CN106834244A - 脯氨酸羟化酶及其应用 - Google Patents

脯氨酸羟化酶及其应用 Download PDF

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CN106834244A
CN106834244A CN201610971207.2A CN201610971207A CN106834244A CN 106834244 A CN106834244 A CN 106834244A CN 201610971207 A CN201610971207 A CN 201610971207A CN 106834244 A CN106834244 A CN 106834244A
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CN106834244B (zh
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洪浩
詹姆斯·盖吉
卢江平
张娜
于文燕
刘芳
李艳君
黄鑫
高娟
张克俭
马玉磊
魏俊璐
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Asymchem Laboratories Fuxin Co Ltd
Asymchem Laboratories Tianjin Co Ltd
Asymchem Laboratories Jilin Co Ltd
Asymchem Life Science Tianjin Co Ltd
Tianjin Asymchem Pharmaceutical Co Ltd
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Asymchem Laboratories Tianjin Co Ltd
Asymchem Laboratories Jilin Co Ltd
Asymchem Life Science Tianjin Co Ltd
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    • C12Y114/11002—Procollagen-proline dioxygenase (1.14.11.2), i.e. proline-hydroxylase

Abstract

本发明提供了一种脯氨酸羟化酶及其应用。脯氨酸羟化酶包括(a)具有SEQ ID NO:2所示的氨基酸序列的蛋白质；(b)SEQ ID NO:2所示的氨基酸序列经过一个或多个氨基酸的突变且具有脯氨酸羟化酶活性的蛋白质；或者(c)保留(b)中的一个或多个氨基酸的突变，具有脯氨酸羟化酶活性，且与(b)中蛋白质的氨基酸序列至少具有78％的同源性的蛋白质。以SEQ ID NO:2所示的氨基酸序列的蛋白质以及利用基因工程手段对其改造得到的突变体，具有比现有的脯氨酸羟化酶更高的催化专一性或者显著提高的催化活性。

Description

脯氨酸羟化酶及其应用
技术领域
本发明涉及基因工程和酶工程，具体而言，涉及一种脯氨酸羟化酶及其应用。
背景技术
脯氨酸的衍生物是药物合成的重要结构单元，尤其羟脯氨酸在自然界中属于稀有氨基酸，是很多重要药物合成的起始原料。根据脯氨酸羟化位置及空间结构的不同，羟脯氨酸共有8种异构体，其中3-羟基-L-脯氨酸和4-羟基-L-脯氨酸作为抗生素、酶抑制剂、抗肿瘤、抗高血压及新型胃药等多种药物的重要原料，是目前生物合成研究的热点。除此之外，羟脯氨酸也可以是天然产物的来源或经化学合成，比如可以是植物材料和胶原蛋白的水解产物，或者以烯丙基溴和二乙基乙酰氨基丙二酸，D-谷氨酸和β-丙氨酸为起始原料进行化学合成。
同样的，羟脯氨酸的衍生物L-羟基哌啶酸也是药物合成的重要结构单元，是β-内酰胺酶抑制剂和TNF-α转化酶抑制剂合成的重要中间体。L-羟基哌啶酸同样可以通过植物或其他自然材料分离或者通过化学合成的方法来制备。但是需要复杂的分离纯化方法或者是复杂的合成步骤，很难实现大规模的工业化生产。而通过羟化酶对哌啶酸进行羟基化是获得羟基哌啶酸的理想方法。目前应用最多的酶为脯氨酸羟化酶，这是一种酮戊二酸依赖性双加氧酶，需要α-酮戊二酸和O2作为共同底物，铁离子作为辅因子。
在很多微生物中都含有脯氨酸羟化酶，如苜蓿中华根瘤菌(Sinorhizobiummeliloti)、链霉菌TH1(Streptomyces sp.strain TH1)及丝状真菌Glarea lozoyensis等。常用的有顺式-脯氨酸-3-羟化酶、顺式-脯氨酸-4-羟化酶和反式-脯氨酸-4-羟化酶三类，分别能催化L-脯氨酸产生顺式-3-羟基-L-脯氨酸、顺式-4-羟基-L-脯氨酸和反式-4-羟基-L-脯氨酸。但在脯氨酸羟化酶对脯氨酸及脯氨酸衍生物的催化反应中，存在一些不容忽视的问题，比如转化率低、催化产生位置异构体等，无法实现大规模的工业生产。来自于苜蓿中华根瘤菌(Sinorhizobium meliloti)的顺式-脯氨酸-4-羟化酶经过基因改造后，对L-哌啶酸羟基化作用的转化率和酶催化的专一性都得到大幅度的提高，但是仍有10％的位置异构体(2S,3R)-3-羟基哌啶-2-羧酸产生(WO2013169725A2)。因此，找到一种能够专一、快速催化脯氨酸及脯氨酸衍生物羟基化作用的酶，对于羟基脯氨酸及其衍生物的工业合成具有重要意义。
发明内容
本发明的主要目的在于提供一种脯氨酸羟化酶及其应用，以解决现有技术中脯氨酸羟化酶催化的选择性较差的问题。
为了实现上述目的，根据本发明的一个方面，提供了一种脯氨酸羟化酶，包括：(a)具有SEQ ID NO:2所示的氨基酸序列的蛋白质；(b)SEQ ID NO:2所示的氨基酸序列经过一个或多个氨基酸的突变且具有脯氨酸羟化酶活性的蛋白质；或者(c)保留(b)中的一个或多个氨基酸的突变，具有脯氨酸羟化酶活性，且与(b)中蛋白质的氨基酸序列至少具有78％的同源性的蛋白质。
进一步地，突变的氨基酸位点选自H14、S16、T25、F26、E27、D30、S33、E34、Y35、S37、I39、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223以及S225中的一个或多个。
进一步地，突变包括如下任意一个或多个：H14R、S16N、T25G、T25R、F26L、E27K、D30S、S33N、E34N、E34G、E34L、E34S、E34D、Y35W、Y35K、S37W、S37F、S37E、S37N、S37T、S37C、I39K、I39R、W40F、K41E、D54G、H55Q、S57L、I58T、I58Y、I58A、I58R、I58V、I58S、I58C、K86P、T91A、F95Y、C97Y、I98V、K106V、K106T、K106Q、F111S、K112E、K112R、S154A、K162E、L166M、I118F、I118V、I118R、H119R、H119F、I120V、K123D、K123N、K123Q、K123S、K123I、K123T、T130N、D134G、V135K、N165H、D173G、K209R、I223V、以及S225A。
进一步地，突变包括如下任意一种组合：E27K+Y35W/K、E27K+I39K/R、E27K+K123D/I/Q/S、E27K+N165H、I39K/R+Y35W/K、I39K/R+K123D/I/Q/S、I39K/R+N165H、K123D+W40F、K123D+Y35W/K、E27K+I39K/R+K123D/I/Q/S、K123D/I/Q/S+N165H、S37C/E/F/N/W/T+I223V、E27K+Y35W/K+I39K/R、E27K+S37C/E/F/N/W/T+I39K/R、E27K+E34N/G/L/D/S+I39K/R、E27K+I39K/R+D30S、E27K+I39K/R+I118F/V/R、E27K+I39K/R+I98V、S37C/E/F/N/W/T+I223V+N165H、Y35W/K+S37C/E/F/N/W/T+W40F、S37C/E/F/N/W/T+I223V+K123D/I/Q/S、E27K+I39K+Y35W/K+S37C/E/F/N/W/T、E27K+I39K/R+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+K106Q+K112E、E27K+I39K/R+Y35W/K+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+S37C/E/F/N/W/T+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I223V+K123D/I/Q/S、S37C/E/F/N/W/T+I39K/R+I223V+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I98V、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I223V、F26L+E27K+I39K/R+K123D/I/Q/S、I223V+S37C/E/F/N/W/T+E27K+I39K/R、I223V+S37C/E/F/N/W/T+E27K+N165H、E27K+S37C/E/F/N/W/T+I39K/R+I98V+K123D/I/Q/S+I223V、K106Q+K112E+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K123D/I/Q/S、E27K+I39K/R+K123D/I/Q/S+N165H、H14R+E34G+K106Q+K112E+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K86P、K123D/I/Q/S+Y35W/K+I120V、E27K+D30S+I39K/R+I58A/C/R/S/T/V/Y+K112E、S37C/E/F/N/W/T+I39K/R+N165H、E27K+E34N/G/L/D/S+I39K/R+I58A/C/R/S/T/V/Y+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+I118F/V/R、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+N165H、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/+D173G+K123D/I/Q/S、H14R+E27K+D30S+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R+N165H、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+Y35W/K+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+K123D/I/Q/S+I223V、H14R+E27K+E34N/G/L/D+I39K/R+I58A/C/R/S/T/V/Y+I98V+K106V/T/Q+K112E/R+I223V以及H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I118F/V/R+I223V，其中，“/”表示“或者”。
为了实现上述目的，根据本发明的一个方面，提供了一种DNA分子，DNA分子编码上述任一种脯氨酸羟化酶。
为了实现上述目的，根据本发明的又一个方面，提供了一种重组质粒，重组质粒连接有上述DNA分子。
进一步地，重组质粒选自如下之一：pET-21b(+)、pET-22b(+)、pET-3a(+)、pET-3d(+)、pET-11a(+)、pET-12a(+)、pET-14b(+)、pET-15b(+)、pET-16b(+)、pET-17b(+)、pET-19b(+)、pET-20b(+)、pET-21a(+)、pET-23a(+)、pET-23b(+)、pET-24a(+)、pET-25b(+)、pET-26b(+)、pET-27b(+)、pET-28a(+)、pET-29a(+)、pET-30a(+)、pET-31b(+)、pET-32a(+)、pET-35b(+)、pET-38b(+)、pET-39b(+)、pET-40b(+)、pET-41a(+)、pET-41b(+)、pET-42a(+)、pET-43a(+)、pET-43b(+)、pET-44a(+)、pET-49b(+)、pQE2、pQE9、pQE30、pQE31、pQE32、pQE40、pQE70、pQE80、pRSET-A、pRSET-B、pRSET-C、pGEX-5X-1、pGEX-6p-1、pGEX-6p-2、pBV220、pBV221、pBV222、pTrc99A、pTwin1、pEZZ18、pKK232-18、pUC-18以及pUC-19。
根据本发明的另一方面，提供了一种宿主细胞，宿主细胞含有上述任一种重组质粒。
进一步地，宿主细胞为原核细胞或真核细胞，优选真核细胞为酵母细胞。
进一步地，宿主细胞为感受态细胞，优选感受态细胞为大肠杆菌BL21细胞或大肠杆菌W3110。
根据本发明的另一方面，提供了一种生产L-羟脯氨酸衍生物的方法，方法包括：以L-脯氨酸衍生物为底物，利用权利要求上述任一种脯氨酸羟化酶催化底物的羟基化，得到通式(I)所示的L-羟脯氨酸衍生物；
其中，R1选自C1-C5的亚烷基或C2-C5亚烯基；R2选自C0-C4亚烷基或C2-C4亚烯
基；R3选自羟基、氨基、C1-C6烷氧基、芳氧基、C1-C6烷硫基或C1-C6芳硫基；R4选
自氢、C1-C6烷基、C2-C6烯基或C2-C6炔基。
进一步地，该方法包括：以α-酮戊二酸和O2为共同底物，以铁离子为辅因子，利用脯氨酸羟化酶催化底物的羟基化，得到L-羟脯氨酸衍生物。
进一步地，L-羟脯氨酸衍生物为顺式-4-羟基-L-脯氨酸或(2S,5S)-5-羟基哌啶-2-羧酸。
进一步地，脯氨酸羟化酶在5～45℃的反应温度，优选5～15℃的反应温度下催化底物的羟基化，得到通式(I)所示的L-羟脯氨酸衍生物。
应用本发明的技术方案，通过选择以SEQ ID NO:2为基础序列，并通过基因工程手段改造后的包含单个或多个氨基酸残基的改变的突变体，或者在保留这些突变的前提下对其他氨基酸位点进行改变，使得改变后的氨基酸序列与(b)中的氨基酸序列具有至少78％同源性的蛋白质，具有比现有的脯氨酸羟化酶更高的催化专一性(即选择性)，或者同时具有比本发明所发现的野生型羟化酶(即氨基酸序列为SEQ ID NO:2的脯氨酸羟化酶)显著提高的催化活性。
附图说明
构成本申请的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。在附图中：
图1为本发明所涉及的脯氨酸羟化酶催化合成(2S,5S)-5-羟基哌啶-2-羧酸(或称顺式-5-羟基哌啶酸)的化学反应过程；以及
图2为本发明所涉及的脯氨酸羟化酶催化合成顺式-4-羟基-L-脯氨酸的化学反应方程式。
具体实施方式
需要说明的是，在不冲突的情况下，本申请中的实施例及实施例中的特征可以相互组合。下面将参考附图并结合实施例来详细说明本发明。
名称解释：
酶的选择性，也称酶的专一性，是指酶在催化生化反应时对底物的选择性。在本发明中，脯氨酸羟化酶的选择性是指脯氨酸羟化酶催化L-脯氨酸衍生物羟基化，得到一种特定构型的L-羟脯氨酸衍生物的程度。在本说明所涉及的反应中，可以通过用反应的非对应异构体过量表征此羟化酶的催化选择性的程度。
转化率：是指某一反应物转化的百分率或分率。转化率是表示反应物反应程度的指标。本发明中L-哌啶酸羟基化作用的转化率是指羟化酶或脯氨酸羟化酶催化L-哌啶酸反应中，生成的产品(2S,5S)-5-羟基哌啶-2-羧酸占体系中L-哌啶酸的百分比。
催化活性：是指每单位容积(或质量)催化剂在单位时间内转化原料反应物的数量。在本发明中，脯氨酸羟化酶的催化活性高低与本发明中所述反应的转化率呈正相关。
进化：用突变或重组等手段去创建分子的多样性，然后对这多样性进行筛选，获得具有新功能的基因或DNA。本发明中，通过突变或重组等手段对羟化酶或脯氨酸羟化酶进行改造，以获得性能提高的羟化酶或脯氨酸羟化酶。
非对应异构体：分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。
非对应异构体过量(diastereomeric excess，缩写为de％)：用来表征两个手性中心时，一个非对映体对其他对映体的过量。即de％＝(R,R+S,S)-(S,R+R,S)/(R,R+S,S+S,R+R,S)。
野生型：是指从大自然中获得的，未经人工诱变或未改造过的。本发明中，野生型脯氨酸羟化酶，是指Genebank中筛选获得的一种天然的，未经人工改造的基因序列编码得到的脯氨酸羟化酶。
同源序列：是指在同一物种不同个体间或不同物种间相同或相似的DNA序列。
本发明中，所涉及到的1wt均指转化1g主原料需要1g脯氨酸羟化酶变体重组湿细胞。
如背景技术部分所提到的，现有技术中的脯氨酸羟化酶存在催化选择性不够高的缺陷，为后续的目的产物的分离增加了困难。为了解决现有技术中脯氨酸羟化酶的选择性不够高的难题，发明人通过基因工程的手段，获得一个催化专一性高的脯氨酸羟化酶。同时，发明人对现有的野生型脯氨酸羟化酶进行各种突变改造，以期获得一种选择性高、且催化活性获得提高的脯氨酸羟化酶。
发明人从Genebank中筛选到了无数条现有脯氨酸羟化酶的氨基酸序列的同源序列，按照同源性从高到底的顺序，对筛选到的各种同源序列进行了不同位置的基因突变，并对各种突变体的羟化酶活性进行了筛选。结果发现，对同源性序列较高的序列进行突变后所得到的突变体的脯氨酸羟化酶活性均与现有的脯氨酸羟化酶无明显区别。最后，仅剩一个与现有的脯氨酸羟化酶的氨基酸序列同源性最低(仅为30％左右)的来源于Kordiajejudonensis的序列，该序列在Genebank中并没有明确的基因功能注释，被注释为是一种假定蛋白。发明人利用基因工程技术对该序列进行了重组表达，出乎意料地发现，该序列编码的蛋白，具有脯氨酸羟化酶活性，并且具有高于现有技术中的脯氨酸羟化酶的选择性。发明人把该蛋白命名为野生型脯氨酸羟化酶。发明人还对该野生型羟化酶的氨基酸序列进行了突变试验，令人更惊喜的是，该来源于Kordia jejudonensis的序列经突变后所获得的突变体不仅具有脯氨酸羟化酶活性，而且其催化活性较野生型脯氨酸羟化酶有显著提高。
进一步地，发明人以该序列为基础序列进行各种突变筛选，最终发现了多个突变位点与脯氨酸羟化酶的活性有关。在保留这些突变位点的前提下，任意改变其他位点的突变对脯氨酸羟化酶的活性无明显影响。
在上述基础上，发明人进行了更深入的研究，发现一些重要的基酸残基位点，该位点突变后，其脯氨酸羟化酶的可溶性表达明显下降或脯氨酸羟化酶催化活性丧失。S3，L94，H105，D107，S131，E132，Y137，M139，W145，H153，N157，V167，D169氨基酸残基位点突变为其它绝大部分氨基酸后，脯氨酸羟化酶的可溶性表达明显下降，有些甚至为零。这些氨基酸的突变影响脯氨酸羟化酶的折叠，使其在E.Coli宿主细胞中的可溶性表达量明显降低，甚至可溶性表达降为零，其突变氨基酸可以选自：Y35A、Y35F、Y35S、W40Y、L94A、L94G、L94S、F95A、F95W、H105R、H105Q、H105E、H105G、R117A、R117P、R117K、P121V、V135A、S131T、H153Y、H153A、H153R、H153K。与此同时，还发现Y32、R93、R117、Y108氨基酸残基位点突变为其它绝大部分氨基酸后，脯氨酸羟化酶催化活性丧失。这些使得脯氨酸羟化酶的催化活性明显降低，甚至丧失活性的突变氨基酸可以选自：Y32N、Y32V、Y32V、Y32Q、Y32E、Y32S、Y32R、Y32D、Y32R、Y32I、Y32P、Y35H、Y35F、Y35A、W40Y、R93K、R93H、R93E、R93A、L94A、F95W、F95A、H105E、H105Q、H105R、H105K、H105A、K106H、D107A、Y108W、Y108A、Y108L、Y108S、Y108P、R117D、R117P、R117N、R117H、R117K、R117A、P121V、H153W、H153F、H153K、H153R、H153A。
对脯氨酸羟化酶的三维结构进行计算机模拟，得到模拟的三维结构图。对该结构图进行分析后，推测上述这些位点多数应该都是参与脯氨酸羟化酶与底物结合或与辅因子结合的重要氨基酸残基，如H105，D107，H153可能参与辅因子与脯氨酸羟化酶的结合，又如Y32、R93、R117可能参与底物与脯氨酸羟化酶的结合，其中R93、R117氨基酸残基可能会与底物形成盐桥。另外，第230-276位之间的氨基酸残基在酶的活性相关区域以外，因此改变或去除该区间的一些氨基酸，对脯氨酸羟化酶的活性影响不会很大。
本发明的突变体是在保持上述对脯氨酸羟化酶的活性影响很大的氨基酸位点不变的情况下，对剩余的氨基酸进行改变而得到的，催化活性或选择性提高的突变体。
在上述研究结果的基础上，发明人提出了本发明的技术方案。在一种典型的实施方式中，提供了一种脯氨酸羟化酶，该脯氨酸羟化酶包括：(a)具有SEQ ID NO:2所示的氨基酸序列的蛋白质；或(b)SEQ ID NO:2所示的氨基酸序列经过一个或多个氨基酸的突变且具有脯氨酸羟化酶活性的蛋白质；或(c)保留所述(b)中的一个或多个氨基酸的突变，具有脯氨酸羟化酶活性，且与(b)中的蛋白质的氨基酸序列至少具有78％的同源性的蛋白质。
上述脯氨酸羟化酶，通过选择以SEQ ID NO:2为基础序列，并通过基因工程手段改造后的包含单个或多个氨基酸残基的改变的突变体，或者在保留这些突变的前提下对其他氨基酸位点进行改变，使得改变后的氨基酸序列与(b)中的氨基酸序列具有至少78％同源性的蛋白质，具有比现有的羟化酶更高的催化专一性(即选择性)，或者同时具有比本发明所发现的野生型羟化酶(即氨基酸序列为SEQ ID NO:2的羟化酶)显著提高的催化活性。
上述脯氨酸羟化酶的催化活性比SEQ ID NO:2编码原始野生型羟化酶至少提高了1倍、2倍、3倍、4倍、5倍、或更高。另外，上述脯氨酸羟化酶的选择性比现有技术也显著提高，其催化产生的(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量(diastereomericexcess)大于90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高。
在某些实施方案中，上述脯氨酸羟化酶在SEQ ID NO:2的氨基酸序列经过如下一个或几个氨基酸位点的突变且具有脯氨酸羟化酶活性的蛋白质：H14、S16、T25、F26、E27、D30、S33、E34、Y35、S37、I39、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223以及S225。其中，与羟化酶催化活性相关的氨基酸位点有：H14、S16、F26、E27、D30、S33、E34、S37、I39、W40、K41、T91、F95、C97、I98、K106、F111、K112、I118、H119、I120、K123、T130、D134、V135、N165、K209以及I223。另外，Y35和S57氨基酸位点涉及到羟化酶的选择性。上述一个或几个氨基酸位点突变对野生型脯氨酸羟化酶的活性或选择性有显著影响，能显著提高突变体的脯氨酸羟化酶催化活性和/或选择性。
对上述脯氨酸羟化酶的三维结构模拟图进一步分析，推测本发明中的脯氨酸羟化酶的活性位点位于β折叠相关区域内，该区域由位于N-端和C-端的α螺旋结构所固定。底物结合位点位于β折叠区域的中心，与辅因子结合区域相邻。我们对这些位点进行分析，发现本发明中涉及脯氨酸羟化酶的上述突变氨基酸残基在脯氨酸羟化酶的三维结构模拟图中主要位于底物结合位点或与辅因子结合相关区域内，例如：E27、D30、S33、E34、Y35、S37、I39、W40、K41、H55、S57、I58、F95、C97、I98、F111、K112、I118、H119、I120氨基酸均可能处于底物结合位点附近，这些氨基酸的改变可能提高了底物结合的特异性，从而使酶的活性或催化选择得到提高。如K106、L166、K123、D134、S154、N165氨基酸均可能处于辅因子结合位点附近，这些氨基酸的改变可能提高了辅因子的结合，对氧的利用和传递起协调作用，从而使酶的活性得到提高。
