Leu Lys Pro Asp Met Ile Ser Cys Ala Lys Gln
275 280 285
Leu Ser Ala Ala Tyr Met Pro Ile Ser Ala Leu Met Ile Asn Ala Lys
290 295 300
Ile Ala Asp Ala Leu Val Asp Gln Ser Arg Lys Ile Gly Thr Phe Ser
305 310 315 320
His Gly Phe Thr Tyr Gly Gly His Pro Val Ala Ala Ala Val Ala Leu
325 330 335
Glu Ala Leu Thr Ile Tyr Glu Glu Ile Asp Ile Val Gly His Val Arg
340 345 350
Ser Val Ala Pro Ala Phe Gln Asp Arg Val Arg Lys Leu Gly Glu His
355 360 365
Pro Leu Ile Gly Glu Ala Arg Gly Val Gly Leu Val Ala Gly Leu Glu
370 375 380
Phe Val Lys Asp Lys Ala Thr Arg Glu Asn Phe Pro Pro Ala Trp Gln
385 390 395 400
Val Ala Asn Gln Ala Gly Lys Phe Ala Thr Ala Arg Gly Val Leu Thr
405 410 415
Arg Gly Leu Gly Asp Met Val Ser Leu Cys Pro Ala Met Ile Ile Asp
420 425 430
Asp Glu Gln Ile Asp Asp Leu Met Thr Arg Met Gly Leu Ala Leu Asp
435 440 445
Asp Thr Leu Ala Trp Ala Arg Ala Gln Gly His Val Gly
450 455 460
<210> 60
<211> 458
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 60
Met Leu Glu Arg Ser Asn Glu Leu Thr Ala Trp Asp Arg Asp His Phe
1 5 10 15
Phe His Pro Ser Thr His Met Gly Met His Ala Arg Gly Glu Thr Pro
20 25 30
Thr Arg Val Ile Gly Gly Gly Glu Gly Val Tyr Ile Thr Asp Ile Ser
35 40 45
Gly Lys Arg Ser Leu Asp Ala Phe Ala Gly Leu Tyr Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Gln Lys Ile Ala Asp Ala Ile Ala Glu Gln Ala Lys
65 70 75 80
Asn Leu Ala Tyr Tyr His Ala Tyr Val Gly His Gly Thr Glu Ala Ser
85 90 95
Ile Arg Leu Ser Lys Met Ile Ile Asp Arg Ala Pro Glu Gly Met Ser
100 105 110
Arg Val Tyr Phe Gly Leu Ser Gly Ser Asp Ala Asn Glu Thr Asn Ile
115 120 125
Lys Leu Ile Trp Tyr Tyr Asn Asn Ile Leu Gly Arg Pro Glu Lys Lys
130 135 140
Lys Ile Ile Ser Arg Trp Arg Gly Tyr His Gly Ser Gly Val Met Thr
145 150 155 160
Gly Ser Leu Thr Gly Leu His Leu Phe His Asn Ala Phe Asp Leu Pro
165 170 175
Arg Ala Pro Ile Leu His Thr Glu Ala Pro Tyr Tyr Phe Arg Arg Pro
180 185 190
Asp Arg Ser Met Ser Glu Glu Gln Phe Ser Gln Tyr Cys Ala Asp Lys
195 200 205
Leu Glu Glu Met Ile Leu Ala Glu Gly Pro Asp Thr Val Ala Ala Phe
210 215 220
Ile Gly Glu Pro Ile Leu Gly Thr Gly Gly Ile Val Pro Pro Pro Lys
225 230 235 240
Gly Tyr Trp Glu Lys Ile Gln Ala Val Leu Gln Lys Tyr Asp Ile Leu
245 250 255
Leu Val Ala Asp Glu Val Val Thr Gly Phe Gly Arg Leu Gly Thr Met
260 265 270
Phe Gly Ser Asp His Tyr Gly Ile Lys Pro Asp Leu Ile Thr Ile Ala
275 280 285
Lys Gly Leu Thr Ser Ala Tyr Ala Pro Leu Ser Gly Thr Ile Val Ser
290 295 300
Asp Lys Leu Trp Gln Val Leu Val Lys Gly Ser Asp Glu Leu Gly Ala
305 310 315 320
Ile Gly His Gly Trp Thr Tyr Ser Ala His Pro Ile Cys Ala Ala Ala
325 330 335
Gly Ile Ala Asn Leu Glu Leu Ile Asp Glu Leu Gly Ile Val Glu Asn
340 345 350
Ala Gly Ser Thr Gly Ala Tyr Phe Arg Ala Glu Leu Gln Lys Ala Val
355 360 365
Gly Asp His Arg His Val Gly Glu Val Arg Gly Asp Gly Leu Met Ala
370 375 380
Ala Ile Glu Phe Val Glu Asp Arg Asp Asp Arg Ala Phe Phe Asp Pro
385 390 395 400
Gly Arg Lys Val Gly Pro Gln Val Ala Ala Ala Leu Leu Glu Arg Gly
405 410 415
Val Ile Gly Arg Ala Met Pro Gln Gly Asp Ile Leu Gly Phe Ala Pro
420 425 430
Pro Leu Cys Leu Thr Arg Asp Glu Ala Asp Ile Val Val Lys Ala Ala
435 440 445
Ala Asp Thr Ile Gln Ser Val Leu Gly Arg
450 455
<210> 61
<211> 422
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 61
Met Thr Ser Leu Thr Asp Arg Lys Asn Ala Ala Ile Ser Arg Gly Val
1 5 10 15
Gly Met Thr Thr Gln Val Tyr Ala Asp Arg Ala Glu Asn Ala Glu Ile
20 25 30
Trp Asp Lys Glu Gly Asn Arg Tyr Ile Asp Phe Ala Ala Gly Ile Ala
35 40 45
Val Val Asn Thr Gly His Arg His Pro Lys Val Ile Ala Ala Val Lys
50 55 60
Ala Gln Leu Asp Arg Phe Thr His Thr Cys His Gln Val Val Pro Tyr
65 70 75 80
Glu Asn Tyr Val His Leu Ala Glu Arg Leu Asn Ala Ile Val Pro Gly
85 90 95
Asp Phe Ala Lys Lys Thr Ile Phe Val Thr Thr Gly Ala Glu Ala Val
100 105 110
Glu Asn Ala Val Lys Ile Ala Arg Ala Ala Thr Gly Arg Gln Ala Val
115 120 125
Val Ala Phe Gly Gly Gly Phe His Gly Arg Thr Phe Met Gly Met Ala
130 135 140
Leu Thr Gly Lys Val Val Pro Tyr Lys Val Gly Phe Gly Ala Met Pro
145 150 155 160
Ala Asp Val Phe His Ala Pro Phe Pro Ile Glu Leu His Gly Val Thr
165 170 175
Val Glu Gln Ser Leu Ser Ala Leu Lys Lys Leu Phe Ala Ala Asp Val
180 185 190
Asp Pro Ala Arg Val Ala Ala Ile Ile Ile Glu Pro Val Gln Gly Glu
195 200 205
Gly Gly Phe Tyr Pro Val Pro Thr Ala Phe Met Lys Ala Leu Arg Glu
210 215 220
Val Cys Asp Gln His Gly Ile Leu Leu Ile Ala Asp Glu Val Gln Thr
225 230 235 240
Gly Phe Ala Arg Thr Gly Lys Leu Phe Ala Met Glu His His Gly Val
245 250 255
Ala Pro Asp Leu Thr Thr Met Ala Lys Ser Leu Ala Gly Gly Phe Pro
260 265 270
Leu Ala Ala Val Thr Gly Arg Ala Glu Ile Met Asp Ala Pro Gly Pro
275 280 285
Gly Gly Leu Gly Gly Thr Tyr Gly Gly Asn Pro Leu Gly Ile Ala Ala
290 295 300
Ala His Ala Val Leu Asp Val Ile Ala Glu Glu Asn Leu Cys Glu Arg
305 310 315 320
Ala Asn Gln Leu Gly Asn Arg Leu Lys Gln Arg Leu Ala Ala Ile Arg
325 330 335
Glu Lys Ala Pro Glu Ile Val Asp Ile Arg Gly Pro Gly Phe Met Asn
340 345 350
Ala Val Glu Phe Asn Asp Val Arg Thr Asn Leu Pro Ser Ala Glu Phe
355 360 365
Ala Asn Lys Val Arg Leu Leu Ala Leu Glu Lys Gly Leu Ile Leu Leu
370 375 380
Thr Cys Gly Val His Gly Asn Val Ile Arg Phe Leu Ala Pro Ile Thr
385 390 395 400
Ile Gln Asp Glu Val Phe Ala Glu Ala Leu Asp Thr Ile Glu Ala Ser
405 410 415
Ile Leu Glu Ala Arg Gly
420
<210> 62
<211> 464
<212> PRT
<213> 庞氏鲁杰氏菌 ATCC 700808
<400> 62
Met Ala Thr Ile Thr Asn His Met Pro Thr Ala Glu Leu Gln Ala Leu
1 5 10 15
Asp Ala Ala His His Leu His Pro Phe Ser Ala Asn Asn Ala Leu Gly
20 25 30
Glu Glu Gly Thr Arg Val Ile Thr Arg Ala Arg Gly Val Trp Leu Asn
35 40 45
Asp Ser Glu Gly Glu Glu Ile Leu Asp Ala Met Ala Gly Leu Trp Cys
50 55 60
Val Asn Ile Gly Tyr Gly Arg Asp Glu Leu Ala Glu Val Ala Ala Arg
65 70 75 80
Gln Met Arg Glu Leu Pro Tyr Tyr Asn Thr Phe Phe Lys Thr Thr His
85 90 95
Val Pro Ala Ile Ala Leu Ala Gln Lys Leu Ala Glu Leu Ala Pro Gly
100 105 110
Asp Leu Asn His Val Phe Phe Ala Gly Gly Gly Ser Glu Ala Asn Asp
115 120 125
Thr Asn Ile Arg Met Val Arg Thr Tyr Trp Gln Asn Lys Gly Gln Pro
130 135 140
Glu Lys Thr Val Ile Ile Ser Arg Lys Asn Ala Tyr His Gly Ser Thr
145 150 155 160
Val Ala Ser Ser Ala Leu Gly Gly Met Ala Gly Met His Ala Gln Ser
165 170 175
Gly Leu Ile Pro Asp Val His His Ile Asn Gln Pro Asn Trp Trp Ala
180 185 190
Glu Gly Gly Asp Met Asp Pro Glu Glu Phe Gly Leu Ala Arg Ala Arg
195 200 205
Glu Leu Glu Glu Ala Ile Leu Glu Leu Gly Glu Asn Arg Val Ala Ala
210 215 220
Phe Ile Ala Glu Pro Val Gln Gly Ala Gly Gly Val Ile Val Ala Pro
225 230 235 240
Asp Ser Tyr Trp Pro Glu Ile Gln Arg Ile Cys Asp Lys Tyr Asp Ile
245 250 255
Leu Leu Ile Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn
260 265 270
Trp Phe Gly Thr Gln Thr Met Gly Ile Arg Pro His Ile Met Thr Ile
275 280 285
Ala Lys Gly Leu Ser Ser Gly Tyr Ala Pro Ile Gly Gly Ser Ile Val
290 295 300
Cys Asp Glu Val Ala His Val Ile Gly Lys Asp Glu Phe Asn His Gly
305 310 315 320
Tyr Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Leu Glu Asn
325 330 335
Leu Arg Ile Leu Glu Glu Glu Asn Ile Leu Asp His Val Arg Asn Val
340 345 350
Ala Ala Pro Tyr Leu Lys Glu Lys Trp Glu Ala Leu Thr Asp His Pro
355 360 365
Leu Val Gly Glu Ala Lys Ile Val Gly Met Met Ala Ser Ile Ala Leu
370 375 380
Thr Pro Asn Lys Ala Ser Arg Ala Lys Phe Ala Ser Glu Pro Gly Thr
385 390 395 400
Ile Gly Tyr Ile Cys Arg Glu Arg Cys Phe Ala Asn Asn Leu Ile Met
405 410 415
Arg His Val Gly Asp Arg Met Ile Ile Ser Pro Pro Leu Val Ile Thr
420 425 430
Pro Ala Glu Ile Asp Glu Met Phe Val Arg Ile Arg Lys Ser Leu Asp
435 440 445
Glu Ala Gln Ala Glu Ile Glu Lys Gln Gly Leu Met Lys Ser Ala Ala
450 455 460
<210> 63
<211> 459
<212> PRT
<213> 百脉根根瘤菌 MAFF303099
<400> 63
Met Leu Asn Gln Ser Asn Glu Leu Asn Ala Trp Asp Arg Asp His Phe
1 5 10 15
Phe His Pro Ser Thr His Met Gly Thr His Ala Arg Gly Glu Ser Pro
20 25 30
Thr Arg Ile Met Ala Gly Gly Glu Gly Val Thr Val Trp Asp Asn Asn
35 40 45
Gly Arg Lys Ser Ile Asp Ala Phe Ala Gly Leu Tyr Cys Val Asn Val
50 55 60
Gly Tyr Gly Arg Gln Lys Ile Ala Asp Ala Ile Ala Thr Gln Ala Lys
65 70 75 80
Asn Leu Ala Tyr Tyr His Ala Tyr Val Gly His Gly Thr Glu Ala Ser
85 90 95
Ile Thr Leu Ala Lys Met Ile Ile Asp Arg Ala Pro Lys Gly Met Ser
100 105 110
Arg Val Tyr Phe Gly Leu Ser Gly Ser Asp Ala Asn Glu Thr Asn Ile
115 120 125
Lys Leu Ile Trp Tyr Tyr Asn Asn Val Leu Gly Arg Pro Glu Lys Lys
130 135 140
Lys Ile Ile Ser Arg Trp Arg Gly Tyr His Gly Ser Gly Val Met Thr
145 150 155 160
Gly Ser Leu Thr Gly Leu Asp Leu Phe His Asn Ala Phe Asp Leu Pro
165 170 175
Arg Ala Pro Val Leu His Thr Glu Ala Pro Tyr Tyr Phe Arg Arg Thr
180 185 190
Asp Arg Ser Met Ser Glu Glu Gln Phe Ser Gln His Cys Ala Asp Lys
195 200 205
Leu Glu Glu Met Ile Leu Ala Glu Gly Pro Glu Thr Ile Ala Ala Phe
210 215 220
Ile Gly Glu Pro Ile Leu Gly Thr Gly Gly Ile Val Pro Pro Pro Ala
225 230 235 240
Gly Tyr Trp Glu Lys Ile Gln Ala Val Leu Lys Lys Tyr Asp Val Leu
245 250 255
Leu Val Ala Asp Glu Val Val Thr Gly Phe Gly Arg Leu Gly Thr Met
260 265 270
Phe Gly Ser Asp His Tyr Gly Ile Lys Pro Asp Leu Ile Thr Ile Ala
275 280 285
Lys Gly Leu Thr Ser Ala Tyr Ala Pro Leu Ser Gly Val Ile Val Ala
290 295 300
Asp Arg Val Trp Gln Val Leu Val Gln Gly Ser Asp Lys Leu Gly Ser
305 310 315 320
Leu Gly His Gly Trp Thr Tyr Ser Ala His Pro Ile Cys Val Ala Ala
325 330 335
Gly Val Ala Asn Leu Glu Leu Ile Asp Glu Met Asp Leu Val Thr Asn
340 345 350
Ala Gly Glu Thr Gly Ala Tyr Phe Arg Ala Glu Leu Ala Lys Ala Val
355 360 365
Gly Gly His Lys Asn Val Gly Glu Val Arg Gly Asp Gly Met Leu Ala
370 375 380
Ala Val Glu Phe Val Ala Asp Lys Asp Asp Arg Val Phe Phe Asp Ala
385 390 395 400
Ser Gln Lys Ile Gly Pro Gln Val Ala Thr Ala Leu Ala Ala Ser Gly
405 410 415
Val Ile Gly Arg Ala Met Pro Gln Gly Asp Ile Leu Gly Phe Ala Pro
420 425 430
Pro Leu Cys Leu Thr Arg Glu Gln Ala Asp Ile Val Val Ser Lys Thr
435 440 445
Ala Asp Ala Val Lys Ser Val Phe Ala Asn Leu
450 455
<210> 64
<211> 310
<212> PRT
<213> 丁香假单胞菌丁香致病变种
<400> 64
Met Ser Lys Lys Phe Ala Leu Thr Ala Glu Gln Arg Ala Ser Phe Glu
1 5 10 15
Lys Asn Gly Phe Ile Gly Pro Phe Asp Ala Tyr Ser Pro Glu Glu Met
20 25 30
Lys Glu Thr Trp Lys Arg Thr Arg Leu Arg Leu Leu Asp Arg Ser Ala
35 40 45
Ala Ala Tyr Gln Asp Leu Asp Ala Ile Ser Gly Gly Thr Asn Ile Ala
50 55 60
Asn Tyr Asp Arg His Leu Asp Asp Asp Phe Leu Ala Ser His Ile Cys
65 70 75 80
Arg Pro Glu Ile Cys Asp Arg Val Glu Ser Ile Leu Gly Pro Asn Val
85 90 95
Leu Cys Trp Arg Thr Glu Phe Phe Pro Lys Tyr Pro Gly Asp Glu Gly
100 105 110
Thr Asp Trp His Gln Ala Asp Thr Phe Ala Asn Ala Ser Gly Lys Pro
115 120 125
Gln Ile Ile Trp Pro Glu Asn Glu Glu Phe Gly Gly Thr Ile Thr Val
130 135 140
Trp Thr Ala Phe Thr Asp Ala Asn Ile Ala Asn Gly Cys Leu Gln Phe
145 150 155 160
Ile Pro Gly Thr Gln Asn Ser Met Asn Tyr Asp Glu Thr Lys Arg Met
165 170 175
Thr Tyr Glu Pro Asp Ala Asn Asn Ser Val Val Lys Asp Gly Val Arg
180 185 190
Arg Gly Phe Phe Gly Tyr Asp Tyr Arg Gln Leu Gln Ile Asp Glu Asn
195 200 205
Trp Lys Pro Asp Glu Ala Ser Ala Val Pro Met Gln Met Lys Ala Gly
210 215 220
Gln Phe Ile Ile Phe Trp Ser Thr Leu Met His Ala Ser Tyr Pro His
225 230 235 240
Ser Gly Glu Ser Gln Glu Met Arg Met Gly Phe Ala Ser Arg Tyr Val
245 250 255
Pro Ser Phe Val His Val Tyr Pro Asp Ser Asp His Ile Glu Glu Tyr
260 265 270
Gly Gly Arg Ile Ser Leu Glu Lys Tyr Gly Ala Val Gln Val Ile Gly
275 280 285
Asp Glu Thr Pro Glu Tyr Asn Arg Leu Val Thr His Thr Thr Arg Gly
290 295 300
Lys Lys Phe Glu Ala Val
305 310
<210> 65
<211> 467
<212> PRT
<213> 类球红细菌 ATCC 17023
<400> 65
Met Thr Arg Asn Asp Ala Thr Asn Ala Ala Gly Ala Val Gly Ala Ala
1 5 10 15
Met Arg Asp His Ile Leu Leu Pro Ala Gln Glu Met Ala Lys Leu Gly
20 25 30
Lys Ser Ala Gln Pro Val Leu Thr His Ala Glu Gly Ile Tyr Val His
35 40 45
Thr Glu Asp Gly Arg Arg Leu Ile Asp Gly Pro Ala Gly Met Trp Cys
50 55 60
Ala Gln Val Gly Tyr Gly Arg Arg Glu Ile Val Asp Ala Met Ala His
65 70 75 80
Gln Ala Met Val Leu Pro Tyr Ala Ser Pro Trp Tyr Met Ala Thr Ser
85 90 95
Pro Ala Ala Arg Leu Ala Glu Lys Ile Ala Thr Leu Thr Pro Gly Asp
100 105 110
Leu Asn Arg Ile Phe Phe Thr Thr Gly Gly Ser Thr Ala Val Asp Ser
115 120 125
Ala Leu Arg Phe Ser Glu Phe Tyr Asn Asn Val Leu Gly Arg Pro Gln
130 135 140
Lys Lys Arg Ile Ile Val Arg Tyr Asp Gly Tyr His Gly Ser Thr Ala
145 150 155 160
Leu Thr Ala Ala Cys Thr Gly Arg Thr Gly Asn Trp Pro Asn Phe Asp
165 170 175
Ile Ala Gln Asp Arg Ile Ser Phe Leu Ser Ser Pro Asn Pro Arg His
180 185 190
Ala Gly Asn Arg Ser Gln Glu Ala Phe Leu Asp Asp Leu Val Gln Glu
195 200 205
Phe Glu Asp Arg Ile Glu Ser Leu Gly Pro Asp Thr Ile Ala Ala Phe
210 215 220
Leu Ala Glu Pro Ile Leu Ala Ser Gly Gly Val Ile Ile Pro Pro Ala
225 230 235 240
Gly Tyr His Ala Arg Phe Lys Ala Ile Cys Glu Lys His Asp Ile Leu
245 250 255
Tyr Ile Ser Asp Glu Val Val Thr Gly Phe Gly Arg Cys Gly Glu Trp
260 265 270
Phe Ala Ser Glu Lys Val Phe Gly Val Val Pro Asp Ile Ile Thr Phe
275 280 285
Ala Lys Gly Val Thr Ser Gly Tyr Val Pro Leu Gly Gly Leu Ala Ile
290 295 300
Ser Glu Ala Val Leu Ala Arg Ile Ser Gly Glu Asn Ala Lys Gly Ser
305 310 315 320
Trp Phe Thr Asn Gly Tyr Thr Tyr Ser Asn Gln Pro Val Ala Cys Ala
325 330 335
Ala Ala Leu Ala Asn Ile Glu Leu Met Glu Arg Glu Gly Ile Val Asp
340 345 350
Gln Ala Arg Glu Met Ala Asp Tyr Phe Ala Ala Ala Leu Ala Ser Leu
355 360 365
Arg Asp Leu Pro Gly Val Ala Glu Thr Arg Ser Val Gly Leu Val Gly
370 375 380
Cys Val Gln Cys Leu Leu Asp Pro Thr Arg Ala Asp Gly Thr Ala Glu
385 390 395 400
Asp Lys Ala Phe Thr Leu Lys Ile Asp Glu Arg Cys Phe Glu Leu Gly
405 410 415
Leu Ile Val Arg Pro Leu Gly Asp Leu Cys Val Ile Ser Pro Pro Leu
420 425 430
Ile Ile Ser Arg Ala Gln Ile Asp Glu Met Val Ala Ile Met Arg Gln
435 440 445
Ala Ile Thr Glu Val Ser Ala Ala His Gly Leu Thr Ala Lys Glu Pro
450 455 460
Ala Ala Val
465
<210> 66
<211> 1183
<212> PRT
<213> 脓肿分枝杆菌 DSM44196
<400> 66
Met Thr Ile Asp Ala Thr Ala Asp Asn Thr Lys Glu Ala Arg Arg Gln
1 5 10 15
Arg Leu Gly Asp Arg Ile Arg Arg Leu Phe Thr Asp Asp Glu Gln Phe
20 25 30
Arg Ala Ala Lys Pro Asp Thr Ala Val Asp Thr Ala Val Ala Gln Pro
35 40 45
Gly Leu Arg Leu Ala Gln Val Val Ala Thr Ile Met Asn Gly Tyr Ala
50 55 60
Asp Arg Pro Ala Leu Gly His Arg Val Gln Glu Leu Val Ala Asp Ala
65 70 75 80
Ala Gly Arg Ser Thr Leu Arg Pro Leu Pro Glu Phe Glu Thr Val Thr
85 90 95
Tyr Gly Glu Leu Trp Gly Met Ala Arg Ala Leu Ala Ser Thr Trp Tyr
100 105 110
His Asp Pro Ala Ala Pro Val Arg Ala Gly Asp Phe Val Ala Met Leu
115 120 125
Gly Phe Thr Ser Val Asp Tyr Thr Ala Val Asp Leu Ala Cys Ile His
130 135 140
Leu Gly Ala Val Ala Val Pro Leu Gln Thr Ser Ala Ser Ala Ser Asn
145 150 155 160
Trp Thr Ala Ile Leu Ala Glu Ser Glu Pro Ala Val Leu Ala Val Ser
165 170 175
Ala Glu Leu Leu Asp Thr Ala Met Glu Ser Val Leu Ala Thr Pro Ser
180 185 190
Leu Arg His Ile Thr Val Phe Asp Tyr His Pro Gly Val Asp Val Gln
195 200 205
Arg Glu Ser Leu Glu Ser Ala Gln His Arg Ile Ala Glu Ala Gly Leu
210 215 220
Pro Ile Ser Val Asp Pro Ile Pro Leu Ala Ile Gly His Gly Arg Ala
225 230 235 240
Leu Pro Asp Ala Pro Leu Phe Thr Ala Glu Glu Gly Thr Asp Pro Leu
245 250 255
Ala Leu Val Ile Tyr Thr Ser Gly Ser Thr Gly Thr Pro Lys Gly Ala
260 265 270
Thr Tyr Ser Glu Lys Met Val Ala Lys Pro Trp Leu Arg Ala Asp Thr
275 280 285
Leu Ser Ser Lys Ala Glu Ile Pro Leu Ile Asn Leu Asn Phe Met Pro
290 295 300
Met Ser His Val Met Gly Arg Gly Ser Leu Val Thr Ala Leu Ala Cys
305 310 315 320
Gly Gly Leu Ala Tyr Phe Ala Ala Ser Ser Asp Met Ser Thr Leu Phe
325 330 335
Glu Asp Ile Thr Leu Thr Arg Pro Thr Val Val Thr Leu Val Pro Arg
340 345 350
Val Cys Asp Met Leu Phe Gln Arg Tyr Arg Asn Glu Val Glu Arg Arg
355 360 365
Thr Gly Leu Asp Pro Ala Ala Asp Leu Ala Thr Leu Asp Ala Asp Val
370 375 380
Lys Thr Asp Ile Arg Glu Asn Leu Phe Gly Gly Arg Val Leu Thr Ile
385 390 395 400
Val Cys Gly Ser Ala Pro Leu Ser Glu Glu Leu Ala Ala Phe Ile Glu
405 410 415
Ser Cys Leu Asp Ala Arg Ile Thr Asp Gly Tyr Gly Ser Thr Glu Ala
420 425 430
Gly Val Ile Val Arg Asn Gly Arg Ile Gln Arg Pro Pro Val Ile Asp
435 440 445
Tyr Lys Leu Val Asp Val Pro Glu Leu Gly Tyr Phe Ser Thr Asp Lys
450 455 460
Pro His Pro Arg Gly Glu Leu Leu Val Lys Ala Glu Ser Val Phe Gly
465 470 475 480
Gly Tyr Phe Lys Arg Pro Asp Val Thr Ala Asp Val Phe Asp Pro Asp
485 490 495
Gly Tyr Tyr Lys Thr Gly Asp Ile Val Ala Glu Leu Glu Pro Asp Lys
500 505 510
Ile Gln Ile Val Asp Arg Arg Asn Asn Val Ile Lys Leu Ser Gln Gly
515 520 525
Glu Phe Val Ala Ile Ala Asn Leu Glu Ala Glu Phe Ala Asn Ser Pro
530 535 540
