申请/专利权人：福尼亚生物处理股份有限公司

申请日：2018-05-10

公开（公告）日：2021-11-19

公开（公告）号：CN110191641B

主分类号：C12N9/18(20060101)

分类号：C12N9/18(20060101);C12N15/55(20060101);A23K20/189(20160101);A23K50/80(20160101)

优先权：["20170803 US 15/668,231"]

专利状态码：有效-授权

法律状态：2021.11.19#授权;2019.09.24#实质审查的生效;2019.08.30#公开

摘要：提供了新的内酯酶。

主权项：1.内酯酶变体，其为图5中表格所示的任一变体。

全文数据：内酯酶及其使用方法对相关申请的交叉引用本申请要求2017年8月3日提交的美国专利申请No.15668,231的优先权，其在此通过引用并入。序列表本申请含有序列表，其已经以ASCII格式电子提交，并且在此通过引用完整并入。在2018年5月8日创建的所述ASCII拷贝命名为114095-5005-WO_ST25.txt，并且大小为367千字节。I.发明背景细菌中的细胞与细胞通信控制通常由基因表达调节的一大批活性，其导致生长期间表型变化和对环境条件的适应。术语“群体感应quorumsensing”或“QS”用来描述单细胞细菌群体以细胞密度依赖性方式使用可扩散小分子作为单一多细胞生物体起作用的能力见Amaraetal.,Chem.Rev.2011,195-208。存在有许多不同类别的“群体感应分子”，即QSM，包括寡肽、二羟基戊二酮DPD衍生物、N-酰基高丝氨酸内酯AHL和其他小分子。信号分子在细菌生长期间产生和分泌；随着群体扩充，QSM的浓度增加。一旦达到阈值水平例如“群体水平quorumlevel”，这些通过调节QS依赖性靶基因表达诱导表型效应。例如，改变的基因表达导致色素沉着、生物膜形成和毒力基因表达，从而导致蛋白酶、裂解酶和外毒素的释放。已经显示群体淬灭quorumquenching，“QQ”酶如AHL降解酶减弱鱼中的细菌性感染参见Caoetal.,J.App.Environ.Microbio.786:18992012。也已经显示了这些酶可用于控制膜的生物膜污染，参见Ohet.al.,Environ.Science&Tech.2012,46:4877-4884和Ladeetal.,Int.J.Biol.Sci.201410:550。本发明提供了新的AHL降解酶和使用方法。II.发明概述因此，本发明提供了包含内酯酶变体酶的组合物，所述内酯酶变体酶与SEQIDNO:1相比包含氨基酸取代，其中所述氨基酸取代在选自17,32,34,35,41,50,62,66,68,69,70,74,76,77,79,80,81,83,84,114,120,122,123,127,132,136,139,155,162,164,184,185,188,189,192,193,195,218,221,222和244的位置处，其中变体内酯酶变体酶保留内酯酶活性并且与SEQIDNO:1至少95％相同。在其他方面，本发明提供了与SEQIDNO:1相比的内酯酶变体酶的组合物，其中氨基酸取代选自L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,D66E,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,E80G,L81P,V83I,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,L136V,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在一些情况下，这些内酯酶变体酶保留内酯酶活性。在一些情况下，它们与SEQIDNO:1至少95％相同。在另一方面，变体酶包含氨基酸取代D66EE80GV83IL136V。在另一方面，所述变体酶任选地还包含选自L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T的氨基酸取代。在另一方面，变体酶包含氨基酸取代D34KD66EE80GL136VA184NG189P。在这方面，变体酶任选地还包含选自LI7V,F32L,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,177V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A185E,S188D,S188N,S188R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T的氨基酸取代。在另一方面，变体酶具有选自D66EQ114H,G35SV41F,G35S,G35SE195A,G189RE195A,L17VG35S,L17VG35SI77VL136VG189R,L17VG35SD66E,L17VD66EL136V,A132E,G189R,L17VD66E,G35SQ114H,Q114HG189R,D66EV83I,D66EE80GV83IL136V,G35SD66EV83IA222T,D66EI77VE80GV83IQ114HA132EG189R,G120S,A69I,A69V,D34E,N74H,G120T,G120H,A69C,D34A,G127A,A69K,A69Q,G120R,D34G,E218T,A69T,D34Q,G120P,A184N,G120A,E218M,E218N,D34K,D34S,E218A,V76S,E244H,S162N,S162C,E244C,S188D,S188R,V76K,A50P,H139T,V76R,H139K,Q123KR164K,E244K,S162H,E244S,Q70S,E244T,E244R,Q70N,Q70Y,V76E,V76L,V76A,E244G,S188N,V76N,L81P,L81PI84V,T62AG79AR164KI193V,G79AL81P,G79AI84VV122LT62AL81PV122II193V,T62AG79AL81P,T62AG79A,L81PA185E,G79AL81PI84VA155D,L68VL81PI84VA155DI193V,L81PI84VI193V,F32LL81PI84VI193V,L68VL81PI84VR164K,G79A,G79AV122IR164K,G79AI84V,V122II193VQ221H,L81PV122I,T62AG79AL81PI193VQ221H,T62AL68VV122IR164K,T62AL81PI84V,V122I,G79AV122I,G79AI193V,D66EE80GV83IL136VD34KA184NG189R,D66EE80GV83IL136VD34KA184NG189RE218T,D66EE80GV83IL136VD34K,D66EE80GV83IL136VD34KA184N,D66EE80GL136VD34K,D66EE80GV83IL136VD34KA184NE218T,D66EE80GL136V,D66EE80GL136VD34KA184NG189RE218T,D66EE80GL136VD34KG189R,D66EE80GV83IL136VR192M,D66EE80GL136VD34KG189RE218T,D66EE80GL136VD34KA184NG189R,D66EE80G^83IL136VA184NG189R和D66EE80GL136VD34KE218T的氨基酸取代组。在另一方面，如上的组合物还包含动物饲料。在另一方面，本发明提供了编码本文的内酯酶变体酶的核酸。在另一方面，本发明提供了包含核酸的表达载体，和包含表达载体的宿主细胞。另一方面，本发明提供了在产生所述酶的条件下培养包含如上的表达载体的宿主细胞，并回收变体酶。III.附图简述图1描绘了如实施例5中测定的SEQIDNO:1的G1P野生型内酯酶的热活性概况。图2描绘了如实施例5中测定的SEQIDNO:1的G1P野生型内酯酶的pH概况。图3描绘了如实施例5中测定的SEQIDNO:1的G1P野生型内酯酶的热稳定性概况。图4A至4C描绘了显示一些第一代变体内酯酶及其活性、pH和热稳定性的表。CL00023198是G1P亲本，其氨基酸序列是SEQIDNO:1，如图7所示。大多数变体是单或双氨基酸变体，如“AA突变”栏中所示，其显示与G1P相比的氨基酸取代例如，对于CL00025408，在位置66处从D到E的取代和位置114处从Q到H的取代。如实施例6至8中所示测定该表的值。图5描绘了显示一些第二代变体内酯酶及其pH和热稳定性的表。CL00025686是G2P亲本，其氨基酸序列是SEQIDNO:33，如图7所示。也就是说，图5中的所有变体也含有D66EE80GV83IL136VG1P变体。如实施例5至9中所示测定该表的值。图6描绘了如实施例9中测定的许多变体内酯酶包括G2P和G3P的热稳定性概况。图7显示了G1P、G2P和G3P的氨基酸序列和比对。图8描绘了显示本发明的一些实施方案中的一些优选变体的变体表。如本文所述，这些可以以任何组合以及与如本文概述的变体组来组合。图9描绘了变体内酯酶G2P的核酸和氨基酸序列。图10描绘了变体内酯酶G3P的核酸和氨基酸序列。图11描绘了野生型内酯酶有时在本文中称为G1P的核酸和氨基酸序列。图12描绘了一些额外的群体感应分子和底物。IV.发明详述A.简介细菌使用可扩散小分子作为信号通过群体感应彼此通信，所述信号在阈值水平触发QS依赖性基因表达，这可导致宿主生物体细菌性感染、生物膜的形成等。这些分子的浓度在纳摩尔至毫摩尔范围内，扩散范围为4至80mm，这取决于系统。这些事件中的一些可以通过使用分解这些分子的群体淬灭酶破坏群体感应系统来解决。由于群体淬灭不杀死细菌或限制细胞生长，但是关闭致病性基因的表达，此类对群体感应系统的干扰可以替换传统抗生素并导致更少的副作用。此类方法可用于农业以对抗细菌性植物病原体，从而用作具有明显缺点的杀虫剂的替代物。类似地，此方法可用于水产养殖，其中可以在不使用可在人消费者中有活性的抗生素的情况下治疗养殖鱼类中的细菌性感染特别是气单胞菌属种Aeromonasspp.感染。此外，大多数可以杀死气单胞菌属种的抗生素被禁止在水产养殖中定期使用，并且无论如何许多气单胞菌属种对常用的抗生素有抗性。鱼类中的气单胞菌感染引起败血症，导致鱼类的鳍和尾腐烂，以及溃疡综合症，从而导致由于气单胞菌感染所致的超过1亿美元损失。此外，食用感染了气单胞菌的鱼类可导致人体肠道或皮肤伤口感染。如本文所讨论的，革兰氏阴性细菌中的主要类别的QS分子之一是N-酰基高丝氨酸内酯AHL，其均包含脱水环酯，并且通常含有不同的链长和取代基。内酯酶是水解酶，其裂解AHL的内酯环，由于阻止它们与靶转录调节剂结合，使得它们在生物学上无活性。代表性方案如下所示：通常，n是至少1的整数，且R是取代基。由于不同的细菌具有不同的特异性，存在有许多可能的AHL底物。用于测试内酯酶活性的通用底物是N-3-氧代辛酰基-L-高丝氨酸内酯N-3-Oxooctanoyl-L-homoserinelactone，但可以使用许多其他底物，包括但不限于图12中所示的那些。然而，通常，天然内酯酶具有有限的pH范围、有限的热活性范围和有限的热稳定性范围。因此，饲料中的内酯酶可以由胃肠道中的温度和或低pH失活，因此可能难以利用。因此，本发明涉及新的变体内酯酶变体酶，其用作用于多种应用的抗细菌剂和抗微生物剂。B.定义本文的“修饰”指多肽序列中的氨基酸取代、插入和或缺失或与蛋白质化学连接的部分的改变。例如，修饰可以是附着于蛋白质的改变的碳水化合物或PEG结构。本文的“氨基酸修饰”指多肽序列中的氨基酸取代、插入和或缺失。为清楚起见，除非另有说明，否则氨基酸修饰始终是由DNA编码的氨基酸，例如含有DNA和RNA的密码子的20种氨基酸。本文的“氨基酸取代”或“取代”指用不同氨基酸替换亲本多肽序列中特定位置处的氨基酸。特别地，在一些实施方案中，取代指不天然存在于特定位置，不天然存在于生物体内或任何生物体中的氨基酸。例如，取代D66E指变体多肽，在本申请中是内酯酶，其中位置66处的天冬氨酸用谷氨酸取代。为清楚起见，已经经过工程化改造而改变核酸编码序列但不改变起始氨基酸例如将CGG编码精氨酸更换为CGA仍编码精氨酸以增加宿主生物体表达水平的蛋白质不是“氨基酸取代”；也就是说，尽管创建编码相同蛋白质的新基因，但是若蛋白质在其起始的特定位置处具有相同的氨基酸，则它不是氨基酸取代。如本文所用，“氨基酸插入”或“插入”指在亲本多肽序列中的特定位置处添加氨基酸序列。例如，-233E或233E表示在位置233之后和位置234之前插入谷氨酸。另外，-233ADE或A233ADE表示在位置233之后和位置234之前插入AlaAspGlu。如本文所用，“氨基酸缺失”或“缺失”指除去亲本多肽序列中特定位置处的氨基酸序列。例如，E233-或E233#、E233Q或E233del表示位置233处的谷氨酸缺失。另外，EDA233-或EDA233#表示在位置233处开始的序列GluAspAla的缺失。如本文所用，“亲本多肽”指随后被修饰以产生变体的起始多肽。亲本多肽可以是天然存在的多肽，或天然存在的多肽的变体或工程化形式。亲本多肽可以指多肽本身、包含亲本多肽的组合物、或者编码它的氨基酸序列。在本申请中，一些实施方案使用GIP、G2P或G3P作为亲本多肽，其中前者是优选的。如本文所用，“变体蛋白质”或“蛋白质变体”或“变体”指凭借至少一个氨基酸修饰而不同于亲本蛋白质序列的蛋白质。蛋白质变体可以指蛋白质本身、包含蛋白质的组合物、或编码它的氨基酸序列。优选地，与亲本蛋白质相比，蛋白质变体具有至少一个氨基酸修饰，例如与亲本相比，约1至约70个氨基酸修饰，优选约1至约5个氨基酸修饰。如下所述，在一些实施方案中，亲本多肽是野生型序列，例如本文称为“GIP”的野生型内酯酶，其是来自砖红色微杆菌Microbacteriumtestaceum的内酯酶。如下文进一步讨论的，本文的蛋白质变体序列优选与亲本蛋白质序列拥有至少约80％的同一性，最优选至少约90％同一性，更优选至少约95-98-99％同一性。变体蛋白质可以指变体蛋白质本身、包含蛋白质变体的组合物、或编码它的DNA序列。因此，本文的“变体内酯酶”指与亲本内酯酶相比在氨基酸序列中具有至少一个氨基酸修饰的新的内酯酶。如本文所讨论的，在一些情况下，亲本内酯酶是第二代或更高代的变体；也就是说，如图6所示，与野生型GIP亲本相比，G2P内酯酶具有4个氨基酸取代。然而，如图7所示，与G2P亲本相比，G3P具有4个氨基酸取代，但与GIP相比总共有8个氨基酸取代。除非另有说明或从上下文中显而易见，否则通常将本发明的变体内酯酶与野生型GIP序列进行比较。另外，除非另有说明，否则本发明的变体内酯酶具有酶活性，即，使用实施例6中所述的内酯酶测定法，使用不含温度处理的测定法可检测的内酯酶活性。如本文所用，“蛋白质”在本文中指至少两个共价连接的氨基酸，其包括蛋白质、多肽、寡肽和肽。肽基通常包含天然存在的氨基酸和肽键。另外，多肽可以包括一个或多个侧链或末端的合成衍生化、糖基化、PEG化、环状置换、环化、与其他分子的接头、与蛋白质或蛋白质域的融合、以及肽标签或标记物的添加。如本文所用，“残基”指蛋白质中的位置及其相关的氨基酸身份。例如，天冬氨酸66也称为Asp66或D66是GIP亲本酶中位置66处的残基。如本文所用，“非天然存在的修饰”指在亲本例如GIP酶中未发现的氨基酸修饰。如本文所用，“氨基酸”和“氨基酸身份”指20种由DNA和RNA编码的天然存在的氨基酸之一。如本文所用，“位置”指蛋白质序列中的位置。通常，位置编号其在下文更全面讨论是相对于成熟内酯酶序列例如排除信号肽的第一个氨基酸。“内酯酶”在本文中指具有内酯酶活性的蛋白质。“内酯酶活性”在本文中指酶催化N-3-氧代辛酰基-L-高丝氨酸内酯的水解，其释放质子，所述质子然后使用pH敏感性比色测定法、MOPSBTB测定法检测，其中在底物水解后释放质子改变微弱缓冲测定法的pH值，因此改变pH敏感性染料BTB的质子化状态；见Yangetal.,Anal.Biochem.3562006:297-299，其在此通过引用明确且特别针对其中描述的测定法并入本文。认为在下文和实施例6中概述的具有可检测活性的酶是本文的内酯酶。本文中就两个序列而言的“同一性”指考虑到比对，相同的氨基酸位于相同的位置。本发明的氨基酸序列“发明序列”与权利要求中提及的亲本氨基酸序列例如对于GIP，SEQIDNO:1之间的同一性程度以两个序列的比对中的精确匹配的数目除以“发明序列”的长度，或SEQIDNO:1的长度以最短者为准计算。结果以百分比同一性表示，如下文计算。为了本发明的目的，使用SEQIDNO:1中公开的成熟多肽来确定本发明的另一种内酯酶中的相应氨基酸残基。另一种内酯酶的氨基酸序列与SEQIDNO:1中公开的成熟多肽比对，并且基于比对，对应于SEQIDNO:1中公开的成熟多肽中的任何氨基酸残基的氨基酸位置编号使用Needleman-Wunsch算法NeedlemanandWunsch,1970,J.Mol.Biol.48:443-453测定，如EMBOSS包EMBOSS:TheEuropeanMolecularBiologyOpenSoftwareSuite,Riceetal.,2000,TrendsGenet.16:276-277，优选5.0.0或更高版本的Needle程序中执行。使用的参数是缺口开放罚分10、缺口延伸罚分0.5，以及EBLOSUM62BLOSUM62的EMBOSS版本取代矩阵。可以通过使用几种计算机程序使用它们各自的默认参数对多个多肽序列进行比对来确定另一种内酯酶中相应的氨基酸残基的鉴定，所述计算机程序包括但不限于MUSCLE通过对数期望的多序列比较；3.5或更高版本；Edgar,2004,NucleicAcidsResearch32:1792-1797，MAFFT6.857或更高版本；KatohandKuma,2002,NucleicAcidsResearch30:3059-3066；Katohetal.,2005,NucleicAcidsResearch33:511-518；KatohandToh,2007,Bioinformatics23:372-374；Katohetal.,2009,MethodsinMolecularBiology537:39-64；KatohandToh,2010,Bioinformatics26:1899-1900，采用ClustalW的EMBOSSEMMA1.83或更高版本；Thompsonetal.,1994,NucleicAcidsResearch22:4673-4680和采用ClustalOmega的EMBL-EBISieversandHiggins,2014,MethodsMolBiol.2014；1079:105-16。当另一种酶与SEQIDNO:1的多肽趋异，使得基于序列的传统比较未能检测它们的关系LindahlandElofsson,2000,J.Mol.Biol.295:613-615时，可以使用其他成对序列比较算法。可以使用利用多肽家族的概率表示概况来搜索数据库的搜索程序来获得基于序列的搜索的更大灵敏度。例如，PSI-BLAST程序通过迭代数据库搜索过程生成概况，并且能够检测远程同源物Atschuletal.,1997,NucleicAcidsRes.25:3389-3402。若多肽的家族或超家族在蛋白质结构数据库中具有一个或多个呈现，则可以实现甚至更大的灵敏度。诸如GenTHREADERJones,1999,J.Mol.Biol.287:797-815；McGuffinandJones,2003,Bioinformatics19:874-881等程序利用来自多种来源PSI-BLAST、二级结构预测、结构比对概况和溶剂化电位的信息作为预测查询序列的结构折叠的神经网络的输入。类似地，Goughetal.,2000,J.Mol.Biol.313:903-919的方法可用于比对未知结构的序列与SCOP数据库中存在的超家族模型。这些比对可以继而用于产生多肽的同源性模型，并且可以使用为所述目的开发的多种工具评估此类模型的准确性。对于已知结构的蛋白质，几种工具和资源可用于检索和产生结构比对。例如，SCOP蛋白质超家族已经在结构上比对，并且那些比对是可访问的且可下载的。可以使用多种算法比对两种或更多种蛋白质结构，例如距离比对矩阵HolmandSander,1998,Proteins33:88-96或组合延伸ShindyalovandBourne,1998,ProteinEngineering11:739-747，并且这些算法的实现可以另外用于查询具有感兴趣结构的结构数据库，以发现可能的结构同源物例如HolmandPark,2000,Bioinformatics16:566-567。在描述本发明的变体时，下面描述的命名法为了便于参考而采用。采用公认的IUPAC单字母或三字母氨基酸缩写。对于氨基酸取代，本文使用以下命名法：原始氨基酸，位置，取代的氨基酸。因此，用异亮氨酸取代位置83处的缬氨酸称为“Val83Ile”或“V83I”。多个突变由正斜线标记“”分开，例如“D66EE80GV83IL136V”，分别代表位置66、80、83和136处的取代。