在上述位点发生突变的基础上，发明人通过将这些位点突变成不同的氨基酸并检测其脯氨酸羟化酶活性的变化，发现当这些氨基酸位点突变为如下中的任意一种或几种的组合之后，羟化酶的活性和/或选择性进一步得到提高。突变包括如下任意一个或几个：H14R、S16N、T25G、T25R、F26L、E27K、D30S、S33N、E34N、E34G、E34L、E34S、E34D、Y35W、Y35K、S37W、S37F、S37E、S37N、S37T、S37C、I39K、I39R、W40F、K41E、D54G、H55Q、S57L、I58T、I58Y、I58A、I58R、I58V、I58S、I58C、K86P、T91A、F95Y、C97Y、I98V、K106V、K106T、K106Q、F111S、K112E、K112R、S154A、K162E、L166M、I118F、I118V、I118R、H119R、H119F、I120V、K123D、K123N、K123Q、K123S、K123I、K123T、T130N、D134G、V135K、N165H、D173G、K209R、I223V以及S225A。更优选地，突变包括如下任意一个或几个：H14R、E27K、E34N、E34G、E34L、E34D、Y35W、Y35K、S37W、S37F、S37E、S37N、S37T、S37C、I39K、I39R、I58T、I58Y、I58A、I58R、I58V、I58S、I58C、K123D、K123N、K123Q、K123S、K123I以及K123T。其中，与脯氨酸羟化酶的选择性相关的氨基酸突变有：Y35W、S57L和S57V。
某些氨基酸的突变可以提高脯氨酸羟化酶在细菌细胞中的可溶性表达量，尤其是在E.Coli宿主细胞中的可溶性表达，这些突变氨基酸可以选自：E27K、D30S、Y35W、Y35K、S37W、S37F、S37E、S37N、S37T、S37C、I39K、I39R、W40F、I58T、I58Y、I58A、I58R、I58V、I58S、I58C、I98V、K106V、K106T、K106Q、H119R、H119F、K123D、K123N、K123Q、K123S、K123I、K123T、N165H、I223V。一般而言，外源基因在原核表达系统中表达时，只有正确折叠形成的可溶性蛋白才具有活性，形成的包涵体是无活性的。可溶性蛋白表达量增加，从而总酶活也会随之增加。
在一种更优选的实施例中，上述突变包括如下任意一种组合：E27K+Y35W/K、E27K+I39K/R、E27K+K123D/I/Q/S、E27K+N165H、I39K/R+Y35W/K、I39K/R+K123D/I/Q/S、I39K/R+N165H、K123D+W40F、K123D+Y35W/K、E27K+I39K/R+K123D/I/Q/S、K123D/I/Q/S+N165H、S37C/E/F/N/W/T+I223V、E27K+Y35W/K+I39K/R、E27K+S37C/E/F/N/W/T+I39K/R、E27K+E34N/G/L/D/S+I39K/R、E27K+I39K/R+D30S、E27K+I39K/R+I118F/V/R、E27K+I39K/R+I98V、S37C/E/F/N/W/T+I223V+N165H、Y35W/K+S37C/E/F/N/W/T+W40F、S37C/E/F/N/W/T+I223V+K123D/I/Q/S、E27K+I39K+Y35W/K+S37C/E/F/N/W/T、E27K+I39K/R+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+K106Q+K112E、E27K+I39K/R+Y35W/K+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+S37C/E/F/N/W/T+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I223V+K123D/I/Q/S、S37C/E/F/N/W/T+I39K/R+I223V+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I98V、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I223V、F26L+E27K+I39K/R+K123D/I/Q/S、I223V+S37C/E/F/N/W/T+E27K+I39K/R、I223V+S37C/E/F/N/W/T+E27K+N165H、E27K+S37C/E/F/N/W/T+I39K/R+I98V+K123D/I/Q/S+I223V、K106Q+K112E+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K123D/I/Q/S、E27K+I39K/R+K123D/I/Q/S+N165H、H14R+E34G+K106Q+K112E+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K86P、K123D/I/Q/S+Y35W/K+I120V、E27K+D30S+I39K/R+I58A/C/R/S/T/V/Y+K112E、S37C/E/F/N/W/T+I39K/R+N165H、E27K+E34N/G/L/D/S+I39K/R+I58A/C/R/S/T/V/Y+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+I118F/V/R、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+N165H、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/+D173G+K123D/I/Q/S、H14R+E27K+D30S+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R+N165H、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+Y35W/K+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+K123D/I/Q/S+I223V、H14R+E27K+E34N/G/L/D+I39K/R+I58A/C/R/S/T/V/Y+I98V+K106V/T/Q+K112E/R+I223V以及H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I118F/V/R+I223V。上述突变组合中“/”表示或者的意思。在上述突变组合中，涉及到Y35W突变的组合，与脯氨酸羟化酶的选择性相关；其它组合与脯氨酸羟化酶的催化活性相关。
上述优选实施例中，脯氨酸羟化酶的催化活性信息是基于该酶对L-哌啶酸的催化活性而进行的筛选，这些更优选实施例中的脯氨酸羟化酶的催化活性结果如表1和表2所示，DNA序列的序列号为奇数，氨基酸序列的序列号为偶数。本发明所述的突变氨基酸是以SEQ ID NO：2的氨基酸序列为基础获得的突变，SEQ ID NO：2是来源于Kordiajejudonensis的假定蛋白的序列。本发明中野生型脯氨酸羟化酶的活性用“1”表示，突变体的脯氨酸羟化酶的活性用“+”表示：“+”表示活性为野生型SEQ ID NO:2的1～2倍，“++”表示活性为野生型SEQ ID NO:2的2～3倍，“+++”表示活性为野生型SEQ ID NO:2的3～4倍，“++++”表示活性为野生型SEQ ID NO:2的4～5倍。
表1.单一位点突变后的脯氨酸羟化酶活性比较
表1中提供了基于SEQ ID NO:2的序列进行不同位点的氨基酸改变对突变的脯氨酸羟化酶催化活性的影响。其中，突变的脯氨酸羟化酶表达自E.coli BL21细胞，催化活性是基于该酶对L-哌啶酸的转化效率。上催化反应是在10mL的反应体现中进行的，该反应体现中含有：L-哌啶酸30g/L，5～10wt的重组酶(1wt指转化1g主原料需要1g脯氨酸羟化酶变体重组湿细胞。)，37.3g/Lα-酮戊二酸，6.1g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。
表2.脯氨酸羟化酶多点突变的活性比较
表2中提供了多个位点的氨基酸改变对脯氨酸羟化酶催化活性的影响。该脯氨酸羟化酶表达自E.Coli BL21细胞，催化活性是基于该酶对L-哌啶酸的转化效率。该催化过程是在20mL的反应体系中进行的，其中含有L-哌啶酸50g/L，2～5wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。
在一些实施方案中，选定与脯氨酸羟化酶活性相关的一个或多个氨基酸残基作为核心保持不变，而在其他氨基酸残基位置引入新的突变，可以产生具有改进性质的脯氨酸羟化酶。因此，上述优选实施例中的任意一个脯氨酸羟化酶都可以作为利用基因工程手段合成其他脯氨酸羟化酶突变体的母本氨基酸序列。例如通过几轮进化而得到氨基酸残基与表1和表2中的氨基酸序列有区别的新突变体。
以上述所公开的任何一个脯氨酸羟化酶，或者在上述所公开的氨酸羟化酶及其变体基础上在其他氨基酸残基位置上进行一个或多个氨基酸残基的突变所得到的新的具有脯氨酸羟化酶活性的突变体都在本发明的保护范围内。此处可以举例说明，但不限于此，含有E27氨基酸残基突变的脯氨酸羟化酶突变体，能够进一步的进行一个或多个其他氨基酸的突变，例如：H14、S16、T25、F26、D30、S33、E34、Y35、S37、I39、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223、S225。另一个例子是，含有I39氨基酸残基突变的脯氨酸羟化酶突变体，能够进一步的进行一个或多个其他氨基酸的突变，例如：H14、S16、T25、F26、E27、D30、S33、E34、Y35、S37、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223、S225。又一个例子是，含有I58氨基酸残基突变的脯氨酸羟化酶突变体，能够进一步的进行一个或多个其他氨基酸的突变，例如：H14、S16、T25、F26、E27、D30、S33、E34、Y35、S37、I39、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223、S225。
上述羟化酶具有脯氨酸羟化酶活性，同时能够催化(2S)-哌啶-2-羧酸转化为(2S,5S)-5-羟基哌啶-2-羧酸，并通过基因工程方法对酶的催化活性进行改进，在某些实施方案中突变体的羟化酶活性比SEQ ID NO：2自身编码的羟化酶的活性提高了1倍、2倍、3倍、4倍、5倍或更高。
在本发明另一种典型的实施方式中，还提供了一种DNA分子，该DNA分子编码上述任一种羟化酶。编码的上述羟化酶具有专一性高和催化活性显著提高的优势。
在本发明又一种典型的实施方式中，还提供了一种重组质粒，该重组质粒连接有上述DNA分子。该DNA分子能够编码上述任一种选择性高，且/或催化活性显著提高的脯氨酸羟化酶。具体序列选自表1和表2中的编号为奇数的序列或者与这些序列在保持上述氨基酸位点变化的前提下，其他位点的氨基酸序列发生取代、添加或缺失突变的核苷酸序列。
上述重组质粒中，任意能够用于表达上述羟化酶的DNA分子的重组质粒均适用于本发明。在本发明的优选实施例中，重组质粒选自如下之一：pET-22b(+)、pET-21b(+)、pET-3a(+)、pET-3d(+)、pET-11a(+)、pET-12a(+)、pET-14b(+)、pET-15b(+)、pET-16b(+)、pET-17b(+)、pET-19b(+)、pET-20b(+)、pET-21a(+)、pET-23a(+)、pET-23b(+)、pET-24a(+)、pET-25b(+)、pET-26b(+)、pET-27b(+)、pET-28a(+)、pET-29a(+)、pET-30a(+)、pET-31b(+)、pET-32a(+)、pET-35b(+)、pET-38b(+)、pET-39b(+)、pET-40b(+)、pET-41a(+)、pET-41b(+)、pET-42a(+)、pET-43a(+)、pET-43b(+)、pET-44a(+)、pET-49b(+)、pQE2、pQE9、pQE30、pQE31、pQE32、pQE40、pQE70、pQE80、pRSET-A、pRSET-B、pRSET-C、pGEX-5X-1、pGEX-6p-1、pGEX-6p-2、pBV220、pBV221、pBV222、pTrc99A、pTwin1、pEZZ18、pKK232-18、pUC-18以及pUC-19。
在本发明再一种典型的实施方式中，还提供了一种宿主细胞，该宿主细胞含有上述任一种重组质粒。具体的宿主细胞可以为原核细胞或真核细胞，优选真核细胞为酵母细胞。更优选地，上述宿主细胞为感受态细胞，进一步优选感受态细胞为大肠杆菌BL21细胞或大肠杆菌W3110。
在本发明的另一种典型的实施方式中，还提供了一种生产L-羟脯氨酸衍生物的方法，该方法包括：以L-脯氨酸衍生物为底物，利用上述任一种羟化酶催化底物的羟基化，得到通式(I)所示的L-羟脯氨酸衍生物；
其中，R1选自(C1-C5)的亚烷基或(C2-C5)亚烯基；R2选自(C0-C4)亚烷基或(C2-C4)亚烯基；R3选自羟基、氨基、(C1-C6)烷氧基、芳氧基、(C1-C6)烷硫基和芳硫基；R4选自氢、(C1-C6)烷基、(C2-C6)烯基和(C2-C6)炔基。
利用本发明的上述脯氨酸羟化酶，将L-脯氨酸衍生物催化为其羟化物，不仅能够提高催化效率，提高L-脯氨酸衍生物的转化率，而且提高了催化的专一性，使得到的L-羟脯氨酸衍生物的目的产物纯度得到提高，减少了后续分离的步骤。
上述利用脯氨酸羟化酶催化合成L-羟脯氨酸衍生物的化学反应方程式如图2所示。其中，是α-酮戊二酸，在脯氨酸羟化酶催化反应中，α-酮戊二酸和O2作为共同底物。在铁离子为辅因子共同作用下，催化底物的羟基化，得到L-羟脯氨酸衍生物；是维生素C，又名抗坏血酸，在脯氨酸羟化酶催化反应中，主要起循环铁离子的作用。
上述脯氨酸羟化酶为一种典型的双加氧酶，催化反应与现有的脯氨酸羟化酶一样，均需要α-酮戊二酸和O2存在，且需要铁离子作为辅因子。因而，上述方法包括以α-酮戊二酸和O2为共同底物，以铁离子为辅因子，在脯氨酸羟化酶的催化下催化底物的羟基化，得到L-羟脯氨酸衍生物。
具体的反应条件可以在现有的脯氨酸羟化酶反应体系的基础上进行适当调整得到。比如，可以对还原剂(例如抗坏血酸)的浓度，洗涤剂浓度，pH，温度，缓冲液，溶剂系统，基底装载，多肽加载，压力和反应时间等进行适当调整。在某些实施方案中，具体的反应条件如下：底物15～120g/L，1.5g～48g/L的羟化酶，1～2.5eq(eq：代表所用物料的质量与主原料质量的比值；)α-酮戊二酸，0.1～0.3eqL-抗坏血酸，1mM～10mM硫酸亚铁铵，反应pH6～8，反应温度5～30℃，反应时间6～96h。在某些实施方案中，合适的反应条件包括向反应液中通入2～5L/h的氧气，及0.5～2％的消泡剂。
在本发明一种优选的实施例中，上述脯氨酸羟化酶能够在5～45℃的反应温度下催化底物的羟基化，得到通式(I)所示的L-羟脯氨酸衍生物。更优选在5～15℃的反应温度下进行上述催化反应。本发明所提高的脯氨酸羟化酶不仅能够在更低的温度下将底物催化为其羟化物，而且能够提高催化专一性(即选择性)，提高产物的纯度。
上述酶能够对多种L-脯氨酸衍生物进行羟基化。在本发明一种优选的实施例中，对L-脯氨酸或-L哌啶酸进行羟基化，得到顺式-4-羟基-L-脯氨酸或(2S,5S)-5-羟基哌啶-2-羧酸。对上述两种底物进行羟基化的专一性最高，可以将底物100％转化为顺式-4-羟基-L-脯氨酸或(2S,5S)-5-羟基哌啶-2-羧酸。
下面将结合具体的实施例来进一步说明本发明的有益效果。下述实验方法如无特别说明，均为常规方法，所使用的实验材料如无特别说明，均可容易地从商业公司获取。
实施例1：脯氨酸羟化酶的重组表达
将源于Kordia jejudonensis的注释为假定蛋白的DNA编码序列SEQ ID NO:1进行密码子优化(按照大肠杆菌密码子偏好性及简并性进行密码子改进，由苏州金唯智生物科技有限公司设计完成)，得到优化后的DNA序列SEQ ID NO:3，编码的脯氨酸羟化酶多肽序列为SEQ ID NO：4。将SEQ ID NO:3的编码序列连接到pET22b(+)表达载体(购买于Novagen，产品编号69744)中，然后转化到大肠杆菌BL21(DE3)中，涂布于含有50μg/ml氨苄青霉素的LB培养皿中，37℃培养过夜。挑取上述培养皿上的单克隆于试管中活化后，接种于500ml含50μg/ml氨苄青霉素的LB液体培养基中，37℃振荡培养至OD600＝0.6时，加入IPTG至终浓度为1mM，在25℃下进行诱导表达。诱导16h后，6000g离心10min收集菌体。菌体用超声破碎仪(JY92-2D，宁波新芝生物科技股份有限公司)破碎细胞，4℃，10000g离心20min获得上清液，用于野生型脯氨酸羟化酶活性检测，作为突变体活性筛选的对照。
实施例2：脯氨酸羟化酶变体的制备
以含SEQ ID NO：3序列的pET22b(+)表达载体为模版，以带有突变位点的引物通过全质粒PCR获得完整的线性片段，将上述PCR产物经DPnⅠ消化除去母本模版后，转化到大肠杆菌BL21(DE3)中，并涂布于含有50μg/ml氨苄青霉素的LB培养皿中，37℃培养过夜。获得含有脯氨酸羟化酶变体氨基酸序列的单克隆，再通过试诱导和基因测序确定突变位点。最终获得的带有单一突变位点的突变体，然后再以单一突变位点的突变为突变母本，再用其他位点带突变的引物进行再次全质粒PCR，再次确定突变位点。
将突变体菌种活化后，接种于500ml含50μg/ml氨苄青霉素的LB液体培养基中，37℃振荡培养至OD600＝0.6时，加入IPTG至终浓度为1mM，在25℃下进行诱导表达。诱导16h后，6000g离心10min收集菌体。菌体用超声破碎仪(JY92-2D，宁波新芝生物科技股份有限公司)破碎细胞，4℃，10000g离心20min获得上清液，用于脯氨酸羟化酶变体活性检测。
实施例3：脯氨酸羟化酶变体的活性筛选
与SEQ ID NO：2相比存在一个氨基酸残基差异的脯氨酸羟化酶变体的活性筛选采用如下10mL的反应液进行筛选，10mL的反应液中含有：L-哌啶酸30g/L，5～10wt(1wt指转化1g主原料需要1g脯氨酸羟化酶变体重组湿细胞。)的重组粗酶，37.3g/Lα-酮戊二酸，6.1g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取200μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率，活性筛选结果如表1所示(表1是根据所有转化率数据比较后得到的活性筛选结果)。
与SEQ ID NO：2相比存在多个氨基酸残基差异的脯氨酸羟化酶变体的活性筛选采用如下20mL的反应液进行筛选。20mL的反应液中含有：L-哌啶酸50g/L，2～5wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率，活性筛选结果如表2所示(表2是根据所有转化率数据比较后得到的活性筛选结果)。
实施例4：脯氨酸羟化酶突变体的克隆与表达
为了便于羟化酶突变体的表达以及鉴定，在其基因的5’和3’末端设计了兼容的限制性酶切位点。可以采用NdeⅠ和XhoⅠ分别将目的基因和pET-22b(+)(其他可在大肠杆菌中表达蛋白质的表达质粒也可使用)分别同时进行酶切，酶切后的目的基因和质粒的较大片段用T4DNA连接酶进行连接反应，将连接产物转化到大肠杆菌DH5α菌株的感受态细胞中，然后将转化后的感受态细胞涂布于含有50μg/ml氨苄青霉素的LB培养平板上，37℃培养过夜。
挑取上述培养皿上长出的单菌落接种于含有50μg/ml氨苄青霉素的LB液体培养基中，37℃振荡培养过夜，收集菌体进行质粒提取、PCR鉴定和双酶切鉴定后，将正确的克隆载体命名为pET22b(+)-R-M并转化到大肠杆菌BL21(DE3)中，将转化的大肠杆菌BL21(DE3)涂布于含有50μg/ml氨苄青霉素的LB培养平板上，37℃培养过夜。挑取上述培养平板上长出的单菌落并接种于5ml含50μg/ml氨苄青霉素的LB液体培养基中，利用菌落PCR进行鉴定，将含有正确的表达载体的大肠杆菌进行后续的诱导表达。将上述菌液转接于500ml含50μg/ml氨苄青霉素的LB液体培养基中，37℃振荡培养至OD600＝0.5～0.6时，加入IPTG至终浓度分别为0.2～1.0mM，在18～25℃下进行诱导表达10～16h后，取出菌液，6000g离心10min收集菌体，于-20℃冻存备用。菌体用超声破碎仪(JY92-2D，宁波新芝生物科技股份有限公司)破碎细胞，4℃，10000g离心20min获得上清液和沉淀，上清液用垂直电泳仪进行SDS-PAGE检测。表达的羟化酶突变体在SDS-PAGE上呈现的分子量约为30KD。
实施例5：表1和表2中的羟化酶与野生型脯氨酸羟化酶的性能比较
按照图1所示的羟化酶催化合成(2S,5S)-5-羟基哌啶-2-羧酸(或称顺式-5-羟基哌啶酸)的化学反应过程进行如下实验：
SEQ ID NO:2编码的野生型脯氨酸羟化酶催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，10wt的羟化酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为98.47％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.36％。
SEQ ID NO:16编码的E27K氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，10wt/8wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为10wt和8wt的反应体系中，40h时，转化率分别为100％，94.53％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.32％。
SEQ ID NO:122编码的N165H氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，10wt和6.67wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为10wt和6.67wt的反应体系中，40h时，转化率分别为100％，90.41％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为99.89％。
又一更优的单点突变体，SEQ ID NO:104编码的K123D氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，8wt/5wt/4wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为8wt，5wt和4wt的反应体系中，40h时，转化率分别为100％，97.83％，89.09％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.68％。
多点突变体，SEQ ID NO:180编码的S37C+I223V氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，8wt/5wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为8wt和5wt的反应体系，40h时，转化率分别为100％和93.99％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.25％。
多点突变体，SEQ ID NO:178编码的I39R+Y35W氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，5wt/3wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为5wt和3wt的反应体系中，40h时，转化率分别为100％和94.45％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为75.02％。虽然该突变体的选择性差一些，但其催化活性明显高于野生型脯氨酸羟化酶。