Leu Val His Gln Ile Cys Val Tyr Gly Ser Ser Glu Arg Ser Tyr Leu
545 550 555 560
Leu Ala Val Val Val Pro Thr Ala Glu Ala Tyr Glu Gln Ser Gly Gly
565 570 575
Asp Glu Asp Leu Leu Lys Arg Leu Ile Ala Asp Ser Leu Ala Gln Val
580 585 590
Ala Arg Glu Ala Gln Leu Gln Ser Tyr Glu Val Pro Arg Asp Phe Leu
595 600 605
Leu Glu Thr Glu Pro Phe Thr Ala Ala Asn Gly Leu Leu Thr Gly Ile
610 615 620
Ala Lys Leu Ala Arg Pro Lys Leu His Glu Lys Tyr Gly Ala Arg Leu
625 630 635 640
Glu Gln Leu Tyr Ser Asp Ile Ala Ala Ala Gln Ala Leu Glu Leu Gln
645 650 655
Ala Leu His Ser Ala Gly His Glu Asp Lys Pro Val Leu Asp Thr Val
660 665 670
Gln Arg Ala Val Thr Ala Leu Leu Gly Leu Ser Ala Ala Glu Val Ser
675 680 685
Pro Asp Ala His Phe Ile Asp Leu Gly Gly Asp Ser Leu Ser Ala Leu
690 695 700
Ala Phe Ser Asp Leu Leu Arg Asp Ile Phe Thr Val Glu Val Pro Val
705 710 715 720
Gly Asp Ile Val Ser Ala Ala Asn Asp Leu Thr Ala Ile Ala Arg Ile
725 730 735
Val Glu Arg His Arg Glu Ala Asp Gly His Ser Val Thr Pro Thr Ala
740 745 750
Glu Ser Val His Gly Ala Gly His Arg Glu Ile Arg Ala Ala Asp Leu
755 760 765
Thr Leu Asp Lys Phe Ile Asp Ala Asp Thr Leu Arg Ala Ala Pro Ala
770 775 780
Leu Ser Thr Phe Thr Gly Thr Pro His Thr Val Leu Leu Thr Gly Ala
785 790 795 800
Asn Gly Tyr Leu Gly Arg Phe Leu Ala Leu Glu Trp Leu Glu Arg Leu
805 810 815
Asp Lys Thr Asp Gly Lys Leu Ile Ala Ile Val Arg Gly Lys Asn Ala
820 825 830
Glu Ala Ala Tyr Arg Arg Leu Glu Glu Ala Phe Asp Thr Gly Asp Thr
835 840 845
Gln Leu Leu Ala His Phe Arg Ser Leu Ala Asp Lys His Leu Glu Val
850 855 860
Leu Ala Gly Asp Ile Gly Asp Pro Asn Leu Gly Leu Asp Ala Asp Thr
865 870 875 880
Trp Gln Arg Leu Ala Glu Thr Val Asp Val Ile Val His Pro Ala Ala
885 890 895
Leu Val Asn His Val Leu Pro Tyr Ser Gln Leu Phe Gly Pro Asn Val
900 905 910
Val Gly Thr Ala Glu Ile Ile Lys Leu Ala Leu Thr Thr Lys Ile Lys
915 920 925
Pro Ile Thr Tyr Leu Ser Thr Val Ala Val Ala Ile Ser Val Asp Pro
930 935 940
Lys Val Phe Asp Glu Asp Ser Asp Ile Arg Thr Ile Ser Ala Val Arg
945 950 955 960
Pro Ile Asn Asp Gly Tyr Ala Asn Gly Tyr Gly Asn Ala Lys Trp Ala
965 970 975
Gly Glu Val Leu Leu Arg Glu Ala His Asp Leu Cys Gly Leu Pro Val
980 985 990
Ala Val Phe Arg Ser Asp Met Ile Leu Ala His Ser Arg Tyr Thr Gly
995 1000 1005
Gln Leu Asn Val Pro Asp Gln Phe Thr Arg Leu Ile Leu Ser Leu
1010 1015 1020
Ile Ala Thr Gly Val Ala Pro Gly Ser Phe Tyr Gln Ala His Ala
1025 1030 1035
Thr Gly Glu Arg Pro Leu Ala His Tyr Asp Gly Leu Pro Ala Asp
1040 1045 1050
Phe Thr Ala Ser Ala Ile Thr Ala Leu Gly Pro Ile Glu Glu Phe
1055 1060 1065
His Thr Tyr Asp Ser Val Asn Pro His Ala Asp Gly Ile Ser Leu
1070 1075 1080
Asp Asn Phe Val Asp Trp Leu Ile Glu Ala Gly Tyr Pro Ile Ala
1085 1090 1095
Arg Ile Asp Asn Tyr Thr Glu Trp Phe Thr Arg Phe Asp Thr Ala
1100 1105 1110
Ile Arg Ser Leu Pro Glu Lys Gln Lys Gln His Ser Leu Leu Pro
1115 1120 1125
Leu Leu His Ala Tyr Arg His Pro Gln His Pro His Asn Gly Ala
1130 1135 1140
Phe Leu Pro Ala Ile Arg Phe Ser Glu Gly Val Gln Ala His Leu
1145 1150 1155
Asn Ala Asp Ile Pro His Leu Thr Arg Glu Leu Ile Ala Lys Tyr
1160 1165 1170
Ala Ala Asp Leu Lys Gln Leu Gly Leu Leu
1175 1180
<210> 67
<211> 1163
<212> PRT
<213> 脓肿分枝杆菌 DSM44196
<400> 67
Met Thr Ala Gly Ala Ala Ala Arg Val Ala Lys Leu Phe Glu Ser Asp
1 5 10 15
Pro Gln Phe Arg Ala Ala Met Pro Asp Pro Ala Val Met Asp Ser Leu
20 25 30
Leu Ala Pro Gly Leu Arg Leu Ser Gln Val Leu His Ala Leu Leu Ser
35 40 45
Gly Tyr Ala Glu Arg Pro Val Met Gly Phe Arg Ser Arg Glu Ser Val
50 55 60
Val Asp Thr Ala Thr Gly Arg Thr Val Asp Arg Leu Leu Pro Ala Phe
65 70 75 80
Glu Thr Ile Thr Tyr Gly Gln Leu Leu Glu Asp Ile Ser Ala Ile Leu
85 90 95
Ala Glu Trp Gln His Gly Asp Ile Pro Met Gly Ala Gly Asp Phe Ile
100 105 110
Ala Thr Ile Gly Phe Ser Ser Pro Asp Tyr Val Thr Leu Asp Leu Ala
115 120 125
Thr Leu Met Asn Gly Ser Val Ser Ile Pro Leu Gln His Asn Thr Ser
130 135 140
Val Ala Gln Leu Arg Met Met Leu Glu Glu Thr Ser Pro Arg Leu Val
145 150 155 160
Ala Ala Ser Ala Asp Cys Leu Asp Leu Ala Val Glu Ala Ala Val Gly
165 170 175
Leu Thr Asp Leu Arg Arg Val Val Val Phe Asp Tyr Arg Ala Glu Thr
180 185 190
Asp Asp His Arg Glu Lys Leu Ala Thr Ala Arg Glu Arg Leu His Ala
195 200 205
Ala Gly Met Asp Val Val Val Glu Pro Leu Ala Glu Val Ile Gly Arg
210 215 220
Gly Arg Asp Leu Pro Glu Pro Val Leu Tyr Thr Ala Gly Asp Asp Gln
225 230 235 240
Arg Thr Ala Leu Ile Met Tyr Thr Ser Gly Ser Thr Gly Ala Pro Lys
245 250 255
Gly Ala Met Phe Thr Glu Trp Thr Val Thr Arg Phe Trp Ser Ser Gly
260 265 270
Ala Ala Pro Asn Arg Asp Thr Pro Ile Ile Asn Val Asn Phe Leu Pro
275 280 285
Leu Asn His Leu Ala Gly Arg Val Gly Leu Leu Thr Ala Phe Ile Pro
290 295 300
Gly Gly Thr Cys Tyr Phe Val Pro Glu Ser Asp Leu Ser Thr Leu Phe
305 310 315 320
Glu Asp Trp Gln Leu Ala Arg Pro Thr His Met Gly Val Val Pro Arg
325 330 335
Val Val Asp Met Leu Phe Gln His Tyr Gln Thr Arg Val Asp Ala Leu
340 345 350
Met Ala Gly Gly Thr Asp Val Asp Thr Ala Asp Arg Leu Ala Lys Thr
355 360 365
Glu Leu Arg Glu Asp Val Leu Gly Gly Arg Val Val Ala Gly Met Leu
370 375 380
Ala Thr Ala Pro Leu Ser Pro Glu Met Lys Ala Phe Leu Glu Ser Ser
385 390 395 400
Leu Asp Phe His Leu Leu Asp Leu Tyr Gly Leu Thr Glu Val Gly Gly
405 410 415
Val Phe Arg Asp Gly Lys Ile Ser Arg Pro Pro Val Leu Asp Tyr Lys
420 425 430
Leu Val Asp Val Pro Glu Leu Gly Tyr Tyr Thr Thr Asp Lys Pro His
435 440 445
Pro Arg Gly Glu Leu Leu Val Lys Ser Ala Thr Ala Thr Pro Gly Tyr
450 455 460
Tyr Lys Arg Pro Asp Val Thr Ala Glu Val Phe Asp Ala Asp Gly Tyr
465 470 475 480
Tyr Arg Thr Gly Asp Val Met Ala Glu Val Ala Pro Asp Gln Leu Val
485 490 495
Tyr Val Asp Arg Arg Asn Asn Val Ile Lys Leu Ala Gln Gly Glu Phe
500 505 510
Val Ala Val Ala Asn Leu Glu Thr Val Tyr Val Gly Ala Pro Leu Val
515 520 525
Arg Gln Ile Phe Val Tyr Gly Asn Ser Glu Arg Ala Tyr Leu Leu Ala
530 535 540
Val Val Val Pro Thr Glu Glu Ala Leu Arg Ala His Pro Asp Pro Val
545 550 555 560
Glu Leu Lys Asn Ser Ile Arg Glu Ser Leu Gln Arg Thr Ala Arg Ser
565 570 575
Asn His Leu His Ser Tyr Glu Leu Pro Ala Asp Phe Ile Ile Glu Thr
580 585 590
Thr Pro Phe Thr Ile Glu Ser Gly Met Leu Ala Ala Val Gly Lys Pro
595 600 605
Ile Arg Pro Lys Met Ile Glu His Tyr Gly Asp Arg Leu Glu Gln Leu
610 615 620
Tyr Val Asp Leu Ala Glu Ala Arg Val Gln Glu Leu Arg Gln Leu Arg
625 630 635 640
Asp Thr Ala Gln Gln Arg Pro Val Leu Asp Thr Val Thr Glu Ala Ala
645 650 655
Gln Ala Leu Leu Gly Met Ser Ala Asp Ala Val Arg Pro Asp His His
660 665 670
Phe Ile Asp Leu Gly Gly Asp Ser Leu Ser Ala Leu Thr Phe Ser Asn
675 680 685
Leu Leu Arg Asp Leu Phe Asp Val Glu Val Pro Val Gly Val Ile Thr
690 695 700
Gly Pro Ala Ala Asp Leu Arg Lys Leu Ala Ala Tyr Ile Gln His Glu
705 710 715 720
Arg Glu His Ser Thr Ala Thr Ala Ala Ser Val His Gly Leu Asp Thr
725 730 735
Thr Val Ile Ser Ala Thr Glu Leu Thr Leu Asp Lys Phe Ile Asp Ala
740 745 750
Glu Thr Leu His Asn Ala Ser Gln Leu Asp Val Pro Ala Gly Ala Val
755 760 765
Ala Thr Val Leu Leu Thr Gly Ala Asn Gly Tyr Leu Gly Arg Phe Leu
770 775 780
Cys Leu Glu Trp Leu Gln Arg Leu Ser Gln Thr Gly Gly Gln Leu Ile
785 790 795 800
Cys Leu Val Arg Gly Asp Asn Ala Asp Gln Ala Leu Ala Arg Leu Val
805 810 815
Ala Ala Tyr Gly Asp Thr Asp Arg Thr Leu Leu Glu Glu Phe His Thr
820 825 830
Leu Ala Arg Arg His Leu Arg Val Ile Ala Ala Asp Ile Ala Gln Pro
835 840 845
Arg Phe Gly Val Asp Asp Ala Thr Trp Glu Gln Leu Ala Arg Asp Val
850 855 860
Asp Lys Ile Val His Pro Ala Ala Leu Val Asn His Val Leu Pro Tyr
865 870 875 880
Asn Gln Leu Phe Gly Pro Asn Val Phe Gly Thr Ala Glu Val Ile Arg
885 890 895
Leu Ala Leu Thr Thr Arg Ile Lys Pro Val Thr Tyr Leu Ser Thr Met
900 905 910
Ala Val Ala Met Thr Val Pro Asp Phe Asp Glu Asp Gly Asp Ile Arg
915 920 925
Thr Val Ser Pro Thr Arg His Ile Asp Pro Gly Tyr Ala Asn Gly Tyr
930 935 940
Ala Asn Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His Asp
945 950 955 960
Ile Cys Gly Leu Pro Val Ser Val Phe Arg Ser Asp Met Ile Leu Thr
965 970 975
His Arg Arg Tyr Ser Gly Gln Leu Asn Val Thr Asp Ala Phe Thr Arg
980 985 990
Met Leu Leu Ser Leu Val Leu Thr Gly Ile Ala Pro Arg Ser Phe Tyr
995 1000 1005
Gln Gly Asp Gly Ser Gly Ala Arg Pro Arg Ala His Tyr Glu Gly
1010 1015 1020
Leu Pro Val Asp Phe Val Thr Glu Ala Ile Thr Ser Leu Gly Leu
1025 1030 1035
Ser Ser Ser Glu Gly Phe Arg Ser Tyr Asp Val Met Asn Pro His
1040 1045 1050
Asp Asp Gly Ile Ser Val Asp Thr Phe Val Asp Trp Leu Met Glu
1055 1060 1065
Asp Gly His Ser Ile Asp Ile Ile Asp Asn Tyr Asp Glu Trp Leu
1070 1075 1080
Ser Arg Phe Glu Thr Ala Leu Arg Gly Leu Pro Asp Glu Gln Arg
1085 1090 1095
Arg Ala Ser Val Leu Pro Leu Leu Asp Ala Tyr Arg Ile Pro Gly
1100 1105 1110
Asn Pro Arg Arg Ala Ala Ala Thr Pro Asn His Val Phe Arg Lys
1115 1120 1125
Ala Val Gln Glu Asn Asn Ile Gly Gly Asp Gly Ala Asp Ile Pro
1130 1135 1140
Gln Ile Asp Arg Ala Leu Ile Ala Lys Tyr Ile Ala Asp Leu Arg
1145 1150 1155
Ala His Arg Leu Leu
1160
<210> 68
<211> 1174
<212> PRT
<213> 海洋分枝杆菌 ATCC BAA-535
<400> 68
Met Ser Pro Ile Thr Arg Glu Glu Arg Leu Glu Arg Arg Ile Gln Asp
1 5 10 15
Leu Tyr Ala Asn Asp Pro Gln Phe Ala Ala Ala Lys Pro Ala Thr Ala
20 25 30
Ile Thr Ala Ala Ile Glu Arg Pro Gly Leu Pro Leu Pro Gln Ile Ile
35 40 45
Glu Thr Val Met Thr Gly Tyr Ala Asp Arg Pro Ala Leu Ala Gln Arg
50 55 60
Ser Val Glu Phe Val Thr Asp Ala Gly Thr Gly His Thr Thr Leu Arg
65 70 75 80
Leu Leu Pro His Phe Glu Thr Ile Ser Tyr Gly Glu Leu Trp Asp Arg
85 90 95
Ile Ser Ala Leu Ala Asp Val Leu Ser Thr Glu Gln Thr Val Lys Pro
100 105 110
Gly Asp Arg Val Cys Leu Leu Gly Phe Asn Ser Val Asp Tyr Ala Thr
115 120 125
Ile Asp Met Thr Leu Ala Arg Leu Gly Ala Val Ala Val Pro Leu Gln
130 135 140
Thr Ser Ala Ala Ile Thr Gln Leu Gln Pro Ile Val Ala Glu Thr Gln
145 150 155 160
Pro Thr Met Ile Ala Ala Ser Val Asp Ala Leu Ala Asp Ala Thr Glu
165 170 175
Leu Ala Leu Ser Gly Gln Thr Ala Thr Arg Val Leu Val Phe Asp His
180 185 190
His Arg Gln Val Asp Ala His Arg Ala Ala Val Glu Ser Ala Arg Glu
195 200 205
Arg Leu Ala Gly Ser Ala Val Val Glu Thr Leu Ala Glu Ala Ile Ala
210 215 220
Arg Gly Asp Val Pro Arg Gly Ala Ser Ala Gly Ser Ala Pro Gly Thr
225 230 235 240
Asp Val Ser Asp Asp Ser Leu Ala Leu Leu Ile Tyr Thr Ser Gly Ser
245 250 255
Thr Gly Ala Pro Lys Gly Ala Met Tyr Pro Arg Arg Asn Val Ala Thr
260 265 270
Phe Trp Arg Lys Arg Thr Trp Phe Glu Gly Gly Tyr Glu Pro Ser Ile
275 280 285
Thr Leu Asn Phe Met Pro Met Ser His Val Met Gly Arg Gln Ile Leu
290 295 300
Tyr Gly Thr Leu Cys Asn Gly Gly Thr Ala Tyr Phe Val Ala Lys Ser
305 310 315 320
Asp Leu Ser Thr Leu Phe Glu Asp Leu Ala Leu Val Arg Pro Thr Glu
325 330 335
Leu Thr Phe Val Pro Arg Val Trp Asp Met Val Phe Asp Glu Phe Gln
340 345 350
Ser Glu Val Asp Arg Arg Leu Val Asp Gly Ala Asp Arg Val Ala Leu
355 360 365
Glu Ala Gln Val Lys Ala Glu Ile Arg Asn Asp Val Leu Gly Gly Arg
370 375 380
Tyr Thr Ser Ala Leu Thr Gly Ser Ala Pro Ile Ser Asp Glu Met Lys
385 390 395 400
Ala Trp Val Glu Glu Leu Leu Asp Met His Leu Val Glu Gly Tyr Gly
405 410 415
Ser Thr Glu Ala Gly Met Ile Leu Ile Asp Gly Ala Ile Arg Arg Pro
420 425 430
Ala Val Leu Asp Tyr Lys Leu Val Asp Val Pro Asp Leu Gly Tyr Phe
435 440 445
Leu Thr Asp Arg Pro His Pro Arg Gly Glu Leu Leu Val Lys Thr Asp
450 455 460
Ser Leu Phe Pro Gly Tyr Tyr Gln Arg Ala Glu Val Thr Ala Asp Val
465 470 475 480
Phe Asp Ala Asp Gly Phe Tyr Arg Thr Gly Asp Ile Met Ala Glu Val
485 490 495
Gly Pro Glu Gln Phe Val Tyr Leu Asp Arg Arg Asn Asn Val Leu Lys
500 505 510
Leu Ser Gln Gly Glu Phe Val Thr Val Ser Lys Leu Glu Ala Val Phe
515 520 525
Gly Asp Ser Pro Leu Val Arg Gln Ile Tyr Ile Tyr Gly Asn Ser Ala
530 535 540
Arg Ala Tyr Leu Leu Ala Val Ile Val Pro Thr Gln Glu Ala Leu Asp
545 550 555 560
Ala Val Pro Val Glu Glu Leu Lys Ala Arg Leu Gly Asp Ser Leu Gln
565 570 575
Glu Val Ala Lys Ala Ala Gly Leu Gln Ser Tyr Glu Ile Pro Arg Asp
580 585 590
Phe Ile Ile Glu Thr Thr Pro Trp Thr Leu Glu Asn Gly Leu Leu Thr
595 600 605
Gly Ile Arg Lys Leu Ala Arg Pro Gln Leu Lys Lys His Tyr Gly Glu
610 615 620
Leu Leu Glu Gln Ile Tyr Thr Asp Leu Ala His Gly Gln Ala Asp Glu
625 630 635 640
Leu Arg Ser Leu Arg Gln Ser Gly Ala Asp Ala Pro Val Leu Val Thr
645 650 655
Val Cys Arg Ala Ala Ala Ala Leu Leu Gly Gly Ser Ala Ser Asp Val
660 665 670
Gln Pro Asp Ala His Phe Thr Asp Leu Gly Gly Asp Ser Leu Ser Ala
675 680 685
Leu Ser Phe Thr Asn Leu Leu His Glu Ile Phe Asp Ile Glu Val Pro
690 695 700
Val Gly Val Ile Val Ser Pro Ala Asn Asp Leu Gln Ala Leu Ala Asp
705 710 715 720
Tyr Val Glu Ala Ala Arg Lys Pro Gly Ser Ser Arg Pro Thr Phe Ala
725 730 735
Ser Val His Gly Ala Ser Asn Gly Gln Val Thr Glu Val His Ala Gly
740 745 750
Asp Leu Ser Leu Asp Lys Phe Ile Asp Ala Ala Thr Leu Ala Glu Ala
755 760 765
Pro Arg Leu Pro Ala Ala Asn Thr Gln Val Arg Thr Val Leu Leu Thr
770 775 780
Gly Ala Thr Gly Phe Leu Gly Arg Tyr Leu Ala Leu Glu Trp Leu Glu
785 790 795 800
Arg Met Asp Leu Val Asp Gly Lys Leu Ile Cys Leu Val Arg Ala Lys
805 810 815
Ser Asp Thr Glu Ala Arg Ala Arg Leu Asp Lys Thr Phe Asp Ser Gly
820 825 830
Asp Pro Glu Leu Leu Ala His Tyr Arg Ala Leu Ala Gly Asp His Leu
835 840 845
Glu Val Leu Ala Gly Asp Lys Gly Glu Ala Asp Leu Gly Leu Asp Arg
850 855 860
Gln Thr Trp Gln Arg Leu Ala Asp Thr Val Asp Leu Ile Val Asp Pro
865 870 875 880
Ala Ala Leu Val Asn His Val Leu Pro Tyr Ser Gln Leu Phe Gly Pro
885 890 895
Asn Ala Leu Gly Thr Ala Glu Leu Leu Arg Leu Ala Leu Thr Ser Lys
900 905 910
Ile Lys Pro Tyr Ser Tyr Thr Ser Thr Ile Gly Val Ala Asp Gln Ile
915 920 925
Pro Pro Ser Ala Phe Thr Glu Asp Ala Asp Ile Arg Val Ile Ser Ala
930 935 940
Thr Arg Ala Val Asp Asp Ser Tyr Ala Asn Gly Tyr Ser Asn Ser Lys
945 950 955 960
Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His Asp Leu Cys Gly Leu
965 970 975
Pro Val Ala Val Phe Arg Cys Asp Met Ile Leu Ala Asp Thr Thr Trp
980 985 990
Ala Gly Gln Leu Asn Val Pro Asp Met Phe Thr Arg Met Ile Leu Ser
995 1000 1005
Leu Ala Ala Thr Gly Ile Ala Pro Gly Ser Phe Tyr Glu Leu Ala
1010 1015 1020
Ala Asp Gly Ala Arg Gln Arg Ala His Tyr Asp Gly Leu Pro Val
1025 1030 1035
Glu Phe Ile Ala Glu Ala Ile Ser Thr Leu Gly Ala Gln Ser Gln
1040 1045 1050
Asp Gly Phe His Thr Tyr His Val Met Asn Pro Tyr Asp Asp Gly
1055 1060 1065
Ile Gly Leu Asp Glu Phe Val Asp Trp Leu Asn Glu Ser Gly Cys
1070 1075 1080
Pro Ile Gln Arg Ile Ala Asp Tyr Gly Asp Trp Leu Gln Arg Phe
1085 1090 1095
Glu Thr Ala Leu Arg Ala Leu Pro Asp Arg Gln Arg His Ser Ser
1100 1105 1110
Leu Leu Pro Leu Leu His Asn Tyr Arg Gln Pro Glu Arg Pro Val
1115 1120 1125
Arg Gly Ser Ile Ala Pro Thr Asp Arg Phe Arg Ala Ala Val Gln
1130 1135 1140
Glu Ala Lys Ile Gly Pro Asp Lys Asp Ile Pro His Val Gly Ala
1145 1150 1155
Pro Ile Ile Val Lys Tyr Val Ser Asp Leu Arg Leu Leu Gly Leu
1160 1165 1170
Leu
<210> 69
<211> 1180
<212> PRT
<213> 海洋分枝杆菌 ATCC BAA-535
<400> 69
Met Ser Ile Thr Cys Val Asp Thr Arg Ala Gln Arg Ser Ala Arg Arg
1 5 10 15
Ile Glu Gln Leu Tyr Ser Thr Asp Ala Gln Phe Ala Ala Ala Arg Pro
20 25 30
Ser Thr Ala Val Gly Ile Ala Ile Ser Lys Ser Gly Leu Gly Leu Pro
35 40 45
Gln Ile Ile Gln Thr Val Met Asp Gly Tyr Pro Gln Arg Pro Ala Leu
50 55 60
Gly Gln Arg Ala Thr Arg Val Val Thr Asp Pro Asn Thr Gly Arg Ser
65 70 75 80
Ser Ala Gln Leu Leu Ala Glu Phe Glu Thr Ile Thr Tyr Arg Glu Leu
85 90 95
Trp Asn Arg Thr Asn Ala Leu Thr Asn Ala Phe Ala Ala Glu Ala Leu
100 105 110
Ala Asp Arg Gly Gln Arg Val Cys Val Leu Gly Phe Ala Ser Ile Asp
115 120 125
Tyr Ala Thr Ile Asp Leu Ala Leu Met Leu Leu Gly Ala Val Ser Val
130 135 140
Pro Leu Pro Thr Asn Ala Ala Arg Ala Gln Leu Cys His Ile Val Ser
145 150 155 160
Glu Thr Gln Pro Ser Leu Ile Ala Ser Ser Thr Glu Asn Leu Pro Asp
165 170 175
Ala Ile Ser Leu Val Leu Ser His Arg Ala Pro His Arg Val Val Val
180 185 190
Phe Asp Tyr Arg Pro Glu Leu Asp Ala His Arg Glu Ala Leu Glu Ala
195 200 205
Ala Arg Ala Arg Leu Ala Ala Ile Pro Val Thr Val Glu Thr Leu Thr
210 215 220
Ala Ile Ile Ala Arg Gly Arg Thr Val Arg Pro Ala Glu Ala Asp Cys
225 230 235 240
Gly Ala Gln Ser Ala Asp Ala Pro Ala Leu Leu Ile Tyr Thr Ser Gly
245 250 255
Ser Thr Gly Ala Pro Lys Gly Val Val Tyr Thr Arg Asn Arg Val Ala
260 265 270
Asp Phe Trp Arg Thr Ser Lys Ala Glu Val Glu Ala Thr Glu Gln Arg
275 280 285
Thr Ala Pro Ser Ile Thr Leu Asn Phe Met Pro Met Ser His Ala Asn
290 295 300
Gly Arg Gln Val Leu Tyr Gly Thr Leu Ser Asn Gly Gly Thr Ala Tyr
305 310 315 320
Phe Thr Ala Arg Ser Asp Leu Ser Thr Leu Phe Asp Asp Leu Ala Leu
325 330 335
Val Arg Pro Thr Glu Leu Gly Phe Pro Pro Arg Ile Trp Asp Met Leu