然而，应当注意，变体的叙述顺序无关紧要；“D66EE80GV83IL136V”与“L136VE80GD66EV83I”属于相同变体。如本文所用，术语“分离的”指根据SDS-PAGE测定，至少20％纯的，优选至少40％纯的，更优选至少60％纯的，甚至更优选至少80％纯的，最优选至少90％纯的，甚至最优选至少95-98％纯的多肽。特别地，优选多肽是“基本上纯的形式”，即，多肽制备物基本上不含与其天然相关的其他多肽物质。例如，这可以通过公知的重组方法或通过经典的纯化方法制备多肽来实现。“重组酶”在本文中指通过重组技术产生酶，并且编码本发明的变体酶的核酸与至少一种外源例如，不是亲本内酯酶天然的序列可操作地连接，所述序列包括例如启动子、终止子、信号序列等，如下面更全面概述的。术语“核酸构建体”指单链或双链的核酸分子，其从天然存在的基因中分离或以在其它情况下不天然存在的方式修饰而包含核酸区段或者是合成的，并且包含一个或多个控制序列。术语“可操作连接”指将控制序列相对于多核苷酸的编码序列置于适当位置处，使得控制序列指导编码序列的表达的构造。V.本发明的内酯酶因此，本发明提供了具有改善的活性的变体内酯酶，其可以用于多种应用，包括动物和人类营养和饲料产品，以及用作作物植物中的杀虫剂，用于在一些实施方案中特别感兴趣的鱼类的饲料原料中。通常，与野生型亲本内酯酶，“内酯酶G1P”或“G1P”例如“第1代亲本”，本文中SEQIDNO:1如图7所示相比，本发明的变体内酯酶具有经修饰的、改善的生物化学性质。本文中可以改善的变体内酯酶的生物化学性质包括但不限于pH活性、pH稳定性、热稳定性、比活性、制剂稳定性包括液体、固体和颗粒、在鱼类和动物和或鱼饲料中的性能。与G1P相比，本发明的变体内酯酶具有一种或多种改善的性质“。改善的”在本文中指至少一种生物化学性质的期望变化。“改善的功能”可以以特定活性的百分比增加或减少，或作为“倍数”变化测量，其中期望性质例如pH稳定性，热稳定性增加或不期望性质例如蛋白酶敏感性降低。也就是说，与G1P相比，变体内酯酶的热稳定性可以增加10％。或者，变体内酯酶可以具有pH稳定性的2倍增加。通常，百分比变化用于描述小于100％的生物化学活性变化，并且倍数变化用于描述大于100％的生物化学活性的变化与亲本酶相比，在许多情况下为G1P。在本发明中，可以实现至少约10％，20％，30％，40％，50％，60％，70％，80％，90％，95％，98％99％的生物化学活性的百分比变化通常增加。在本发明中，与起始酶或亲本酶相比，测量“倍数增加”或减少。例如，如图所示，与G1P相比，G2P具有2.5倍增加的温度耐受性：这通过[变体活性亲本活性]计算。在许多实施方案中，改善为至少1.5倍1.5倍，2倍，3倍，4倍，5倍，6倍，7倍，8倍，9倍或10倍或更高。通常，在针对G1P酶攻击变体内酯酶的条件下，使用内酯酶活性测定法与G1P酶野生型相比测量改善。A.内酯酶活性可以以多种方式测试变体内酯酶，然而，所有变体内酯酶将在某种水平保留内酯酶活性，如本文所讨论的。基本的内酯酶测定法如实施例5和如下所示进行：在适当的温度，pH等条件在下面讨论下攻击后，使样品达到0.05MTris-HCl，pH7.2的终浓度。在4％DMSO中制备的底物N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃温育对于基线活性，其他温度如50℃和60℃至测定法通用热活性，200rpm，30分钟。然后，使用75μl含有3.5MNaOH：2M羟胺盐酸盐::1:1的淬灭剂使反应进行淬灭。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度，作为由于反应引起的质子释放的指示；见Yanget.al.，同上，对于测定法和测定条件，通过引用并入本文。酶可以是纯化的样品、发酵样品或原始样品。本发明的变体内酯酶可以具有许多生物化学性质中的一种或多种的改善，包括但不限于pH活性、pH稳定性、热稳定性、比活性、制剂稳定性包括液体、固体和团粒、在动物和或动物饲料中的性能。B.热稳定性在许多实施方案中，本发明的变体内酯酶具有增加的热稳定性，特别是在用于生产动物饲料的条件下，例如，所述条件在造粒过程期间经常使用高温达一段时间，所述时间段应使野生型内酯酶失活。在此上下文中的“热稳定性”指在相同的热攻击条件下变体酶比亲本内酯酶例如G1P更稳定，也就是说，变体的活性在相同条件通常使用如本文所概述的和如实施例7中所示的内酯酶测定法下高于GIP的活性。在一个实施方案中，当暴露于50℃，55℃，55.5℃，58℃和或60℃的温度达一段时间时，变体内酯酶比亲本内酯酶更稳定，根据使用变体内酯酶的最终条件，所述时间段的范围一般为约1分钟、2分钟、3分钟、4分钟、5分钟、7分钟、8分钟、9分钟或10分钟或更长时间，一些实施方案利用5分钟至10分钟、5分钟至15分钟、5分钟至60分钟、10分钟至60分钟的热攻击时间，在本发明中均可使用。在一些实施方案中，使用55.5℃和10分钟的攻击，并且在一些实施方案中，使用55.5℃和60分钟的攻击。因此，在一些实施方案中，与亲本内酯酶，特别是GIP相比，变体内酯酶具有增加的热稳定性，在50℃持续至少5分钟，在55℃持续至少5-10分钟，在55.5℃持续至少5-10分钟，且在一些实施方案中在55.5℃持续至少60分钟。此外，pH也可以是热稳定性的考虑因素。因此，在一些实施方案中，变体内酯酶与亲本内酯酶相比具有增加的热稳定性，在55.5℃，pH7.2持续至少5-10至60分钟。因此，如图4、5和6所示，本发明的许多变体内酯酶表现出增加的热稳定性。C.pH稳定性在许多实施方案中，本发明的变体内酯酶在较低pH具有增加的pH稳定性。在此上下文中“增加的pH稳定性”意味着在相同的pH攻击条件下变体酶比亲本内酯酶例如GIP更稳定，也就是说，在相同条件通常使用如本文概述的和如实施例5中所示的内酯酶测定法下，变体的活性高于GIP的活性。VI.特定的变体内酯酶因此，本发明提供了许多特定的变体内酯酶，其具有改善的活性，特别是热稳定性和或pH稳定性，特别是如本文概述的特定pH下的热稳定性。在一些实施方案中，变体内酯酶在选自17,32,34,35,41,50,62,66,68,69,70,74,76,77,79,80,81,83,84,114,120,122,123,127,132,136,139,155,162,164,184,185,188,189,192,193,195,218,221,222和244的位置相对于G1P处具有一个或多个氨基酸取代。在一些实施方案中，变体内酯酶具有选自这些位置的1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19或20个氨基酸取代，在一些实施方案中1至10得到应用。如本领域技术人员所理解的，与G1P相比，变体内酯酶可具有许多氨基酸取代，尽管应当认为是变体内酯酶，但它必须在如本文所述的内酯酶测定法中仍具有可检测的活性，并且它必须与G1P充分相同以在如本文所讨论的序列比对程序中比对。在一些实施方案中，变体内酯酶具有选自以下的一个或多个氨基酸取代：L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,D66E,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,E80G,L81P,V83I,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,L136V,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在一些实施方案中，变体内酯酶具有选自这些位置的1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19或20个氨基酸取代，在一些实施方案中1至10得到应用。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置17处具有亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是L17V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置32处具有苯丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是F32L。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置34处具有天冬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自D34A,D34E,D34G,D34K,D34Q和D34S。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置35处具有甘氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是G35S。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置41处具有缬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是V41F。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置50处具有丙氨酸的氨基酸取代。在一些实施方案中，取代用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是A50P。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置62处具有苏氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是T62A。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置66处具有天冬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是D66E。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置68处具有亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是L68V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置69处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自A69C,A69I,A69K,A69Q,A69T和A69V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置70处具有谷氨酰胺的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自Q70N，Q70S和Q70Y。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置74处具有天冬酰胺的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是N74H。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置76处具有缬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自V76A,V76E,V76K,V76L,V76N,V76R和V76S。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置77处具有异亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是I77V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置79处具有甘氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是G79A。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置80处具有谷氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是E80G。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置81处具有亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是L81P。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置83处具有缬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是V83I。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置84处具有异亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是I84V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置114处具有谷氨酰胺的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是Q114H。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置120处具有甘氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自G120A，G120H，G120P，G120R，G120S和G120T。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置122处具有缬氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是V122I。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置17处具有谷氨酰胺的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是Q123K。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置127处具有甘氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是G127A。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置132处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是A132E。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置136处具有亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是L136V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置139处具有组氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自H139K和H139T。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置155处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是A155D。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置162处具有丝氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自A162C，A162H和A162N。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置164处具有精氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是R164K。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置184处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是A184N。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置185处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应的实施方案。在一些实施方案中，氨基酸取代是A185E。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置188处具有丝氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自S188D，S188N和S188R。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置189处具有甘氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是G189R。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置192处具有精氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是R192M。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置193处具有异亮氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是I193V。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置195处具有谷氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案中不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是E195A。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置218处具有谷氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫键形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代选自E218A，E218M，E218N和E218T。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置221处具有谷氨酰胺的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是Q221H。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置222处具有丙氨酸的氨基酸取代。在一些实施方案中，取代是用其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，谷氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。在一些实施方案中，氨基酸取代是A222T。在一些实施方案中，变体内酯酶在SEQIDNO:1的位置244处具有谷氨酸的氨基酸取代。在一些实施方案中，取代是用任何其他19种天然存在的氨基酸中的任一种，即谷氨酰胺，丝氨酸，亮氨酸，苏氨酸，天冬酰胺，赖氨酸，精氨酸，组氨酸，天冬氨酸，半胱氨酸，甘氨酸，脯氨酸，丙氨酸，异亮氨酸，甲硫氨酸，苯丙氨酸，色氨酸，缬氨酸和酪氨酸，一些实施方案不利用半胱氨酸由于可能的二硫化物形成或脯氨酸由于空间效应。。在一些实施方案中，氨基酸取代选自E244C,E244G,E244H,E244K,E244R,E244S和E244T。在一些实施方案中，变体内酯酶包含G2P变体D66EE80GV83IL136V和上文概述的另外的单一氨基酸变体中的至少一种，包括但不限于L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,VI221,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在实施方案中，变体内酯酶可以具有1,2,3,4,5或6个另外的氨基酸变体。