一更优的组合突变体，SEQ ID NO:140编码的I39R+K123D氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，4wt/3wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为4wt和3wt的反应体系中，40h时，转化率分别为97.35％和92.08％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为99.26％。
又一更优的组合突变体，SEQ ID NO:160编码的S37C+I39K+I223V+K123D氨基酸残基突变的脯氨酸羟化酶突变体催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，3wt/2wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为3wt和2wt的反应体系中，40h时，转化率分别为95.23％和88.41％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为99.90％。
实施例6：
发明人自主设计并构建的氨基酸序列PH1(见序列表中SEQ ID NO:289)与SEQ IDNO:2同源性78％，该蛋白用于催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸。采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，9wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为98.56％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.45％。
实施例7：
1)SEQ ID NO:2氨基酸序列中I39K+S33N氨基酸残基突变后，与SEQ ID NO:2具有99％的同源性，其氨基酸序列见序列表中的290号。该序列编码的突变酶用于催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸。采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，8wt/6wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。重组酶用量分别为8wt和6wt的反应体系中，40h时，转化率分别为100％，91.62％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.89％。
2)SEQ ID NO:2氨基酸序列中E27K+I39K+F28L+S31A氨基酸残基突变后，与SEQ IDNO:2具有98.6％的同源性，其氨基酸序列见序列表中的SEQ ID NO:291号。该序列编码的突变酶用于催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸。采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，5wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为98.75％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.30％。
3)以实施例6中构建的与SEQ ID NO:2同源性为78％的SEQ ID NO:289序列为基础进行突变，将该序列的E27K氨基酸残基突变后，得到序列表中的SEQ ID NO:292所示的氨基酸序列，其与SEQ ID NO:2具有97.8％的同源性。该序列编码的突变脯氨酸羟化酶用于催化L-哌啶酸制备(2S,5S)-5-羟基哌啶-2-羧酸。采用如下20mL的反应液。20mL的反应液中含有：L-哌啶酸50g/L，7wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为97.25％，(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为98.40％。
对比例1：
现有技术中使用一种来源于Sinorhizobium meliloti菌的重组脯氨酸羟化酶制备(2S,5S)-5-羟基哌啶-2-羧酸过程中，最高的(2S,5S)-5-羟基哌啶-2-羧酸的非对应异构体过量为90.7％，有9.3％的位置异构体(2S,3R)-3-羟基哌啶-2-羧酸产生(WO2013169725A2)。
与实施例5相比，SEQ ID NO:2的羟化酶的非对应异构体过量为98.36％，而对比例1的非对应异构体过量为90.7％。可见，本发明中的SEQ ID NO:2的羟化酶的选择性优于现有技术。
实施例8：
表1和表2中的羟化酶在顺式-4-羟基-L-脯氨酸制备中的应用，其反应过程如图2所示。
SEQ ID NO:104编码的K123D氨基酸残基突变的脯氨酸羟化酶突变体催化L-脯氨酸制备顺式-4-羟基-L-脯氨酸，采用如下20mL的反应液进行制备。20mL的反应液中含有：L-脯氨酸50g/L，5wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为92.36％，非对应异构体过量为99.30％。
SEQ ID NO:160编码的S37C+I39K+I223V+K123D氨基酸残基突变的脯氨酸羟化酶突变体L-脯氨酸制备顺式-4-羟基-L-脯氨酸采用如下20mL的反应液。20mL的反应液中含有：L-脯氨酸50g/L，4wt的重组酶，62.2g/Lα-酮戊二酸，10.2g/L的L-抗坏血酸，5mM硫酸亚铁铵，反应pH6.5，反应温度10℃，反应时间40h。反应结束取100μL反应体系，加入200μL乙腈，混匀后再加入3000μL纯化水，10000rpm离心5min收集上清液用于HPLC检测转化率。40h时，转化率为99.47％，非对应异构体过量为99.56％。
可见，表1和表2中的很多突变体羟化酶还可以应用在顺式-4-羟基-L-脯氨酸制备中，还可以得到高纯度和高选择性的顺式-4-羟基-L-脯氨酸。
从以上的描述中，可以看出，本发明上述的实施例实现了如下技术效果：通过采用以SEQ ID NO:2作为突变的脯氨酸羟化酶筛选的基础序列，通过基因工程的手段，得到了多个脯氨酸羟化酶催化活性及选择性显著提高的羟化酶。这些羟化酶具有使底物转化率高以及催化专一位置异构体少，能特异性的催化脯氨酸衍生物的羟基化作用。尤其是能够催化L-哌啶酸的羟化化而产生的(2S,5S)-5-羟基哌啶-2-羧酸(或称顺式-5-羟基哌啶酸)以及L-脯氨酸的羟基化而产生顺式-4-羟基-L-脯氨酸。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
SEQUENCE LISTING
<110> 凯莱英医药集团（天津）股份有限公司;
凯莱英生命科学技术（天津）有限公司；
天津凯莱英制药有限公司；
凯莱英医药化学（阜新）技术有限公司；
吉林凯莱英医药化学有限公司;
<120> 脯氨酸羟化酶及其应用
<130> PN47502KLY
<160> 292
<170> PatentIn version 3.5
<210> 1
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<212> DNA
<213> Kordia jejudonensis
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<213> Kordia jejudonensis
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Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
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<220>
<223> 密码子优化的DNA序列
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<220>
<223> 来源于Kordia jejudonensis的密码子优化后的氨基酸
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<210> 5
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<213> Artificial Sequence
<220>
<223> H14R突变体的基因序列
<400> 5
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 6
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R突变体的氨基酸序列
<400> 6
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 7
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S16N突变体的基因序列
<400> 7
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgaatga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 8
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S16N突变体的氨基酸序列
<400> 8
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Asn
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 9
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> T25G突变体的基因序列
<400> 9
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acggtttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 10
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> T25G突变体的氨基酸序列
<400> 10
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Gly Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 11
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> T25R突变体的基因序列
<400> 11
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg accgtttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 12
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> T25R突变体的氨基酸序列
<400> 12
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Arg Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 13
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> F26L突变体的基因序列
<400> 13
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactctgga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 14
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> F26L突变体的氨基酸序列
<400> 14
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Leu Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 15
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K突变体的基因序列
<400> 15
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 16
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K突变体的氨基酸序列
<400> 16
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 17
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> D30S突变体的基因序列
<400> 17
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacagc agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 18
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> D30S突变体的氨基酸序列
<400> 18
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Ser Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 19
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S33N突变体的基因序列
<400> 19
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacaatg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 20
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S33N突变体的氨基酸序列
<400> 20
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Asn Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 21
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E34N突变体的基因序列
<400> 21
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagca attacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 22
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E34N突变体的氨基酸序列
<400> 22
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Asn Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 23
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E34G突变体的基因序列
<400> 23
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg gttacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 24
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E34G突变体的氨基酸序列
<400> 24
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 25
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E34L突变体的基因序列
<400> 25
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcc tgtacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 26
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E34L突变体的氨基酸序列
<400> 26
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Leu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 27
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E34D突变体的基因序列
<400> 27
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg attacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 28
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E34D突变体的氨基酸序列
<400> 28
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Asp Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 29
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E34S突变体的基因序列
<400> 29
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagca gctacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 30
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E34S突变体的氨基酸序列
<400> 30
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Ser Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 31
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> Y35W突变体的基因序列
<400> 31
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatgggcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 32
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> Y35W突变体的氨基酸序列
<400> 32
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 33
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> Y35K突变体的基因序列
<400> 33
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aaaaagcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 34
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> Y35K突变体的氨基酸序列
<400> 34
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Lys Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 35
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37W突变体的基因序列
<400> 35
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg gggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 36
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37W突变体的氨基酸序列
<400> 36
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Trp Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 37
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37F突变体的基因序列
<400> 37
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttt tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 38
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37F突变体的氨基酸序列
<400> 38
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 39
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37E突变体的基因序列
<400> 39
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgctga aggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 40
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37E突变体的氨基酸序列
<400> 40
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Glu Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 41
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37N突变体的基因序列
<400> 41
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgctaa tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 42
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37N突变体的氨基酸序列
<400> 42
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Asn Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 43