340 345 350
Leu Glu Arg Phe Gly Arg Glu Val Asp Arg Arg Leu Arg Asp Gly Thr
355 360 365
Ala Glu Gly Ala Asp Pro Gly Ala Leu Lys Ala Arg Val Ala Ala Asp
370 375 380
Leu Arg Gln Val Leu Leu Gly Gly Arg Tyr Ala Leu Ala Met Met Gly
385 390 395 400
Ser Ala Pro Ile Ser Glu Gln Met Lys Ala Ser Val Glu Ser Leu Leu
405 410 415
Asp Leu Asp Val Met Glu Gly Tyr Gly Ser Thr Glu Ala Gly Thr Val
420 425 430
Ile Ile Asn Asn Glu Val Gln Arg Pro Gln Val Ile Asp Tyr Lys Leu
435 440 445
Val Asp Val Ala Glu Leu Gly Tyr Phe Leu Thr Asp Arg Pro Tyr Pro
450 455 460
Arg Gly Glu Leu Leu Val Lys Thr Arg Thr Leu Phe Ser Gly Tyr Tyr
465 470 475 480
Arg Asp Pro Glu Asp Gly Ala Gln Val Phe Asp Pro Asp Gly Phe Tyr
485 490 495
Arg Thr Gly Asp Ile Met Ala Gln Val Gly Pro Asp Arg Leu Ala Tyr
500 505 510
Leu Asp Arg Arg Asn Asn Val Leu Lys Leu Ser Gln Gly Glu Phe Val
515 520 525
Ala Val Ser Arg Leu Glu Ala Ile Phe Ala Asn Ser Pro Leu Val Arg
530 535 540
Gln Ile Phe Val Tyr Ala Asn Gly Ala Arg Ala Tyr Pro Leu Ala Val
545 550 555 560
Val Val Pro Thr Gln Asp Ala Gln Ser Arg His Gly Arg Ala Glu Leu
565 570 575
Lys Ala Glu Leu His Thr Ser Leu His Arg Val Ala Met Ser Ala Gly
580 585 590
Leu Ala Pro Tyr Glu Ile Pro Arg Asp Phe Ile Val Glu Thr Thr Pro
595 600 605
Phe Thr Pro Gln Asn Gly Leu Leu Thr Ala Ile His Lys Leu Ala Arg
610 615 620
Pro His Leu Thr Gln Arg Tyr Gly Ala Arg Leu Glu Leu Leu Tyr Thr
625 630 635 640
Glu Leu Ala Asp Ser Gln Thr Arg Arg Leu His Arg Leu Arg Gln Thr
645 650 655
Gly Gly Arg Leu Pro Ala Leu Glu Thr Ile Arg Arg Ala Ala Gly Ala
660 665 670
Leu Leu Gly Thr Glu Thr Thr Glu Pro Arg Pro Glu Ala His Phe Lys
675 680 685
Asp Leu Gly Gly Asp Ser Val Ser Ala Val Thr Phe Ser Asn Leu Leu
690 695 700
His Asp Ile Tyr Gly Phe Asp Val Pro Val Gly Val Ile Leu Gly Pro
705 710 715 720
Ala Thr Asp Leu Arg Ala Leu Ala Ser His Val Glu Ser Arg Arg Gly
725 730 735
Ala Gly Trp Ser Gly Pro Ser Phe Ala Ser Val His Val Pro Arg Ala
740 745 750
Thr Ser Val His Ala Gly Asp Leu Lys Leu Ala Lys Phe Leu Asp Thr
755 760 765
Lys Thr Leu Ala Ala Ala Thr Ser Leu Pro Ala Ala Asp Ala Arg Ala
770 775 780
Arg Thr Val Leu Leu Thr Gly Ala Thr Gly Phe Leu Gly Arg Tyr Leu
785 790 795 800
Val Leu Glu Trp Leu Arg Arg Leu Arg Ala Val Gly Gly Lys Leu Ile
805 810 815
Cys Leu Val Arg Ala Ala Ser Asp Glu Gln Ala Arg Val Arg Leu Asp
820 825 830
Thr Ala Phe Asp Ser Gly Asp Pro Gln Leu Pro Glu His Phe Arg Gln
835 840 845
Leu Ala Val Asp Arg Leu Glu Val Leu Ala Gly Asp Lys Ser Glu Pro
850 855 860
Gly Leu Gly Leu Asp Gly Pro Thr Trp Gln Arg Leu Ala Asp Thr Val
865 870 875 880
Asp Leu Ile Val Asp Pro Ala Thr Leu Val Asn His Val Leu Ser Tyr
885 890 895
Arg Gln Leu Phe Ala Pro Asn Val Ala Gly Thr Ala Glu Leu Leu Arg
900 905 910
Leu Ala Leu Thr Thr Lys Arg Lys Pro Tyr Ala Tyr Val Ser Thr Val
915 920 925
Ser Val Ala Asn Gln Ile Glu Pro Ser Ala Phe Thr Glu Asp Ala Asp
930 935 940
Ile Arg Glu Ile Ser Arg Thr Arg Thr Ile Asp Asp Ser Phe Ala Asn
945 950 955 960
Gly Tyr Thr Thr Ser Lys Trp Ala Ser Glu Val Leu Leu Arg Glu Ala
965 970 975
His Asp Leu Cys Gly Leu Pro Val Thr Val Phe Arg Cys Asp Met Ile
980 985 990
Leu Ala Asp Thr Ser Tyr Ala Gly Gln Leu Asn Leu Ala Asp Thr Phe
995 1000 1005
Thr Arg Leu Met Leu Ser Val Ala Ala Thr Gly Ile Ala Pro Ala
1010 1015 1020
Ser Phe Tyr Arg Leu Gly Pro Asp Gly Lys Arg Gln Pro Ala His
1025 1030 1035
Phe Asp Gly Leu Pro Val Glu Phe Ile Ala Glu Ala Val Ala Thr
1040 1045 1050
Leu Gly Ala Arg Arg His Asp Gly Phe Gln Val His His Val Ala
1055 1060 1065
Asn Pro His His Asp Gly Val Gly Leu Asp Glu Tyr Val Asp Trp
1070 1075 1080
Leu Val Asp Ala Gly Cys Pro Ile Arg Arg Ile Pro Asp Tyr Asp
1085 1090 1095
Glu Trp Leu Ser Arg Phe Glu Thr Ala Leu His Ala Leu Pro Asp
1100 1105 1110
Arg Lys Arg Arg His Ser Leu Leu Pro Leu Leu Gln Asn Tyr Arg
1115 1120 1125
Glu Pro Ala Glu Pro Ile Arg Gly Gly Ile Ala Pro Ala Pro Arg
1130 1135 1140
Phe Arg Gly Ala Val Arg Gln Ala Lys Ile Gly Arg Asp Asn Asp
1145 1150 1155
Ile Pro His Val Gly Pro Ala Ile Ile Ala Lys Tyr Ala Ser Asp
1160 1165 1170
Leu Gln Leu Leu Gly Leu Ala
1175 1180
<210> 70
<211> 1168
<212> PRT
<213> 耻垢分枝杆菌 ATCC 700084
<400> 70
Met Thr Ile Glu Thr Arg Glu Asp Arg Phe Asn Arg Arg Ile Asp His
1 5 10 15
Leu Phe Glu Thr Asp Pro Gln Phe Ala Ala Ala Arg Pro Asp Glu Ala
20 25 30
Ile Ser Ala Ala Ala Ala Asp Pro Glu Leu Arg Leu Pro Ala Ala Val
35 40 45
Lys Gln Ile Leu Ala Gly Tyr Ala Asp Arg Pro Ala Leu Gly Lys Arg
50 55 60
Ala Val Glu Phe Val Thr Asp Glu Glu Gly Arg Thr Thr Ala Lys Leu
65 70 75 80
Leu Pro Arg Phe Asp Thr Ile Thr Tyr Arg Gln Leu Ala Gly Arg Ile
85 90 95
Gln Ala Val Thr Asn Ala Trp His Asn His Pro Val Asn Ala Gly Asp
100 105 110
Arg Val Ala Ile Leu Gly Phe Thr Ser Val Asp Tyr Thr Thr Ile Asp
115 120 125
Ile Ala Leu Leu Glu Leu Gly Ala Val Ser Val Pro Leu Gln Thr Ser
130 135 140
Ala Pro Val Ala Gln Leu Gln Pro Ile Val Ala Glu Thr Glu Pro Lys
145 150 155 160
Val Ile Ala Ser Ser Val Asp Phe Leu Ala Asp Ala Val Ala Leu Val
165 170 175
Glu Ser Gly Pro Ala Pro Ser Arg Leu Val Val Phe Asp Tyr Ser His
180 185 190
Glu Val Asp Asp Gln Arg Glu Ala Phe Glu Ala Ala Lys Gly Lys Leu
195 200 205
Ala Gly Thr Gly Val Val Val Glu Thr Ile Thr Asp Ala Leu Asp Arg
210 215 220
Gly Arg Ser Leu Ala Asp Ala Pro Leu Tyr Val Pro Asp Glu Ala Asp
225 230 235 240
Pro Leu Thr Leu Leu Ile Tyr Thr Ser Gly Ser Thr Gly Thr Pro Lys
245 250 255
Gly Ala Met Tyr Pro Glu Ser Lys Thr Ala Thr Met Trp Gln Ala Gly
260 265 270
Ser Lys Ala Arg Trp Asp Glu Thr Leu Gly Val Met Pro Ser Ile Thr
275 280 285
Leu Asn Phe Met Pro Met Ser His Val Met Gly Arg Gly Ile Leu Cys
290 295 300
Ser Thr Leu Ala Ser Gly Gly Thr Ala Tyr Phe Ala Ala Arg Ser Asp
305 310 315 320
Leu Ser Thr Phe Leu Glu Asp Leu Ala Leu Val Arg Pro Thr Gln Leu
325 330 335
Asn Phe Val Pro Arg Ile Trp Asp Met Leu Phe Gln Glu Tyr Gln Ser
340 345 350
Arg Leu Asp Asn Arg Arg Ala Glu Gly Ser Glu Asp Arg Ala Glu Ala
355 360 365
Ala Val Leu Glu Glu Val Arg Thr Gln Leu Leu Gly Gly Arg Phe Val
370 375 380
Ser Ala Leu Thr Gly Ser Ala Pro Ile Ser Ala Glu Met Lys Ser Trp
385 390 395 400
Val Glu Asp Leu Leu Asp Met His Leu Leu Glu Gly Tyr Gly Ser Thr
405 410 415
Glu Ala Gly Ala Val Phe Ile Asp Gly Gln Ile Gln Arg Pro Pro Val
420 425 430
Ile Asp Tyr Lys Leu Val Asp Val Pro Asp Leu Gly Tyr Phe Ala Thr
435 440 445
Asp Arg Pro Tyr Pro Arg Gly Glu Leu Leu Val Lys Ser Glu Gln Met
450 455 460
Phe Pro Gly Tyr Tyr Lys Arg Pro Glu Ile Thr Ala Glu Met Phe Asp
465 470 475 480
Glu Asp Gly Tyr Tyr Arg Thr Gly Asp Ile Val Ala Glu Leu Gly Pro
485 490 495
Asp His Leu Glu Tyr Leu Asp Arg Arg Asn Asn Val Leu Lys Leu Ser
500 505 510
Gln Gly Glu Phe Val Thr Val Ser Lys Leu Glu Ala Val Phe Gly Asp
515 520 525
Ser Pro Leu Val Arg Gln Ile Tyr Val Tyr Gly Asn Ser Ala Arg Ser
530 535 540
Tyr Leu Leu Ala Val Val Val Pro Thr Glu Glu Ala Leu Ser Arg Trp
545 550 555 560
Asp Gly Asp Glu Leu Lys Ser Arg Ile Ser Asp Ser Leu Gln Asp Ala
565 570 575
Ala Arg Ala Ala Gly Leu Gln Ser Tyr Glu Ile Pro Arg Asp Phe Leu
580 585 590
Val Glu Thr Thr Pro Phe Thr Leu Glu Asn Gly Leu Leu Thr Gly Ile
595 600 605
Arg Lys Leu Ala Arg Pro Lys Leu Lys Ala His Tyr Gly Glu Arg Leu
610 615 620
Glu Gln Leu Tyr Thr Asp Leu Ala Glu Gly Gln Ala Asn Glu Leu Arg
625 630 635 640
Glu Leu Arg Arg Asn Gly Ala Asp Arg Pro Val Val Glu Thr Val Ser
645 650 655
Arg Ala Ala Val Ala Leu Leu Gly Ala Ser Val Thr Asp Leu Arg Ser
660 665 670
Asp Ala His Phe Thr Asp Leu Gly Gly Asp Ser Leu Ser Ala Leu Ser
675 680 685
Phe Ser Asn Leu Leu His Glu Ile Phe Asp Val Asp Val Pro Val Gly
690 695 700
Val Ile Val Ser Pro Ala Thr Asp Leu Ala Gly Val Ala Ala Tyr Ile
705 710 715 720
Glu Gly Glu Leu Arg Gly Ser Lys Arg Pro Thr Tyr Ala Ser Val His
725 730 735
Gly Arg Asp Ala Thr Glu Val Arg Ala Arg Asp Leu Ala Leu Gly Lys
740 745 750
Phe Ile Asp Ala Lys Thr Leu Ser Ala Ala Pro Gly Leu Pro Arg Ser
755 760 765
Gly Thr Glu Ile Arg Thr Val Leu Leu Thr Gly Ala Thr Gly Phe Leu
770 775 780
Gly Arg Tyr Leu Ala Leu Glu Trp Leu Glu Arg Met Asp Leu Val Asp
785 790 795 800
Gly Lys Val Ile Cys Leu Val Arg Ala Arg Ser Asp Asp Glu Ala Arg
805 810 815
Ala Arg Leu Asp Ala Thr Phe Asp Thr Gly Asp Ala Thr Leu Leu Glu
820 825 830
His Tyr Arg Ala Leu Ala Ala Asp His Leu Glu Val Ile Ala Gly Asp
835 840 845
Lys Gly Glu Ala Asp Leu Gly Leu Asp His Asp Thr Trp Gln Arg Leu
850 855 860
Ala Asp Thr Val Asp Leu Ile Val Asp Pro Ala Ala Leu Val Asn His
865 870 875 880
Val Leu Pro Tyr Ser Gln Met Phe Gly Pro Asn Ala Leu Gly Thr Ala
885 890 895
Glu Leu Ile Arg Ile Ala Leu Thr Thr Thr Ile Lys Pro Tyr Val Tyr
900 905 910
Val Ser Thr Ile Gly Val Gly Gln Gly Ile Ser Pro Glu Ala Phe Val
915 920 925
Glu Asp Ala Asp Ile Arg Glu Ile Ser Ala Thr Arg Arg Val Asp Asp
930 935 940
Ser Tyr Ala Asn Gly Tyr Gly Asn Ser Lys Trp Ala Gly Glu Val Leu
945 950 955 960
Leu Arg Glu Ala His Asp Trp Cys Gly Leu Pro Val Ser Val Phe Arg
965 970 975
Cys Asp Met Ile Leu Ala Asp Thr Thr Tyr Ser Gly Gln Leu Asn Leu
980 985 990
Pro Asp Met Phe Thr Arg Leu Met Leu Ser Leu Val Ala Thr Gly Ile
995 1000 1005
Ala Pro Gly Ser Phe Tyr Glu Leu Asp Ala Asp Gly Asn Arg Gln
1010 1015 1020
Arg Ala His Tyr Asp Gly Leu Pro Val Glu Phe Ile Ala Glu Ala
1025 1030 1035
Ile Ser Thr Ile Gly Ser Gln Val Thr Asp Gly Phe Glu Thr Phe
1040 1045 1050
His Val Met Asn Pro Tyr Asp Asp Gly Ile Gly Leu Asp Glu Tyr
1055 1060 1065
Val Asp Trp Leu Ile Glu Ala Gly Tyr Pro Val His Arg Val Asp
1070 1075 1080
Asp Tyr Ala Thr Trp Leu Ser Arg Phe Glu Thr Ala Leu Arg Ala
1085 1090 1095
Leu Pro Glu Arg Gln Arg Gln Ala Ser Leu Leu Pro Leu Leu His
1100 1105 1110
Asn Tyr Gln Gln Pro Ser Pro Pro Val Cys Gly Ala Met Ala Pro
1115 1120 1125
Thr Asp Arg Phe Arg Ala Ala Val Gln Asp Ala Lys Ile Gly Pro
1130 1135 1140
Asp Lys Asp Ile Pro His Val Thr Ala Asp Val Ile Val Lys Tyr
1145 1150 1155
Ile Ser Asn Leu Gln Met Leu Gly Leu Leu
1160 1165
<210> 71
<211> 835
<212> PRT
<213> 耻垢分枝杆菌 ATCC 700084
<400> 71
Met Trp Asp Met Leu Phe Gly Thr Phe Gln Ser Glu Val Arg Arg Arg
1 5 10 15
Ser Ala Asp Gln Gly Ala His Gly Asp Leu Glu Glu Leu Ala Ala Glu
20 25 30
Val Met Thr Asp Leu Ala Gln His Leu Leu Gly Gly Arg Tyr Leu Leu
35 40 45
Ala Met Thr Gly Ser Ala Pro Ile Ser Asp Glu Asn Lys Ala Phe Val
50 55 60
Glu Glu Leu Leu Asp Leu His Leu Ile Glu Gly Tyr Gly Ser Thr Glu
65 70 75 80
Ala Gly Met Val Phe Val Asp Gly Thr Val Arg Arg Pro Ala Val Ile
85 90 95
Asp Tyr Lys Leu Ala Asp Val Pro Asp Leu Gly Tyr Phe His Thr Asp
100 105 110
Gln Pro His Pro Arg Gly Glu Leu Leu Val Lys Thr Arg Asp Leu Phe
115 120 125
Pro Gly Tyr Tyr Lys Arg Pro Glu Val Thr Ala Ala Val Phe Asp Pro
130 135 140
Asp Gly Phe Tyr His Thr Gly Asp Val Val Ala Glu Val Gly Pro Asp
145 150 155 160
Gln Leu Val Tyr Leu Asp Arg Arg Asn Asn Val Leu Lys Leu Ser Gln
165 170 175
Gly Glu Phe Val Thr Val Ser Lys Leu Glu Pro Leu Phe Gly Asp Ser
180 185 190
Pro Leu Val Arg Gln Ile Tyr Ile Tyr Gly Asn Ser Ala Arg Ala Tyr
195 200 205
Leu Leu Ala Val Val Val Pro Thr Glu Asp Ala Leu Arg His Ser Gly
210 215 220
Gly Asn Ile Asp Gln Leu Lys Pro Ala Ile Ser Ala Ser Phe Gln Asp
225 230 235 240
Ile Ala Arg Ala Ala Gly Leu Gln Ser Tyr Glu Ile Pro Arg Asp Phe
245 250 255
Leu Val Glu Thr Thr Pro Phe Ser Leu Glu Asn Gly Leu Leu Thr Gly
260 265 270
Ile Arg Lys Leu Ala Arg Pro Lys Leu Lys Ala His Tyr Ala Glu Arg
275 280 285
Leu Glu Ala Leu Tyr Ala Glu Leu Ala Asp Lys Gln Ala Asp Glu Leu
290 295 300
Ala Ala Leu Arg Arg Asn Gly Ala Gln Arg Pro Val Leu Glu Thr Val
305 310 315 320
Ser Arg Ala Ala Gly Ala Leu Leu Gly Ala Thr Ala Asp Thr Leu Ala
325 330 335
Pro Asp Ala His Phe Thr Asp Leu Gly Gly Asp Ser Leu Ser Ala Leu
340 345 350
Thr Phe Gly Asn Leu Leu Arg Glu Val Phe Gly Val Asp Val Pro Val
355 360 365
Gly Val Ile Val Ser Pro Ala Ser Asp Leu Arg Ser Ile Ala Asn Tyr
370 375 380
Val Glu Ala Gln Arg Gly Ser Gly Ala Ala Gly Pro Ser Phe Ala Ser
385 390 395 400
Val His Gly Arg Asn Ala Thr Gln Val Arg Ala Val Asp Leu Thr Leu
405 410 415
Glu Lys Phe Leu Asp Ser Gln Thr Leu Thr Ala Pro Arg Ser Ala Pro
420 425 430
Ser Ala Asp Val Arg Thr Val Leu Leu Thr Gly Ala Thr Gly Phe Leu
435 440 445
Gly Arg Tyr Leu Ala Leu Glu Trp Leu Glu Arg Met Ser Glu Val Gly
450 455 460
Gly Thr Val Ile Cys Leu Val Arg Ala Lys Asp Asp Ala Ser Ala Arg
465 470 475 480
Arg Arg Leu Asp Arg Thr Phe Asp Ser Gly Asp Pro Glu Leu Leu His
485 490 495
His Tyr Leu Asp Leu Ala Asp Arg His Leu Arg Val Ile Ala Gly Ala
500 505 510
Lys Gly Glu Ala Asn Leu Gly Leu Asp Pro Gly Thr Trp Gln Gln Leu
515 520 525
Ala Asp Thr Val Asp Val Ile Val Asp Pro Ala Ala Leu Val Asn His
530 535 540
Val Leu Pro Tyr Gln Glu Leu Phe Gly Pro Asn Val Val Gly Thr Ala
545 550 555 560
Glu Leu Ile Arg Leu Ala Leu Thr Thr Lys Pro Lys Pro Tyr Thr Tyr
565 570 575
Val Ser Thr Ile Gly Val Gly Asp Gln Ile Glu Pro Ser Met Phe Thr
580 585 590
Glu Asp Ala Asp Ile Arg Thr Val Ser Ala Val Arg Ala Ile Asp Asp
595 600 605
Ser Tyr Ala Asn Gly Tyr Gly Asn Ser Lys Trp Ala Gly Glu Val Leu
610 615 620
Leu Arg Glu Ala His Asp Arg Cys Gly Leu Pro Val Ala Val Phe Arg
625 630 635 640
Cys Asp Met Ile Leu Ala Asp Thr Thr Trp Ala Gly Gln Leu Asn Val
645 650 655
Pro Asp Met Phe Thr Arg Leu Met Leu Ser Leu Val Ala Thr Gly Ile
660 665 670
Ala Pro Gly Ser Phe Tyr Glu Leu Asp Ala His Gly Asn Arg Gln Arg
675 680 685
Ala His Tyr Asp Gly Leu Pro Val Gly Phe Ile Ala Glu Ala Ile Ala
690 695 700
Thr Leu Gly Ala Ala Ala Ala Asp Gly Phe Ser Thr Tyr His Val Met
705 710 715 720
Asn Pro His Asp Asp Gly Ile Gly Leu Asp Glu Phe Val Asp Trp Leu
725 730 735
Asp Glu Ala Gly Tyr Pro Ile Glu Arg Ile Pro Asp Tyr Ala Ala Trp
740 745 750
Leu Gln Arg Phe Asp Thr Ala Leu His Ala Leu Pro Asp Ala Gln Arg
755 760 765
Gln Ala Ser Leu Leu Pro Leu Leu His Asn Tyr Gln His Pro Glu Lys
770 775 780
Pro Ile Arg Gly Ser Gln Ala Pro Thr Asp Arg Phe Arg Ala Ala Val
785 790 795 800
Arg Gln Val Lys Ile Gly Ala Gly Glu Gly Ser Arg Asp Ile Pro Gln
805 810 815
Val Ser Ala Pro Ile Ile Val Lys Tyr Val Thr Asp Leu Gln Lys Val
820 825 830
Gly Leu Leu
835
<210> 72
<211> 1164
<212> PRT
<213> 巴西诺卡氏菌 NBRC 14402
<400> 72
Met Glu Arg Lys Ala Glu Val Leu Ala Ala Arg Arg Val Glu Asp Leu
1 5 10 15
Ile Glu Arg Asp Ala Gln Val Arg Ala Ala Ile Pro Asp Pro Val Val
20 25 30
Thr Ala Glu Leu Glu Arg Ala Asp Gly Ser Leu Ala Gln Thr Val Ala
35 40 45
Arg Val Met Ala Gly Tyr Ala Asp Arg Pro Ala Leu Gly Gln Arg Ala
50 55 60
Val Glu Phe Val Pro Asp Glu Asn Gly Cys Arg Arg Thr Arg Leu Leu
65 70 75 80
Pro Trp Phe Asp Thr Ile Thr Phe Gly Glu Leu Trp Gln Arg Val Gly
85 90 95
Thr Val Ala Ala Val Trp Gln Ser Arg Ala Glu Arg Ser Val Arg Ala
100 105 110
Gly Asp Phe Val Ala Val Leu Gly Ser Thr Gly Ile Asp Tyr Thr Val
115 120 125
Val Asp Leu Ala Cys Thr Tyr Ser Gly Ala Val Pro Val Pro Leu Gln
130 135 140
Ala Gly Ala Ser Pro Thr Gln Leu Ala Pro Ile Val Arg Glu Val Glu
145 150 155 160
Pro Lys Val Leu Ala Thr Asp Val Gly Gln Leu Glu Val Ala Val Asp
165 170 175
Leu Ala Ser Ala Gly Asp Ser Val Arg Ser Leu Leu Ile Phe Gly Leu
180 185 190
Tyr Ala Glu Asp Asp Glu His Cys Ala Ala Val Glu Ser Ala Arg Arg
195 200 205
Arg Leu Ala Asp Thr Pro Val Val Val Glu Thr Ile Val Glu Leu Leu
210 215 220
Ala Ser Gly Gln Asp Arg Pro Ala Ala Ser Leu His Ala Ser Ala Asp
225 230 235 240
Pro Asp Glu Leu Ala Leu Leu Ile Tyr Thr Ser Gly Ser Thr Gly Thr
245 250 255
Pro Lys Gly Ala Met Tyr Thr Gln Arg Leu Leu Thr Asn Ala Trp Cys
260 265 270
Ala Ala Gly Ala Ser Pro Met Pro Ser Ile Ala Leu Ser Tyr Leu Pro
275 280 285
Met Ser His Thr Met Ala Arg Gln Leu Leu Leu Thr Gly Leu Ala Arg
290 295 300
Gly Gly Thr Val Tyr Phe Ala Ala Arg Asn Asp Met Ser Thr Leu Phe
305 310 315 320
Asp Asp Phe Ala Leu Ala Arg Pro Thr Leu Leu Gly Phe Val Pro Arg
325 330 335
Val Cys Asp Met Val Leu Gln Arg Phe Gln Ser Glu Met Ala Arg Arg
340 345 350
Val Gly Ala Asp Asp Glu Pro Thr Val Val Glu Gln Glu Val Lys Thr
355 360 365
Glu Leu Arg Glu Gln Phe Leu Gly Gly Arg Phe Leu Val Ala Ser Ile
370 375 380
Gly Ser Ala Pro Leu Ser Ala Asp Met Arg Glu Phe Met Gln Ser Val
385 390 395 400
Leu Gly Ile Ala Leu Ile Asp Ala Tyr Gly Ser Thr Glu Thr Gly Gly
405 410 415
Val Leu Met Asn Asn Lys Val Val Arg Thr Ala Ile Leu Asp Tyr Lys
420 425 430
Leu Val Asp Val Pro Glu Leu Gly Tyr Phe Ser Thr Asp Arg Pro His
435 440 445
Pro Arg Gly Glu Leu Leu Ile Lys Ser Arg Thr Leu Ile Pro Gly Tyr
450 455 460
Tyr Lys Arg Pro Glu Leu Asn Ser Gln Phe Phe Asp Ala Glu Gly Phe
465 470 475 480
Tyr Arg Thr Gly Asp Val Met Ala Gln Thr Gly Phe Asp Glu Leu Val
485 490 495