在一些实施方案中，变体内酯酶包含G3P变体D34KI83VA184NG189R和上文概述的另外的单一氨基酸变体中的至少一种，包括但不限于L17V,F32L,G35S,V41F,A50P,T62A,D66E,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,E80G,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,VI221,Q123K,G127A,A132E,L136V,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A185E,S188D,S188N,S188R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在该实施方案中，变体内酯酶可以具有1,2,3,4,5或6个另外的氨基酸变体。在一些实施方案中，变体内酯酶包含G2P和G3P变体D34KD66EE80GI83VL136VA184NG189R和上文概述的另外的单一氨基酸变体中的至少一种，包括但不限于L17V,F32L,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,VI221,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A185E,S188D,S188N,S188R,R192M,I193V,El95A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在该实施方案中，变体内酯酶可以具有1,2,3,4,5或6个另外的氨基酸变体。在一些实施方案中，变体内酯酶包含G2P和G3P变体D34KD66EE80GV83IL136VA184NG189R和上文概述的另外的单一氨基酸变体中的至少一种，包括但不限于L17V,F32L,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A185E,S188D,S188N,S188R,R192M,I193V,El95A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。在实施方案中，变体内酯酶可具有1,2,3,4,5或6个另外的氨基酸变体。在一些实施方案中，变体内酯酶包含G2P和G3P变体D34KD66EE80GV83IL136VA184NG189R，并且在其他位置处可以具有另外的氨基酸取代。在该实施方案中，变体内酯酶可以具有1,2,3,4,5或6个另外的氨基酸变体；此外，在该实施方案中，变体内酯酶在如本文所概述的内酯酶测定法中保留可检测的活性。在一些实施方案中，变体内酯酶包含G2P和G3P变体D34KD66EE80GI83VL136VA184NG189R，并且在其他位置处可以具有另外的氨基酸取代。在该实施方案中，变体内酯酶可以具有1,2,3,4,5或6个另外的氨基酸变体；此外，在实施方案中，变体内酯酶在如本文所述的内酯酶测定法中保留可检测的活性。本发明的一些具体实施方案是与SEQIDNO:1相比的内酯酶变体，其具有选自以下的氨基酸取代组:D66EQ114H,G35SV41F,G35S,G35SE195A,G189RE195A,L17VG35S,L17VG35SI77VL136VG189R,L17VG35SD66E,L17VD66EL136V,A132E,G189R,L17VD66E,G35SQ114H,Q114HG189R,D66EV83I,D66EE80GV83IL136V,G35SD66EV83IA222T,D66EI77VE80GV83IQ114HA132EG189R,G120S,A69I,A69V,D34E,N74H,G120T,G120H,A69C,D34A,G127A,A69K,A69Q,G120R,D34G,E218T,A69T,D34Q,G120P,A184N,G120A,E218M,E218N,D34K,D34S,E218A,V76S,E244H,S162N,S162C,E244C,S188D,S188R,V76K,A50P,H139T,V76R,H139K,Q123KR164K,E244K,S162H,E244S,Q70S,E244T,E244R,Q70N,Q70Y,V76E,V76L,V76A,E244G,S188N,V76N,L81P,L81PI84V,T62AG79AR164KI193V,G79AL81P,G79AI84VV122I,T62AL81PV12211193V,T62AG79AL81P,T62AG79A,L81PA185E,G79AL81PI84VA155D,L68VL81PI84VA155DI193V,L81PI84VI193V,F32LL81PI84VI193V,L68VL81PI84VR164K,G79A,G79AV122IR164K,G79AI84V,VI2211193VQ221H,L81PV122I,T62AG79AL81PI193VQ221H,T62AL68VV122IR164K,T62AL81PI84V,V122I,G79AV122I,G79AI193V,D66EE80GV83IL136VD34KA184NG189R,D66EE80GV83IL136VD34KA184NG189RE218T,D66EE80GV83IL136VD34K,D66EE80GV83IL136VD34KA184N,D66EE80GL136VD34K,D66EE80GV83IL136VD34KA184NE218T,D66EE80GL136V,D66EE80GL136VD34KA184NG189RE218T,D66EE80GL136VD34KG189R,D66EE80GV83IL136VRl92M,D66EE80GL136VD34KG189RE218T,D66EE80GL136VD34KA184NG189R,D66EE80GV83IL136VA184NG189R和D66EE80GL136VD34KE218T。VII.本发明的核酸本发明另外提供了编码本发明的变体内酯酶的核酸。如本领域技术人员所理解的，由于遗传密码的简并性，可以制备极大量的核酸，所有核酸都编码本发明的变体内酯酶。因此，鉴定出特定的氨基酸序列后，本领域技术人员可以通过简单地以不改变蛋白质氨基酸序列的方式修饰一个或多个密码子的序列来制备任何数量的不同核酸。因此，提供氨基酸序列允许产生非常大量的编码蛋白质的不同核酸序列。在一些实施方案中，特定变体内酯酶由特定核酸序列编码，如序列表中所列。如本领域所知，编码本发明变体内酯酶的核酸可以掺入表达载体中，如本领域已知的并且取决于用于产生本发明内酯酶的宿主细胞。通常，核酸与任何数量的调节元件启动子、复制起点、选择标志物、核糖体结合位点、诱导物等可操作连接。表达载体可以是染色体外或整合载体。然后，将本发明的核酸和或表达载体转化到本领域公知的任何数量的不同类型的宿主细胞中，包括哺乳动物、细菌、酵母、昆虫和或真菌细胞，细菌和真菌在许多实施方案中得到应用。A.变体的制备编码本发明的变体内酯酶的核酸可以使用本领域中已知的任何诱变程序制备，例如定点诱变和合成基因构建，如本领域技术人员公知的。合成基因构建需要体外合成设计的多核苷酸分子以编码感兴趣的多肽。可以利用许多技术进行基因合成，例如Tianetal.2004,Nature432:1050-1054描述的基于多重微芯片的技术和类似技术，其中将寡核苷酸合成并且在光可编程微流体芯片上组装。优选的技术是i调节序列本发明还涉及核酸构建体，其包含与一个或多个控制序列可操作连接的编码本发明变体的多核苷酸，所述控制序列与与控制序列相容的条件下指导编码序列在合适的宿主细胞中的表达。可以以多种方式操作多核苷酸以提供变体的表达。根据表达载体，在将多核苷酸插入载体之前对其进行操作可能是期望的或必需的。利用重组DNA方法修饰多核苷酸的技术在本领域是公知的。控制序列可以是启动子，即由宿主细胞识别以表达多核苷酸的多核苷酸。启动子含有介导变体表达的转录控制序列。启动子可以是在宿主细胞中显示转录活性的任何多核苷酸，包括突变体、截短的启动子和杂合启动子，并且可以从编码与宿主细胞同源或异源的胞外或胞内多肽的基因获得。用于指导本发明的核酸构建体在丝状真菌宿主细胞中转录的合适启动子的实例是从构巢曲霉Aspergillusnidulans乙酰胺酶、黑曲霉Aspergillusniger中性α-淀粉酶、黑曲霉酸稳定性α-淀粉酶、黑曲霉ox泡盛曲霉Aspergillusawamori内酯酶glaA、米曲霉AspergillusoryzaeTAKA淀粉酶、米曲霉碱性蛋白酶、米曲霉丙糖磷酸异构酶、尖孢镰孢Fusariumoxysporum胰蛋白酶样蛋白酶WO9600787、Fusariumvenenatum淀粉葡糖苷酶WO0056900、FusariumvenenatumDariaWO0056900、FusariumvenenatumQuinnWO0056900、米赫根毛霉Rhizomucormiehei脂肪酶、米赫根毛霉天冬氨酸蛋白酶、里氏木霉Trichodermareeseiβ-葡萄糖苷酶、里氏木霉纤维二糖水解酶I、里氏木霉纤维二糖水解酶II、里氏木霉内切葡聚糖酶I、里氏木霉内切葡聚糖酶II、里氏木霉内切葡聚糖酶III、里氏木霉内切葡聚糖酶IV、里氏木霉内切葡聚糖酶V、里氏木霉木聚糖酶I、里氏木霉木聚糖酶II、里氏木霉β-木糖苷酶的基因获得的启动子，以及NA2-tpi启动子来自曲霉中性α-淀粉酶基因的修饰启动子，其中非翻译前导序列已经被来自曲霉磷酸丙糖异构酶基因的非翻译前导序列取代；非限制性实例包括来自黑曲霉中性α-淀粉酶基因的修饰启动子，其中非翻译前导序列已被来自构巢曲霉或米曲霉磷酸丙糖异构酶基因的非翻译前导序列取代；和其突变体、截短的启动子和杂合启动子。在酵母宿主中，从酿酒酵母烯醇化酶ENO-1、酿酒酵母半乳糖激酶GAL1、酿酒酵母醇脱氢酶甘油醛-3-磷酸脱氢酶ADH1，ADH2GAP、酿酒酵母丙糖磷酸异构酶TPI、酿酒酵母金属硫蛋白CUP1和酿酒酵母3-磷酸甘油酸激酶的基因获得有用的启动子。Romanoseta,1992,Yeast8:423-488描述了酵母宿主细胞的其他有用启动子。控制序列也可以是转录终止子，其被宿主细胞识别以终止转录。终止子序列与编码变体的多核苷酸的3’末端可操作地连接。可以使用在宿主细胞中有功能的任何终止子。在一些实施方案中，丝状真菌宿主细胞的终止子获自构巢曲霉邻氨基苯甲酸合酶、黑曲霉内酯酶，黑曲霉α-葡糖苷酶，米曲霉TAKA淀粉酶和尖孢镰孢胰蛋白酶样蛋白酶的基因。在一些实施方案中，酵母宿主细胞的终止子获自酿酒酵母烯醇化酶、酿酒酵母细胞色素CCYC1和酿酒酵母甘油醛-3-磷酸脱氢酶的基因。Romanosetal.,1992，同上描述了酵母宿主细胞的其他有用的终止子。控制序列还可以是启动子下游和基因编码序列上游的mRNA稳定子区域，其增加基因的表达。合适的mRNA稳定子区域的实例获自苏云金芽孢杆菌crylllA基因WO9425612和枯草芽孢杆菌SP82基因Hueetal,1995,JournalofBacteriology111:3465-3471。控制序列也可以是前导序列，对宿主细胞的翻译重要的mRNA的非翻译区。前导序列与编码变体的多核苷酸的5’末端可操作地连接。可以使用在宿主细胞中有功能的任何前导序列。在一些实施方案中，丝状真菌宿主细胞的前导序列获自米曲霉TAKA淀粉酶和构巢曲霉丙糖磷酸异构酶的基因。在一些实施方案中，酵母宿主细胞的合适前导序列获自酿酒酵母烯醇化酶ENO-1、酿酒酵母3-磷酸甘油酸激酶、酿酒酵母α-因子和酿酒酵母醇脱氢酶甘油醛-3-磷酸脱氢酶ADH2GAP的基因。控制序列还可以是多腺苷酸化序列，其是与变体编码序列的3’末端可操作连接的序列，并且当转录时被宿主细胞识别为将聚腺苷残基添加至转录的mRNA的信号。可以使用在宿主细胞中有功能的任何聚腺苷酸化序列。在一些实施方案中，丝状真菌宿主细胞的聚腺苷酸化序列获自构巢曲霉邻氨基苯甲酸合酶、黑曲霉内酯酶、黑曲霉α-葡糖苷酶、米曲霉TAKA淀粉酶和尖孢镰孢胰蛋白酶样蛋白酶的基因。GuoandSherman,1995,Mol.CellularBiol.15:5983-5990描述了用于酵母宿主细胞的有用的多腺苷酸化序列。控制序列也可以是信号肽编码区，其编码与变体的N端连接的信号肽，并指导变体葡糖淀粉酶表达到细胞分泌途径中。多核苷酸的编码序列的5’末端可以固有地含有信号肽编码序列，所述信号肽编码序列与编码变体葡糖淀粉酶的编码序列的片段以符合翻译读码框的方式天然连接。或者，编码序列的5’端可以含有对编码序列而言外源的信号肽编码序列。可以需要外源信号肽编码序列，其中编码序列不天然含有信号肽编码序列。或者，外源信号肽编码序列可以简单地替换天然信号肽编码序列，以增强变体葡糖淀粉酶的分泌。然而，可以使用任何信号肽编码序列，其指导表达的变体进入宿主细胞的分泌途径中。用于丝状真菌宿主细胞的有效信号肽编码序列是从黑曲霉中性淀粉酶、黑曲霉内酯酶、米曲霉TAKA淀粉酶、特异腐质霉Humicolainsolens纤维素酶、特异腐质霉内切葡聚糖酶V、柔毛腐质霉Humicolalanuginosa脂肪酶和米赫根毛霉天冬氨酸蛋白酶的基因获得的信号肽编码序列。还可能期望添加相对于宿主细胞生长调节变体的表达的调节序列。调节系统的实例是响应于化学或物理刺激包括调节化合物的存在而开启或关闭基因表达的那些。原核系统中的调节系统包括lac、tac和trp操纵子系统。在酵母中，可以使用ADH2系统或GAL1系统。在丝状真菌中，可以使用黑曲霉内酯酶启动子、米曲霉TAKAα-淀粉酶启动子和米曲霉内酯酶启动子。调节序列的其他实例是允许基因扩增的那些。在真核系统中，这些调节序列包括在甲氨蝶呤存在下扩增的二氢叶酸还原酶基因，并且用重金属扩增金属硫蛋白基因。在这些情况下，编码变体的多核苷酸将与调节序列可操作连接。1.表达载体本发明还涉及重组表达载体，其包含编码本发明变体的多核苷酸、启动子和转录和翻译终止信号。可以将各种核苷酸和控制序列连接在一起以产生重组表达载体，其可以包括一个或多个方便的限制性位点，以允许在此类位点处插入或取代编码变体的多核苷酸。或者，可以通过将多核苷酸或包含多核苷酸的核酸构建体插入适合于表达的载体中来表达多核苷酸。在创建表达载体时，编码序列位于载体中，使得编码序列与适合于表达的控制序列可操作连接。重组表达载体可以是任何载体例如质粒或病毒，其可以方便地进行重组DNA程序，并且可以实现多核苷酸的表达。载体的选择通常取决于载体与接受载体导入的宿主细胞的相容性。载体可以是线性或闭合的环状质粒。载体可以是自主复制载体，即，作为染色体外实体存在的载体，其复制不依赖于染色体复制，例如质粒、染色体外元件、小染色体、或人工染色体。载体可以含有用于确保自身复制的任何手段。或者，载体可以是当导入宿主细胞中时整合到基因组中并且与已经接受其整合的染色体一起复制的载体。此外，可以使用单个载体或质粒或一起含有待引入宿主细胞的基因组中的总DNA的两个或更多个载体或质粒，或者转座子。预期用于本发明方法的载体包括整合和非整合型载体两者。在一些实施方案中，载体含有一个或多个选择标志物，其允许容易地选择经转化的、经转染的、经转导的细胞或类似细胞。选择标志物是下述的基因，其产物提供杀生物剂或病毒抗性、对重金属的抗性、营养缺陷型的原养型等等。用于酵母宿主细胞的合适标志物包括但不限于ADE2、HIS3、LEU2、LYS2、MET3、TRP1和URA3。用于丝状真菌宿主细胞的选择标志物包括但不限于amdS乙酰胺酶、argB鸟氨酸氨基甲酰转移酶、bar膦丝菌素乙酰基转移酶、hph潮霉素磷酸转移酶、niaD硝酸盐还原酶、pyrG乳清苷-5’-磷酸脱羧酶、sC硫酸盐腺苷酸转移酶和trpC邻氨基苯甲酸合成酶，以及它们的等价物。优选用于曲霉属细胞的是构巢曲霉或米曲霉amdS和pyrG基因以及吸水链霉菌Streptomyceshygroscopicusbar基因。在一些实施方案中，载体含有一种或多种元件，其允许将载体整合到宿主细胞的基因组中，或者允许载体不依赖于基因组在细胞中自主复制。为了整合到宿主细胞基因组中，载体可以依赖于编码变体的多核苷酸序列或载体的任何其他元件，以通过同源或非同源重组整合到基因组中。或者，载体可以含有额外的多核苷酸，用于在染色体中的精确位置处指导通过同源重组整合到宿主细胞的基因组中。为了增加在精确位置处整合的可能性，整合元件应当包含足够数量的核酸，例如100至10,000个碱基对，400至10,000个碱基对，以及800至10,000个碱基对，其与相应的靶序列具有高度的序列同一性程度以增强同源重组的概率。整合元件可以是与宿主细胞的基因组中的靶序列同源的任何序列。此外，整合元件可以是非编码或编码多核苷酸。另一方面，可以通过非同源重组将载体整合到宿主细胞的基因组中。对于自主复制，载体可以进一步包含复制起点，使载体能够在所讨论的宿主细胞中自主复制。复制起点可以是介导在细胞中发挥功能的自主复制的任何质粒复制子。术语“复制起点”或“质粒复制子”指使质粒或载体能够在体内复制的多核苷酸。用于酵母宿主细胞的复制起点的实例是2微米复制起点、ARS1、ARS4、ARS1和CEN3的组合、以及ARS4和CEN6的组合。可用于丝状真菌细胞的复制起点的实例是AMA1和ANSIGemsetal.,1991,Gene98:61-67；Cullenetal.,1987,NucleicAcidsRes.15:9163-9175；WO0024883。可以根据WO0024883中公开的方法完成AMA1基因的分离和包含该基因的质粒或载体的构建。可以将本发明的多核苷酸的超过一个拷贝插入宿主细胞中以增加变体的产生。通过将序列的至少一个另外的拷贝整合到宿主细胞基因组中或通过与多核苷酸一起包含可扩增的选择标志物基因其中含有选择标志物基因的扩增拷贝以及因此含有多核苷酸的额外拷贝的细胞可以通过在适当的选择剂存在下培养细胞来选择来获得多核苷酸的拷贝数的增加。用于连接上述元件以构建本发明的重组表达载体的程序对于本领域技术人员是公知的参见例如Sambrooketal,1989，同上。2.密码子优化可以对本发明的任何变体内酯酶多肽进行密码子优化，以优化所用的宿主细胞中的表达。此类方法在本领域中是公知的，并且描述于例如WO2007142954中。在异源表达系统中，优化步骤可以改善宿主产生期望的变体内酯酶多肽的能力。蛋白质表达受许多因素控制，包括那些影响转录、mRNA加工、稳定性和翻译起始的。多核苷酸优化步骤可以包括改善宿主产生外源蛋白质的能力的步骤以及帮助研究人员有效设计表达构建体的步骤。优化策略可以包括例如翻译起始区的修饰、mRNA结构元件的改变、以及不同密码子偏倚的使用。以下段落讨论了可能导致异源蛋白质表达减少的潜在问题，并且可以克服这些问题的技术。在一些实施方案中，减少的异源蛋白质表达源自罕见的密码子诱导的翻译暂停。罕见的密码子诱导的翻译暂停包括感兴趣的多核苷酸中存在宿主生物体中很少使用的密码子，由于它们在可用的tRNA库中的稀缺性而对蛋白质翻译具有负面影响。一种改善宿主生物体中的最佳翻译的方法包括进行包括进行密码子优化，这可以导致在合成的多核苷酸序列中修饰罕见的宿主密码子。在一些实施方案中，通过交替的翻译起始产生减少的异源蛋白质表达。交替的翻译起始可以包括无意中含有能够起核糖体结合位点RBS功能的基序的合成多核苷酸序列。这些位点可以导致从基因内部位点开始翻译截短的蛋白质。降低产生截短蛋白其在纯化过程中可能难以除去的可能性的一种方法包括从优化的多核苷酸序列修饰推定的内部RBS序列。在一些实施方案中，通过重复诱导的聚合酶滑动发生减少的异源蛋白质表达。重复诱导的聚合酶滑动涉及核苷酸序列重复，其已经显示导致DNA聚合酶的滑动或打滑，这可导致移码突变。此类重复也可导致RNA聚合酶的滑动。在具有高G+C含量偏倚的生物体中，可以有更高程度的由G或C核苷酸重复构成的重复。因此，一种降低诱导RNA聚合酶滑动的可能性的方法包括改变G或C核苷酸的扩充重复。在一些实施方案中，通过干扰二级结构发生减少的异源蛋白质表达。二级结构可以隔离RBS序列或起始密码子，并且与蛋白质表达的降低相关。茎环结构也可以参与转录暂停和减弱。优化的多核苷酸序列可以在核苷酸序列的RBS和基因编码区中含有最小的二级结构，以允许改善的转录和翻译。在一些实施方案中，限制性位点可以影响异源蛋白质表达。通过修饰可以干扰转录单元随后亚克隆到宿主表达载体中的限制性位点，可以优化多核苷酸序列。优化DNA序列可以负面或正面影响基因表达或蛋白质产生。例如，用更常见的密码子修饰不太常见的密码子可以影响mRNA的半衰期或通过引入干扰信使翻译的二级结构改变其结构。在某些情况下，可能有必要改变优化的信使。可以优化AUG或基因的一部分。在一些实施方案中，通过优化基本上整个基因来实现期望的表达调节。在其他实施方案中，通过优化基因的部分但不是全部来实现期望的调节。可以调整任何编码序列的密码子选择以实现期望的性质，例如特定细胞类型中的高水平表达。此类优化的起点可以是具有100％共同密码子的编码序列，或含有共同和非共同密码子的混合物的编码序列。可以产生在其密码子选择上不同的两个或更多个候选序列并且测试以确定它们是否拥有期望的特性。可以通过使用计算机来搜索调节元件例如沉默子或增强子的存在，并且搜索可以通过密码子选择变化转化成此类调节元件的编码序列区域的存在来评估候选序列。另外的标准可以包括富集特定核苷酸，例如A，C，G或U，特定氨基酸的密码子偏倚，或特定mRNA二级或三级结构的存在或缺乏。