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37T突变体的基因序列
<400> 43
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgctac cggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 44
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37T突变体的氨基酸序列
<400> 44
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Thr Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 45
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C突变体的基因序列
<400> 45
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 46
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C突变体的氨基酸序列
<400> 46
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 47
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39K突变体的基因序列
<400> 47
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 48
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39K突变体的氨基酸序列
<400> 48
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 49
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39R突变体的基因序列
<400> 49
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 50
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39R突变体的氨基酸序列
<400> 50
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 51
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> W40F突变体的基因序列
<400> 51
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtattttt 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 52
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> W40F突变体的氨基酸序列
<400> 52
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Phe Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 53
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K41E突变体的基因序列
<400> 53
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
gaaacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 54
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K41E突变体的氨基酸序列
<400> 54
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Glu Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 55
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> D54G突变体的基因序列
<400> 55
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg gtcatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 56
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> D54G突变体的氨基酸序列
<400> 56
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Gly His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 57
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H55Q突变体的基因序列
<400> 57
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accagctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 58
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H55Q突变体的氨基酸序列
<400> 58
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp Gln Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 59
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S57L突变体的基因序列
<400> 59
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgct gattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 60
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S57L突变体的氨基酸序列
<400> 60
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Leu Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 61
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58T突变体的基因序列
<400> 61
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc taccgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 62
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58T突变体的氨基酸序列
<400> 62
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Thr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 63
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58Y突变体的基因序列
<400> 63
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 64
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58Y突变体的氨基酸序列
<400> 64
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 65
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58A突变体的基因序列
<400> 65
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgcagaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 66
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58A突变体的氨基酸序列
<400> 66
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ala Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 67
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58R突变体的基因序列
<400> 67
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tcgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 68
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58R突变体的氨基酸序列
<400> 68
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Arg Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 69
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58V突变体的基因序列
<400> 69
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgttgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 70
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58V突变体的氨基酸序列
<400> 70
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Val Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 71
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58S突变体的基因序列
<400> 71
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tagcgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 72
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58S突变体的氨基酸序列
<400> 72
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ser Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 73
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I58C突变体的基因序列
<400> 73
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 74
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I58C突变体的氨基酸序列
<400> 74
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Cys Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 75
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K86P突变体的基因序列
<400> 75
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagccgga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 76
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K86P突变体的氨基酸序列
<400> 76
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Pro Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 77
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> T91A突变体的基因序列
<400> 77
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag gcagtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 78
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> T91A突变体的氨基酸序列
<400> 78
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Ala Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 79
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> F95Y突变体的基因序列
<400> 79
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgtatatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 80
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> F95Y突变体的氨基酸序列
<400> 80
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Tyr Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 81
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> C97Y突变体的基因序列
<400> 81
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgta tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 82
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> C97Y突变体的氨基酸序列
<400> 82
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Tyr Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 83
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I98V突变体的基因序列
<400> 83
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 84
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I98V突变体的氨基酸序列
<400> 84
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 85
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K106V突变体的基因序列
<400> 85
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacgttga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 86
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K106V突变体的氨基酸序列
<400> 86
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Val Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 87
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K106T突变体的基因序列
<400> 87
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaccga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 88
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K106T突变体的氨基酸序列
<400> 88
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Thr Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 89
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K106Q突变体的基因序列
<400> 89
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 90
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K106Q突变体的氨基酸序列
<400> 90
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 91
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> F111S突变体的基因序列
<400> 91
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa agcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 92
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> F111S突变体的氨基酸序列
<400> 92
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Ser Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 93
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K112E突变体的基因序列
<400> 93
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 94
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K112E突变体的氨基酸序列
<400> 94
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 95
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I118F突变体的基因序列
<400> 95
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg ttttcacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 96
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I118F突变体的氨基酸序列
<400> 96
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Phe His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 97
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H119R突变体的基因序列
<400> 97
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccgtatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 98
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H119R突变体的氨基酸序列
<400> 98
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile Arg Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 99
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H119F突变体的基因序列
<400> 99
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatctttatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 100
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H119F突变体的氨基酸序列
<400> 100
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile Phe Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 101
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I120V突变体的基因序列
<400> 101
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacgtt 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 102
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I120V突变体的氨基酸序列
<400> 102
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Val Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 103
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123D突变体的基因序列