Tyr Val Asp Arg Arg Asn Ser Val Leu Lys Leu Ser Gln Gly Glu Phe
500 505 510
Val Ala Val Ser Lys Leu Glu Ala Ile Phe Val Gly Ser Pro Leu Val
515 520 525
Glu Gln Ile Tyr Val Tyr Gly Ser Ser Glu Arg Ala Tyr Leu Leu Ala
530 535 540
Val Ile Val Pro Val Ala Glu Ala Ile Ala Thr His Ala Gly Thr Ala
545 550 555 560
Glu Leu Lys Ala Ala Ile Ser Glu Ser Leu Arg Gln Ile Ala Lys Asp
565 570 575
Ala Glu Leu Ser Ser Tyr Glu Val Pro Arg Asp Phe Leu Leu Glu Ser
580 585 590
Glu Pro Phe Thr Val Asp Asn Gly Leu Leu Ala Gly Leu Ser Lys Leu
595 600 605
Leu Arg Pro Ser Leu Lys Glu Arg Tyr Gly Glu Arg Leu Glu Ala Leu
610 615 620
Tyr Arg Val Ala Glu Ala Ala Gln Thr Gln Glu Leu Ala Ala Leu Arg
625 630 635 640
Gln Gln Ala Gly Met Leu Pro Val Leu Glu Thr Val Ser Arg Ala Ala
645 650 655
Gln Ala Val Leu Gly Val Ser Ala Ala Glu Leu Arg Pro Asp Ala His
660 665 670
Phe Thr Asp Leu Gly Gly Asp Ser Leu Ala Ala Leu Ser Phe Ser Thr
675 680 685
Leu Leu Gln Glu Leu Leu Gly Val Gln Val Pro Val Gly Val Ile Ala
690 695 700
Ser Ser Ala Asn Asp Leu Arg Arg Ile Ala Asn Tyr Ala Val Ala Glu
705 710 715 720
Arg Ser Ala Gly Ser Leu Arg Pro Thr Ser Thr Gly Val His Gly Thr
725 730 735
Gly Ser Gln Leu Arg Ala Ile Asp Leu Arg Leu Asp Lys Phe Ile Asp
740 745 750
Pro Ser Thr Leu Ala Ala Ala Thr Thr Leu Pro Arg Ala Gly Glu Pro
755 760 765
Arg Thr Val Leu Leu Thr Gly Ala Asn Gly Tyr Leu Gly Arg Phe Leu
770 775 780
Cys Leu Glu Trp Leu Gln Arg Leu His His Ser Gly Gly Thr Leu Ile
785 790 795 800
Cys Val Val Arg Gly Ile Asp Ala Val Ala Ala Arg Glu Arg Leu Asp
805 810 815
Glu Val Phe Asp Ser Gly Asp Pro Glu Leu Leu His Arg Tyr Arg Glu
820 825 830
Leu Ala Glu Gly Thr Leu Glu Val Leu Cys Gly Asp Ile Ser Glu Pro
835 840 845
Gly Leu Gly Leu Ser Glu Arg Asp Trp Arg Arg Leu Ala Asp Thr Val
850 855 860
Glu Leu Ile Val His Ala Ala Ala Leu Val Asn His Val Leu Pro Tyr
865 870 875 880
Gly Gln Leu Phe Gly Pro Asn Val Val Gly Thr Ala Glu Ile Ile Arg
885 890 895
Leu Ala Met Thr Thr Arg Ile Lys Pro Val Thr Tyr Leu Ser Thr Val
900 905 910
Ala Val Ala Ala Gln Val Ala Pro Glu Gln Phe Thr Glu Asp Gly Asp
915 920 925
Ile Arg Glu Ile Ser Ala Val Arg Ser Leu Asp Glu Gly Tyr Ala Asp
930 935 940
Gly Tyr Gly Thr Ser Lys Trp Ala Gly Glu Val Leu Leu Arg Glu Ala
945 950 955 960
Tyr Asp Leu Cys Gly Leu Pro Val Ala Val Phe Arg Ser Asp Met Ile
965 970 975
Leu Ala His Ser Arg Phe Ala Gly Gln Leu Asn Val Ser Asp Met Phe
980 985 990
Thr Arg Leu Val Leu Ser Val Leu Ala Thr Gly Val Ala Pro Lys Ser
995 1000 1005
Phe Tyr Glu Thr Asp Arg Met Gly Asn Arg Gln Arg Ala His Tyr
1010 1015 1020
Asp Gly Leu Pro Val Asp Phe Thr Ala Gln Ala Ile Thr Val Leu
1025 1030 1035
Gly Ser Gln Val Val Ser Gly Phe Glu Thr Phe Asp Val Leu Asn
1040 1045 1050
Pro His Asp Asp Gly Leu Ser Leu Asp Glu Phe Val Asp Trp Leu
1055 1060 1065
Ile Ala Ala Gly His Ser Ile Asp Arg Ile Asp Gly Tyr Ala Glu
1070 1075 1080
Trp Leu Ser Arg Phe Gly Thr Ala Leu Arg Val Leu Ser Glu Arg
1085 1090 1095
Gln Arg Gln His Ser Val Leu Pro Leu Leu His Ala Tyr Arg Arg
1100 1105 1110
Ala Ala Val Pro Ile Pro Gly Ala Ala Leu Pro Ala Lys Lys Phe
1115 1120 1125
Gln Ala Ala Val Gln Asp Ala Gln Leu Gly Pro Gly Arg Asp Ile
1130 1135 1140
Pro His Leu Thr Pro Asp Leu Ile Glu Lys Tyr Val Ser Asp Leu
1145 1150 1155
Lys Leu Arg Asn Leu Leu
1160
<210> 73
<211> 1165
<212> PRT
<213> 巴西诺卡氏菌 NBRC 14402
<400> 73
Met Thr Asp Val Glu Val Ala Arg Arg Ile Asp Asp Leu Ser Ala Thr
1 5 10 15
Asp Glu Val Leu Ala Gly Ala Leu Pro Asp Pro Ala Val Asn Ala Arg
20 25 30
Ile Gln Asp Pro Ala Val Gly Leu Ala Glu Leu Ile Arg Ile Val Phe
35 40 45
Asp Ala Tyr Gly Gly Arg Pro Ala Val Gly Ala Arg Ala Thr Arg Leu
50 55 60
Val Thr Asp Pro Val Thr Gly Arg Thr Thr Leu His Val Leu Pro Glu
65 70 75 80
Tyr Glu Thr Thr Thr Tyr Ala Glu Leu Gly Arg Gln Val Ala Ala Val
85 90 95
Ala Ala Gly Leu Arg Gly Glu Ala Ala Asp Gly Gly Trp Val Arg Pro
100 105 110
Gly Glu His Val Ala Met Leu Gly Phe Thr Ser Val Glu Tyr Thr Val
115 120 125
Leu Asp Leu Ala Ala Thr Leu Ala Gly Ala Ile Ala Val Pro Met Gln
130 135 140
Ser Asn Ala Pro Ala Val Gln Leu Arg Pro Ile Leu Ala Glu Thr Gln
145 150 155 160
Pro Arg Val Leu Ala Ala Gly Val Asp Gln Leu Ala Glu Ala Leu Asp
165 170 175
Leu Ala Asp Gly Glu His Arg Pro Gly Leu Ile Val Val Leu Asp His
180 185 190
Arg Ala Glu Val Asp Ala His Arg Glu Ala Tyr Glu Ala Ala Ala Ala
195 200 205
Arg Leu Ala Gly Thr Gly Val Ala Leu Lys Thr Leu Ala Glu Leu Ala
210 215 220
Asp Ser Gly Lys Ala Gly Glu Val Ala Pro Glu Ala Ala Asp Ser Glu
225 230 235 240
Arg Ile Val Leu Leu Ile Tyr Thr Ser Gly Ser Thr Gly Thr Pro Lys
245 250 255
Gly Ala Met Tyr Ser Glu Arg Leu Val Ala Asn Leu Trp Gln Gly Lys
260 265 270
Phe Gly Leu Gly Glu Gly Pro His Ala Ala Gly Pro Trp Ile Thr Leu
275 280 285
Asn Phe Met Pro Met Ser His Leu Met Gly Arg Tyr Thr Leu Tyr Gly
290 295 300
Thr Leu Ala Arg Gly Gly Ile Ala Tyr Phe Ala Ala Ser Ser Asp Leu
305 310 315 320
Ser Thr Phe Leu Glu Asp Leu Ser Leu Val Arg Pro Thr Gln Leu Gln
325 330 335
Phe Val Pro Arg Val Trp Asp Leu Leu Tyr Gln Glu Tyr Gln Arg Ala
340 345 350
Leu Asp Gly Val Gly Pro Ala Glu Arg Ser Asp Arg Ala Pro Glu Leu
355 360 365
Leu Ala Glu Leu Arg Arg Asp Leu Leu Gly Gly Arg Ala Leu Gly Ala
370 375 380
Val Thr Gly Ser Ala Pro Ile Ser Val Glu Val Arg Glu Phe Val Asp
385 390 395 400
Ala Leu Leu Gly Phe His Leu Pro Asp Gly Tyr Gly Ser Thr Glu Ala
405 410 415
Gly Gly Ile Thr Val Asp Gly Lys Val Ala Arg Pro Pro Val Leu Asp
420 425 430
Tyr Lys Leu Ala Asp Val Pro Glu Leu Gly Tyr Tyr Arg Thr Asp Arg
435 440 445
Pro Tyr Pro Arg Gly Glu Leu Leu Val Lys Thr Gln Asn Ile Phe Pro
450 455 460
Gly Tyr Tyr Arg Arg Pro Glu Val Thr Ala Glu Val Phe Asp Pro Asp
465 470 475 480
Gly Tyr Tyr Arg Thr Gly Asp Ile Val Ala Glu Val Gln Pro Asp Glu
485 490 495
Leu Arg Tyr Leu Asp Arg Arg Asn Asn Val Leu Lys Leu Ser Gln Gly
500 505 510
Glu Phe Val Thr Val Ser Lys Leu Glu Ala Ala Tyr Gly Ala Ser Pro
515 520 525
Leu Ile Gln Gln Ile Tyr Ile Tyr Gly Asn Ser Ser Arg Pro Tyr Leu
530 535 540
Leu Ala Val Ile Val Pro Asp Asp Ala Val Leu Glu Arg Val Ser Gly
545 550 555 560
Asp Val Gly Ala Ile Gly Pro Leu Leu Thr Glu Ala Leu Arg Ala Val
565 570 575
Ala Ala Asp Ala Gly Leu Gln Ser Tyr Glu Ile Pro Arg Asp Phe Leu
580 585 590
Val Glu Thr Thr Pro Phe Thr Val Glu Asn Gly Leu Leu Thr Gly Ile
595 600 605
Arg Lys Leu Ala Arg Pro Lys Leu Arg Gln Arg Tyr Gly Glu Leu Leu
610 615 620
Glu Gln Leu Tyr Gln Arg Leu Ser Asp Gly Gln Ala Glu Glu Leu Arg
625 630 635 640
Ala Leu Arg Thr Gly Ala Ala Asp Arg Pro Val Leu Glu Thr Val Thr
645 650 655
Arg Val Ala Ala Ala Leu Leu Gly Ile Pro Ala Ala Asp Val His Pro
660 665 670
Ala Ala His Phe Thr Glu Leu Gly Gly Asp Ser Leu Ser Ala Leu Thr
675 680 685
Phe Gly Asn Ala Leu His Asp Ile Leu Gly Val Glu Val Pro Val Gly
690 695 700
Val Ile Val Ser Pro Ala Thr Asp Leu Arg Ala Leu Ala Thr Phe Leu
705 710 715 720
Glu Ser Gly Ala Gln Ala Thr Arg Pro Thr Phe Glu Ser Val His Gly
725 730 735
Pro Glu Ala Thr Glu Val Arg Ala Ala Asp Leu Thr Leu Glu Lys Phe
740 745 750
Leu Asp Ala Ala Thr Leu Asp Gly Ala Ala Ala Leu Pro Ala Pro Asn
755 760 765
Tyr Ala Val Arg Thr Val Leu Leu Thr Gly Ala Thr Gly Phe Leu Gly
770 775 780
Arg Phe Leu Ala Leu Ala Leu Leu Glu Arg Leu Gly Pro Val Asp Gly
785 790 795 800
Thr Leu Ile Cys Leu Val Arg Ala Thr Asp Asp Ala Ala Ala Arg Gln
805 810 815
Arg Leu Asp Ala Val Phe Asp Ser Gly Asp Pro Gln Leu Leu Thr His
820 825 830
Tyr Arg Arg Leu Ala Glu Arg His Leu Glu Val Val Ala Gly Asp Lys
835 840 845
Gly Asp Leu Asp Leu Gly Leu Asp Arg Ala Thr Trp Gln Gln Leu Ala
850 855 860
Glu Arg Val Asp Leu Ile Val Asp Cys Ala Ala Leu Val Asn His Val
865 870 875 880
Leu Pro Tyr Arg Gln Leu Phe Gly Pro Asn Val Val Gly Thr Ala Glu
885 890 895
Leu Ile Arg Leu Ala Leu Thr Gly Lys Gln Lys Ile Phe Asp Tyr Ile
900 905 910
Ser Thr Val Gly Val Gly Asp Gln Ile Ala Ala Gly Gln Phe Val Glu
915 920 925
Asp Ala Asp Ile Arg Thr Ile Ser Ala Thr Arg Ser Val Asp Glu Arg
930 935 940
Tyr Ala Asn Gly Tyr Gly Asn Ser Lys Trp Ala Gly Glu Val Leu Leu
945 950 955 960
Arg Ala Ala His Glu Arg Tyr Gly Leu Pro Val Ser Val Phe Arg Cys
965 970 975
Asp Met Ile Met Val Asp Gly Arg Tyr Val Gly Gln Leu Asn Val Pro
980 985 990
Asp Met Phe Thr Arg Leu Met Leu Ser Leu Val Ala Thr Gly Val Ala
995 1000 1005
Pro Gly Ser Phe Tyr Gln Thr Asp Ala Ala Gly Gln Arg Gln Arg
1010 1015 1020
Ala His Tyr Asp Gly Leu Pro Val Asp Phe Ile Ala Glu Ala Val
1025 1030 1035
Thr Glu Leu Gly Val His Glu Gly Phe Ser Thr Tyr His Val Met
1040 1045 1050
Asn Pro His His Asp Gly Ile Gly Leu Asp Glu Phe Val Asp Trp
1055 1060 1065
Leu Val Ala Ala Gly Tyr Pro Ile Ala Arg Val Ala Asp Tyr Gln
1070 1075 1080
Ala Trp Leu Glu Gln Phe Gly Thr Lys Leu Arg Ala Leu Pro Glu
1085 1090 1095
Gln Gln Arg Arg His Ser Leu Leu Pro Leu Leu His Ser Tyr Ala
1100 1105 1110
His Pro Gln Pro Pro Val Glu Gly Ser Val Ala Pro Ala Asp Arg
1115 1120 1125
Phe Arg Ala Ala Val Gln Asp Ala Asn Ile Gly Pro Asp Lys Asp
1130 1135 1140
Ile Pro His Leu Thr Glu Ser Thr Ile Leu Lys Tyr Val Thr Asn
1145 1150 1155
Leu Glu His Leu Gly Leu Leu
1160 1165
<210> 74
<211> 264
<212> PRT
<213> 炭疽芽孢杆菌
<400> 74
Met His Met Gln Leu Lys Gly Lys Thr Ala Leu Val Thr Gly Ser Thr
1 5 10 15
Ala Gly Ile Gly Lys Ala Ile Ala Thr Ser Leu Val Ala Glu Gly Ala
20 25 30
Asn Val Leu Ile Asn Gly Arg Arg Glu Glu Asn Val Asn Glu Thr Ile
35 40 45
Lys Glu Ile Arg Ala Gln Tyr Pro Asp Ala Ile Leu Gln Pro Val Val
50 55 60
Ala Asp Leu Gly Thr Glu Gln Gly Cys Gln Asp Val Ile Glu Lys Tyr
65 70 75 80
Pro Lys Val Asp Ile Leu Ile Asn Asn Leu Gly Ile Phe Glu Pro Val
85 90 95
Glu Tyr Phe Asp Ile Pro Asp Glu Asp Trp Phe Lys Leu Phe Glu Val
100 105 110
Asn Ile Met Ser Gly Val Arg Leu Thr Arg Ser Tyr Leu Lys Lys Met
115 120 125
Ile Glu Arg Lys Glu Gly Arg Val Ile Phe Ile Ala Ser Glu Ala Ala
130 135 140
Ile Met Pro Ser Gln Glu Met Ala His Tyr Ser Ala Thr Lys Thr Met
145 150 155 160
Gln Leu Ser Leu Ser Arg Ser Leu Ala Glu Leu Thr Thr Gly Thr Asn
165 170 175
Val Thr Val Asn Thr Ile Met Pro Gly Ser Thr Leu Thr Glu Gly Val
180 185 190
Glu Thr Met Leu Asn Ser Leu Tyr Pro Asn Glu Gln Leu Thr Ile Glu
195 200 205
Glu Ala Glu Lys Arg Phe Met Lys Glu Asn Arg Pro Thr Ser Ile Ile
210 215 220
Gln Arg Leu Ile Arg Pro Glu Glu Ile Ala His Leu Val Thr Phe Leu
225 230 235 240
Ser Ser Pro Leu Ser Ser Ala Ile Asn Gly Ser Ala Leu Arg Ile Asp
245 250 255
Gly Gly Leu Val Arg Ser Val Phe
260
<210> 75
<211> 254
<212> PRT
<213> 炭疽芽孢杆菌
<400> 75
Met Lys Glu Lys Val Val Ile Ile Thr Gly Gly Ser Ser Gly Met Gly
1 5 10 15
Lys Gly Met Ala Thr Arg Phe Ala Lys Glu Gly Ala Arg Val Val Ile
20 25 30
Thr Gly Arg Thr Lys Glu Lys Leu Glu Glu Ala Lys Leu Glu Ile Glu
35 40 45
Gln Phe Pro Gly Gln Ile Leu Thr Val Gln Met Asp Val Arg Asn Thr
50 55 60
Asp Asp Ile Gln Lys Met Ile Glu Gln Ile Asp Glu Lys Phe Gly Arg
65 70 75 80
Ile Asp Ile Leu Ile Asn Asn Ala Ala Gly Asn Phe Ile Cys Pro Ala
85 90 95
Glu Asp Leu Ser Val Asn Gly Trp Asn Ser Val Ile Asn Ile Val Leu
100 105 110
Asn Gly Thr Phe Tyr Cys Ser Gln Ala Ile Gly Lys Tyr Trp Ile Glu
115 120 125
Lys Gly Ile Lys Gly Asn Ile Ile Asn Met Val Ala Thr Tyr Ala Trp
130 135 140
Asp Ala Gly Pro Gly Val Ile His Ser Ala Ala Ala Lys Ala Gly Val
145 150 155 160
Leu Ala Met Thr Lys Thr Leu Ala Val Glu Trp Gly Arg Lys Tyr Gly
165 170 175
Ile Arg Val Asn Ala Ile Ala Pro Gly Pro Ile Glu Arg Thr Gly Gly
180 185 190
Ala Asp Lys Leu Trp Ile Ser Glu Glu Met Ala Lys Arg Thr Ile Gln
195 200 205
Ser Val Pro Leu Gly Arg Leu Gly Thr Pro Glu Glu Ile Ala Gly Leu
210 215 220
Ala Tyr Tyr Leu Cys Ser Asp Glu Ala Ala Tyr Ile Asn Gly Thr Cys
225 230 235 240
Met Thr Met Asp Gly Gly Gln His Leu His Gln Tyr Pro Phe
245 250
<210> 76
<211> 257
<212> PRT
<213> 副百日咳博德特氏菌 ATCC BAA-587
<400> 76
Met Ala Asp His Ser Ile Gln Gly Lys Val Ala Leu Ile Ala Gly Gly
1 5 10 15
Ala Lys Asn Leu Gly Gly Leu Ile Ala Arg Asp Leu Ala Ala His Gly
20 25 30
Ala Arg Ala Ile Ala Ile His Tyr Asn Ser Pro Ala Ser Lys Ala Asp
35 40 45
Ala Asp Ala Thr Val Ala Ala Leu Gln Ala Ala Gly Ala Gln Ala Val
50 55 60
Ala Leu Gln Gly Asp Leu Thr Thr Ala Ala Ala Val Glu Lys Leu Tyr
65 70 75 80
Ala Asp Ala Leu Ala Ala Val Gly Arg Pro Asp Ile Ala Ile Asn Thr
85 90 95
Val Gly Lys Val Leu Lys Lys Pro Phe Ser Glu Ile Ser Glu Ala Glu
100 105 110
Tyr Asp Glu Met Ala Ala Val Asn Ala Lys Ser Ala Phe Phe Phe Leu
115 120 125
Lys Glu Ala Gly Arg His Val Asn Asp His Gly Lys Ile Val Thr Leu
130 135 140
Val Thr Ser Leu Leu Gly Ala Phe Thr Pro Phe Tyr Ser Ala Tyr Ala
145 150 155 160
Gly Thr Lys Ala Pro Val Glu His Tyr Thr Arg Ala Ala Ala Lys Glu
165 170 175
Phe Gly Ala Arg Gly Ile Ser Val Thr Ala Val Gly Pro Gly Pro Met
180 185 190
Asp Thr Pro Phe Phe Tyr Gly Gln Glu Ser Pro Glu Ala Val Ala Tyr
195 200 205
His Gln Ser Ala Ala Ala Leu Ser Pro His Ser Pro Thr Gly Leu Thr
210 215 220
His Ile Glu Asp Val Val Pro Phe Ile Arg His Leu Val Ser Asp Gly
225 230 235 240
Trp Trp Ile Thr Gly Gln Thr Leu Leu Ile Asn Gly Gly Tyr Thr Thr
245 250 255
Lys
<210> 77
<211> 253
<212> PRT
<213> 紫色色杆菌ATCC 12472
<400> 77
Met Thr Met Lys Thr Glu Phe Ala Gln Asp Ala Leu Ala Gly Arg Val
1 5 10 15
Ile Leu Ile Thr Gly Ala Ser Gln Gly Ile Gly Arg Glu Ala Ala Leu
20 25 30
Thr Phe Ala Arg His Gly Ala Thr Val Val Leu Leu Ser Arg Ser Val
35 40 45
Lys Gly Leu Glu Lys Val Tyr Asp Glu Ile Val Ala Ala Gly Gly Pro
50 55 60
Glu Pro Ala Ala Val Pro Leu Asp Leu Leu Asn Ala Gly Glu Asn Glu
65 70 75 80
Phe Asn Gln Leu Ala Leu Thr Ile Gln Arg Glu Phe Gly Arg Leu Asp
85 90 95
Gly Ile Ala His Cys Ala Ser His Phe Tyr Ala Leu Ser Pro Leu Thr
100 105 110
Asn Gln Thr Ile Glu Glu Trp Met Asn Gln Tyr Arg Ile Asn Thr Val
115 120 125
Ala Pro Phe Ala Leu Thr Arg Ala Cys Leu Pro Leu Leu Lys Glu Ala
130 135 140
Pro Asp Ala Ser Val Leu Phe Val Gly Glu His His Ala Leu His Pro
145 150 155 160
Ala Ala Tyr Trp Gly Gly Phe Gly Ala Ser Asn Ala Gly Leu Pro Tyr
165 170 175
Leu Thr Lys Val Ala Ala Asp Glu Trp Glu Met Leu Pro Ser Leu Arg
180 185 190
Val Asn Leu Leu Leu Pro Gly Pro Val Asn Ser Pro Gln Arg Asn Arg
195 200 205
Thr His Pro Gly Glu Asp Lys Ser Glu Arg Ala Asp Leu Ala Asp Leu
210 215 220
Met Pro His Phe Leu Tyr Trp Leu Gly Ala Asp Ser Arg Gly Arg Ser
225 230 235 240
Gly Glu Val Val Glu Leu Asp Leu Arg Lys Pro Arg Gly
245 250
<210> 78
<211> 380
<212> PRT
<213> 大肠杆菌 P12b
<400> 78
Met Pro His Leu Ala Leu Leu Ile Ser Lys Gly Ala Ile Met Asp Arg
1 5 10 15
Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp Val Ile Asn
20 25 30
Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp Leu Val Val
35 40 45
Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val Glu Lys Ser
50 55 60
Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe Gly Gly Glu
65 70 75 80
Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala Glu Thr Ala
85 90 95
Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr Leu Asp Thr
100 105 110
Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala Ile Ala Pro
115 120 125
Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser Val Ile Tyr
130 135 140
Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro Asn Asn Pro
145 150 155 160
Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala Pro Ala Arg
165 170 175
Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp Phe Glu Ala
180 185 190
Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly Gly Lys Cys
195 200 205
Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn Thr Leu Leu
210 215 220
Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His Val Val Thr
225 230 235 240
Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu Ser Gly Val
245 250 255
Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val His Asn Gly
260 265 270
Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly Glu Lys Val
275 280 285
Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala Pro Val Glu
290 295 300
Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly Leu Pro Ile
305 310 315 320
Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala Lys Met Arg
325 330 335
Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile His Asn Met
340 345 350
Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu Leu Val Ala
355 360 365
Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
370 375 380
<210> 79
<211> 383
<212> PRT
<213> 大肠杆菌
<400> 79
Met Met Ala Asn Arg Met Ile Leu Asn Glu Thr Ala Trp Phe Gly Arg
1 5 10 15
Gly Ala Val Gly Ala Leu Thr Asp Glu Val Lys Arg Arg Gly Tyr Gln
20 25 30
Lys Ala Leu Ile Val Thr Asp Lys Thr Leu Val Gln Cys Gly Val Val
35 40 45