可以基于许多此类标准来对候选序列进行调节。构建有希望的候选序列并通过实验进行评估。可以彼此独立地评估多个候选物，或者过程可以是迭代的，通过使用最有希望的候选物作为新的起始点，或者通过组合两个或更多个候选物的区域以产生新的混合物进行。可以包括进一步的修饰和评估。修饰候选序列的密码子选择可以导致正或负元件的创建或破坏。通常，正元件指其从候选序列的改变或除去可以导致治疗性蛋白质的表达降低，或其创建可以导致治疗性蛋白质的表达增加的任何元件。例如，正元件可以包括增强子、启动子、下游启动子元件、正调节子例如转录激活子的DNA结合位点、或负责赋予或修饰mRNA二级或三级结构的序列。负元件指其从候选序列的改变或除去可以导致治疗性蛋白质的表达增加，或其创建将导致治疗性蛋白质的表达降低的任何元件。负元件包括沉默子、负调节剂例如转录阻遏物的DNA结合位点、转录暂停位点、或负责赋予或修饰mRNA二级或三级结构的序列。通常，负元件比正元件更频繁产生。因此，导致蛋白质表达增加的密码子选择的任何变化更可能源自负元件的破坏而非正元件的创建。此外，候选序列的改变更可能破坏正元件而非创建正元件。在一些实施方案中，选择和修饰候选序列以增加治疗性蛋白质的产生。可以修饰候选序列，例如通过顺序改变密码子或通过随机改变候选序列中的密码子进行。然后，通过测定所得治疗性蛋白质的表达水平或通过评估另一项参数例如，与表达水平相关的参数来评估修饰的候选序列。选择与未改变的候选序列相比产生增加水平的治疗性蛋白质的候选序列。在一些实施方案中，可以修饰一个或一组密码子，例如在不参考蛋白质或信使结构的情况下，并且进行测试。或者，可以在信使水平特性上选择一个或多个密码子，例如，预定区域中的位置，例如高或低GC含量，具有诸如增强子或沉默子等结构的区域中的位置，可以经修饰而引入诸如增强子或沉默子等结构的区域中的位置，具有或预测具有二级或三级结构例如链内配对，链间配对的区域中的位置，缺乏或预测缺乏二级或三级结构例如链内或链间配对的区域中的位置。若特定的修饰的区域产生期望的结果，则选择该特定的修饰的区域。系统性产生候选序列的方法是有用的。例如，在合成核酸序列的各个位置处的一个或一组例如，连续的密码子块可以用常见的密码子或用非常见的密码子，在例如起始序列已被优化时修饰，并且对所得的序列进行评估。可以通过如下来生成候选物：优化或去优化序列中的密码子的给定“窗口”以生成第一候选物，并且将窗口移动到序列中的新位置，并优化或去优化窗口下的新位置中的密码子以提供第二候选物。候选物可以通过确定它们提供的表达水平来评估，或者通过评估另一个参数，例如与表达水平相关的参数来评估。一些参数可以通过检查或计算来评估，例如，拥有或缺乏高或低GC含量；序列元件，如增强子或沉默子；二级或三级结构，例如链内或链间配对。在一些实施方案中，优化的核酸序列可以以由尚未优化的核酸序列表达的水平的至少110％，150％，200％，500％，1,000％，5,000％或甚至10,000％的水平表达本发明的变体内酯酶多肽。以变体内酯酶的氨基酸序列开始，可以设计候选DNA序列。在合成DNA序列的设计过程中，可以将密码子选择的频率与宿主表达生物体的密码子选择进行比较，并且可以在合成序列中修饰罕见的宿主密码子。另外，可以修饰合成候选DNA序列以除去不期望的酶限制性位点并添加或改变任何期望的信号序列、接头或非翻译区。可以对合成的DNA序列分析可能干扰翻译过程的二级结构的存在，例如GC重复和茎环结构。在合成候选DNA序列之前，可以检查优化的序列设计以验证序列正确编码期望的氨基酸序列。最后，可以使用DNA合成技术，例如本领域已知的那些技术合成候选DNA序列。在一些实施方案中，可以利用宿主生物体中的一般密码子选择，例如本文所述的那些密码子选择中的任一种，来优化宿主生物体中的异源多核苷酸序列的表达。可以评估很少会被认为对于宿主表达系统中的特定氨基酸优选的密码子的百分比和分布。使用5％和10％选择的值作为罕见密码子测定的截留值。VIII.宿主细胞和生产菌株如本领域技术人员所理解的，存在多种用于重组产生本发明的变体内酯酶的生产宿主生物，包括但不限于细菌细胞和包括酵母的真菌细胞。此外，虽然G1P亲本内酯酶是未糖基化的，但是在酵母和真菌中通过产生的糖基化不会对内酯酶活性产生不利影响。本发明还涉及重组宿主细胞，其包含与一个或多个指导本发明的变体产生的控制序列可操作连接的编码本发明的变体葡糖淀粉酶的多核苷酸。将包含多核苷酸的构建体或载体导入宿主细胞中，使得将构建体或载体维持为染色体整合体或如前所述的自我复制的染色体外载体。由于在复制期间发生的突变，术语“宿主细胞”包括与亲本细胞不同的亲本细胞的任何后代。宿主细胞的选择在很大程度上取决于编码变体的基因及其来源。在一些实施方案中，宿主细胞表现出变体葡糖淀粉酶的瞬时表达。在一些实施方案中，宿主细胞是稳定转染的宿主或宿主细胞，其稳定地即，永久地表达变体内酯酶。在一些实施方案中，宿主细胞是生产宿主细胞。宿主细胞可以是可用于重组产生变体的任何细胞，例如原核生物或真核生物。此类宿主细胞包括但不限于细菌、真菌和酵母细胞。宿主细胞也可以是真核生物，例如植物，酵母或真菌细胞。宿主细胞可以是真菌细胞。如本文所用，“真菌”包括门子囊菌纲Ascomycota，担子菌纲Basidiomycota，壶菌亚门Chytridiomycota和接合菌亚纲Zygomycota以及卵菌门Oomycota和所有有丝分裂真菌mitosporicfungi如Hawksworthetal,于AinsworthandBisby'sDictionaryofTheFungi,第8版,1995,CABInternational,UniversityPress,Cambridge,UK定义。真菌宿主细胞可以是酵母细胞。如本文所用的，“酵母”包括产子囊酵母ascosporogenousyeast内孢霉目Endomycetales、产担子酵母basidiosporogenousyeast和属于半知真菌FungiImperfecti芽生菌目Blastomycetes的酵母。由于酵母的分类在将来可能会改变，为了本发明的目的，酵母应当如BiologyandActivitiesofYeastSkinner,Passmore,andDavenport编,Soc.App.Bacteriol.SymposiumSeries.9,1980所述定义。酵母宿主细胞可以是假丝酵母属Candida，汉逊酵母属Hansenula，克鲁维酵母属Kluyveromyces，毕赤酵母属Pichia，酵母属Saccharomyces，裂殖酵母属Schizosaccharomyces或耶氏酵母属Yarrowia细胞，例如乳酸克鲁维酵母Kluyveromyceslactis，卡尔酵母Saccharomycescarlsbergensis，酿酒酵母Saccharomycescerevisiae，糖化酵母Saccharomycesdiastaticus，道格拉氏酵母Saccharomycesdouglasii，克鲁氏酵母Saccharomyceskluyveri，诺氏酵母Saccharomycesnorbensis，卵形酵母Saccharomycesoviformis或解脂耶氏酵母Yarrowialipolytica细胞。真菌宿主细胞可以是丝状真菌细胞。“丝状真菌”包括分部真菌门Eumycota和卵菌门的所有丝状形式如Hawksworth等，1995，同上所定义。丝状真菌的特征通常在于由几丁质，纤维素，葡聚糖，壳聚糖，甘露聚糖和其他复合多糖构成的菌丝体壁。营养生长通过菌丝延伸进行，并且碳分解代谢是专性有氧的。相反，酵母例如酿酒酵母的营养生长通过单细胞原植体的出芽进行，并且碳分解代谢可以是发酵的。丝状真菌宿主细胞可以是支顶孢属Acremonium，曲霉属Aspergillus，短梗霉属Aureobasidium，烟管菌属Bjerkandera，拟蜡菌属Ceriporiopsis，金小孢子属Chrysosporium，鬼伞属Coprinus，云芝属Coriolus，隐球菌属Cryptococcus，Filibasidium，镰孢属Fusarium，腐质霉属Humicola，Magnaporthe，毛霉属Mucor，毁丝霉属Myceliophthora，新丽鞭毛菌属Neocallimastix，链孢霉属Neurospora，拟青霉属Paecilomyces，青霉属Penicillium，毛平革菌属Phanerochaete，射脉菌属Phlebia，单鞭毛菌属Piromyces，侧耳属Pleurotus，裂折菌属Schizophyllum，踝节菌属Talaromyces，热子囊菌属Thermoascus，梭孢壳属Thielavia，弯颈霉属Tolypocladium，槐耳属Trametes，或木霉属Trichoderma细胞。例如，丝状真菌宿主细胞可以是泡盛曲霉Aspergillusawamori，臭曲霉Aspergillusfoetidus，烟曲霉Aspergillusfumigatus，日本曲霉Aspergillusjaponicus，构巢曲霉Aspergillusnidulans，黑曲霉Aspergillusniger，米曲霉Aspergillusoryzae，黑刺烟管菌Bjerkanderaadusta，干拟蜡菌Ceriporiopsisaneirina，Ceriporiopsiscaregiea，Ceriporiopsisgilvescens，Ceriporiopsispannocinta，Ceriporiopsisrivulosa，Ceriporiopsissubrufa，虫拟蜡菌Ceriporiopsissubvermispora，Chrysosporiuminops，嗜角质金孢子菌Chrysosporiumkeratinophilum，Chrysosporiumlucknowense，Chrysosporiummerdarium，毡金孢子菌Chrysosporiumpannicola，Chrysosporiumqueenslandicum，热带金孢子菌Chrysosporiumtropicum，Chrysosporiumzonatum，灰盖鬼伞Coprinuscinereus，毛革盖菌Coriolushirsutus，杆孢状镰孢Fusariumbactridioides，禾谷镰孢Fusariumcerealis，克鲁克威尔镰孢Fusariumcrookwellense，大刀镰孢Fusariumculmorum，禾谷镰孢Fusariumgraminearum，禾赤镰孢Fusariumgraminum，异孢镰孢Fusariumheterosporum，合欢木镰孢Fusariumnegundi、尖孢镰孢Fusariumoxysporum、多枝镰孢Fusariumreticulatum、粉红镰孢Fusariumroseum、接骨木镰孢Fusariumsambucinum、肤色镰孢Fusariumsarcochroum、拟分枝孢镰孢Fusariumsporotrichioides、硫色镰孢Fusariumsulphureum、圆镰孢Fusariumtorulosum、拟丝孢镰孢Fusariumtrichothecioides、Fusariumvenenatum，特异腐质霉Humicolainsolens，疏棉状腐质霉Humicolalanuginosa，米黑毛霉Mucormiehei，嗜热毁丝霉Myceliophthorathermophila，粗糙脉孢菌Neurosporacrassa，产紫青霉Penicilliumpurpurogenum，黄孢平革菌Phanerochaetechrysosporium，辐射射脉菌Phlebiaradiata，刺芹侧耳Pleurotuseryngii，土生梭孢霉Thielaviaterrestris，长绒毛栓菌Trametesvillosa，变色栓菌Trametesversicolor，哈茨木霉Trichodermaharzianum，康宁木霉Trichodermakoningii，长枝木霉Trichodermalongibrachiatum，里氏木霉Trichodermareesei或绿色木霉Trichodermaviride。真菌细胞可以通过涉及原生质体形成，原生质体转化和细胞壁再生的方法以本身已知的方式转化。用于转化曲霉属和木霉属宿主细胞的合适方法描述于EP238023,Yeltonetal,1984,Proc.Natl.Acad.Sci.USA81:1470-1474和Christensenetal,1988,BiolTechnology6:1419-1422。适合于转化镰孢物种的方法由Malardieretal.,1989,Gene78:147-156和WO9600787'描述。酵母可以使用由BeckerandGuarente,InAbelson,J.N.andSimon,M.I.编,GuidetoYeastGeneticsandMolecularBiology,MethodsinEnzymology,第194卷,第182-187页,AcademicPress,Inc.,NewYork；Itoetal,1983,J.Bacteriol.153:163；和Hinnenetal,1978,Proc.Natl.Acad.Sci.USA75:1920描述的程序转化。IX.组合物本发明还提供了包含变体内酯酶的组合物。在一些实施方案中，组合物包含载体和或赋形剂。在一些实施方案中，组合物富含本发明的变体内酯酶多肽。术语“富含”表示组合物的内酯酶活性已经增加，例如，富集因子至少为1。在一些实施方案中，配制组合物以提供期望的特征，例如低颜色，低气味和可接受的贮存稳定性。在一些实施方案中，组合物包含本发明的变体内酯酶多肽作为主要酶组分，例如单组分组合物。此组合物可用作鱼饲料原料的添加剂。X.生产方法本发明还涉及产生变体内酯酶多肽的方法，包括：a在适于表达变体内酯酶多肽的条件下培养本发明的宿主细胞；并且b任选地回收变体内酯酶多肽。使用本领域已知的方法在适于产生变体内酯酶多肽的营养培养基中培养宿主细胞。例如，可以通过摇瓶培养，或在合适的培养基和允许表达和或分离变体的条件下进行的实验室或工业发酵罐中的小规模或大规模发酵包括连续、分批、补料-分批或固态发酵培养细胞。使用本领域已知的程序，在包含碳源和氮源和无机盐的合适营养培养基中进行培养。合适的培养基可从商业供应商获得，或者可根据公开的组合物例如，在美国典型培养物保藏中心的目录中制备。若变体内酯酶多肽分泌到营养培养基中，则可以直接从培养基中回收变体内酯酶多肽。若不分泌变体，则可以从细胞裂解物中回收。可以使用对变体特异性的本领域已知的方法检测变体内酯酶多肽。这些检测方法包括但不限于使用特异性抗体，酶产物的形成或酶底物的消失。例如，酶测定法可用于测定变体内酯酶多肽的活性。可以使用本领域已知的方法回收变体内酯酶多肽。例如，可通过常规程序从营养培养基中回收变体内酯酶多肽，包括但不限于收集、离心、过滤、提取、喷雾干燥、蒸发或沉淀。可以通过本领域中已知的多种程序纯化变体，包括但不限于层析例如，离子交换，亲和，疏水，色谱聚焦和尺寸排阻，电泳程序例如，制备等电聚焦，差异溶解度例如，硫酸铵沉淀，SDS-PAGE或提取参见例如ProteinPurification,JansonandRyden编,VCHPublishers,NewYork,1989以获得基本上纯的变体。在另一个方面，不回收变体，而是使用表达变体的本发明的宿主细胞作为变体的来源。在一个具体实施方案中，不回收本发明的变体内酯酶，并且宿主细胞是酵母宿主细胞。特别地，酵母是假丝酵母属、汉逊酵母属、克鲁维酵母属、毕赤酵母属、酵母属，裂殖酵母属或耶氏酵母属细胞，例如乳酸克鲁维酵母、卡尔酵母、酿酒酵母，糖化酵母，道格拉氏酵母，克鲁氏酵母，诺氏酵母，卵形酵母或解脂耶氏酵母细胞。在一些实施方案中，酵母是酿酒酵母。XI.内酯酶制剂和用途如本文所讨论的，在动物饲料中使用内酯酶具有许多益处，包括抗菌性质，以及对肠生物群系的改善，导致一般的健康益处。在一些实施方案中，将本发明的变体内酯酶配制并添加到饲料或可以作为饲料的组分制备。在前一种情况下，可以通过在载体饲料如小麦粉上配制内酯酶来完成内酯酶的饲料原料添加。如本领域技术人员所理解的，本发明的变体内酯酶的制剂取决于其最终用途和相关条件。用于本发明的变体内酯酶的合适制剂包括液体制剂，干燥制剂包括喷雾干燥制剂，粉末制剂，颗粒制剂和造粒制剂。在一些实施方案中，本发明的酶组合物即，多肽组合物可以是适合使用的任何形式，诸如例如除去或不除去细胞的粗制发酵液，具有或不具有细胞碎片的细胞裂解物，半纯化或纯化的酶组合物，或宿主细胞，作为酶来源。在一些实施方案中，酶组合物可以是干粉或颗粒，无尘颗粒，液体，稳定液体或稳定的受保护酶。例如，液体酶组合物可以通过根据已建立的方法添加稳定剂如糖，糖醇或另一种多元醇和或乳酸或另一种有机酸来稳定。在一些实施方案中，本发明的多肽组合物的剂量和使用该组合物的其他条件可以基于本领域已知的方法确定。在一个实施方案中，将乳酸酶添加到动物饲料原料中并如本领域已知的那样造粒，使得形成其中含有内酯酶的饲料。在其他实施方案中，可以将内酯酶以液体形式喷雾或给料到动物饲料中。XII.实施例A.实施例1：基因合成和克隆内酯酶的起始基因UniProKB登录号：C6L862由GenScript合成。将合成的基因克隆到pET-20b+载体NovagenEMDMillipore，USA：目录号69739中。B.实施例2：突变体收集设计和构建在第一代改善中，使用起始内酯酶基因作为亲本GIP。为了改善GIP的热稳定性和pH耐受性，基于GIP蛋白质序列设计了4个突变体集合。设计包括每个变体的1至多个特定突变。使用Lightning试剂盒AgilentTechnologies，SantaClara，California构建突变体集合，随后克隆到pET-20b+载体NovagenEMDMillipore，USA：目录号69739中。在第二代改善中，使用来自第一代的1个改善的变体作为第2代亲本G2P。为了进一步改善G2P的热稳定性和pH耐受性，基于第一代中鉴定的有利突变设计了1个突变体集合。设计包括每个变体的1个至多个特定突变。随后，使用Lightning试剂盒AgilentTechnologies，SantaClara，California构建突变体集合。C.实施例3：含有HTP内酯酶的湿细胞团粒的制备将来自单菌落的包含重组内酯酶编码基因的BL21DE3大肠杆菌细胞ThermoFisherScientific，USA：目录号C6000-03接种到96孔浅孔微量滴定板的各个孔中，所述孔包含含有1％葡萄糖和100μgmL氨苄青霉素的180μlLB。将培养物在30℃，200rpm和85％湿度下培养过夜。将来自每个孔的20μl过夜培养物转移到含有380mLTerrificBrothTB和100μgmL氨苄青霉素的96深孔板的相应孔中。将深孔板在37℃，250rpm和85％湿度下温育2.5-3.0小时OD6000.6-0.8。然后，通过到终浓度1mM的IPTG诱导细胞培养物，并在与最初使用的相同条件下温育过夜。然后，在4℃以4000rpm离心10分钟沉淀细胞。弃去上清液，并且将团粒在-80℃冷冻，然后裂解。D.实施例4：HTP内酯酶板的裂解如上所述，将150μLB-PER细菌蛋白质提取试剂ThermoFisherScientific，USA：目录号78248加入到每孔中的细胞糊中。将细胞在室温在台式振荡器上振荡裂解1.5小时。将板在4000rpm和4℃离心10分钟。将澄清的上清液用于进行生物化学测定法以确定活性、pH耐受性和热稳定性。E.实施例5：候选物的初始筛选：活性pH热稳定性概况1.热活性概况：对于G1P的热活性概况，使用0.05MTris-HCl,pH7.2将来自实施例4的裂解物稀释40倍。在96孔浅微量滴定板中，将25μl稀释的裂解物添加到在4％DMSO中制备的25μlN-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应，pH7.2中的终浓度8mM底物。将反应在30℃，50℃和60℃，200rpm温育30分钟。用75μl含有3.5MNaOH：2M羟胺盐酸盐::1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在图1中显示了结果。2.pH值概况：对于G1P的pH概况，将来自实施例4的裂解物在0.05M缓冲液pH3-8pH3：柠檬酸盐，pH4和5：乙酸钠，pH6：磷酸钠，pH7和8：Tris-HCl下稀释40倍。在96孔浅微量滴定板中，将25μl经稀释的裂解物添加到在4％DMSO中制备的25μlN-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19NO4，FW：241.3至其相应pH缓冲液的反应中的终浓度8mM底物。将反应在30℃，200rpm温育30分钟。用75μl含有3.5MNaOH：2M羟胺盐酸盐::1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在图2中显示了结果。3.热稳定性概况：使用0.05MTris-HCl，pH7.2将来自实施例4的裂解物稀释40倍。将70μl稀释的裂解物转移至PCR板并在30-60℃在热循环仪中加热10分钟。