<400> 103
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 104
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123D突变体的氨基酸序列
<400> 104
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 105
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123N突变体的基因序列
<400> 105
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 106
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123N突变体的氨基酸序列
<400> 106
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asn Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 107
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123Q突变体的基因序列
<400> 107
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgcaga tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 108
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123Q突变体的氨基酸序列
<400> 108
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Gln Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 109
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123S突变体的基因序列
<400> 109
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgagca tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 110
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123S突变体的氨基酸序列
<400> 110
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ser Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 111
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123I突变体的基因序列
<400> 111
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgatta tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 112
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123I突变体的氨基酸序列
<400> 112
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ile Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 113
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123T突变体的基因序列
<400> 113
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgacca tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 114
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123T突变体的氨基酸序列
<400> 114
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Thr Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 115
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> T130N突变体的基因序列
<400> 115
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgaat tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 116
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> T130N突变体的氨基酸序列
<400> 116
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Asn Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 117
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> D134G突变体的基因序列
<400> 117
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag gtgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 118
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> D134G突变体的氨基酸序列
<400> 118
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Gly Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 119
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> V135K突变体的基因序列
<400> 119
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag ataaagttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 120
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> V135K突变体的氨基酸序列
<400> 120
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Lys Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 121
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> N165H突变体的基因序列
<400> 121
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 122
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> N165H突变体的氨基酸序列
<400> 122
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 123
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K209R突变体的基因序列
<400> 123
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatccgtggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 124
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K209R突变体的氨基酸序列
<400> 124
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Arg Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 125
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I223V突变体的基因序列
<400> 125
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 126
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I223V突变体的氨基酸序列
<400> 126
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 127
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ K123D突变体的基因序列
<400> 127
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 128
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+ K123D突变体的氨基酸序列
<400> 128
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 129
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ I39K突变体的基因序列
<400> 129
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 130
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K突变体的氨基酸序列
<400> 130
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 131
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39K+K123Q突变体的基因序列
<400> 131
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacaa aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 132
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39K+K123Q突变体的氨基酸序列
<400> 132
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Lys Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 133
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39K+ K123D突变体的基因序列
<400> 133
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 134
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39K+ K123D突变体的氨基酸序列
<400> 134
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 135
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C+I223V+ K123D突变体的基因序列
<400> 135
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 136
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C+I223V+ K123D突变体的氨基酸序列
<400> 136
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 137
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123D+N165H突变体的基因序列
<400> 137
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 138
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123D+N165H突变体的氨基酸序列
<400> 138
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 139
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39R+K123D突变体的基因序列
<400> 139
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 140
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39R+K123D突变体的氨基酸序列
<400> 140
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 141
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123D突变体的基因序列
<400> 141
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 142
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123D突变体的氨基酸序列
<400> 142
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 143
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ I39R突变体的基因序列
<400> 143
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 144
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+ I39R突变体的氨基酸序列
<400> 144
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 145
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39R+N165H突变体的基因序列
<400> 145
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 146
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39R+N165H突变体的氨基酸序列
<400> 146
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 147
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39K+N165H突变体的基因序列
<400> 147
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 148
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39K+N165H突变体的氨基酸序列
<400> 148
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 149
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ I39K+K123D突变体的基因序列
<400> 149
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 150
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+ I39K+K123D突变体的氨基酸序列
<400> 150
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 151
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C+I223V+N165H突变体的基因序列
<400> 151
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 152
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C+I223V+N165H突变体的氨基酸序列
<400> 152
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 153
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123D+W40F突变体的基因序列
<400> 153
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtattttt 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 154
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123D+W40F突变体的氨基酸序列
<400> 154
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Phe Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 155
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+N165H突变体的基因序列
<400> 155
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 156
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+N165H突变体的氨基酸序列
<400> 156
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 157
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ S37C+I223V+ K123D突变体的基因序列
<400> 157
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 158
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+ S37C+I223V+ K123D突变体的氨基酸序列
<400> 158
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 159
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C+I39K+I223V+ K123D突变体的基因序列
<400> 159
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 160
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C+I39K+I223V+ K123D突变体的氨基酸序列
<400> 160
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 161
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+K123D+I98V突变体的基因序列
<400> 161
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacattc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 162
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+K123D+I98V突变体的氨基酸序列
<400> 162
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 163
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+K123D+I223V突变体的基因序列
<400> 163
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 164
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+K123D+I223V突变体的氨基酸序列
<400> 164
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 165
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> F26L+E27K+I39K+K123D突变体的基因序列
<400> 165
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactctgaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 166
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> F26L+E27K+I39K+K123D突变体的氨基酸序列
<400> 166
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Leu Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 167
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I223V+S37C+E27K+I39K突变体的基因序列
<400> 167
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 168
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I223V+S37C+E27K+I39K突变体的氨基酸序列
<400> 168
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 169
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I223V+S37C+E27K+N165H突变体的基因序列