Ala Lys Val Thr Asp Lys Met Asp Ala Ala Gly Leu Ala Trp Ala Ile
50 55 60
Tyr Asp Gly Val Val Pro Asn Pro Thr Ile Thr Val Val Lys Glu Gly
65 70 75 80
Leu Gly Val Phe Gln Asn Ser Gly Ala Asp Tyr Leu Ile Ala Ile Gly
85 90 95
Gly Gly Ser Pro Gln Asp Thr Cys Lys Ala Ile Gly Ile Ile Ser Asn
100 105 110
Asn Pro Glu Phe Ala Asp Val Arg Ser Leu Glu Gly Leu Ser Pro Thr
115 120 125
Asn Lys Pro Ser Val Pro Ile Leu Ala Ile Pro Thr Thr Ala Gly Thr
130 135 140
Ala Ala Glu Val Thr Ile Asn Tyr Val Ile Thr Asp Glu Glu Lys Arg
145 150 155 160
Arg Lys Phe Val Cys Val Asp Pro His Asp Ile Pro Gln Val Ala Phe
165 170 175
Ile Asp Ala Asp Met Met Asp Gly Met Pro Pro Ala Leu Lys Ala Ala
180 185 190
Thr Gly Val Asp Ala Leu Thr His Ala Ile Glu Gly Tyr Ile Thr Arg
195 200 205
Gly Ala Trp Ala Leu Thr Asp Ala Leu His Ile Lys Ala Ile Glu Ile
210 215 220
Ile Ala Gly Ala Leu Arg Gly Ser Val Ala Gly Asp Lys Asp Ala Gly
225 230 235 240
Glu Glu Met Ala Leu Gly Gln Tyr Val Ala Gly Met Gly Phe Ser Asn
245 250 255
Val Gly Leu Gly Leu Val His Gly Met Ala His Pro Leu Gly Ala Phe
260 265 270
Tyr Asn Thr Pro His Gly Val Ala Asn Ala Ile Leu Leu Pro His Val
275 280 285
Met Arg Tyr Asn Ala Asp Phe Thr Gly Glu Lys Tyr Arg Asp Ile Ala
290 295 300
Arg Val Met Gly Val Lys Val Glu Gly Met Ser Leu Glu Glu Ala Arg
305 310 315 320
Asn Ala Ala Val Glu Ala Val Phe Ala Leu Asn Arg Asp Val Gly Ile
325 330 335
Pro Pro His Leu Arg Asp Val Gly Val Arg Lys Glu Asp Ile Pro Ala
340 345 350
Leu Ala Gln Ala Ala Leu Asp Asp Val Cys Thr Gly Gly Asn Pro Arg
355 360 365
Glu Ala Thr Leu Glu Asp Ile Val Glu Leu Tyr His Thr Ala Trp
370 375 380
<210> 80
<211> 417
<212> PRT
<213> 大肠杆菌
<400> 80
Met Leu Glu Gln Met Gly Ile Ala Ala Lys Gln Ala Ser Tyr Lys Leu
1 5 10 15
Ala Gln Leu Ser Ser Arg Glu Lys Asn Arg Val Leu Glu Lys Ile Ala
20 25 30
Asp Glu Leu Glu Ala Gln Ser Glu Ile Ile Leu Asn Ala Asn Ala Gln
35 40 45
Asp Val Ala Asp Ala Arg Ala Asn Gly Leu Ser Glu Ala Met Leu Asp
50 55 60
Arg Leu Ala Leu Thr Pro Ala Arg Leu Lys Gly Ile Ala Asp Asp Val
65 70 75 80
Arg Gln Val Cys Asn Leu Ala Asp Pro Val Gly Gln Val Ile Asp Gly
85 90 95
Gly Val Leu Asp Ser Gly Leu Arg Leu Glu Arg Arg Arg Val Pro Leu
100 105 110
Gly Val Ile Gly Val Ile Tyr Glu Ala Arg Pro Asn Val Thr Val Asp
115 120 125
Val Ala Ser Leu Cys Leu Lys Thr Gly Asn Ala Val Ile Leu Arg Gly
130 135 140
Gly Lys Glu Thr Cys Arg Thr Asn Ala Ala Thr Val Ala Val Ile Gln
145 150 155 160
Asp Ala Leu Lys Ser Cys Gly Leu Pro Ala Gly Ala Val Gln Ala Ile
165 170 175
Asp Asn Pro Asp Arg Ala Leu Val Ser Glu Met Leu Arg Met Asp Lys
180 185 190
Tyr Ile Asp Met Leu Ile Pro Arg Gly Gly Ala Gly Leu His Lys Leu
195 200 205
Cys Arg Glu Gln Ser Thr Ile Pro Val Ile Thr Gly Gly Ile Gly Val
210 215 220
Cys His Ile Tyr Val Asp Glu Ser Val Glu Ile Ala Glu Ala Leu Lys
225 230 235 240
Val Ile Val Asn Ala Lys Thr Gln Arg Pro Ser Thr Cys Asn Thr Val
245 250 255
Glu Thr Leu Leu Val Asn Lys Asn Ile Ala Asp Ser Phe Leu Pro Ala
260 265 270
Leu Ser Lys Gln Met Ala Glu Ser Gly Val Thr Leu His Ala Asp Ala
275 280 285
Ala Ala Leu Ala Gln Leu Gln Ala Gly Pro Ala Lys Val Val Ala Val
290 295 300
Lys Ala Glu Glu Tyr Asp Asp Glu Phe Leu Ser Leu Asp Leu Asn Val
305 310 315 320
Lys Ile Val Ser Asp Leu Asp Asp Ala Ile Ala His Ile Arg Glu His
325 330 335
Gly Thr Gln His Ser Asp Ala Ile Leu Thr Arg Asp Met Arg Asn Ala
340 345 350
Gln Arg Phe Val Asn Glu Val Asp Ser Ser Ala Val Tyr Val Asn Ala
355 360 365
Ser Thr Arg Phe Thr Asp Gly Gly Gln Phe Gly Leu Gly Ala Glu Val
370 375 380
Ala Val Ser Thr Gln Lys Leu His Ala Arg Gly Pro Met Gly Leu Glu
385 390 395 400
Ala Leu Thr Thr Tyr Lys Trp Ile Gly Ile Gly Asp Tyr Thr Ile Arg
405 410 415
Ala
<210> 81
<211> 252
<212> PRT
<213> 大肠杆菌
<400> 81
Met His Tyr Gln Pro Lys Gln Asp Leu Leu Asn Asp Arg Ile Ile Leu
1 5 10 15
Val Thr Gly Ala Ser Asp Gly Ile Gly Arg Glu Ala Ala Met Thr Tyr
20 25 30
Ala Arg Tyr Gly Ala Thr Val Ile Leu Leu Gly Arg Asn Glu Glu Lys
35 40 45
Leu Arg Gln Val Ala Ser His Ile Asn Glu Glu Thr Gly Arg Gln Pro
50 55 60
Gln Trp Phe Ile Leu Asp Leu Leu Thr Cys Thr Ser Glu Asn Cys Gln
65 70 75 80
Gln Leu Ala Gln Arg Ile Ala Val Asn Tyr Pro Arg Leu Asp Gly Val
85 90 95
Leu His Asn Ala Gly Leu Leu Gly Asp Val Cys Pro Met Ser Glu Gln
100 105 110
Asn Pro Gln Val Trp Gln Asp Val Met Gln Val Asn Val Asn Ala Thr
115 120 125
Phe Met Leu Thr Gln Ala Leu Leu Pro Leu Leu Leu Lys Ser Asp Ala
130 135 140
Gly Ser Leu Val Phe Thr Ser Ser Ser Val Gly Arg Gln Gly Arg Ala
145 150 155 160
Asn Trp Gly Ala Tyr Ala Ala Ser Lys Phe Ala Thr Glu Gly Met Met
165 170 175
Gln Val Leu Ala Asp Glu Tyr Gln Gln Arg Leu Arg Val Asn Cys Ile
180 185 190
Asn Pro Gly Gly Thr Arg Thr Ala Met Arg Ala Ser Ala Phe Pro Thr
195 200 205
Glu Asp Pro Gln Lys Leu Lys Thr Pro Ala Asp Ile Met Pro Leu Tyr
210 215 220
Leu Trp Leu Met Gly Asp Asp Ser Arg Arg Lys Thr Gly Met Thr Phe
225 230 235 240
Asp Ala Gln Pro Gly Arg Lys Pro Gly Ile Ser Gln
245 250
<210> 82
<211> 386
<212> PRT
<213> 乔斯特红球菌 RHA1
<400> 82
Met Thr Ala His Leu Ser Leu Pro Arg Phe Ala Arg Ile Gly Ala Gly
1 5 10 15
Ala Val Asp Glu Ile Gly Ser Val Val Asp Gln Leu Gly Leu Gln Arg
20 25 30
Pro Val Leu Val Thr Asp Ser Tyr Leu Thr Gly Thr Gly Ala Ala Asp
35 40 45
Arg Ile Met Ser Leu Leu Arg Ser Ala Gly Lys Glu Pro Ala Leu Phe
50 55 60
Ser Gly Thr Val Pro Asp Pro Thr Thr Asp Ser Leu Glu Glu Gly Leu
65 70 75 80
Arg Ile Val Ala Glu His Asn Ala Asp Ser Val Ile Gly Phe Gly Gly
85 90 95
Gly Ser Pro Met Asp Thr Ala Lys Ala Leu Ala Val Leu Ser Ala Asn
100 105 110
Gly Gly Glu Met Arg Ser Tyr Lys Ala Pro His Val Tyr Ala Gly Lys
115 120 125
Ala Leu Pro Ile Ile Ala Ile Pro Thr Thr Ala Gly Ser Gly Ser Glu
130 135 140
Ala Thr Gln Phe Thr Val Ile Ser Asp Ser Ala Ser Asp Glu Lys Met
145 150 155 160
Leu Cys Pro Gly Leu Ser Phe Leu Pro Ile Ala Ala Ile Val Asp Phe
165 170 175
Glu Leu Thr Met Ser Met Pro Pro Arg Leu Thr Ala Asp Thr Gly Ile
180 185 190
Asp Ala Leu Thr His Ala Ile Glu Ala Tyr Val Ser Lys Lys Ala Gln
195 200 205
Pro Phe Thr Asp Gly Leu Ala Leu Ser Ala Ile Arg Thr Ile Gly Lys
210 215 220
His Ile Gly Thr Ala Tyr Ala Asp Gly Glu Asp Arg Thr Ala Arg Glu
225 230 235 240
Ala Met Met Val Ala Ser Met Gln Ala Gly Met Ala Phe Ser Asn Ser
245 250 255
Ser Val Ala Leu Val His Gly Met Ser Arg Pro Ile Gly Ala His Phe
260 265 270
His Val Ala His Gly Leu Ser Asn Ala Met Leu Leu Pro Ala Ile Thr
275 280 285
Ala Phe Ser Val Asn Gly Ala Glu Ser Arg Tyr Ala Asp Cys Ala Arg
290 295 300
Ala Phe Gly Ala Ala Pro Glu Ser Ala Gly Asp Ser Val Ala Ala Gln
305 310 315 320
Phe Leu Val Glu Thr Leu Ser Thr Leu Cys His Asp Leu Lys Val Pro
325 330 335
Thr Pro Leu Ala Tyr Gly Ile Asp Arg Asp Ala Trp Asp Gly Leu Ile
340 345 350
Pro Leu Met Ala Glu Gln Ala Leu Ala Ser Gly Ser Pro Gly Asn Asn
355 360 365
Pro Val Val Pro Thr Ala Gly Gln Ile Glu Thr Leu Tyr Ser Glu Val
370 375 380
Phe Ser
385
<210> 83
<211> 354
<212> PRT
<213> 乔斯特红球菌菌株 RHA1
<400> 83
Met Arg Pro Phe Val His Ser Gly Ser Pro Ala Arg Ile Val Phe Gly
1 5 10 15
Arg Gly Ala Arg Asp Gly Val Lys Glu Glu Val Glu Arg Leu Gly Ala
20 25 30
Arg Arg Ala Val Val Leu Ser Thr Ala Gln Gln Arg Glu Ser Ala Gly
35 40 45
Arg Leu Ala Asp Asp Leu Gly Ala Leu Gly Val Gly Val Ile Asp Arg
50 55 60
Ala Thr Met His Thr Pro Val Glu Val Thr Ala Gln Ala Leu Val Glu
65 70 75 80
Ile Glu Gln Leu Asp Ala Asp Cys Val Val Ala Leu Gly Gly Gly Ser
85 90 95
Thr Thr Gly Leu Gly Lys Ala Ile Ala Ala Arg Thr Gly Leu Pro Gln
100 105 110
Val Ala Val Pro Thr Thr Tyr Ala Gly Ser Glu Val Thr Pro Ile Leu
115 120 125
Gly Glu Thr Glu Asn Gly Val Lys Thr Thr Arg Arg Gly Pro Glu Ile
130 135 140
Leu Pro Glu Thr Val Val Tyr Asp Pro Ala Leu Thr Asp Ser Leu Pro
145 150 155 160
Ile Pro Leu Ser Ile Ala Ser Gly Leu Asn Ala Met Ala His Ala Ala
165 170 175
Glu Gly Val Tyr Ala Arg Asp Gly Ser Pro Ile Phe Thr Leu Met Ala
180 185 190
Leu Glu Gly Leu Ala Ala Leu Arg Asp Ala Leu Arg Glu Leu Thr Ser
195 200 205
Asp Pro Ala Asp His Asp Ala Arg Asp Arg Ala Leu Tyr Gly Ala Trp
210 215 220
Leu Cys Gly Thr Val Leu Gly Gly Val Gly Met Ser Val His His Lys
225 230 235 240
Leu Cys His Thr Leu Gly Gly Ala Leu Asp Leu Pro His Ala Gln Thr
245 250 255
His Ala Ile Leu Leu Pro His Thr Ile Gly Phe Val Glu Glu Thr Glu
260 265 270
Pro Thr Val Leu Ser Pro Val Ala Asp Leu Phe Gly Gly Ser Ala Gly
275 280 285
Arg Gly Leu His Asp Phe Ala Val Glu Val Gly Ala Pro Leu Arg Leu
290 295 300
Ser Glu Leu Gly Val Thr Pro Glu Gln Leu Asp Arg Val Ala Asp Leu
305 310 315 320
Ala Leu Ala Ala Pro Tyr Trp Ser Pro Arg Pro Leu Asp Arg Asp Ser
325 330 335
Ile Arg Arg Leu Leu Gln His Ala Tyr Asp Gly Thr Arg Pro Glu Ser
340 345 350
Thr Pro
<210> 84
<211> 358
<212> PRT
<213> 乔斯特红球菌 RHA1
<400> 84
Met Thr Phe Thr Ser Ser Phe His Tyr Gln Ala Leu Pro Met Arg Val
1 5 10 15
Ser Phe Glu Val Gly Ala Val Thr Lys Ile Gly Glu Glu Leu Asp Arg
20 25 30
Leu Gly Leu Ser Arg Ala Leu Val Leu Cys Thr Pro Glu Gln Lys His
35 40 45
Leu Ala Asp Leu Val Met Asp His Leu Gly Asp Arg Gly Ala Gly Ile
50 55 60
Phe Asp Gln Ala Arg Met His Val Pro Val Glu Thr Ala Asp Arg Ala
65 70 75 80
Arg Asp Arg Ala Arg Lys Leu Gly Ala Asp Ser Cys Val Val Ile Gly
85 90 95
Gly Gly Ser Ser Ile Gly Leu Gly Lys Ala Ile Ala Leu Glu His Gly
100 105 110
Leu Pro Ile Ile Ala Ile Pro Thr Thr Tyr Ala Gly Ser Glu Met Thr
115 120 125
Pro Ile Trp Gly Leu Thr Arg Asp Gly Arg Lys Gln Thr Gly Arg Asp
130 135 140
Ala Lys Val Leu Pro Gln Ser Val Ile Tyr Asp Pro Gln Leu Thr Ala
145 150 155 160
Thr Leu Pro Ala Glu Ile Ser Ala Ala Ser Gly Leu Asn Ala Val Ala
165 170 175
His Ala Val Glu Ala Leu Tyr Ala Pro Asp Ala Ser Pro Ile Ile Ser
180 185 190
Leu Met Ala Glu Glu Gly Val Arg Ala Phe Ala Glu Ser Leu Pro Ala
195 200 205
Val Val Ala Asp Gly Asn Asp Leu Asp Ala Arg Ser Lys Ala Leu Tyr
210 215 220
Gly Ala Trp Leu Cys Gly Ala Cys Leu Gly Ala Thr Thr Met Ser Leu
225 230 235 240
His His Lys Leu Cys His Val Leu Gly Gly Ser Leu Asp Leu Pro His
245 250 255
Ala Gln Thr His Ala Ile Val Leu Pro His Val Leu Gly Tyr Asn Glu
260 265 270
Ser His Ala Pro Ala Ala Arg Ala Ala Leu Gln Arg Ala Leu Gly Thr
275 280 285
Asp Gly Asp Pro Ser Thr Ala Leu Trp Glu Leu Glu Gln Gln Leu Gln
290 295 300
Ile Pro His Ser Leu Glu Glu Leu Gly Val Arg Glu Thr Gln Leu Pro
305 310 315 320
Ala Val Ile Asp Glu Val Leu Ser Asn Pro Tyr Ser Asn Pro Thr Pro
325 330 335
Val Thr Glu Asp Gly Leu Arg His Leu Leu Gly Gln Ala Leu Arg Gly
340 345 350
Asp Arg Pro Thr Arg Arg
355
<210> 85
<211> 253
<212> PRT
<213> 阿维链霉菌 ATCC 31267
<400> 85
Met Asp Leu Ser Asn Arg Thr Val Leu Ile Val Gly Gly Thr Ser Gly
1 5 10 15
Ile Gly Arg Glu Leu Ala Arg Arg Phe Ala Ala Ala Gly Ser Thr Val
20 25 30
Ala Val Gly Gly Arg Ser Pro Glu Ala Leu Ser Glu Leu Ala Gly Glu
35 40 45
Gly Phe Gly Thr Ile Ser Ile Asp Val Thr Asp Ser Ala Ser Val Ala
50 55 60
Ser Ala Arg Asp Ala Val Leu Ala Arg Tyr Pro Glu Leu Asp Thr Val
65 70 75 80
Val Thr Met Pro Gly Ile Met Leu Leu Glu Asp Leu Arg Asp Pro Ala
85 90 95
His Phe Glu Ala Ala Ala Thr Thr Ile Asp Thr Asn Leu Val Gly Thr
100 105 110
Ile Arg Val Ile Asp Ala Phe Thr Pro His Leu Val Arg Arg Gly Ala
115 120 125
Gly Thr Phe Ile Thr Val Thr Ser Gly Ile Ala Phe Leu Pro Phe Pro
130 135 140
Pro Met Pro Thr Tyr Ala Ala Ser Lys Ala Ala Val His Ala Tyr Ser
145 150 155 160
Glu Ala Leu Arg Ala Gln Leu Asp Gly Thr Gly Val Ser Val Val Glu
165 170 175
Leu Val Pro Pro Ala Val Ala Thr Ala Gly Gln Glu Lys Val Asn Pro
180 185 190
His Ala Leu Pro Leu Asp Asp Phe Ala Thr Glu Val Val Gln Leu Leu
195 200 205
Ser Glu Asp Pro Thr Pro Arg Glu Ile Leu Val Lys Ala Ala Leu Met
210 215 220
His Arg Trp Ala Glu Arg Asp Gly Thr Tyr Asp Asp Leu Val Ala Gln
225 230 235 240
Arg Ser Gln Ala Leu Ala Met Leu Pro Ser Arg Glu Gly
245 250
<210> 86
<211> 265
<212> PRT
<213> 天蓝色链霉菌 ATCC BAA-471
<400> 86
Met Thr Glu Arg Leu Arg Ser Phe Arg Met Thr Ala Leu Thr Asn Lys
1 5 10 15
Thr Ala Leu Val Thr Gly Gly Ser Arg Gly Ile Gly Arg Ala Ile Ala
20 25 30
Gln Arg Leu Gly Lys Glu Gly Ala Leu Val Ala Leu Thr Tyr Ser Ser
35 40 45
Asp Glu Ala Ala Ala Lys Glu Thr Val His Ser Ile Glu Ala Ala Gly
50 55 60
Gly Arg Ala Phe Ala Phe Arg Ser Gln Leu Gly Val Pro Gly Asp Ala
65 70 75 80
Glu Ala Leu Trp Gln Ala Phe Asp Ala Gln Ile Gly Gln Tyr Ala Asp
85 90 95
Gly Gly Ser Gly Leu Asp Ile Leu Val Asn Asn Ala Gly Ile Ala Gly
100 105 110
Pro Gly Leu Ile His Glu Val Glu Glu Ala Glu Tyr Asp Lys Val Phe
115 120 125
Ala Val Asn Ala Lys Ala Pro Phe Phe Ile Ile Gln Lys Gly Leu Glu
130 135 140
Arg Leu Arg Asp Gly Gly Arg Ile Val Asn Ile Ser Ser Gly Val Thr
145 150 155 160
Lys Val Ala Phe Pro Gly Met Thr Ser Tyr Ala Ala Ala Lys Gly Ala
165 170 175
Val Glu Val Leu Thr Leu Thr Leu Ala Gln Thr Leu Gly Ser Arg Gly
180 185 190
Ile Thr Val Asn Ala Val Ser Pro Gly Thr Ile Glu Thr Asp Ile His
195 200 205
Pro Trp Met Ala Asp Pro Ala Ala Lys Ala His Ala Ala Gly Phe Ser
210 215 220
Val Phe Asn Arg Val Gly Gln Pro Asp Asp Val Ala Asp Val Val Ala
225 230 235 240
Phe Leu Ala Ser Asp Asp Ala Arg Trp Val Thr Gly Gln Asn Ile Asp
245 250 255
Ala Ser Gly Gly Ser Gly Leu Gly Leu
260 265
<210> 87
<211> 294
<212> PRT
<213> 肠道沙门氏菌亚种肠道血清型甲型副伤寒B菌株 ATCC BAA-1585
<400> 87
Met Ser His Leu Tyr Asp Pro Thr Thr Gln Tyr Tyr Thr Gly Glu Tyr
1 5 10 15
Pro Lys Gln Lys Gln Pro Ala Pro Gly Val Gln Ala Lys Met Thr Pro
20 25 30
Val Pro Asp Cys Gly Glu Lys Ser Tyr Val Gly Ser Gly Arg Leu Lys
35 40 45
Asp Arg Lys Ala Leu Val Thr Gly Gly Asp Ser Gly Ile Gly Arg Ala
50 55 60
Ala Ala Ile Ala Tyr Ala Arg Glu Gly Ala Asp Val Ala Ile Asn Tyr
65 70 75 80
Leu Pro Ala Glu Glu Glu Asp Ala Gln Gln Val Lys Ala Leu Ile Glu
85 90 95
Glu Cys Gly Arg Lys Ala Val Leu Leu Pro Gly Asp Leu Ser Asp Glu
100 105 110
Ser Phe Ala Arg Ser Leu Val His Lys Ala Arg Glu Ala Leu Gly Gly
115 120 125
Leu Asp Ile Leu Ala Leu Val Ala Gly Lys Gln Thr Ala Ile Pro Glu
130 135 140
Ile Lys Asp Leu Thr Ser Glu Gln Phe Gln Gln Thr Phe Ala Val Asn
145 150 155 160
Val Phe Ala Leu Phe Trp Ile Thr Gln Glu Ala Ile Pro Leu Leu Pro
165 170 175
Lys Gly Ala Ser Ile Ile Thr Thr Ser Ser Ile Gln Ala Tyr Gln Pro
180 185 190
Ser Pro His Leu Leu Asp Tyr Ala Ala Thr Lys Ala Ala Ile Leu Asn
195 200 205
Tyr Ser Arg Gly Leu Ala Lys Gln Val Ala Glu Lys Gly Ile Arg Val
210 215 220
Asn Ile Val Ala Pro Gly Pro Ile Trp Thr Ala Leu Gln Ile Ser Gly
225 230 235 240
Gly Gln Thr Gln Asp Lys Ile Pro Gln Phe Gly Gln Gln Thr Pro Met
245 250 255
Lys Arg Ala Gly Gln Pro Ala Glu Leu Ala Pro Val Tyr Val Tyr Leu
260 265 270
Ala Ser Gln Glu Ser Ser Tyr Val Thr Ala Glu Val His Gly Val Cys
275 280 285
Gly Gly Glu His Leu Gly
290
<210> 88
<211> 245
<212> PRT
<213> 野油菜黄单胞菌野油菜致病变种 ATCC 33913
<400> 88
Met Thr Asp Ser Ser Lys Val Val Leu Ile Thr Gly Ala Ala Arg Arg
1 5 10 15
Ile Gly Ala Gln Ile Ala Thr Thr Leu His Gly Ala Gly Tyr Arg Val
20 25 30
Ala Leu His Ala His Arg Ser Gly Asp Ala Leu Ala Ala Arg Val Ala
35 40 45
Ala Leu Cys Ala Gln Arg Ala Gly Ser Ala Cys Ala Leu Gln Ala Asp
50 55 60
Leu Arg Thr Pro Glu Ala Pro Ala Gln Leu Val Asp Ala Cys Val Ala
65 70 75 80
Ala Phe Gly Arg Leu Asp Ala Val Val Asn Asn Ala Ser Ala Phe Tyr
85 90 95
Pro Thr Val Leu Gly Glu Ala Thr Pro Ala Gln Trp Asp Glu Leu Phe
100 105 110
Ala Val Asn Ala Arg Ala Pro Phe Phe Ile Ala Gln Ala Ala Ala Ala
115 120 125
Gln Leu Arg Ala His His Gly Ala Ile Val Asn Leu Thr Asp Leu His
130 135 140
Ala Glu Gln Pro Met Arg Gln His Pro Leu Tyr Gly Ala Ser Lys Ser
145 150 155 160
Ala Leu Glu Met Leu Thr Arg Ser Leu Ala Leu Glu Leu Ala Pro Gln
165 170 175
Val Arg Val Asn Ala Val Ala Pro Gly Ala Ile Leu Trp Pro Glu Asp
180 185 190
Gly Lys Ala Asp Ala Ala Lys Gln Ala Leu Leu Ala Arg Thr Pro Leu
195 200 205
Ala Arg Ile Gly Thr Pro Glu Glu Val Ala Glu Ala Val Arg Trp Leu
210 215 220
Leu Asp Asp Ala Ser Phe Ile Thr Gly His Thr Leu Arg Val Asp Gly
225 230 235 240
Gly Arg Arg Leu Ser
245
<210> 89
<211> 434
<212> PRT
<213> 炭疽芽孢杆菌
<400> 89
Met Val Val Lys Phe Thr Lys Ser Glu Ala Leu His Lys Glu Ala Leu
1 5 10 15
Glu His Ile Val Gly Gly Val Asn Ser Pro Ser Arg Ser Phe Lys Ala
20 25 30
Val Gly Gly Gly Ala Pro Ile Ala Met Glu Arg Gly Lys Gly Ala Tyr
35 40 45
Phe Trp Asp Val Asp Gly Asn Lys Tyr Ile Asp Tyr Leu Ala Ala Tyr
50 55 60
Gly Pro Ile Ile Thr Gly His Ala His Pro His Ile Thr Lys Ala Ile
65 70 75 80
Thr Thr Ala Ala Glu Asn Gly Val Leu Tyr Gly Thr Pro Thr Ala Leu
85 90 95
Glu Val Lys Phe Ala Lys Met Leu Lys Glu Ala Met Pro Ala Leu Asp
100 105 110
Lys Val Arg Phe Val Asn Ser Gly Thr Glu Ala Val Met Thr Thr Ile
115 120 125
Arg Val Ala Arg Ala Tyr Thr Gly Arg Thr Lys Ile Met Lys Phe Ala