攻击后，将25μl稀释的裂解物添加25μl在4％DMSO中制备的N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃，200rpm温育30分钟。使用75μl含有3.5MNaOH：2M羟胺盐酸盐：1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在图3中显示了结果。F.实施例6：未处理的酶促测定法对于未处理的测定法，使用0.05MTris-HCl，pH7.2将来自实施例4的裂解物稀释40倍。在96孔浅微量滴定板中，将25μl稀释的裂解物添加25μl在4％DMSO中制备的N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃，200rpm温育30分钟。使用75μl含有3.5MNaOH：2M羟胺盐酸盐：1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在相同条件下将变体的酶活性与亲本进行比较以确定活性改善。在图4-5中显示了结果。G.实施例7：具有温度攻击的酶促测定法使用0.05MTris-HCl，pH7.2将来自实施例4的裂解物稀释40倍。将70μl经稀释的裂解物转移至PCR板，并且在55.5℃在热循环仪中加热10分钟以分别筛选第1代G1和第2代G2变体。攻击后，将25μl稀释的裂解物添加25μl在4％DMSO中制备的N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃，200rpm温育30分钟。使用75μl含有3.5MNaOH：2M羟胺盐酸盐：1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在相同条件下将变体的酶活性与亲本进行比较以确定活性改善。在图4-5中显示了结果。H.实施例8：具有pH攻击的酶促测定法对于pH攻击测定法，使用0.1M乙酸钠，pH4.8将来自实施例4的裂解物稀释40倍。在96孔浅微量滴定板中，将25μl稀释的裂解物添加25μl在4％DMSO中制备的N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃，200rpm温育30分钟。使用75μl含有3.5MNaOH：2M羟胺盐酸盐：1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。在相同条件下将变体的酶活性与亲本进行比较以确定活性改善。在图4-5中显示了结果。I.实施例9：验证来自两轮酶改善的最佳变体的热稳定性概况使用0.05MTris-HCl，pH7.2将来自实施例4的裂解物稀释40倍。将70μl经稀释的裂解物转移至PCR板，并在30-70℃在热循环仪中加热10分钟。攻击后，将25μl稀释的裂解物添加25μl在4％DMSO中制备的N-3-氧代辛酰基-L-高丝氨酸内酯底物C12H19N04，FW：241.3至反应中的终浓度8mM底物，pH7.2。将反应在30℃，200rpm温育30分钟。使用75μl含有3.5MNaOH：2M羟胺盐酸盐：1:1的淬灭剂淬灭反应。淬灭后，添加75μl着色试剂。通过混合添加等体积的4MHCl中的10％FeCl3至95％EtOH来新鲜制备着色试剂。将板摇动1-2分钟后，将它们以4000rpm离心2分钟。然后在530nm处读取板吸光度。通过将活性与在室温放置10分钟的酶进行比较来进行残留活性计算。图6中显示了结果。序列表福尼亚生物处理股份有限公司Banerjee,GoutamiYang,JieOo,KhinZhang,XiyunHu,EricLinFukushima,Tatsuya内酯酶及其使用方法114095-5005-WOUS15668,2312017-08-03220PatentInversion3.51251PRT人工序列合成内酯酶CL00023198G1P1MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg2452502753DNA人工序列合成内酯酶CL00023198G1P2atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc7533251PRT人工序列合成内酯酶CL000254083MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuHisLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg2452504753DNA人工序列合成内酯酶CL000254084atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccacctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc7535251PRT人工序列合成内酯酶CL000254175MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgPheAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg2452506753DNA人工序列合成内酯酶CL000254176atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtacgg120ttcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc7537251PRT人工序列合成内酯酶CL000254817MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg2452508753DNA人工序列合成内酯酶CL000254818atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc7539251PRT人工序列合成内酯酶CL000255009MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProAlaAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525010753DNA人工序列合成内酯酶CL0002550010atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggccgacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75311251PRT人工序列合成内酯酶CL0002553511MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProAlaAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525012753DNA人工序列合成内酯酶CL0002553512atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggccgacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75313251PRT人工序列合成内酯酶CL0002555013MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015ValHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525014753DNA人工序列合成内酯酶CL0002555014atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcgtccacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75315251PRT人工序列合成内酯酶CL0002556715MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015ValHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValValProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525016753DNA人工序列合成内酯酶CL0002556716atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcgtccacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtggtccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75317251PRT人工序列合成内酯酶CL0002558317MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015ValHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525018753DNA人工序列合成内酯酶CL0002558318atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcgtccacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75319251PRT人工序列合成内酯酶CL0002559419MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015ValHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525020753DNA人工序列合成内酯酶CL0002559420atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcgtccacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75321251PRT人工序列人工序列CL0002562221MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyGluAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525022753DNA人工序列合成内酯酶CL0002562222atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggaggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75323251PRT人工序列合成内酯酶CL0002563023MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525024753DNA人工序列合成内酯酶CL0002563024atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtacgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75325251PRT人工序列合成内酯酶CL0002563425MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015ValHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525026753DNA人工序列合成内酯酶CL0002563426atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcgtccacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75327251PRT人工序列合成内酯酶CL0002563527MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuHisLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525028753DNA人工序列合成内酯酶CL0002563528atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccacctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75329251PRT人工序列合成内酯酶CL0002565929MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuHisLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525030753DNA人工序列合成内酯酶CL0002565930atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccacctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75331251PRT人工序列人工内酯酶CL0002566631MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525032753DNA人工序列合成内酯酶CL0002566632atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75333251PRT人工序列合成内酯酶CL00025686G2P33MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525034753DNA人工序列合成内酯酶CL00025686G2P34atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75335251PRT人工序列合成内酯酶CL0002568735MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspSerPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnThrValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525036753DNA人工序列合成内酯酶CL0002568736atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgactccttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660cagaccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75337251PRT人工序列合成内酯酶CL0002568937MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValValProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuHisLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyGluAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525038753DNA人工序列合成内酯酶CL0002568938atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtacgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtggtccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccacctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggaggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75339251PRT人工序列合成内酯酶CL0002571739MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnSerGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525040753DNA人工序列合成内酯酶CL0002571740atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagtct360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75341251PRT人工序列合成内酯酶CL0002572141MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuIleGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525042753DNA人工序列合成内酯酶CL0002572142atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcattcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75343251PRT人工序列合成内酯酶CL0002572843MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuValGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525044753DNA人工序列合成内酯酶CL0002572844atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgttcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75345251PRT人工序列合成内酯酶CL0002574845MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrGluGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525046753DNA人工序列合成内酯酶CL0002574846atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75347251PRT人工序列合成内酯酶CL0002575447MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluHisAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525048753DNA人工序列合成内酯酶CL0002575448atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagcatgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75349251PRT人工序列合成内酯酶CL0002576249MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnThrGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525050753DNA人工序列合成内酯酶CL0002576250atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagact360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75351251PRT人工序列合成内酯酶CL0002579151MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnHisGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525052753DNA人工序列合成内酯酶CL0002579152atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagcat360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75353251PRT人工序列合成内酯酶CL0002592453MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuCysGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525054753DNA人工序列合成内酯酶CL0002592454atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctctgtcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75355251PRT人工序列合成内酯酶CL0002593555MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAlaGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525056753DNA人工序列合成内酯酶CL0002593556atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgcgggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75357251PRT人工序列合成内酯酶CL0002595557MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluAlaMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525058753DNA人工序列合成内酯酶CL0002595558atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggcgatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75359251PRT人工序列合成内酯酶CL0002595859MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuLysGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525060753DNA人工序列合成内酯酶CL0002595860atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcaagcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75361251PRT人工序列合成内酯酶CL0002596161MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuGlnGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525062753DNA人工序列合成内酯酶CL0002596162atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctccagcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75363251PRT人工序列合成内酯酶CL0002596363MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnArgGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525064753DNA人工序列合成内酯酶CL0002596364atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagcgt360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75365251PRT人工序列合成内酯酶CL0002598465MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrGlyGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525066753DNA人工序列合成内酯酶CL0002598466atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgggggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75367251PRT人工序列合成内酯酶CL0002601767MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525068753DNA人工序列合成内酯酶CL0002601768atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggacgatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75369251PRT人工序列合成内酯酶CL0002602069MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuThrGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525070753DNA人工序列合成内酯酶CL0002602070atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcactcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75371251PRT人工序列合成内酯酶CL0002604171MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrGlnGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525072753DNA人工序列合成内酯酶CL0002604172atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcacccagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75373251PRT人工序列合成内酯酶CL0002605273MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnProGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525074753DNA人工序列合成内酯酶CL0002605274atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagccg360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75375251PRT人工序列合成内酯酶CL0002605475MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525076753DNA人工序列合成内酯酶CL0002605476atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaatgcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75377251PRT人工序列合成内酯酶CL0002606177MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnAlaGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525078753DNA人工序列合成内酯酶CL0002606178atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcaggcg360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75379251PRT人工序列合成内酯酶CL0002608879MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgMetIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525080753DNA人工序列合成内酯酶CL0002608880atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggatgatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75381251PRT人工序列合成内酯酶CL0002608981MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgAsnIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525082753DNA人工序列合成内酯酶CL0002608982atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaatatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75383251PRT人工序列合成内酯酶CL0002610283MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525084753DNA人工序列合成内酯酶CL0002610284atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75385251PRT人工序列合成内酯酶CL0002610485MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrSerGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525086753DNA人工序列合成内酯酶CL0002610486atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcacctctggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75387251PRT人工序列合成内酯酶CL0002611487MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgAlaIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525088753DNA人工序列合成内酯酶CL0002611488atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggcgatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75389251PRT人工序列合成内酯酶CL0002624189MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspSerIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525090753DNA人工序列合成内酯酶CL0002624190atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgactctatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75391251PRT人工序列合成内酯酶CL0002628991MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValHisArgValGluAlaPheValArg24525092753DNA人工序列合成内酯酶CL0002628992atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtccatcgcgtcgaggctttcgtccgc75393251PRT人工序列合成内酯酶CL0002629693MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgAsnProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525094753DNA人工序列合成内酯酶CL0002629694atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggaatccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75395251PRT人工序列合成内酯酶CL0002629795MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgCysProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg24525096753DNA人工序列合成内酯酶CL0002629796atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtgtccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc75397251PRT人工序列合成内酯酶CL0002630897MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValCysArgValGluAlaPheValArg24525098753DNA人工序列合成内酯酶CL0002630898atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtctgtcgcgtcgaggctttcgtccgc75399251PRT人工序列合成内酯酶CL0002631099MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValAspGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250100753DNA人工序列合成内酯酶CL00026310100atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtggatggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753101251PRT人工序列合成内酯酶CL00026313101MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValArgGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250102753DNA人工序列合成内酯酶CL00026313102atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgaggggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753103251PRT人工序列合成内酯酶CL00026320103MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspLysIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250104753DNA人工序列合成内酯酶CL00026320104atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacaagatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753105251PRT人工序列合成内酯酶CL00026328105MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerProAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250106753DNA人工序列合成内酯酶CL00026328106atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcacctgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753107251PRT人工序列合成内酯酶CL00026344107MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleThrGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250108753DNA人工序列合成内酯酶CL00026344108atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatcacgggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753109251PRT人工序列合成内酯酶CL00026349109MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspArgIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250110753DNA人工序列合成内酯酶CL00026349110atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacaggatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753111251PRT人工序列合成内酯酶CL00026369111MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleLysGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250112753DNA人工序列合成内酯酶CL00026369112atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatcaagggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753113251PRT人工序列合成内酯酶CL00026396113MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValLysGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProLysGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250114753DNA人工序列合成内酯酶CL00026396114atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgaagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaagcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753115251PRT人工序列合成内酯酶CL00026398115MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValLysArgValGluAlaPheValArg245250116753DNA人工序列合成内酯酶CL00026398116atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcaagcgcgtcgaggctttcgtccgc753117