<400> 169
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 170
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I223V+S37C+E27K+N165H突变体的氨基酸序列
<400> 170
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 171
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123D+Y35W突变体的基因序列
<400> 171
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatgggcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 172
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123D+Y35W突变体的氨基酸序列
<400> 172
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 173
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K123D+Y35W+I120V突变体的基因序列
<400> 173
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatgggcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacgtg 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 174
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K123D+Y35W+I120V突变体的氨基酸序列
<400> 174
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Val Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 175
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+Y35W突变体的基因序列
<400> 175
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatgggcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 176
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+Y35W突变体的氨基酸序列
<400> 176
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 177
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I39R+Y35W突变体的基因序列
<400> 177
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatgggcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 178
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39R+Y35W突变体的氨基酸序列
<400> 178
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 179
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C+I223V突变体的基因序列
<400> 179
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttg tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 180
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C+I223V突变体的氨基酸序列
<400> 180
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 181
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+Y35W突变体的基因序列
<400> 181
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatgggcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 182
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+Y35W突变体的氨基酸序列
<400> 182
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 183
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+I98V+K123D+I223V突变体的基因序列
<400> 183
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 184
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37C+I39K+I98V+K123D+I223V突变体的氨基酸序列
<400> 184
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Cys Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 185
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123D+N165H突变体的基因序列
<400> 185
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctggata tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tccatctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 186
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123D+N165H突变体的氨基酸序列
<400> 186
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Asp Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile His Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 187
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39R+Y35W突变体的基因序列
<400> 187
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatgggcttc tggtcgttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 188
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39R+Y35W突变体的氨基酸序列
<400> 188
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Trp Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 189
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> S37C+I39K+N165H突变体的基因序列
<400> 189
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatta tcactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcggtaaa 480
gtgaaacgta tcgttctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 190
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> S37C+I39K+N165H突变体的氨基酸序列
<400> 190
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Ile Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Gly Lys
145 150 155 160
Val Lys Arg Ile Val Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 191
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+ I39K+D30S突变体的基因序列
<400> 191
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacagc agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 192
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+ I39K+D30S突变体的氨基酸序列
<400> 192
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Ser Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 193
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34N突变体的基因序列
<400> 193
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagca attacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 194
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34N突变体的氨基酸序列
<400> 194
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Asn Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 195
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34G突变体的基因序列
<400> 195
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 196
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34G突变体的氨基酸序列
<400> 196
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 197
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37W突变体的基因序列
<400> 197
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttg gggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 198
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37W突变体的氨基酸序列
<400> 198
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Trp Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 199
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37E突变体的基因序列
<400> 199
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctga aggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 200
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37E突变体的氨基酸序列
<400> 200
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Glu Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 201
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+Y35K突变体的基因序列
<400> 201
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aaaaagcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 202
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+Y35K突变体的氨基酸序列
<400> 202
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Lys Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 203
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N突变体的基因序列
<400> 203
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 204
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N突变体的氨基酸序列
<400> 204
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 205
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123Q突变体的基因序列
<400> 205
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgcaga tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 206
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123Q突变体的氨基酸序列
<400> 206
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Gln Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 207
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123S突变体的基因序列
<400> 207
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgagca tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 208
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123S突变体的氨基酸序列
<400> 208
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ser Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 209
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K106Q+K112E突变体的基因序列
<400> 209
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 210
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K106Q+K112E突变体的氨基酸序列
<400> 210
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 211
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> I98V+E27K+I39K突变体的基因序列
<400> 211
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 212
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I98V+E27K+I39K突变体的氨基酸序列
<400> 212
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 213
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+I118F突变体的基因序列
<400> 213
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg ttttcacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 214
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+I118F突变体的氨基酸序列
<400> 214
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Phe His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 215
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37T突变体的基因序列
<400> 215
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctac cggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 216
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37T突变体的氨基酸序列
<400> 216
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Thr Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 217
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123I突变体的基因序列
<400> 217
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgatta tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 218
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+K123I突变体的氨基酸序列
<400> 218
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ile Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 219
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> K106Q+K112E+I223V突变体的基因序列
<400> 219
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg aatacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 220
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> K106Q+K112E+I223V突变体的氨基酸序列
<400> 220
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 221
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E34G+K106Q+K112E+I223V突变体的基因序列
<400> 221
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcga gttcaacgac agctacagcg gttacgcttc tggtatttgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 222
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E34G+K106Q+K112E+I223V突变体的氨基酸序列
<400> 222
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 223
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34L突变体的基因序列
<400> 223
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcc tgtacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 224
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+E34L突变体的氨基酸序列
<400> 224
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Leu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 225
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58T突变体的基因序列
<400> 225
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc taccgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 226
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58T突变体的氨基酸序列