130 135 140
Gly Cys Tyr His Gly His Ser Asp Leu Val Leu Val Ala Ala Gly Ser
145 150 155 160
Gly Pro Ser Thr Leu Gly Thr Pro Asp Ser Ala Gly Val Pro Gln Ser
165 170 175
Ile Ala Gln Glu Val Ile Thr Val Pro Phe Asn Asn Val Glu Thr Leu
180 185 190
Lys Glu Ala Leu Asp Lys Trp Gly His Glu Val Ala Ala Ile Leu Val
195 200 205
Glu Pro Ile Val Gly Asn Phe Gly Ile Val Glu Pro Lys Pro Gly Phe
210 215 220
Leu Glu Lys Val Asn Glu Leu Val His Glu Ala Gly Ala Leu Val Ile
225 230 235 240
Tyr Asp Glu Val Ile Thr Ala Phe Arg Phe Met Tyr Gly Gly Ala Gln
245 250 255
Asp Leu Leu Gly Val Thr Pro Asp Leu Thr Ala Leu Gly Lys Val Ile
260 265 270
Gly Gly Gly Leu Pro Ile Gly Ala Tyr Gly Gly Lys Lys Glu Ile Met
275 280 285
Glu Gln Val Ala Pro Leu Gly Pro Ala Tyr Gln Ala Gly Thr Met Ala
290 295 300
Gly Asn Pro Ala Ser Met Ala Ser Gly Ile Ala Cys Leu Glu Val Leu
305 310 315 320
Gln Gln Glu Gly Leu Tyr Glu Lys Leu Asp Glu Leu Gly Ala Met Leu
325 330 335
Glu Lys Gly Ile Leu Glu Gln Ala Ala Lys His Asn Ile Asp Ile Thr
340 345 350
Leu Asn Arg Leu Lys Gly Ala Leu Thr Val Tyr Phe Thr Thr Asn Thr
355 360 365
Ile Glu Asp Tyr Asp Ala Ala Gln Asp Thr Asp Gly Glu Met Phe Gly
370 375 380
Lys Phe Phe Lys Leu Met Leu Gln Glu Gly Val Asn Leu Ala Pro Ser
385 390 395 400
Lys Tyr Glu Ala Trp Phe Leu Thr Thr Glu His Thr Lys Glu Asp Ile
405 410 415
Glu Tyr Thr Ile Glu Ala Val Gly Arg Ala Phe Ala Ala Leu Ala Asp
420 425 430
Asn Lys
<210> 90
<211> 383
<212> PRT
<213> 炭疽芽孢杆菌
<400> 90
Met Gln Leu Phe His Lys Thr Val Asn Arg Arg Gly Thr His Ser Ile
1 5 10 15
Lys Trp Asp Thr Tyr Lys Asn Glu Glu Leu Ile His Ala Trp Ile Ala
20 25 30
Asp Met Asp Phe Glu Val Pro Gln Pro Ile Gln Thr Ala Leu Lys Lys
35 40 45
Arg Ile Glu His Pro Ile Phe Gly Tyr Thr Leu Pro Pro Glu Asn Ile
50 55 60
Gly Asp Ile Ile Cys Asn Trp Thr Lys Lys Gln Tyr Asn Trp Asp Ile
65 70 75 80
Gln Lys Glu Trp Ile Val Phe Ser Ala Gly Ile Val Pro Ala Leu Ser
85 90 95
Thr Ser Ile Gln Ala Phe Thr Lys Glu Asn Glu Ser Val Leu Val Gln
100 105 110
Pro Pro Ile Tyr Pro Pro Phe Phe Glu Met Val Thr Thr Asn Asn Arg
115 120 125
Gln Leu Cys Val Ser Pro Leu Gln Lys Gln Asn Asp Thr Tyr Ala Ile
130 135 140
Asp Phe Glu His Leu Glu Lys Gln Phe Gln Gln Gly Val Lys Leu Met
145 150 155 160
Leu Leu Cys Ser Pro His Asn Pro Ile Gly Arg Val Trp Lys Lys Glu
165 170 175
Glu Leu Thr Lys Leu Gly Ser Leu Cys Thr Lys Tyr Asn Val Ile Val
180 185 190
Val Ala Asp Glu Ile His Ser Asp Ile Ile Tyr Ala Asp His Thr His
195 200 205
Thr Pro Phe Ala Ser Leu Ser Glu Glu Leu Ala Ala Arg Thr Ile Thr
210 215 220
Cys Met Ala Pro Ser Lys Thr Phe Asn Ile Ala Gly Leu Gln Ala Ser
225 230 235 240
Ile Ile Ile Ile Pro Asn Glu Lys Leu Arg Gln Ala Phe Thr Ser Ile
245 250 255
Gln Tyr Arg Gln Gly Phe His Gly Leu Asn Ile Phe Ala Tyr Thr Ala
260 265 270
Met Gln Ser Ala Tyr Thr Glu Cys Asn Asp Trp Leu Asn Glu Ile Arg
275 280 285
Phe Tyr Ile Glu Asp Asn Ala Lys Phe Ala Cys Glu Tyr Ile Lys Asp
290 295 300
His Ile Pro Thr Leu Ser Val Met Lys Pro Glu Gly Ser Phe Leu Leu
305 310 315 320
Trp Ile Asp Cys Ser Ala Leu Asn Leu Ser Gln Asp Glu Arg Thr Lys
325 330 335
Leu Leu Glu Glu Lys Gly Lys Ile Ile Val Glu Pro Gly Glu Lys Tyr
340 345 350
Gly Leu Gly Gly Glu Glu His Ile Gly Ile Asn Ile Gly Cys Pro Arg
355 360 365
Ser Val Leu Glu Glu Ile Leu Asn Arg Leu Arg His Thr Phe Ser
370 375 380
<210> 91
<211> 425
<212> PRT
<213> 除烃海杆菌菌株 ATCC 700491
<400> 91
Met Ser Asn Lys Glu Leu Gln Ala Leu Lys Glu Arg Tyr Val Ala Ala
1 5 10 15
Gly Ala Ala Ser Pro Asn Asp Gln Phe Ala Asp His Ala Leu Asn Ala
20 25 30
Glu Val Trp Asp Ala Asp Gly Lys Arg Met Ile Asp Phe Ala Gly Gly
35 40 45
Ile Gly Val Leu Asn Ile Gly His Arg His Pro Arg Val Val Glu Ala
50 55 60
Val Lys Ala Gln Leu Asp Lys Leu Met His Thr Cys Gln Thr Val Met
65 70 75 80
Pro Tyr Glu Gly Tyr Val Lys Leu Ala Gln Lys Leu Ser Glu Val Val
85 90 95
Pro Val Lys Gly His Gly Lys Val Met Leu Ala Asn Ser Gly Ala Glu
100 105 110
Ala Leu Glu Asn Ala Val Lys Ile Ala Arg Ala Ala Thr Gly Lys Asn
115 120 125
Asn Val Ile Cys Phe Asp Gly Gly Tyr His Gly Arg Thr Phe Phe Thr
130 135 140
Met Ala Met Asn Gly Lys Ala Ala Pro Tyr Gln Thr Ser Phe Gly Pro
145 150 155 160
Met Pro Gly Leu Val Tyr Arg Ala Pro Tyr Pro Val Pro Tyr His Gly
165 170 175
Ile Ser Glu Glu Glu Ser Leu Arg Gly Leu Ala Met Thr Met Lys Ala
180 185 190
Asp Ser Pro Ala His Glu Thr Ala Ala Ile Val Leu Glu Pro Val Leu
195 200 205
Gly Glu Gly Gly Phe Tyr Pro Ala Pro Pro Ser Phe Leu Lys Ala Ile
210 215 220
Arg Lys Ile Cys Asp Glu Asn Asp Ile Met Met Ile Val Asp Glu Val
225 230 235 240
Gln Ser Gly Phe Gly Arg Thr Gly Lys Leu Phe Ala Ile Glu His Ser
245 250 255
Gly Ile Glu Pro Asp Ile Met Thr Met Ala Lys Ser Met Ala Asp Gly
260 265 270
Met Pro Ile Ser Ala Ile Val Gly Thr Asp Lys Val Met Asp Ser Ser
275 280 285
Gly Pro Asn Ser Leu Gly Gly Thr Tyr Thr Gly Ser Pro Thr Ala Cys
290 295 300
Ala Ala Ala Leu Ala Val Phe Asp Val Ile Lys Glu Glu Asp Ile Leu
305 310 315 320
Gly Arg Ser Gln Arg Leu Gly Glu Lys Leu Arg Ala Arg Phe Asp Gln
325 330 335
Trp Gln Glu Gln Phe Thr His Val Asp Asn Ala Arg Asn Leu Gly Pro
340 345 350
Met Ala Ala Phe Glu Leu Val Glu Ser Lys Asp Ser His Lys Pro Met
355 360 365
Pro Glu Leu Ala Gly Ala Ile Thr Lys Lys Ala Lys Glu Lys Gly Leu
370 375 380
Ile Leu Leu Ser Cys Gly Met His Gly Asn Thr Leu Arg Phe Leu Met
385 390 395 400
Pro Val Thr Ile Glu Asp Glu Val Leu Glu Glu Gly Leu Asn Ile Val
405 410 415
Glu Glu Cys Leu Lys Glu Val Gly Ala
420 425
<210> 92
<211> 460
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 92
Met Thr Ala Gln Leu Asn Pro Gln Arg Asp Thr Arg Asp Tyr Gln Gln
1 5 10 15
Leu Asp Ala Ala His His Ile His Ala Phe Leu Asp Gln Lys Ala Leu
20 25 30
Asn Arg Glu Gly Pro Arg Val Met Val Arg Gly Asp Gly Leu Gln Leu
35 40 45
Trp Asp Asn Asp Gly Lys Arg Tyr Leu Asp Gly Met Ser Gly Leu Trp
50 55 60
Cys Thr Asn Leu Gly Tyr Gly Arg Gln Asp Leu Ala Ala Ala Ala Ser
65 70 75 80
Arg Gln Leu Glu Gln Leu Pro Tyr Tyr Asn Met Phe Phe His Thr Thr
85 90 95
His Pro Ala Val Val Glu Leu Ser Glu Met Leu Phe Ser Leu Leu Pro
100 105 110
Asp His Tyr Ser His Ala Ile Tyr Thr Asn Ser Gly Ser Glu Ala Asn
115 120 125
Glu Val Leu Ile Arg Thr Val Arg Arg Tyr Trp Gln Ile Leu Gly Lys
130 135 140
Pro Gln Lys Lys Ile Met Ile Gly Arg Trp Asn Gly Tyr His Gly Ser
145 150 155 160
Thr Leu Gly Ser Thr Ala Leu Gly Gly Met Lys Phe Met His Glu Met
165 170 175
Gly Gly Met Leu Pro Asp Phe Ala His Ile Asp Glu Pro Tyr Trp Tyr
180 185 190
Ala Asn Gly Gly Glu Leu Ser Pro Ala Glu Phe Gly Arg Arg Ala Ala
195 200 205
Leu Gln Leu Glu Glu Lys Ile Leu Glu Leu Gly Ala Glu Asn Val Ala
210 215 220
Ala Phe Val Ala Glu Pro Phe Gln Gly Ala Gly Gly Met Ile Phe Pro
225 230 235 240
Pro Gln Ser Tyr Trp Pro Glu Ile Gln Arg Ile Cys Arg Gln Tyr Asp
245 250 255
Val Leu Leu Cys Ala Asp Glu Val Ile Gly Gly Phe Gly Arg Thr Gly
260 265 270
Glu Trp Phe Ala His Glu His Phe Gly Phe Gln Pro Asp Thr Leu Ser
275 280 285
Ile Ala Lys Gly Leu Thr Ser Gly Tyr Ile Pro Met Gly Gly Leu Val
290 295 300
Leu Gly Lys Arg Ile Ala Glu Val Leu Val Glu Gln Gly Gly Val Phe
305 310 315 320
Ala His Gly Leu Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala
325 330 335
Ile Ala Asn Leu Lys Ala Leu Arg Asp Glu Gly Val Val Thr Arg Val
340 345 350
Arg Glu Glu Thr Gly Pro Tyr Leu Gln Arg Cys Leu Arg Glu Val Phe
355 360 365
Gly Asp His Pro Leu Val Gly Glu Val Gln Gly Ala Gly Phe Val Ala
370 375 380
Ala Leu Gln Phe Ala Glu Asp Lys Val Thr Arg Lys Arg Phe Ala Asn
385 390 395 400
Glu Asn Asp Leu Ala Trp Arg Cys Arg Thr Ile Gly Phe Glu Glu Gly
405 410 415
Val Ile Ile Arg Ser Thr Leu Gly Arg Met Ile Met Ala Pro Ala Leu
420 425 430
Val Ala Gly Arg Ala Glu Ile Asp Glu Leu Ile Asp Lys Thr Arg Ile
435 440 445
Ala Val Asp Arg Thr Ala Arg Glu Ile Gly Val Leu
450 455 460
<210> 93
<211> 468
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 93
Met Asn Ala Arg Leu His Ala Thr Ser Pro Leu Gly Asp Ala Asp Leu
1 5 10 15
Val Arg Ala Asp Gln Ala His Tyr Met His Gly Tyr His Val Phe Asp
20 25 30
Asp His Arg Val Asn Gly Ser Leu Asn Ile Ala Ala Gly Asp Gly Ala
35 40 45
Tyr Ile Tyr Asp Thr Ala Gly Asn Arg Tyr Leu Asp Ala Val Gly Gly
50 55 60
Met Trp Cys Thr Asn Ile Gly Leu Gly Arg Glu Glu Met Ala Arg Thr
65 70 75 80
Val Ala Glu Gln Thr Arg Leu Leu Ala Tyr Ser Asn Pro Phe Cys Asp
85 90 95
Met Ala Asn Pro Arg Ala Ile Glu Leu Cys Arg Lys Leu Ala Glu Leu
100 105 110
Ala Pro Gly Asp Leu Asp His Val Phe Leu Thr Thr Gly Gly Ser Thr
115 120 125
Ala Val Asp Thr Ala Ile Arg Leu Met His Tyr Tyr Gln Asn Cys Arg
130 135 140
Gly Lys Arg Ala Lys Lys His Val Ile Thr Arg Ile Asn Ala Tyr His
145 150 155 160
Gly Ser Thr Phe Leu Gly Met Ser Leu Gly Gly Lys Ser Ala Asp Arg
165 170 175
Pro Ala Glu Phe Asp Phe Leu Asp Glu Arg Ile His His Leu Ala Cys
180 185 190
Pro Tyr Tyr Tyr Arg Ala Pro Glu Gly Leu Gly Glu Ala Glu Phe Leu
195 200 205
Asp Gly Leu Val Asp Glu Phe Glu Arg Lys Ile Leu Glu Leu Gly Ala
210 215 220
Asp Arg Val Gly Ala Phe Ile Ser Glu Pro Val Phe Gly Ser Gly Gly
225 230 235 240
Val Ile Val Pro Pro Ala Gly Tyr His Arg Arg Met Trp Glu Leu Cys
245 250 255
Gln Arg Tyr Asp Val Leu Tyr Ile Ser Asp Glu Val Val Thr Ser Phe
260 265 270
Gly Arg Leu Gly His Phe Phe Ala Ser Gln Ala Val Phe Gly Val Gln
275 280 285
Pro Asp Ile Ile Leu Thr Ala Lys Gly Leu Thr Ser Gly Tyr Gln Pro
290 295 300
Leu Gly Ala Cys Ile Phe Ser Arg Arg Ile Trp Glu Val Ile Ala Glu
305 310 315 320
Pro Asp Lys Gly Arg Cys Phe Ser His Gly Phe Thr Tyr Ser Gly His
325 330 335
Pro Val Ala Cys Ala Ala Ala Leu Lys Asn Ile Glu Ile Ile Glu Arg
340 345 350
Glu Gly Leu Leu Ala His Ala Asp Glu Val Gly Arg Tyr Phe Glu Glu
355 360 365
Arg Leu Gln Ser Leu Arg Asp Leu Pro Ile Val Gly Asp Val Arg Gly
370 375 380
Met Arg Phe Met Ala Cys Val Glu Phe Val Ala Asp Lys Ala Ser Lys
385 390 395 400
Ala Leu Phe Pro Glu Ser Leu Asn Ile Gly Glu Trp Val His Leu Arg
405 410 415
Ala Gln Lys Arg Gly Leu Leu Val Arg Pro Ile Val His Leu Asn Val
420 425 430
Met Ser Pro Pro Leu Ile Leu Thr Arg Glu Gln Val Asp Thr Val Val
435 440 445
Arg Val Leu Arg Glu Ser Ile Glu Glu Thr Val Glu Asp Leu Val Arg
450 455 460
Ala Gly His Arg
465
<210> 94
<211> 444
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 94
Met Thr Met Asn Asp Glu Pro Gln Ser Ser Ser Leu Asp Asn Phe Trp
1 5 10 15
Met Pro Phe Thr Ala Asn Arg Gln Phe Lys Ala Arg Pro Arg Leu Leu
20 25 30
Glu Ser Ala Glu Gly Ile His Tyr Ile Ala Gln Gly Gly Arg Arg Ile
35 40 45
Leu Asp Gly Thr Ala Gly Leu Trp Cys Cys Asn Ala Gly His Gly Arg
50 55 60
Arg Glu Ile Ser Glu Ala Val Ala Arg Gln Ile Ala Thr Leu Asp Tyr
65 70 75 80
Ala Pro Pro Phe Gln Met Gly His Pro Leu Pro Phe Glu Leu Ala Ala
85 90 95
Arg Leu Thr Glu Ile Ala Pro Pro Ser Leu Asn Lys Val Phe Phe Thr
100 105 110
Asn Ser Gly Ser Glu Ser Ala Asp Thr Ala Leu Lys Ile Ala Leu Ala
115 120 125
Tyr Gln Arg Ala Ile Gly Gln Gly Thr Arg Thr Arg Leu Ile Gly Arg
130 135 140
Glu Leu Gly Tyr His Gly Val Gly Phe Gly Gly Leu Ser Val Gly Gly
145 150 155 160
Met Val Asn Asn Arg Lys Ala Phe Ser Ala Asn Leu Leu Pro Gly Val
165 170 175
Asp His Leu Pro His Thr Leu Asp Val Ala Arg Asn Ala Phe Thr Val
180 185 190
Gly Leu Pro Glu His Gly Val Glu Lys Ala Glu Glu Leu Glu Arg Leu
195 200 205
Val Thr Leu His Gly Ala Glu Asn Ile Ala Ala Val Ile Val Glu Pro
210 215 220
Met Ser Gly Ser Ala Gly Val Val Leu Pro Pro Lys Gly Tyr Leu Gln
225 230 235 240
Arg Leu Arg Glu Ile Thr Arg Lys His Gly Ile Leu Leu Ile Phe Asp
245 250 255
Glu Val Ile Thr Gly Phe Gly Arg Val Gly Glu Ala Phe Ala Ala Gln
260 265 270
Arg Trp Gly Val Val Pro Asp Leu Leu Thr Cys Ala Lys Gly Leu Thr
275 280 285
Asn Gly Ser Ile Pro Met Gly Ala Val Phe Val Asp Glu Lys Ile His
290 295 300
Ala Ala Phe Met Gln Gly Pro Gln Gly Ala Ile Glu Phe Phe His Gly
305 310 315 320
Tyr Thr Tyr Ser Gly His Pro Val Ala Cys Ala Ala Ala Leu Ala Thr
325 330 335
Leu Asp Ile Tyr Arg Arg Asp Asp Leu Phe Gln Arg Ala Val Glu Leu
340 345 350
Glu Gly Tyr Trp Gln Asp Ala Leu Phe Ser Leu Arg Asp Leu Pro Asn
355 360 365
Val Val Asp Ile Arg Ala Val Gly Leu Val Gly Gly Val Gln Leu Ala
370 375 380
Pro His Ala Asp Gly Pro Gly Lys Arg Gly Tyr Asp Val Phe Glu Arg
385 390 395 400
Cys Phe Trp Glu His Asp Leu Met Val Arg Val Thr Gly Asp Ile Ile
405 410 415
Ala Met Ser Pro Pro Leu Ile Ile Asp Lys Pro His Ile Asp Gln Ile
420 425 430
Val Glu Arg Leu Ala Gln Ala Ile Arg Ala Ser Val
435 440
<210> 95
<211> 452
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 95
Met Ser Glu Gln Asn Ser Gln Thr Leu Ala Trp Gln Thr Met Ser Arg
1 5 10 15
Asp His His Leu Ala Pro Phe Ser Asp Val Arg Gln Leu Ala Glu Lys
20 25 30
Gly Pro Arg Ile Ile Thr Ser Ala Lys Gly Val Tyr Leu Trp Asp Ser
35 40 45
Glu Gly Asn Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Ala
50 55 60
Val Gly Tyr Gly Arg Glu Glu Leu Ala Glu Val Ala Ser Gln Gln Met
65 70 75 80
Lys Gln Leu Pro Tyr Tyr Asn Leu Phe Phe Gln Thr Ala His Pro Pro
85 90 95
Ala Leu Glu Leu Ala Lys Ala Ile Ala Glu Val Ala Pro Gln Gly Met
100 105 110
Asn His Val Phe Phe Thr Gly Ser Gly Ser Glu Gly Asn Asp Thr Val
115 120 125
Leu Arg Met Val Arg His Tyr Trp Ala Leu Lys Gly Gln Lys Asn Lys
130 135 140
Lys Val Ile Ile Gly Arg Ile Asn Gly Tyr His Gly Ser Thr Val Ala
145 150 155 160
Gly Ala Ala Leu Gly Gly Met Ser Gly Met His Gln Gln Gly Gly Val
165 170 175
Ile Pro Asp Val Val His Ile Pro Gln Pro Tyr Trp Phe Gly Glu Gly
180 185 190
Gly Asp Met Thr Glu Ala Asp Phe Gly Val Trp Ala Ala Glu Gln Leu
195 200 205
Glu Lys Lys Ile Leu Glu Val Gly Val Asp Asn Val Ala Ala Phe Ile
210 215 220
Ala Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Gln Thr
225 230 235 240
Tyr Trp Pro Lys Ile Lys Glu Ile Leu Ala Arg Tyr Asp Ile Leu Phe
245 250 255
Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp Phe
260 265 270
Gly Thr Asp Tyr Tyr Asp Leu Lys Pro Asp Leu Met Thr Ile Ala Lys
275 280 285
Gly Leu Thr Ser Gly Tyr Ile Pro Met Gly Gly Val Ile Val Arg Asp
290 295 300
Glu Val Ala Lys Val Ile Ser Glu Gly Gly Asp Phe Asn His Gly Phe
305 310 315 320
Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Gly Leu Glu Asn Leu
325 330 335
Arg Ile Leu Arg Asp Glu Gln Ile Ile Gln Gln Val His Asp Lys Thr
340 345 350
Ala Pro Tyr Leu Gln Gln Arg Leu Arg Glu Leu Ala Asp His Pro Leu
355 360 365
Val Gly Glu Val Arg Gly Leu Gly Met Leu Gly Ala Ile Glu Leu Val
370 375 380
Lys Asp Lys Ala Thr Arg Ala Arg His Glu Gly Lys Gly Val Gly Met
385 390 395 400
Ile Cys Arg Gln His Cys Phe Asp Asn Gly Leu Ile Met Arg Ala Val
405 410 415
Gly Asp Thr Met Ile Ile Ala Pro Pro Leu Val Ile Ser Ile Glu Glu
420 425 430
Ile Asp Glu Leu Val Glu Lys Ala Arg Lys Cys Leu Asp Leu Thr Tyr
435 440 445
Glu Ala Val Arg
450
<210> 96
<211> 470
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 96
Met Ala Thr Pro Ser Lys Ala Phe Ala Ile Ala His Asp Pro Leu Val
1 5 10 15
Glu Ala Asp Lys Ala His Tyr Met His Gly Tyr His Val Phe Asp Glu
20 25 30
His Arg Glu Gln Gly Ala Leu Asn Ile Val Ala Gly Glu Gly Ala Tyr
35 40 45
Ile Arg Asp Thr His Gly Asn Arg Phe Leu Asp Ala Val Gly Gly Met
50 55 60
Trp Cys Thr Asn Ile Gly Leu Gly Arg Glu Glu Met Ala Leu Ala Ile
65 70 75 80
Val Asp Gln Val Arg Gln Leu Ala Tyr Ser Asn Pro Phe Ser Asp Met
85 90 95
Ala Asn Asp Val Ala Ile Glu Leu Cys Gln Lys Leu Ala Gln Leu Ala
100 105 110
Pro Gly Asp Leu Asn His Val Phe Leu Thr Thr Gly Gly Ser Thr Ala
115 120 125
Val Asp Thr Ala Tyr Arg Leu Ile Gln Tyr Tyr Gln Asn Cys Arg Gly
130 135 140
Lys Pro His Lys Lys His Ile Ile Ala Arg Tyr Asn Ala Tyr His Gly
145 150 155 160
Ser Thr Thr Leu Thr Met Ser Ile Gly Asn Lys Ala Ala Asp Arg Val
165 170 175
Pro Glu Phe Asp Tyr His His Asp Leu Ile His His Val Ser Asn Pro
180 185 190
Asn Pro Tyr Arg Ala Pro Asp Asp Met Asp Glu Ala Glu Phe Leu Asp
195 200 205
Phe Leu Val Ala Glu Phe Glu Asp Lys Ile Leu Ser Leu Gly Ala Asp
210 215 220
Asn Val Ala Ala Phe Phe Ala Glu Pro Ile Met Gly Ser Gly Gly Val
225 230 235 240
Ile Ile Pro Pro Glu Gly Tyr Phe Gln Arg Met Trp Gln Leu Cys Gln
245 250 255
Thr Tyr Asp Ile Leu Phe Val Ala Asp Glu Val Val Thr Ser Phe Gly
260 265 270
Arg Leu Gly Thr Phe Phe Ala Ser Glu Glu Leu Phe Gly Val Thr Pro
275 280 285
Asp Ile Ile Thr Thr Ala Lys Gly Leu Thr Ser Ala Tyr Leu Pro Leu
290 295 300
Gly Ala Cys Ile Phe Ser Glu Arg Ile Trp Gln Val Ile Ala Glu Pro
305 310 315 320
Gly Lys Gly Arg Cys Phe Thr His Gly Phe Thr Tyr Ser Gly His Pro
325 330 335
Val Cys Cys Thr Ala Ala Leu Lys Asn Ile Glu Ile Ile Glu Arg Glu
340 345 350
Gln Leu Leu Asp His Val Asn Asp Val Gly Ser Tyr Leu Glu Gln Arg
355 360 365
Leu Gln Ser Leu Arg Asp Leu Pro Leu Val Gly Asp Val Arg Cys Met