251PRT人工序列合成内酯酶CL00026451117MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgHisProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250118753DNA人工序列合成内酯酶CL00026451118atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggcatccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753119251PRT人工序列合成内酯酶CL00026455119MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValSerArgValGluAlaPheValArg245250120753DNA人工序列合成内酯酶CL00026455120atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtctcgcgcgtcgaggctttcgtccgc753121251PRT人工序列合成内酯酶CL00026470121MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaSerLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250122753DNA人工序列合成内酯酶CL00026470122atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgagtctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753123251PRT人工序列合成内酯酶CL00026476123MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValThrArgValGluAlaPheValArg245250124753DNA人工序列合成内酯酶CL00026476124atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcactcgcgtcgaggctttcgtccgc753125251PRT人工序列合成内酯酶CL00026535125MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValArgArgValGluAlaPheValArg245250126753DNA人工序列合成内酯酶CL00026535126atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtccggcgcgtcgaggctttcgtccgc753127251PRT人工序列合成内酯酶CL00026540127MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaAsnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250128753DNA人工序列合成内酯酶CL00026540128atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgaatctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753129251PRT人工序列合成内酯酶CL00026559129MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaTyrLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250130753DNA人工序列合成内酯酶CL00026559130atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgtatctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753131251PRT人工序列合成内酯酶CL00026564131MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspGluIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250132753DNA人工序列合成内酯酶CL00026564132atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgagatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753133251PRT人工序列合成内酯酶CL00026641133MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspLeuIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250134753DNA人工序列合成内酯酶CL00026641134atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacctgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753135251PRT人工序列合成内酯酶CL00026661135MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspAlaIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250136753DNA人工序列合成内酯酶CL00026661136atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgctatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753137251PRT人工序列合成内酯酶CL00026679137MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGlyArgValGluAlaPheValArg245250138753DNA人工序列合成内酯酶CL00026679138atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgggcgcgtcgaggctttcgtccgc753139251PRT人工序列合成内酯酶CL00026695139MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValAsnGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250140753DNA人工序列合成内酯酶CL00026695140atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgaatggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753141251PRT人工序列合成内酯酶CL00026696141MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspAsnIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250142753DNA人工序列合成内酯酶CL00026696142atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacaatatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753143251PRT人工序列合成内酯酶CL00028441143MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250144753DNA人工序列合成内酯酶CL00028441144atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753145251PRT人工序列合成内酯酶CL00028451145MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250146753DNA人工序列合成内酯酶CL00028451146atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753147251PRT人工序列合成内酯酶CL00028468147MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProLysGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250148753DNA人工序列合成内酯酶CL00028468148atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaagcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753149251PRT人工序列合成内酯酶CL00028526149MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250150753DNA人工序列合成内酯酶CL00028526150atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753151251PRT人工序列合成内酯酶CL00028549151MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250152753DNA人工序列合成内酯酶CL00028549152atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753153251PRT人工序列合成内酯酶CL00028564153MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250154753DNA人工序列合成内酯酶CL00028564154atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753155251PRT人工序列合成内酯酶CL00028574155MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250156753DNA人工序列合成内酯酶CL00028574156atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753157251PRT人工序列合成内酯酶CL00028598157MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250158753DNA人工序列合成内酯酶CL00028598158atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753159251PRT人工序列合成内酯酶CL00028616159MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaGluLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250160753DNA人工序列合成内酯酶CL00028616160atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggagctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753161251PRT人工序列合成内酯酶CL00028623161MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAspArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250162753DNA人工序列合成内酯酶CL00028623162atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggatcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753163251PRT人工序列合成内酯酶CL00028629163MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrValAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAspArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250164753DNA人工序列合成内酯酶CL00028629164atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccgtcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggatcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753165251PRT人工序列合成内酯酶CL00028657165MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250166753DNA人工序列合成内酯酶CL00028657166atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753167251PRT人工序列合成内酯酶CL00028669167MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspLeu202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250168753DNA人工序列合成内酯酶CL00028669168atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatctgaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753169251PRT人工序列合成内酯酶CL00028670169MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrValAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProLysGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250170753DNA人工序列合成内酯酶CL00028670170atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccgtcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaagcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753171251PRT人工序列合成内酯酶CL00028687171MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250172753DNA人工序列合成内酯酶CL00028687172atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753173251PRT人工序列合成内酯酶CL00028741173MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProLysGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250174753DNA人工序列合成内酯酶CL00028741174atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaagcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753175251PRT人工序列合成内酯酶CL00028745175MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250176753DNA人工序列合成内酯酶CL00028745176atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753177251PRT人工序列合成内酯酶CL00028752177MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProHisAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250178753DNA人工序列合成内酯酶CL00028752178atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660cacgccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753179251PRT人工序列合成内酯酶CL00028754179MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250180753DNA人工序列合成内酯酶CL00028754180atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753181251PRT人工序列合成内酯酶CL00028773181MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580ProProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProHisAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250182753DNA人工序列合成内酯酶CL00028773182atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240cccccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660cacgccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753183251PRT人工序列合成内酯酶CL00028835183MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrValAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProLysGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250184753DNA人工序列合成内酯酶CL00028835184atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccgtcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaagcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753185251PRT人工序列合成内酯酶CL00028844185MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaAlaAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580ProProValValValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250186753DNA人工序列合成内酯酶CL00028844186atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcggccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240cccccggtggtcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753187251PRT人工序列合成内酯酶CL00028864187MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250188753DNA人工序列合成内酯酶CL00028864188atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccggggaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753189251PRT人工序列合成内酯酶CL00028867189MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnIleGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250190753DNA人工序列合成内酯酶CL00028867190atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360cagatccagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753191251PRT人工序列合成内酯酶CL00028868191MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisAspThrLeuAlaGlnLeuAlaGluAsnAspValIleProAlaGlu65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProLeuPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190ValProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250192753DNA人工序列合成内酯酶CL00028868192atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgacaccctcgcgcagctcgccgagaacgacgtgatccccgccgaa240ctgccggtgatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccgctgttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgagggtcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753193251PRT人工序列合成内酯酶CL00028915193MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250194753DNA人工序列合成内酯酶CL00028915194atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753195251PRT人工序列合成内酯酶CL00028922195MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250196753DNA人工序列合成内酯酶CL00028922196atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaccatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753197251PRT人工序列合成内酯酶CL00028925197MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250198753DNA人工序列合成内酯酶CL00028925198atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753199251PRT人工序列合成内酯酶CL00028945199MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250200753DNA人工序列合成内酯酶CL00028945200atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753201251PRT人工序列合成内酯酶CL00028948201MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250202753DNA人工序列合成内酯酶CL00028948202atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753203251PRT人工序列合成内酯酶CL00028964203MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250204753DNA人工序列合成内酯酶CL00028964204atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaccatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753205251PRT人工序列合成内酯酶CL00028973205MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250206753DNA人工序列合成内酯酶CL00028973206atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753207251PRT人工序列合成内酯酶CL00028979207MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250208753DNA人工序列合成内酯酶CL00028979208atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaccatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753209251PRT人工序列合成内酯酶CL00029127209MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250210753DNA人工序列合成内酯酶CL00029127210atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753211251PRT人工序列合成内酯酶CL00029160211MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrMet180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250212753DNA人工序列合成内酯酶CL00029160212atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgatgatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753213251PRT人工序列合成内酯酶CL00029162213MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250214753DNA人工序列合成内酯酶CL00029162214atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaccatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753215251PRT人工序列合成内酯酶CL00029199G3P215MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250216753DNA人工序列合成内酯酶CL00029199G3P216atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753217251PRT人工序列合成内酯酶CL00029214217MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrAspGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProIleIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAsnAlaLeuValSerArgLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgGluIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250218753DNA人工序列合成内酯酶CL00029214218atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccgacggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccgatcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactgaacgcgctcgtgagccgcctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcgggagatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753219251PRT人工序列合成内酯酶CL00029220219MetIleLeuAlaHisAspValSerGlySerGlyProLeuLeuValLeu151015LeuHisGlyIleThrGluAspArgArgSerTrpAspProValAspPhe202530ThrLysGlyPheThrValValArgValAspLeuArgGlyHisGlyAla354045SerAlaAlaGluGluProTyrAspIleProThrLeuAlaThrAspVal505560HisGluThrLeuAlaGlnLeuAlaGluAsnAspValIleProGlyGly65707580LeuProValIleValGlyHisSerMetGlyGlyIleValAlaThrAla859095TyrGlyAlaLeuPheProAlaArgAlaIleValAsnValAspGlnPro100105110LeuGlnLeuAlaGlyMetGlnGlyGlnValGlnGlnAlaGluGlyMet115120125LeuArgGlyAlaAspPheProValPheIleHisGlyMetPheAlaGln130135140MetAlaGlyGlyLeuAspAlaGluGluLeuAlaArgValAsnGlyIle145150155160ArgSerProArgGlnAspValValLeuGlyMetTrpArgProLeuLeu165170175GluAspSerProGluGluLeuAlaAlaLeuValSerGlyLeuThrArg180185190IleProGluAspValProTyrLeuValIleThrGlyLeuAspAlaGly195200205ProGluTyrAlaAlaTrpLeuGlnArgThrIleProGlnAlaValGln210215220GluValTrpGlnProProThrHisTyrProHisLeuValAspProAla225230235240ArgPheValGluArgValGluAlaPheValArg245250220753DNA人工序列合成内酯酶CL00029220220atgatcctcgcccacgacgtgtcgggctccggcccgctgctggtcctcctgcacggcatc60accgaagaccgccgcagctgggatccggtcgatttcaccaagggcttcacggtcgtgcgg120gtcgacctgcgcgggcacggggcatcagccgccgaagaaccgtacgacatccccacgctc180gcgaccgacgtgcacgagaccctcgcgcagctcgccgagaacgacgtgatccccgggggc240ctgccggtcatcgtcggccactcgatgggcgggatcgtcgcgacggcgtacggcgcgctc300ttccccgcgcgggcgatcgtcaacgtggaccagcctctccagctcgcgggcatgcagggc360caggtgcagcaggcggaggggatgctccgcggggcggacttcccggtcttcatccacggc420atgttcgcgcagatggcgggcggcctggatgccgaggagctggcgcgggtgaatggcatc480cggtctccgaggcaggacgtcgtcctcgggatgtggcggccgcttctcgaggactcaccc540gaagaactggcggcgctcgtgagcggtctgacgaggatcccggaggacgtcccgtacctc600gtgatcacgggtctcgatgccgggccagagtacgcggcgtggctgcagcggaccatcccg660caggccgtccaggaggtctggcagccgccgacccactacccgcacctcgtcgacccggca720cggttcgtcgagcgcgtcgaggctttcgtccgc753

权利要求：1.包含内酯酶变体酶的组合物，所述内酯酶变体酶包含与SEQIDNO:1相比的氨基酸取代，其中所述氨基酸取代在选自17,32,34,35,41,50,62,66,68,69,70,74,76,77,79,80,81,83,84,114,120,122,123,127,132,136,139,155,162,164,184,185,188,189,192,193,195,218,221,222和244的位置处，其中所述内酯酶变体酶保留内酯酶活性并且与SEQIDNO:1至少95％相同。2.根据权利要求1的组合物，其中所述氨基酸取代选自L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,D66E,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,E80G,L81P,V83I,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,L136V,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T。3.根据权利要求1的组合物，其中所述变体酶包含氨基酸取代D66EE80GV83IL136V。4.根据权利要求3的组合物，其中所述变体酶还包含选自L17V,F32L,D34A,D34E,D34G,D34K,D34Q,D34S,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,184V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122I,Q123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A184N,A185E,S188D,S188N,S188R,G189R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T的氨基酸取代。5.根据权利要求1的组合物，其中所述变体酶包含氨基酸取代D34KD66EE80GL136VA184NG189P。6.根据权利要求5的组合物，其中所述变体酶还包含选自LI7V,F32L,G35S,V41F,A50P,T62A,L68V,A69C,A69I,A69K,A69Q,A69T,A69V,Q70N,Q70S,Q70Y,N74H,V76A,V76E,V76K,V76L,V76N,V76R,V76S,I77V,G79A,L81P,I84V,Q114H,G120A,G120H,G120P,G120R,G120S,G120T,V122LQ123K,G127A,A132E,H139K,H139T,A155D,S162C,S162H,S162N,R164K,A185E,S188D,S188N,S188R,R192M,I193V,E195A,E218A,E218M,E218N,E218T,Q221H,A222T,E244C,E244G,E244H,E244K,E244R,E244S和E244T的氨基酸取代。7.根据权利要求1的组合物，其中所述变体酶具有选自D66EQ114H,G35SV41F,G35S,G35SE195A,G189RE195A,L17VG35S,L17VG35SI77VL136VG189R,L17VG35SD66E,L17VD66EL136V,A132E,G189R,L17VD66E,G35SQ114H,Q114HG189R,D66EV83I,D66EE80GV83IL136V,G35SD66EV83IA222T,D66EI77VE80GV83IQ114HA132EG189R,G120S,A69I,A69V,D34E,N74H,G120T,G120H,A69C,D34A,G127A,A69K,A69Q,G120R,D34G,E218T,A69T,D34Q,G120P,A184N,G120A,E218M,E218N,D34K,D34S,E218A,V76S,E244H,S162N,S162C,E244C,S188D,S188R,V76K,A50P,H139T,V76R,H139K,Q123KR164K,E244K,S162H,E244S,Q70S,E244T,E244R,Q70N,Q70Y,V76E,V76L,V76A,E244G,S188N,V76N,L81P,L81PI84V,T62AG79AR164KI193V,G79AL81P,G79AI84VV122I,T62AL81PV122II193V,T62AG79AL81P,T62AG79A,L81PA185E,G79AL81PI84VA155D,L68VL81PI84VA155DI193V,L81PI84VI193V,F32LL81PI84VI193V,L68VL81PI84VR164K,G79A,G79AV122IR164K,G79AI84V,V122II193VQ221H,L81PV122I,T62AG79AL81PI193VQ221H,T62AL68VV122IR164K,T62AL8IP184V,V122I,G79AV122I,G79AI193V,D66EE80GV83IL136VD34KA184NG189R,D66EE80GV83IL136VD34KA184NG189RE218T,D66EE80GV83IL136VD34K,D66EE80GV83IL136VD34KA184N,D66EE80GL136VD34K,D66EE80GV83IL136VD34KA184NE218T,D66EE80GL136V,D66EE80GL136VD34KA184NG189RE218T,D66EE80GL136VD34KG189R,D66EE80GV83IL136VR192M,D66EE80GL136VD34KG189RE218T,D66EE80GL136VD34KA184NG189R,D66EE80GV83IL136VA184NG189R和D66EE80GL136VD34KE218T的氨基酸取代组。8.根据权利要求1的组合物，其还包含动物饲料。9.编码权利要求1的内酯酶变体酶的核酸。10.表达载体，其包含权利要求9的核酸。11.宿主细胞，其包含权利要求10的表达载体。12.制备内酯酶变体酶的方法，所述方法包括在产生所述酶的条件下培养权利要求11的宿主细胞，并回收所述酶。

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