<400> 226
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Thr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 227
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58Y突变体的基因序列
<400> 227
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 228
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58Y突变体的氨基酸序列
<400> 228
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 229
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58A突变体的基因序列
<400> 229
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgcagaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 230
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58A突变体的氨基酸序列
<400> 230
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ala Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 231
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58R突变体的基因序列
<400> 231
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tcgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 232
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58R突变体的氨基酸序列
<400> 232
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Arg Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 233
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58V突变体的基因序列
<400> 233
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgttgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 234
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58V突变体的氨基酸序列
<400> 234
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Val Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 235
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58S突变体的基因序列
<400> 235
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tagtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 236
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37F+I58S突变体的氨基酸序列
<400> 236
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ser Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 237
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58C突变体的基因序列
<400> 237
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 238
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58C突变体的氨基酸序列
<400> 238
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Cys Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 239
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+D30S+I39K+I58R+K112E突变体的基因序列
<400> 239
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacagc agctacagcg aatacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tcgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcgagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 240
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+D30S+I39K+I58R+K112E突变体的氨基酸序列
<400> 240
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Ser Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Arg Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 241
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+E34N+I39K+I58Y+I223V突变体的基因序列
<400> 241
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagca attacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 242
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+E34N+I39K+I58Y+I223V突变体的氨基酸序列
<400> 242
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Asn Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 243
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37N+I39K+I58Y+D173G突变体的基因序列
<400> 243
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagca attacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccgtata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 244
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37N+I39K+I58Y+D173G突变体的氨基酸序列
<400> 244
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Asn Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Tyr Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 245
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37F+I39K+I58Y+D173G突变体的基因序列
<400> 245
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagct tttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccgtata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 246
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37F+I39K+I58Y+D173G突变体的氨基酸序列
<400> 246
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Phe Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Tyr Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 247
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58A突变体的基因序列
<400> 247
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgcagaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 248
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58A突变体的氨基酸序列
<400> 248
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ala Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 249
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58R+K123Q突变体的基因序列
<400> 249
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg aatacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tcgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgcaga tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 250
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+S37N+I58R+K123Q突变体的氨基酸序列
<400> 250
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Arg Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Gln Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 251
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+S37F+I39K+I58Y+D173G+I118R突变体的基因序列
<400> 251
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagct tttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tcgtcacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccgtata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 252
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+S37F+I39K+I58Y+D173G+I118R突变体的氨基酸序列
<400> 252
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Phe Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Arg His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Tyr Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 253
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58R突变体的基因序列
<400> 253
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcc tgtacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tcgtgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 254
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58R突变体的氨基酸序列
<400> 254
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Leu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Arg Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 255
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58Y+D173G突变体的基因序列
<400> 255
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcc tgtacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tcgtcacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 256
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58Y+D173G突变体的氨基酸序列
<400> 256
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Leu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Arg His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 257
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58Y+D173G+K123Q突变体的基因序列
<400> 257
atggaatcca agatcatcgg taaagtcaac ttcgaggagc atctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcc tgtacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc ttatgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tatcaacggc 300
ctgatcatcc cacacaaaga ctacctggaa ttcaagaaag gcttcacccg tcgtcacatc 360
ccgctgcaga tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacatcc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 258
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+E34L+S37N+I39K+I58Y+D173G+K123Q突变体的氨基酸序列
<400> 258
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Leu Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Tyr Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Arg His Ile Pro Leu Gln Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 259
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+D30S+E34G+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 259
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacagc agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 260
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+D30S+E34G+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 260
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Ser Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 261
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34N+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 261
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagca attacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 262
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34N+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 262
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Asn Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 263
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 263
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 264
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 264
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 265
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37W+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 265
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttg gggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 266
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37W+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 266
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Trp Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 267
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37F+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 267
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttt tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 268
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37F+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 268
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Phe Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 269
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37E+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 269
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgctga aggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 270
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37E+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 270
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Glu Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 271
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+Y35K+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 271
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gtaaagcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 272