370 375 380
Lys Leu Met Ala Cys Val Glu Phe Val Ala Asn Lys Ala Ser Lys Ala
385 390 395 400
Leu Phe Ala Asp Glu Val Asn Ile Gly Glu Arg Ile His Ser Lys Ala
405 410 415
Gln Glu Lys Gly Leu Leu Val Arg Pro Ile Met His Leu Asn Val Met
420 425 430
Ser Pro Pro Leu Ile Ile Thr His Ala Gln Val Asp Glu Ile Val Glu
435 440 445
Thr Leu Arg Gln Cys Ile Ile Glu Thr Ala Arg Glu Leu Thr Ala Leu
450 455 460
Gly Leu Tyr Gln Gly Arg
465 470
<210> 97
<211> 453
<212> PRT
<213> 恶臭假单胞菌 KT2440
<400> 97
Met Ser Val Asn Asn Pro Gln Thr Arg Glu Trp Gln Thr Leu Ser Gly
1 5 10 15
Glu His His Leu Ala Pro Phe Ser Asp Tyr Lys Gln Leu Lys Glu Lys
20 25 30
Gly Pro Arg Ile Ile Thr Lys Ala Gln Gly Val His Leu Trp Asp Ser
35 40 45
Glu Gly His Lys Ile Leu Asp Gly Met Ala Gly Leu Trp Cys Val Ala
50 55 60
Val Gly Tyr Gly Arg Glu Glu Leu Val Gln Ala Ala Glu Lys Gln Met
65 70 75 80
Arg Glu Leu Pro Tyr Tyr Asn Leu Phe Phe Gln Thr Ala His Pro Pro
85 90 95
Ala Leu Glu Leu Ala Lys Ala Ile Thr Asp Val Ala Pro Lys Gly Met
100 105 110
Thr His Val Phe Phe Thr Gly Ser Gly Ser Glu Gly Asn Asp Thr Val
115 120 125
Leu Arg Met Val Arg His Tyr Trp Ala Leu Lys Gly Lys Pro His Lys
130 135 140
Gln Thr Ile Ile Gly Arg Ile Asn Gly Tyr His Gly Ser Thr Phe Ala
145 150 155 160
Gly Ala Cys Leu Gly Gly Met Ser Gly Met His Glu Gln Gly Gly Leu
165 170 175
Pro Ile Pro Gly Ile Val His Ile Pro Gln Pro Tyr Trp Phe Gly Glu
180 185 190
Gly Gly Asp Met Thr Pro Asp Glu Phe Gly Val Trp Ala Ala Glu Gln
195 200 205
Leu Glu Lys Lys Ile Leu Glu Val Gly Glu Asp Asn Val Ala Ala Phe
210 215 220
Ile Ala Glu Pro Ile Gln Gly Ala Gly Gly Val Ile Ile Pro Pro Glu
225 230 235 240
Thr Tyr Trp Pro Lys Val Lys Glu Ile Leu Ala Arg Tyr Asp Ile Leu
245 250 255
Phe Val Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Trp
260 265 270
Phe Gly Ser Asp Tyr Tyr Asp Leu Lys Pro Asp Leu Met Thr Ile Ala
275 280 285
Lys Gly Leu Thr Ser Gly Tyr Ile Pro Met Gly Gly Val Ile Val Arg
290 295 300
Asp Thr Val Ala Lys Val Ile Ser Glu Gly Gly Asp Phe Asn His Gly
305 310 315 320
Phe Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Gly Leu Glu Asn
325 330 335
Leu Arg Ile Leu Arg Asp Glu Lys Ile Val Glu Lys Ala Arg Thr Glu
340 345 350
Ala Ala Pro Tyr Leu Gln Lys Arg Leu Arg Glu Leu Gln Asp His Pro
355 360 365
Leu Val Gly Glu Val Arg Gly Leu Gly Met Leu Gly Ala Ile Glu Leu
370 375 380
Val Lys Asp Lys Ala Thr Arg Ser Arg Tyr Glu Gly Lys Gly Val Gly
385 390 395 400
Met Ile Cys Arg Thr Phe Cys Phe Glu Asn Gly Leu Ile Met Arg Ala
405 410 415
Val Gly Asp Thr Met Ile Ile Ala Pro Pro Leu Val Ile Ser His Ala
420 425 430
Glu Ile Asp Glu Leu Val Glu Lys Ala Arg Lys Cys Leu Asp Leu Thr
435 440 445
Leu Glu Ala Ile Gln
450
<210> 98
<211> 413
<212> PRT
<213> 乔斯特红球菌 RHA1
<400> 98
Met Thr Arg Leu Ser Pro Leu Leu Ala Gln Ala Thr Pro Val Thr Val
1 5 10 15
Asp His Gly Glu Gly Cys Tyr Leu Tyr Gly Thr 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 His Val Val Glu Ala Ala Arg Arg Gln Ile Gly Ser Leu Ile His
50 55 60
Gly Gln Tyr Thr Thr Val Met His Gln Pro Met Leu Glu Leu Val Asp
65 70 75 80
Arg Leu Gly Ser Val Leu Pro Ala Gly Leu Asp Ser Leu Phe Phe Ala
85 90 95
Asn Ser Gly Ser Glu Ala Val Glu Ala Ser Leu Arg Leu Ser Arg Gln
100 105 110
Ala Thr Gly Arg Pro Asn Val Ile Val Phe His Gly Gly Phe His Gly
115 120 125
Arg Thr Val Ala Thr Ala Thr Met Thr Thr Ser Gly Thr Arg Phe Ser
130 135 140
Ala Gly Phe Ser Pro Leu Met Gly Gly Val His Val Ala Pro Phe Pro
145 150 155 160
Asn Ala Tyr Arg Tyr Gly Trp Ser Glu Glu Glu Ala Thr Ala Phe Ala
165 170 175
Leu Lys Glu Leu Asp Tyr Ile Phe Ala Thr Leu Thr Ala Pro Asn Glu
180 185 190
Thr Ala Ala Phe Val Val Glu Pro Val Leu Gly Glu Gly Gly Tyr Val
195 200 205
Pro Gly Asn Thr Ala Phe Phe Gln Gly Leu Arg Glu Arg Ala Asp Arg
210 215 220
Tyr Gly Ile Leu Leu Val Ile Asp Glu Ile Gln Thr Gly Phe Gly Arg
225 230 235 240
Thr Gly Lys Phe Phe Gly His Gln His Phe Asp Val Arg Pro Asp Ile
245 250 255
Ile Thr Ile Ala Lys Gly Leu Ala Ser Gly Phe Pro Leu Ser Gly Ile
260 265 270
Ala Ala Ser Glu Ala Leu Met Ala Lys Gly Trp Pro Gly Ser Gln Gly
275 280 285
Gly Thr Tyr Gly Gly Asn Ala Val Ser Cys Ala Ala Ala Val Ala Thr
290 295 300
Leu Glu Val Ile Glu Lys Glu Asp Leu Val Ala Asn Ala Ala Ala Arg
305 310 315 320
Gly Val Gln Leu Leu Asp Gly Ala Arg Thr Arg Ala Ile Asp Gly Ile
325 330 335
Gly Asp Val Arg Gly Leu Gly Leu Leu Val Gly Ser Glu Phe Thr Ala
340 345 350
Ala Asp Gly Ser Ala Asp Arg Thr Lys Ala Ser Ala Ala Gln Gln Leu
355 360 365
Ala Ala Lys Lys Gly Leu Leu Leu Leu Thr Cys Gly Ala His Met Asn
370 375 380
Val Val Arg Met Ile Pro Pro Leu Ile Val Thr Ala Glu Gln Ile Glu
385 390 395 400
Asp Ala Leu Lys Ile Trp Ser Glu Val Leu Asp Glu Val
405 410
<210> 99
<211> 461
<212> PRT
<213> 乔斯特红球菌 RHA1
<400> 99
Met Thr Thr Val Thr Asn Ser Ser Pro Thr Thr Ala His Leu Glu Ala
1 5 10 15
Ala Ala Arg Arg His Leu Trp Gly His Phe Ser Arg His Gly Gln Asn
20 25 30
Ile Thr Pro Pro Ile Ile Thr Arg Gly Glu Gly Ala Arg Ile Trp Asp
35 40 45
Thr Ala Gly Lys Ser Tyr Leu Asp Gly Leu Ser Gly Leu Phe Val Val
50 55 60
Gln Ala Gly His Gly Arg Thr Glu Leu Ala Glu Ala Ala Ala Lys Gln
65 70 75 80
Ala Glu Gln Leu Ala Phe Phe Pro Leu Trp Ser Tyr Ala Thr Glu Pro
85 90 95
Ala Ile Glu Leu Ala Glu Arg Leu Ala Gly Tyr Ala Pro Gly Asp Leu
100 105 110
Asn Arg Val Phe Phe Thr Thr Gly Gly Gly Glu Ala Val Glu Ser Ala
115 120 125
Trp Lys Leu Ala Lys Gln Tyr Phe Lys Lys Val Gly Lys Pro Gly Lys
130 135 140
His Lys Val Ile Ser Arg Ser Ile Ala Tyr His Gly Thr Pro Gln Gly
145 150 155 160
Ala Leu Ala Ile Thr Gly Ile Pro Ala Leu Lys Ala Pro Phe Glu Pro
165 170 175
Leu Thr Pro Gly Ala Phe Arg Val Pro Asn Thr Asn Ile Tyr Arg Ala
180 185 190
Pro Glu Pro Leu Gly Ser Asp Pro Lys Ala Phe Gly Ile Trp Ala Ala
195 200 205
Asp Arg Ile Ala Glu Ala Ile Glu Phe Glu Gly Pro Asp Thr Val Ala
210 215 220
Ala Val Phe Leu Glu Pro Val Gln Asn Ala Gly Gly Cys Phe Pro Pro
225 230 235 240
Pro Pro Gly Tyr Phe Glu Arg Val Arg Gln Ile Cys Asp Glu Tyr Asp
245 250 255
Val Leu Leu Val Ser Asp Glu Val Ile Cys Ala Phe Gly Arg Ile Gly
260 265 270
Ser Met Phe Ala Cys Asp Asp Phe Gly Tyr Val Pro Asp Ile Ile Thr
275 280 285
Cys Ala Lys Gly Leu Thr Ser Gly Tyr Ser Pro Ile Gly Ala Met Ile
290 295 300
Ala Ser Asp Arg Leu Phe Glu Pro Phe Ser Asp Gly Thr Ser Met Phe
305 310 315 320
Ala His Gly Tyr Thr Phe Gly Gly His Pro Val Ser Ala Ala Val Ala
325 330 335
Leu Ala Asn Leu Asp Ile Phe Glu Arg Glu Gly Leu Asn Ala His Val
340 345 350
Ala Glu Gln Ala Pro Ala Phe Arg Ala Thr Leu Asp Lys Leu Thr Asp
355 360 365
Leu Pro Met Val Gly Asp Val Arg Gly Glu Gly Phe Phe Tyr Gly Ile
370 375 380
Glu Leu Val Lys Asp Lys Thr Thr Lys Glu Ser Phe Thr Asp Asp Glu
385 390 395 400
Ala Glu Arg Ile Leu His Gly Phe Leu Ser Thr Ala Leu Phe Asp Ala
405 410 415
Gly Leu Tyr Cys Arg Ala Asp Asp Arg Gly Asp Pro Val Ile Gln Leu
420 425 430
Ala Pro Pro Leu Ile Cys Gly Gln Ala Glu Phe Asp Glu Ile Glu His
435 440 445
Ile Leu Arg Ser Val Leu Thr Glu Ala Trp Thr Leu Leu
450 455 460
<210> 100
<211> 459
<212> PRT
<213> 阿维链霉菌 ATCC 31267
<400> 100
Met Gly Asn Pro Ile Ala Val Ser Lys Asp Leu Ser Arg Thr Ala Tyr
1 5 10 15
Asp His Leu Trp Met His Phe Thr Arg Met Ser Ser Tyr Glu Asn Ala
20 25 30
Pro Val Pro Thr Ile Val Arg Gly Glu Gly Thr Tyr Ile Tyr Asp Asp
35 40 45
Lys Gly Lys Arg Tyr Leu Asp Gly Leu Ser Gly Leu Phe Val Val Gln
50 55 60
Ala Gly His Gly Arg Thr Glu Leu Ala Glu Thr Ala Phe Lys Gln Ala
65 70 75 80
Gln Glu Leu Ala Phe Phe Pro Val Trp Ser Tyr Ala His Pro Lys Ala
85 90 95
Val Glu Leu Ala Glu Arg Leu Ala Asn Tyr Ala Pro Gly Asp Leu Asn
100 105 110
Lys Val Phe Phe Thr Thr Gly Gly Gly Glu Ala Val Glu Thr Ala Trp
115 120 125
Lys Leu Ala Lys Gln Tyr Phe Lys Leu Gln Gly Lys Pro Thr Lys Tyr
130 135 140
Lys Val Ile Ser Arg Ala Val Ala Tyr His Gly Thr Pro Gln Gly Ala
145 150 155 160
Leu Ser Ile Thr Gly Leu Pro Ala Leu Lys Ala Pro Phe Glu Pro Leu
165 170 175
Val Pro Gly Ala His Lys Val Pro Asn Thr Asn Ile Tyr Arg Ala Pro
180 185 190
Leu Phe Gly Asp Asp Pro Glu Ala Phe Gly Arg Trp Ala Ala Asp Gln
195 200 205
Ile Glu Gln Gln Ile Leu Phe Glu Gly Pro Glu Thr Val Ala Ala Val
210 215 220
Phe Leu Glu Pro Val Gln Asn Ala Gly Gly Cys Phe Pro Pro Pro Pro
225 230 235 240
Gly Tyr Phe Gln Arg Val Arg Glu Ile Cys Asp Gln Tyr Asp Val Leu
245 250 255
Leu Val Ser Asp Glu Val Ile Cys Ala Phe Gly Arg Leu Gly Thr Met
260 265 270
Phe Ala Cys Asp Lys Phe Gly Tyr Val Pro Asp Met Ile Thr Cys Ala
275 280 285
Lys Gly Met Thr Ser Gly Tyr Ser Pro Ile Gly Ala Cys Ile Val Ser
290 295 300
Asp Arg Ile Ala Glu Pro Phe Tyr Lys Gly Asp Asn Thr Phe Leu His
305 310 315 320
Gly Tyr Thr Phe Gly Gly His Pro Val Ser Ala Ala Val Gly Val Ala
325 330 335
Asn Leu Asp Leu Phe Glu Arg Glu Gly Leu Asn Gln His Val Leu Asp
340 345 350
Asn Glu Ser Ala Phe Leu Thr Thr Leu Gln Lys Leu His Asp Leu Pro
355 360 365
Ile Val Gly Asp Val Arg Gly Asn Gly Phe Phe Tyr Gly Ile Glu Leu
370 375 380
Val Lys Asp Lys Ala Thr Lys Glu Thr Phe Thr Asp Glu Glu Ser Glu
385 390 395 400
Arg Val Leu Tyr Gly Phe Val Ser Lys Lys Leu Phe Glu Tyr Gly Leu
405 410 415
Tyr Cys Arg Ala Asp Asp Arg Gly Asp Pro Val Ile Gln Leu Ser Pro
420 425 430
Pro Leu Ile Ser Asn Gln Ser Thr Phe Asp Glu Ile Glu Ser Ile Ile
435 440 445
Arg Gln Val Leu Thr Glu Ala Trp Thr Lys Leu
450 455
<210> 101
<211> 458
<212> PRT
<213> 苜蓿根瘤菌 1021
<400> 101
Met Tyr Ser Asn Ser Leu Ile Glu Leu Asp Arg Ala His Leu Ile His
1 5 10 15
Pro Val Ala Ser Tyr Arg Gly His Glu Lys Leu Gly Val Arg Val Leu
20 25 30
Ala Ser Ala Lys Gly Ala Thr Val Thr Asp Ala Ser Gly Arg Gln Leu
35 40 45
Ile Asp Gly Phe Ala Gly Leu Trp Cys Val Asn Ala Gly Tyr Gly Gln
50 55 60
Glu Thr Ile Val Glu Ala Ala Ala Lys Gln Met Arg Glu Leu Ser Tyr
65 70 75 80
Ala Thr Ala Tyr Phe Gly Leu Gly Ser Glu Pro Ala Ile Arg Leu Ala
85 90 95
Ser Glu Leu Ala Glu Arg Ala Pro Gly Asn Leu Asn His Val Tyr Phe
100 105 110
Thr Leu Gly Gly Ser Asp Ala Val Asp Ser Thr Ile Arg Phe Ile Arg
115 120 125
Tyr Tyr Trp Thr Ala Arg Gly Glu Pro Gln Arg Asp Gln Phe Ile Ser
130 135 140
Val Glu Gln Gly Tyr His Gly Ser Ser Thr Val Gly Ala Gly Leu Thr
145 150 155 160
Ala Leu Pro Ala Phe His Thr Gly Phe Gly Ile Pro Phe Asp Trp Gln
165 170 175
His Lys Ile Pro Ser His Tyr Ala Tyr Arg Asn Pro Val Gly Asp Asp
180 185 190
Pro Gln Ala Ile Ile Ala Ala Ser Leu Thr Ala Leu Arg Arg Lys Val
195 200 205
Glu Glu Ile Gly Pro Glu Arg Val Ala Ala Phe Tyr Ala Glu Pro Ile
210 215 220
Gln Gly Ser Gly Gly Val Leu Val Pro Pro Arg Gly Trp Met Lys Ala
225 230 235 240
Met Arg Glu Leu Cys Arg Asp Leu Gly Ile Leu Phe Val Ala Asp Glu
245 250 255
Val Ile Thr Gly Phe Gly Arg Thr Gly Pro Leu Phe Ala Ser Thr Glu
260 265 270
Asn Glu Ile Val Pro Asp Phe Ile Thr Thr Ala Lys Gly Leu Thr Ser
275 280 285
Gly Tyr Val Pro Met Gly Ala Val Phe Met Ala Asp His Ile Tyr Gln
290 295 300
Thr Ile Ala Asp Gly Ala Gly Ala Ser Ala Val Gly His Gly Tyr Thr
305 310 315 320
Tyr Ser Ala His Pro Val Ser Ala Ala Val Gly Leu Glu Val Leu Arg
325 330 335
Leu Tyr Glu Asn Gly Leu Leu Glu Asn Gly Val Lys Ala Gly Ala Arg
340 345 350
Leu Met Glu Gly Leu Gly Ser Leu Arg Asp His Pro Leu Val Gly Asp
355 360 365
Val Arg Gly Arg Gly Met Leu Ala Ala Ile Glu Leu Val Val Asp Lys
370 375 380
Ala Gln Lys Thr Pro Leu Pro Ala Ala Ala Glu Pro Ala Arg Arg Ile
385 390 395 400
Phe Asp Arg Ala Trp Glu Asn Gly Leu Val Ile Arg Ala Phe Ala Asn
405 410 415
Gly Val Leu Gly Tyr Ala Pro Pro Leu Cys Cys Ser Glu Thr Glu Ile
420 425 430
Asp Ala Ile Ile Glu Arg Thr Arg Gln Ser Leu Asp Glu Thr Leu Glu
435 440 445
Asp Pro Asp Val Arg Arg Ala Leu Lys Thr
450 455
<210> 102
<211> 447
<212> PRT
<213> 苜蓿中华根瘤菌
<400> 102
Met Thr His Leu Leu His Arg Ala Ile His Ala Lys Leu Pro Val Ala
1 5 10 15
Val Arg Gly Glu Gly Ile Arg Leu Phe Asp Ala Glu Gly Arg Ala Tyr
20 25 30
Ile Asp Ala Ser Gly Gly Ala Ala Val Ser Cys Leu Gly His Ala His
35 40 45
Pro Asp Val Leu Ala Ala Leu His Ala Gln Leu Asp Arg Ile Ala Tyr
50 55 60
Ala His Thr Gly Phe Phe Thr Thr Glu Ala Ala Glu Arg Leu Ala Asp
65 70 75 80
Arg Leu Val Ala Asp Ala Pro Asp Gly Leu Asp His Val Tyr Leu Val
85 90 95
Ser Gly Gly Ser Glu Ala Val Glu Ala Ala Leu Lys Met Ala Arg Gln
100 105 110
Tyr Phe Val Glu Lys Gly Glu Pro Gln Arg Arg His Ile Ile Ala Arg
115 120 125
Arg Gln Ser Tyr His Gly Asn Thr Leu Gly Ala Leu Ala Ala Gly Gly
130 135 140
Asn Glu Trp Arg Arg Ala Gln Phe Arg Pro Leu Leu Val Glu Thr His
145 150 155 160
His Ile Asp Pro Cys Phe Ala Tyr Arg Leu Gln Gln Ala Gly Glu Ser
165 170 175
Asp Thr Asp Tyr Ala Ala Arg Ala Ala Gly Gln Leu Glu Ala Lys Ile
180 185 190
Leu Glu Leu Gly Ala Asp Gln Val Ile Ala Phe Val Ala Glu Thr Val
195 200 205
Val Gly Ala Thr Ala Gly Ala Val Pro Pro Val Ala Asp Tyr Leu Lys
210 215 220
Arg Val Arg Ala Ile Cys Asp Lys Tyr Gly Val Leu Leu Ile Leu Asp
225 230 235 240
Glu Val Met Cys Gly Met Gly Arg Thr Gly Thr Leu His Ala Cys Glu
245 250 255
Gln Asp Gly Val Val Pro Asp Leu Met Thr Ile Ala Lys Gly Leu Gly
260 265 270
Gly Gly Tyr Gln Pro Val Gly Ala Val Leu Leu Ser Ser Ala Ile Phe
275 280 285
Gln Ala Phe Ser Glu Gly Ser Gly Phe Phe Gln His Gly His Thr Tyr
290 295 300
Met Gly His Pro Met Ala Ala Ala Ala Gly Leu Ala Val Gln Glu Val
305 310 315 320
Ile Arg Arg Asp Gly Leu Leu Glu Asn Val Val Ser Met Gly Asp Tyr
325 330 335
Leu Glu Arg Cys Leu Val Glu Arg Leu Gly Asn His His His Val Gly
340 345 350
Asp Ile Arg Gly Arg Gly Leu Phe Arg Ala Val Glu Leu Val Ala Asp
355 360 365
Arg Ala Thr Lys Ser Pro Phe Asp Pro Gly Leu Lys Leu Asn Ala Arg
370 375 380
Ile Lys Lys Glu Ala Met Ala Arg Gly Leu Met Val Tyr Pro Met Gly
385 390 395 400
Gly Thr Ile Asp Gly Gln Arg Gly Asp His Val Leu Leu Ala Pro Pro
405 410 415
Phe Ile Leu Asn Arg Glu Asp Val Gly Gln Ile Val Glu Arg Leu Gly
420 425 430
Asp Ala Val Asp Ala Ala Ile Ala Ser Leu Thr Lys Ala Ala Gly
435 440 445
<210> 103
<211> 445
<212> PRT
<213> 中慢生根瘤菌属物种 LUK
<400> 103
Met Asn Glu Pro Ile Gly Glu Pro Gly Arg Ser Pro Ala Ser Asp Thr
1 5 10 15
Ala Glu Lys Ala Gln Ala Ile Ala Ala Ala Arg Asn Thr Phe Ala Arg
20 25 30
Asp Asn Pro Val Ser Ala Gly His His Glu Arg Ala Arg Arg Ser Met
35 40 45
Pro Gly Gly Asn Thr Arg Ser Ile Leu Phe His Arg Pro Phe Pro Leu
50 55 60
Val Ile Ala Gln Gly Thr Gly Ser Arg Phe Gln Asp Val Asp Gly His
65 70 75 80
Ala Tyr Val Asn Phe Leu Gly Glu Tyr Thr Ala Gly Leu Phe Gly His
85 90 95
Ser His Pro Val Ile Arg Ala Ala Val Glu Arg Ala Leu Ala Val Gly
100 105 110
Leu Asn Leu Ser Thr Gln Thr Glu Asn Glu Ala Leu Phe Ala Glu Ala
115 120 125
Val Cys Asp Arg Phe Pro Ser Ile Asp Leu Val Arg Phe Thr Asn Ser
130 135 140
Gly Thr Glu Ala Asn Leu Met Ala Leu Ala Thr Ala Thr Ala Ile Thr
145 150 155 160
Gly Arg Lys Thr Val Leu Ala Phe Asp Gly Gly Tyr His Gly Gly Leu
165 170 175
Leu Asn Phe Ala Ser Gly His Ala Pro Thr Asn Ala Pro Tyr His Val
180 185 190
Val Leu Gly Val Tyr Asn Asp Val Glu Gly Thr Ala Asp Leu Leu Lys
195 200 205
Arg His Gly His Asp Cys Ala Ala Ile Leu Val Glu Pro Met Leu Gly
210 215 220
Ala Gly Gly Cys Val Pro Ala Glu Arg Ala Phe Leu Asp Leu Leu Arg
225 230 235 240
Ala Glu Ala Ser Arg Cys Gly Ala Leu Leu Ile Phe Asp Glu Val Met
245 250 255
Thr Ser Arg Leu Ser Gly Gly Gly Ala Gln Glu Met Leu Gly Ile Ser
260 265 270
Ala Asp Leu Thr Thr Leu Gly Lys Tyr Ile Gly Gly Gly Met Ser Phe
275 280 285
Gly Ala Phe Gly Gly Arg Arg Asp Leu Met Glu Arg Phe Asp Pro Ala
290 295 300
Arg Asp Gly Ala Phe Ala His Ala Gly Thr Phe Asn Asn Asn Ile Leu
305 310 315 320
Thr Met Ser Ala Gly His Ala Ala Leu Thr Gln Ile Tyr Thr Arg Gln
325 330 335
Ala Ala Ser Asp Leu Ser Ala Ser Gly Asp Arg Phe Arg Ala Asn Leu
340 345 350
Asn Arg Ile Ala Val Glu Asn Gln Ala Pro Leu Gln Phe Thr Gly Leu
355 360 365
Gly Ser Leu Gly Thr Ile His Phe Ser Arg Ala Pro Ile Arg Ser Ala
370 375 380
Gly Asp Val Arg Ala Ala Asp Gln Gln Leu Lys Glu Leu Phe Phe Phe
385 390 395 400
His Met Leu Arg Lys Gly Ile Tyr Leu Ala Pro Arg Gly Met Tyr Ala
405 410 415
Leu Ser Leu Glu Ile Ala Asp Ala Gly Arg Asp Ala Phe Ala Glu Ala
420 425 430
Leu Ala Asp Phe Ile Gly Glu Gln Arg Ala Leu Leu Met
435 440 445
<210> 104
<211> 496
<212> PRT
<213> 铜绿假单胞菌 ATCC 15692
<400> 104
Met Phe Glu Ser Ala Glu Val Gly His Ser Ile Asp Lys Asp Thr Tyr
1 5 10 15
Glu Lys Ala Val Ile Glu Leu Arg Glu Ala Leu Leu Glu Ala Gln Phe
20 25 30
Glu Leu Lys Gln Gln Ala Arg Phe Pro Val Ile Ile Leu Ile Asn Gly
35 40 45
Ile Glu Gly Ala Gly Lys Gly Glu Thr Val Lys Leu Leu Asn Glu Trp
50 55 60
Met Asp Pro Arg Leu Ile Glu Val Gln Ser Phe Leu Arg Pro Ser Asp
65 70 75 80
Glu Glu Leu Glu Arg Pro Pro Gln Trp Arg Phe Trp Arg Arg Leu Pro
85 90 95
Pro Lys Gly Arg Thr Gly Ile Phe Phe Gly Asn Trp Tyr Ser Gln Met
100 105 110
Leu Tyr Ala Arg Val Glu Gly His Ile Lys Glu Ala Lys Leu Asp Gln
115 120 125
Ala Ile Asp Ala Ala Glu Arg Phe Glu Arg Met Leu Cys Asp Glu Gly
130 135 140
Ala Leu Leu Phe Lys Phe Trp Phe His Leu Ser Lys Lys Gln Leu Lys
145 150 155 160
Glu Arg Leu Lys Ala Leu Glu Lys Asp Pro Gln His Ser Trp Lys Leu
165 170 175
Ser Pro Leu Asp Trp Lys Gln Ser Glu Val Tyr Asp Arg Phe Val His
180 185 190
Tyr Gly Glu Arg Val Leu Arg Arg Thr Ser Arg Asp Tyr Ala Pro Trp
195 200 205
Tyr Val Val Glu Gly Ala Asp Glu Arg Tyr Arg Ala Leu Thr Val Gly
210 215 220
Arg Ile Leu Leu Glu Gly Leu Gln Ala Ala Leu Ala Thr Lys Glu Arg
225 230 235 240
Ala Lys Arg Gln Pro His Ala Ala Pro Leu Val Ser Ser Leu Asp Asn
245 250 255
Arg Gly Leu Leu Asp Ser Leu Asp Leu Gly Gln Tyr Leu Asp Lys Asp
260 265 270
Ala Tyr Lys Glu Gln Leu Ala Ala Glu Gln Ala Arg Leu Ala Gly Leu
275 280 285
Ile Arg Asp Lys Arg Phe Arg Gln His Ser Leu Val Ala Val Phe Glu
290 295 300
Gly Asn Asp Ala Ala Gly Lys Gly Gly Ala Ile Arg Arg Val Thr Asp
305 310 315 320
Ala Leu Asp Pro Arg Gln Tyr His Ile Val Pro Ile Ala Ala Pro Thr
325 330 335
Glu Glu Glu Arg Ala Gln Pro Tyr Leu Trp Arg Phe Trp Arg His Ile
340 345 350
Pro Ala Arg Arg Gln Phe Thr Ile Phe Asp Arg Ser Trp Tyr Gly Arg
355 360 365
Val Leu Val Glu Arg Ile Glu Gly Phe Cys Ala Pro Ala Asp Trp Leu
370 375 380
Arg Ala Tyr Gly Glu Ile Asn Asp Phe Glu Glu Gln Leu Ser Glu Tyr
385 390 395 400
Gly Ile Ile Val Val Lys Phe Trp Leu Ala Ile Asp Lys Gln Thr Gln
405 410 415
Met Glu Arg Phe Lys Glu Arg Glu Lys Thr Pro Tyr Lys Arg Tyr Lys
420 425 430
Ile Thr Glu Glu Asp Trp Arg Asn Arg Asp Lys Trp Asp Gln Tyr Val
435 440 445
Asp Ala Val Gly Asp Met Val Asp Arg Thr Ser Thr Glu Ile Ala Pro
450 455 460
Trp Thr Leu Val Glu Ala Asn Asp Lys Arg Phe Ala Arg Val Lys Val
465 470 475 480
Leu Arg Thr Ile Asn Asp Ala Ile Glu Ala Ala Tyr Lys Lys Asp Lys
485 490 495
<210> 105
<211> 300
<212> PRT
<213> 苜蓿根瘤菌菌株 1021
<400> 105
Met Ala Leu Asp Glu Ala Pro Ala Glu Ala Arg Pro Gly Ser Arg Ala
1 5 10 15
Val Glu Leu Glu Ile Asp Gly Arg Ser Arg Ile Phe Asp Ile Asp Asp
20 25 30
Pro Asp Leu Pro Lys Trp Ile Asp Glu Glu Ala Phe Arg Ser Asp Asp
35 40 45
Tyr Pro Tyr Lys Lys Lys Leu Asp Arg Glu Glu Tyr Glu Glu Thr Leu
50 55 60
Thr Lys Leu Gln Ile Glu Leu Val Lys Val Gln Phe Trp Met Gln Ala
65 70 75 80
Thr Gly Lys Arg Val Met Ala Val Phe Glu Gly Arg Asp Ala Ala Gly
85 90 95
Lys Gly Gly Ala Ile His Ala Thr Thr Ala Asn Met Asn Pro Arg Ser
100 105 110
Ala Arg Val Val Ala Leu Thr Lys Pro Thr Glu Thr Glu Arg Gly Gln
115 120 125
Trp Tyr Phe Gln Arg Tyr Val Ala Thr Phe Pro Thr Ala Gly Glu Phe
130 135 140
Val Leu Phe Asp Arg Ser Trp Tyr Asn Arg Ala Gly Val Glu Pro Val
145 150 155 160
Met Gly Phe Cys Thr Pro Asp Gln Tyr Glu Gln Phe Leu Lys Glu Ala
165 170 175
Pro Arg Phe Glu Glu Met Ile Ala Asn Glu Gly Ile His Leu Phe Lys
180 185 190
Phe Trp Ile Asn Ile Gly Arg Glu Met Gln Leu Lys Arg Phe His Asp
195 200 205
Arg Arg His Asp Pro Leu Lys Ile Trp Lys Leu Ser Pro Met Asp Ile
210 215 220
Ala Ala Leu Ser Lys Trp Asp Asp Tyr Thr Gly Lys Arg Asp Arg Met
225 230 235 240
Leu Lys Glu Thr His Thr Glu His Gly Pro Trp Ala Val Ile Arg Gly
245 250 255
Asn Asp Lys Arg Arg Ser Arg Ile Asn Val Ile Arg His Met Leu Thr
260 265 270
Lys Leu Asp Tyr Asp Gly Lys Asp Glu Ala Ala Ile Gly Glu Val Asp
275 280 285
Glu Lys Ile Leu Gly Ser Gly Pro Gly Phe Leu Arg
290 295 300
<210> 106
<211> 378
<212> PRT
<213> 枯草芽孢杆菌168
<400> 106
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> 107
<211> 454
<212> PRT
<213> 酿酒酵母
<400> 107
Met Ser Glu Pro Glu Phe Gln Gln Ala Tyr Glu Glu Val Val Ser Ser
1 5 10 15
Leu Glu Asp Ser Thr Leu Phe Glu Gln His Pro Glu Tyr Arg Lys Val
20 25 30
Leu Pro Ile Val Ser Val Pro Glu Arg Ile Ile Gln Phe Arg Val Thr
35 40 45
Trp Glu Asn Asp Lys Gly Glu Gln Glu Val Ala Gln Gly Tyr Arg Val
50 55 60
Gln Tyr Asn Ser Ala Lys Gly Pro Tyr Lys Gly Gly Leu Arg Phe His
65 70 75 80
Pro Ser Val Asn Leu Ser Ile Leu Lys Phe Leu Gly Phe Glu Gln Ile
85 90 95
Phe Lys Asn Ser Leu Thr Gly Leu Asp Met Gly Gly Gly Lys Gly Gly
100 105 110
Leu Cys Val Asp Leu Lys Gly Arg Ser Asn Asn Glu Ile Arg Arg Ile
115 120 125
Cys Tyr Ala Phe Met Arg Glu Leu Ser Arg His Ile Gly Gln Asp Thr
130 135 140
Asp Val Pro Ala Gly Asp Ile Gly Val Gly Gly Arg Glu Ile Gly Tyr
145 150 155 160
Leu Phe Gly Ala Tyr Arg Ser Tyr Lys Asn Ser Trp Glu Gly Val Leu
165 170 175
Thr Gly Lys Gly Leu Asn Trp Gly Gly Ser Leu Ile Arg Pro Glu Ala
180 185 190
Thr Gly Tyr Gly Leu Val Tyr Tyr Thr Gln Ala Met Ile Asp Tyr Ala
195 200 205
Thr Asn Gly Lys Glu Ser Phe Glu Gly Lys Arg Val Thr Ile Ser Gly
210 215 220
Ser Gly Asn Val Ala Gln Tyr Ala Ala Leu Lys Val Ile Glu Leu Gly
225 230 235 240
Gly Thr Val Val Ser Leu Ser Asp Ser Lys Gly Cys Ile Ile Ser Glu
245 250 255
Thr Gly Ile Thr Ser Glu Gln Val Ala Asp Ile Ser Ser Ala Lys Val
260 265 270
Asn Phe Lys Ser Leu Glu Gln Ile Val Asn Glu Tyr Ser Thr Phe Ser
275 280 285
Glu Asn Lys Val Gln Tyr Ile Ala Gly Ala Arg Pro Trp Thr His Val
290 295 300
Gln Lys Val Asp Ile Ala Leu Pro Cys Ala Thr Gln Asn Glu Val Ser
305 310 315 320
Gly Glu Glu Ala Lys Ala Leu Val Ala Gln Gly Val Lys Phe Ile Ala
325 330 335
Glu Gly Ser Asn Met Gly Ser Thr Pro Glu Ala Ile Ala Val Phe Glu
340 345 350
Thr Ala Arg Ser Thr Ala Thr Gly Pro Ser Glu Ala Val Trp Tyr Gly
355 360 365
Pro Pro Lys Ala Ala Asn Leu Gly Gly Val Ala Val Ser Gly Leu Glu
370 375 380
Met Ala Gln Asn Ser Gln Arg Ile Thr Trp Thr Ser Glu Arg Val Asp
385 390 395 400
Gln Glu Leu Lys Arg Ile Met Ile Asn Cys Phe Asn Glu Cys Ile Asp
405 410 415
Tyr Ala Lys Lys Tyr Thr Lys Asp Gly Lys Val Leu Pro Ser Leu Val
420 425 430
Lys Gly Ala Asn Ile Ala Ser Phe Ile Lys Val Ser Asp Ala Met Phe
435 440 445
Asp Gln Gly Asp Val Phe
450
<210> 108
<211> 401
<212> PRT
<213> 假单胞菌属物种菌株 101
<220>
<221> 点突变_D222Q
<222> (222)..(222)
<223> D突变为Q
<220>
<221> 点突变_H224N
<222> (224)..(224)
<223> H突变为N
<400> 108
Met Ala Lys Val Leu Cys Val Leu Tyr Asp Asp Pro Val Asp Gly Tyr
1 5 10 15
Pro Lys Thr Tyr Ala Arg Asp Asp Leu Pro Lys Ile Asp His Tyr Pro
20 25 30
Gly Gly Gln Thr Leu Pro Thr Pro Lys Ala Ile Asp Phe Thr Pro Gly
35 40 45
Gln Leu Leu Gly Ser Val Ser Gly Glu Leu Gly Leu Arg Lys Tyr Leu
50 55 60
Glu Ser Asn Gly His Thr Leu Val Val Thr Ser Asp Lys Asp Gly Pro
65 70 75 80
Asp Ser Val Phe Glu Arg Glu Leu Val Asp Ala Asp Val Val Ile Ser
85 90 95
Gln Pro Phe Trp Pro Ala Tyr Leu Thr Pro Glu Arg Ile Ala Lys Ala
100 105 110
Lys Asn Leu Lys Leu Ala Leu Thr Ala Gly Ile Gly Ser Asp His Val
115 120 125
Asp Leu Gln Ser Ala Ile Asp Arg Asn Val Thr Val Ala Glu Val Thr
130 135 140
Tyr Cys Asn Ser Ile Ser Val Ala Glu His Val Val Met Met Ile Leu
145 150 155 160
Ser Leu Val Arg Asn Tyr Leu Pro Ser His Glu Trp Ala Arg Lys Gly
165 170 175
Gly Trp Asn Ile Ala Asp Cys Val Ser His Ala Tyr Asp Leu Glu Ala
180 185 190
Met His Val Gly Thr Val Ala Ala Gly Arg Ile Gly Leu Ala Val Leu
195 200 205
Arg Arg Leu Ala Pro Phe Asp Val His Leu His Tyr Thr Asp Arg His
210 215 220
Arg Leu Pro Glu Ser Val Glu Lys Glu Leu Asn Leu Thr Trp His Ala
225 230 235 240
Thr Arg Glu Asp Met Tyr Pro Val Cys Asp Val Val Thr Leu Asn Cys
245 250 255
Pro Leu His Pro Glu Thr Glu His Met Ile Asn Asp Glu Thr Leu Lys
260 265 270
Leu Phe Lys Arg Gly Ala Tyr Ile Val Asn Thr Ala Arg Gly Lys Leu
275 280 285
Cys Asp Arg Asp Ala Val Ala Arg Ala Leu Glu Ser Gly Arg Leu Ala
290 295 300
Gly Tyr Ala Gly Asp Val Trp Phe Pro Gln Pro Ala Pro Lys Asp His
305 310 315 320
Pro Trp Arg Thr Met Pro Tyr Asn Gly Met Thr Pro His Ile Ser Gly
325 330 335
Thr Thr Leu Thr Ala Gln Ala Arg Tyr Ala Ala Gly Thr Arg Glu Ile
340 345 350
Leu Glu Cys Phe Phe Glu Gly Arg Pro Ile Arg Asp Glu Tyr Leu Ile
355 360 365
Val Gln Gly Gly Ala Leu Ala Gly Thr Gly Ala His Ser Tyr Ser Lys
370 375 380
Gly Asn Ala Thr Gly Gly Ser Glu Glu Ala Ala Lys Phe Lys Lys Ala
385 390 395 400
Val
<210> 109
<211> 1174
<212> PRT
<213> 爱荷华诺卡氏菌
<400> 109
Met Ala Val Asp Ser Pro Asp Glu Arg Leu Gln Arg Arg Ile Ala Gln
1 5 10 15
Leu Phe Ala Glu Asp Glu Gln Val Lys Ala Ala Arg Pro Leu Glu Ala
20 25 30
Val Ser Ala Ala Val Ser Ala Pro Gly Met Arg Leu Ala Gln Ile Ala
35 40 45
Ala Thr Val Met Ala Gly Tyr Ala Asp Arg Pro Ala Ala Gly Gln Arg
50 55 60
Ala Phe Glu Leu Asn Thr Asp Asp Ala Thr Gly Arg Thr Ser Leu Arg
65 70 75 80
Leu Leu Pro Arg Phe Glu Thr Ile Thr Tyr Arg Glu Leu Trp Gln Arg
85 90 95
Val Gly Glu Val Ala Ala Ala Trp His His Asp Pro Glu Asn Pro Leu
100 105 110
Arg Ala Gly Asp Phe Val Ala Leu Leu Gly Phe Thr Ser Ile Asp Tyr
115 120 125
Ala Thr Leu Asp Leu Ala Asp Ile His Leu Gly Ala Val Thr Val Pro
130 135 140
Leu Gln Ala Ser Ala Ala Val Ser Gln Leu Ile Ala Ile Leu Thr Glu
145 150 155 160
Thr Ser Pro Arg Leu Leu Ala Ser Thr Pro Glu His Leu Asp Ala Ala
165 170 175
Val Glu Cys Leu Leu Ala Gly Thr Thr Pro Glu Arg Leu Val Val Phe
180 185 190
Asp Tyr His Pro Glu Asp Asp Asp Gln Arg Ala Ala Phe Glu Ser Ala
195 200 205
Arg Arg Arg Leu Ala Asp Ala Gly Ser Leu Val Ile Val Glu Thr Leu
210 215 220
Asp Ala Val Arg Ala Arg Gly Arg Asp Leu Pro Ala Ala Pro Leu Phe
225 230 235 240
Val Pro Asp Thr Asp Asp Asp Pro Leu Ala Leu Leu Ile Tyr Thr Ser
245 250 255
Gly Ser Thr Gly Thr Pro Lys Gly Ala Met Tyr Thr Asn Arg Leu Ala
260 265 270
Ala Thr Met Trp Gln Gly Asn Ser Met Leu Gln Gly Asn Ser Gln Arg
275 280 285
Val Gly Ile Asn Leu Asn Tyr Met Pro Met Ser His Ile Ala Gly Arg
290 295 300
Ile Ser Leu Phe Gly Val Leu Ala Arg Gly Gly Thr Ala Tyr Phe Ala
305 310 315 320
Ala Lys Ser Asp Met Ser Thr Leu Phe Glu Asp Ile Gly Leu Val Arg
325 330 335
Pro Thr Glu Ile Phe Phe Val Pro Arg Val Cys Asp Met Val Phe Gln
340 345 350
Arg Tyr Gln Ser Glu Leu Asp Arg Arg Ser Val Ala Gly Ala Asp Leu
355 360 365
Asp Thr Leu Asp Arg Glu Val Lys Ala Asp Leu Arg Gln Asn Tyr Leu
370 375 380
Gly Gly Arg Phe Leu Val Ala Val Val Gly Ser Ala Pro Leu Ala Ala
385 390 395 400
Glu Met Lys Thr Phe Met Glu Ser Val Leu Asp Leu Pro Leu His Asp
405 410 415
Gly Tyr Gly Ser Thr Glu Ala Gly Ala Ser Val Leu Leu Asp Asn Gln
420 425 430
Ile Gln Arg Pro Pro Val Leu Asp Tyr Lys Leu Val Asp Val Pro Glu
435 440 445
Leu Gly Tyr Phe Arg Thr Asp Arg Pro His Pro Arg Gly Glu Leu Leu
450 455 460
Leu Lys Ala Glu Thr Thr Ile Pro Gly Tyr Tyr Lys Arg Pro Glu Val
465 470 475 480
Thr Ala Glu Ile Phe Asp Glu Asp Gly Phe Tyr Lys Thr Gly Asp Ile
485 490 495
Val Ala Glu Leu Glu His Asp Arg Leu Val Tyr Val Asp Arg Arg Asn
500 505 510
Asn Val Leu Lys Leu Ser Gln Gly Glu Phe Val Thr Val Ala His Leu
515 520 525
Glu Ala Val Phe Ala Ser Ser Pro Leu Ile Arg Gln Ile Phe Ile Tyr
530 535 540
Gly Ser Ser Glu Arg Ser Tyr Leu Leu Ala Val Ile Val Pro Thr Asp
545 550 555 560
Asp Ala Leu Arg Gly Arg Asp Thr Ala Thr Leu Lys Ser Ala Leu Ala
565 570 575
Glu Ser Ile Gln Arg Ile Ala Lys Asp Ala Asn Leu Gln Pro Tyr Glu
580 585 590
Ile Pro Arg Asp Phe Leu Ile Glu Thr Glu Pro Phe Thr Ile Ala Asn
595 600 605
Gly Leu Leu Ser Gly Ile Ala Lys Leu Leu Arg Pro Asn Leu Lys Glu
610 615 620
Arg Tyr Gly Ala Gln Leu Glu Gln Met Tyr Thr Asp Leu Ala Thr Gly
625 630 635 640
Gln Ala Asp Glu Leu Leu Ala Leu Arg Arg Glu Ala Ala Asp Leu Pro
645 650 655
Val Leu Glu Thr Val Ser Arg Ala Ala Lys Ala Met Leu Gly Val Ala
660 665 670
Ser Ala Asp Met Arg Pro Asp Ala His Phe Thr Asp Leu Gly Gly Asp
675 680 685
Ser Leu Ser Ala Leu Ser Phe Ser Asn Leu Leu His Glu Ile Phe Gly
690 695 700
Val Glu Val Pro Val Gly Val Val Val Ser Pro Ala Asn Glu Leu Arg
705 710 715 720
Asp Leu Ala Asn Tyr Ile Glu Ala Glu Arg Asn Ser Gly Ala Lys Arg
725 730 735
Pro Thr Phe Thr Ser Val His Gly Gly Gly Ser Glu Ile Arg Ala Ala
740 745 750
Asp Leu Thr Leu Asp Lys Phe Ile Asp Ala Arg Thr Leu Ala Ala Ala
755 760 765
Asp Ser Ile Pro His Ala Pro Val Pro Ala Gln Thr Val Leu Leu Thr
770 775 780
Gly Ala Asn Gly Tyr Leu Gly Arg Phe Leu Cys Leu Glu Trp Leu Glu
785 790 795 800
Arg Leu Asp Lys Thr Gly Gly Thr Leu Ile Cys Val Val Arg Gly Ser
805 810 815
Asp Ala Ala Ala Ala Arg Lys Arg Leu Asp Ser Ala Phe Asp Ser Gly
820 825 830
Asp Pro Gly Leu Leu Glu His Tyr Gln Gln Leu Ala Ala Arg Thr Leu
835 840 845
Glu Val Leu Ala Gly Asp Ile Gly Asp Pro Asn Leu Gly Leu Asp Asp
850 855 860
Ala Thr Trp Gln Arg Leu Ala Glu Thr Val Asp Leu Ile Val His Pro
865 870 875 880
Ala Ala Leu Val Asn His Val Leu Pro Tyr Thr Gln Leu Phe Gly Pro
885 890 895
Asn Val Val Gly Thr Ala Glu Ile Val Arg Leu Ala Ile Thr Ala Arg
900 905 910
Arg Lys Pro Val Thr Tyr Leu Ser Thr Val Gly Val Ala Asp Gln Val
915 920 925
Asp Pro Ala Glu Tyr Gln Glu Asp Ser Asp Val Arg Glu Met Ser Ala
930 935 940
Val Arg Val Val Arg Glu Ser Tyr Ala Asn Gly Tyr Gly Asn Ser Lys
945 950 955 960
Trp Ala Gly Glu Val Leu Leu Arg Glu Ala His Asp Leu Cys Gly Leu
965 970 975
Pro Val Ala Val Phe Arg Ser Asp Met Ile Leu Ala His Ser Arg Tyr
980 985 990
Ala Gly Gln Leu Asn Val Gln Asp Val Phe Thr Arg Leu Ile Leu Ser
995 1000 1005
Leu Val Ala Thr Gly Ile Ala Pro Tyr Ser Phe Tyr Arg Thr Asp
1010 1015 1020
Ala Asp Gly Asn Arg Gln Arg Ala His Tyr Asp Gly Leu Pro Ala
1025 1030 1035
Asp Phe Thr Ala Ala Ala Ile Thr Ala Leu Gly Ile Gln Ala Thr
1040 1045 1050
Glu Gly Phe Arg Thr Tyr Asp Val Leu Asn Pro Tyr Asp Asp Gly
1055 1060 1065
Ile Ser Leu Asp Glu Phe Val Asp Trp Leu Val Glu Ser Gly His
1070 1075 1080
Pro Ile Gln Arg Ile Thr Asp Tyr Ser Asp Trp Phe His Arg Phe
1085 1090 1095
Glu Thr Ala Ile Arg Ala Leu Pro Glu Lys Gln Arg Gln Ala Ser
1100 1105 1110
Val Leu Pro Leu Leu Asp Ala Tyr Arg Asn Pro Cys Pro Ala Val
1115 1120 1125
Arg Gly Ala Ile Leu Pro Ala Lys Glu Phe Gln Ala Ala Val Gln
1130 1135 1140
Thr Ala Lys Ile Gly Pro Glu Gln Asp Ile Pro His Leu Ser Ala
1145 1150 1155
Pro Leu Ile Asp Lys Tyr Val Ser Asp Leu Glu Leu Leu Gln Leu
1160 1165 1170
Leu
<210> 110
<211> 206
<212> PRT
<213> 大肠杆菌 K12
<400> 110
Met Lys Thr Thr His Thr Ser Leu Pro Phe Ala Gly His Thr Leu His
1 5 10 15
Phe Val Glu Phe Asp Pro Ala Asn Phe Cys Glu Gln Asp Leu Leu Trp
20 25 30
Leu Pro His Tyr Ala Gln Leu Gln His Ala Gly Arg Lys Arg Lys Thr
35 40 45
Glu His Leu Ala Gly Arg Ile Ala Ala Val Tyr Ala Leu Arg Glu Tyr
50 55 60
Gly Tyr Lys Cys Val Pro Ala Ile Gly Glu Leu Arg Gln Pro Val Trp
65 70 75 80
Pro Ala Glu Val Tyr Gly Ser Ile Ser His Cys Gly Thr Thr Ala Leu
85 90 95
Ala Val Val Ser Arg Gln Pro Ile Gly Ile Asp Ile Glu Glu Ile Phe
100 105 110
Ser Val Gln Thr Ala Arg Glu Leu Thr Asp Asn Ile Ile Thr Pro Ala
115 120 125
Glu His Glu Arg Leu Ala Asp Cys Gly Leu Ala Phe Ser Leu Ala Leu
130 135 140
Thr Leu Ala Phe Ser Ala Lys Glu Ser Ala Phe Lys Ala Ser Glu Ile
145 150 155 160
Gln Thr Asp Ala Gly Phe Leu Asp Tyr Gln Ile Ile Ser Trp Asn Lys
165 170 175
Gln Gln Val Ile Ile His Arg Glu Asn Glu Met Phe Ala Val His Trp
180 185 190
Gln Ile Lys Glu Lys Ile Val Ile Thr Leu Cys Gln His Asp
195 200 205
<210> 111
<211> 224
<212> PRT
<213> 枯草芽孢杆菌菌株168
<400> 111
Met Lys Ile Tyr Gly Ile Tyr Met Asp Arg Pro Leu Ser Gln Glu Glu
1 5 10 15
Asn Glu Arg Phe Met Ser Phe Ile Ser Pro Glu Lys Arg Glu Lys Cys
20 25 30
Arg Arg Phe Tyr His Lys Glu Asp Ala His Arg Thr Leu Leu Gly Asp
35 40 45
Val Leu Val Arg Ser Val Ile Ser Arg Gln Tyr Gln Leu Asp Lys Ser
50 55 60
Asp Ile Arg Phe Ser Thr Gln Glu Tyr Gly Lys Pro Cys Ile Pro Asp
65 70 75 80
Leu Pro Asp Ala His Phe Asn Ile Ser His Ser Gly Arg Trp Val Ile
85 90 95
Cys Ala Phe Asp Ser Gln Pro Ile Gly Ile Asp Ile Glu Lys Thr Lys
100 105 110
Pro Ile Ser Leu Glu Ile Ala Lys Arg Phe Phe Ser Lys Thr Glu Tyr
115 120 125
Ser Asp Leu Leu Ala Lys Asp Lys Asp Glu Gln Thr Asp Tyr Phe Tyr
130 135 140
His Leu Trp Ser Met Lys Glu Ser Phe Ile Lys Gln Glu Gly Lys Gly
145 150 155 160
Leu Ser Leu Pro Leu Asp Ser Phe Ser Val Arg Leu His Gln Asp Gly
165 170 175
Gln Val Ser Ile Glu Leu Pro Asp Ser His Ser Pro Cys Tyr Ile Lys
180 185 190
Thr Tyr Glu Val Asp Pro Gly Tyr Lys Met Ala Val Cys Ala Ala His
195 200 205
Pro Asp Phe Pro Glu Asp Ile Thr Met Val Ser Tyr Glu Glu Leu Leu
210 215 220

Claims (8)

1.一种用于生产己二酸的方法，所述方法包括使用具有2-烯酸酯还原酶活性的2-烯酸酯还原酶将6-碳不饱和二元羧酸酶促转化为己二酸，所述2-烯酸酯还原酶的氨基酸序列如SEQ ID NO:1或2所示。
2.如权利要求1所述的方法，其中所述2-烯酸酯还原酶包含由SEQ ID NO:1或2的氨基酸残基363至382所限定的4Fe-4S簇。
3.如权利要求1所述的方法，其中所述不饱和6-碳二元羧酸是粘康酸或2-己烯二酸。
4.如权利要求1所述的方法，其中所述6-碳不饱和二元羧酸向己二酸的酶促转化发生在被工程化以表达2-烯酸酯还原酶的宿主微生物中。
5.如权利要求4所述的方法，其中所述方法包括在生产己二酸的条件下在基本上有氧的培养基中培养所述宿主微生物足够长的时间以生产己二酸，其中基本上有氧的培养基表示溶解氧的量等于或大于饱和度的10％。
6.如权利要求4或5所述的方法，其中所述宿主微生物为细菌。
7.如权利要求4或5所述的方法，其中所述宿主微生物为大肠杆菌。
8.一种组合物，其包含如权利要求1或2所限定的2-烯酸酯还原酶，或如权利要求4-7中任一项所限定的宿主微生物，以及粘康酸和/或2-己烯二酸。
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