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+Y35K+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 272
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Lys Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 273
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37N+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 273
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgctaa tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 274
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37N+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 274
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Asn Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 275
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123Q+I223V突变体的基因序列
<400> 275
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgcaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 276
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123Q+I223V突变体的氨基酸序列
<400> 276
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Gln Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 277
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123S+I223V突变体的基因序列
<400> 277
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgagca tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 278
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123S+I223V突变体的氨基酸序列
<400> 278
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ser Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 279
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I58A+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 279
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgcagaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 280
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I58A+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 280
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ala Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 281
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+I118F+I223V突变体的基因序列
<400> 281
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tttccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 282
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+I118F+I223V突变体的氨基酸序列
<400> 282
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Phe His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 283
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37T+I39K+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 283
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgctac cggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 284
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+S37T+I39K+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 284
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Thr Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 285
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I58V+I98V+K106Q+K112E+I223V突变体的基因序列
<400> 285
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tgttgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgaaaa tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 286
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I58V+I98V+K106Q+K112E+I223V突变体的氨基酸序列
<400> 286
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Val Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 287
<211> 831
<212> DNA
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123I+I223V突变体的基因序列
<400> 287
atggaatcca agatcatcgg taaagtcaac ttcgaggagc gtctgctgga taaagagctg 60
aaactgatcg acactttcaa attcaacgac agctacagcg gttacgcttc tggtaaatgg 120
aagacttgca tgctgtggaa ccgttctggt cagaaagatg accatctgtc tattgaacat 180
gatacctacg tgaaacctac tgaatacggc aaacagctgg catacgtaaa cgaaatcatc 240
gctaacacct ttaagaaaga acacatcaag acggtgcgtc tgttcatgtg tgttaacggc 300
ctgatcatcc cacaccagga ctacctggaa ttcgaaaaag gcttcacccg tatccacatc 360
ccgctgatta tcaacgaaca cgcactgacc tctgaagaag atgttgttta caacatgcag 420
aaaggtgaaa tttggttcat cgaaggccgt aaaatccaca gcgctgccaa tttcagcaaa 480
gtgaaacgta tcaacctggt catcgacttc gcgccggata ttccgtttga agaactgttc 540
ctgaattctg agaactatca accgaacctg atcccgaaaa tctctcaacg tacccagctg 600
aaagaagaag agctgggtta tatcaaaggt ctgtctaaaa ttatcaatga aatgaacttt 660
gacgacgttc tgtccattct gagcaaaatt cacttctatc gcaacgtttc ctccgaactg 720
gtttttggct ggctggacga aattgccacg gcgtccaaca actataacat tcagcgcaaa 780
gcgcaggaag taaccgatct gctgattcgc aaaggcccga ttaataacta a 831
<210> 288
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> H14R+E27K+E34G+I39K+I98V+K106Q+K112E+K123I+I223V突变体的氨基酸序列
<400> 288
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu Arg Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Gly Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Val Asn Gly Leu Ile Ile Pro His Gln Asp Tyr Leu Glu Phe Glu
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Ile Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Val Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 289
<211> 280
<212> PRT
<213> Artificial Sequence
<220>
<223> PH1的氨基酸序列
<400> 289
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Lys Glu Leu Thr Asp Glu Val Ile Glu Gly Ile
195 200 205
Leu Gly Phe Ser Ile Ile Ile Ser Glu Ala Asn Tyr Arg Glu Ile Val
210 215 220
Ser Ile Leu Ala Lys Leu His Phe Phe Tyr Lys Ala Asp Cys Arg Ser
225 230 235 240
Met Tyr Asp Trp Leu Lys Glu Ile Cys Lys Arg Arg Gly Asp Pro Ala
245 250 255
Leu Ile Glu Lys Thr Ala Ser Leu Glu Arg Phe Phe Leu Gly His Arg
260 265 270
Glu Arg Gly Glu Val Met Thr Tyr
275 280
<210> 290
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> I39K+S33N的氨基酸序列
<400> 290
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Glu Phe Asn Asp Ser Tyr
20 25 30
Asn Glu Tyr Ala Ser Gly Lys Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 291
<211> 276
<212> PRT
<213> Artificial Sequence
<220>
<223> E27K+I39K+F28L+S31A的氨基酸序列
<400> 291
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Leu Asn Asp Ala Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Arg Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Thr Gln Leu Lys Glu Glu Glu Leu Gly Tyr Ile
195 200 205
Lys Gly Leu Ser Lys Ile Ile Asn Glu Met Asn Phe Asp Asp Ile Leu
210 215 220
Ser Ile Leu Ser Lys Ile His Phe Tyr Arg Asn Val Ser Ser Glu Leu
225 230 235 240
Val Phe Gly Trp Leu Asp Glu Ile Ala Thr Ala Ser Asn Asn Tyr Asn
245 250 255
Ile Gln Arg Lys Ala Gln Glu Val Thr Asp Leu Leu Ile Arg Lys Gly
260 265 270
Pro Ile Asn Asn
275
<210> 292
<211> 280
<212> PRT
<213> Artificial Sequence
<220>
<223> PH1突变E27K后的氨基酸序列
<400> 292
Met Glu Ser Lys Ile Ile Gly Lys Val Asn Phe Glu Glu His Leu Leu
1 5 10 15
Asp Lys Glu Leu Lys Leu Ile Asp Thr Phe Lys Phe Asn Asp Ser Tyr
20 25 30
Ser Glu Tyr Ala Ser Gly Ile Trp Lys Thr Cys Met Leu Trp Asn Arg
35 40 45
Ser Gly Gln Lys Asp Asp His Leu Ser Ile Glu His Asp Thr Tyr Val
50 55 60
Lys Pro Thr Glu Tyr Gly Lys Gln Leu Ala Tyr Val Asn Glu Ile Ile
65 70 75 80
Ala Asn Thr Phe Lys Lys Glu His Ile Lys Thr Val Arg Leu Phe Met
85 90 95
Cys Ile Asn Gly Leu Ile Ile Pro His Lys Asp Tyr Leu Glu Phe Lys
100 105 110
Lys Gly Phe Thr Arg Ile His Ile Pro Leu Lys Ile Asn Glu His Ala
115 120 125
Leu Thr Ser Glu Glu Asp Val Val Tyr Asn Met Gln Lys Gly Glu Ile
130 135 140
Trp Phe Ile Glu Gly Arg Lys Ile His Ser Ala Ala Asn Phe Ser Lys
145 150 155 160
Val Lys Arg Ile Asn Leu Val Ile Asp Phe Ala Pro Asp Ile Pro Phe
165 170 175
Glu Glu Leu Phe Leu Asn Ser Glu Asn Tyr Gln Pro Asn Leu Ile Pro
180 185 190
Lys Ile Ser Gln Arg Lys Glu Leu Thr Asp Glu Val Ile Glu Gly Ile
195 200 205
Leu Gly Phe Ser Ile Ile Ile Ser Glu Ala Asn Tyr Arg Glu Ile Val
210 215 220
Ser Ile Leu Ala Lys Leu His Phe Phe Tyr Lys Ala Asp Cys Arg Ser
225 230 235 240
Met Tyr Asp Trp Leu Lys Glu Ile Cys Lys Arg Arg Gly Asp Pro Ala
245 250 255
Leu Ile Glu Lys Thr Ala Ser Leu Glu Arg Phe Phe Leu Gly His Arg
260 265 270
Glu Arg Gly Glu Val Met Thr Tyr
275 280

Claims (14)

1.一种脯氨酸羟化酶，其特征在于，包括：
(a)具有SEQ ID NO:2所示的氨基酸序列的蛋白质；
(b)SEQ ID NO:2所示的氨基酸序列经过一个或多个氨基酸的突变且具有脯氨酸羟化酶活性的蛋白质；或者
(c)保留所述(b)中的一个或多个氨基酸的突变、具有脯氨酸羟化酶活性，且与(b)中蛋白质的氨基酸序列至少具有78％的同源性的蛋白质。
2.根据权利要求1所述的脯氨酸羟化酶，其特征在于，所述突变的氨基酸位点选自H14、S16、T25、F26、E27、D30、S33、E34、Y35、S37、I39、W40、K41、D54、H55、S57、I58、K86、T91、F95、C97、I98、K106、F111、K112、K162、L166、I118、H119、I120、K123、T130、D134、V135、S154、N165、D173、K209、I223以及S225中的一个或多个。
3.根据权利要求2所述的脯氨酸羟化酶，其特征在于，所述突变包括如下任意一个或多个：H14R、S16N、T25G、T25R、F26L、E27K、D30S、S33N、E34N、E34G、E34L、E34S、E34D、Y35W、Y35K、S37W、S37F、S37E、S37N、S37T、S37C、I39K、I39R、W40F、K41E、D54G、H55Q、S57L、I58T、I58Y、I58A、I58R、I58V、I58S、I58C、K86P、T91A、F95Y、C97Y、I98V、K106V、K106T、K106Q、F111S、K112E、K112R、S154A、K162E、L166M、I118F、I118V、I118R、H119R、H119F、I120V、K123D、K123N、K123Q、K123S、K123I、K123T、T130N、D134G、V135K、N165H、D173G、K209R、I223V、以及S225A。
4.根据权利要求1至3中任一项所述的脯氨酸羟化酶，其特征在于，所述突变包括如下任意一种组合：E27K+Y35W/K、E27K+I39K/R、E27K+K123D/I/Q/S、E27K+N165H、I39K/R+Y35W/K、I39K/R+K123D/I/Q/S、I39K/R+N165H、K123D+W40F、K123D+Y35W/K、E27K+I39K/R+K123D/I/Q/S、K123D/I/Q/S+N165H、S37C/E/F/N/W/T+I223V、E27K+Y35W/K+I39K/R、E27K+S37C/E/F/N/W/T+I39K/R、E27K+E34N/G/L/D/S+I39K/R、E27K+I39K/R+D30S、E27K+I39K/R+I118F/V/R、E27K+I39K/R+I98V、S37C/E/F/N/W/T+I223V+N165H、Y35W/K+S37C/E/F/N/W/T+W40F、S37C/E/F/N/W/T+I223V+K123D/I/Q/S、E27K+I39K+Y35W/K+S37C/E/F/N/W/T、E27K+I39K/R+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+K106Q+K112E、E27K+I39K/R+Y35W/K+S37C/E/F/N/W/T+K123D/I/Q/S、E27K+I39K/R+S37C/E/F/N/W/T+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I223V+K123D/I/Q/S、S37C/E/F/N/W/T+I39K/R+I223V+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I98V、E27K+S37C/E/F/N/W/T+I39K/R+K123D/I/Q/S+I223V、F26L+E27K+I39K/R+K123D/I/Q/S、I223V+S37C/E/F/N/W/T+E27K+I39K/R、I223V+S37C/E/F/N/W/T+E27K+N165H、E27K+S37C/E/F/N/W/T+I39K/R+I98V+K123D/I/Q/S+I223V、K106Q+K112E+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K123D/I/Q/S、E27K+I39K/R+K123D/I/Q/S+N165H、H14R+E34G+K106Q+K112E+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+K86P、K123D/I/Q/S+Y35W/K+I120V、E27K+D30S+I39K/R+I58A/C/R/S/T/V/Y+K112E、S37C/E/F/N/W/T+I39K/R+N165H、E27K+E34N/G/L/D/S+I39K/R+I58A/C/R/S/T/V/Y+I223V、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+I118F/V/R、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R、E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+N165H、E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/+D173G+K123D/I/Q/S、H14R+E27K+D30S+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、T25G/R+E27K+S37C/E/F/N/W/T+I39K/R+I58A/C/R/S/T/V/Y+D173G+K123D/I/Q/S+I118F/V/R+N165H、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+S37C/E/F/N/W/T+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+Y35W/K+I39K/R+I98V+K106V/T/Q+K112E/R+I223V、H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+K123D/I/Q/S+I223V、H14R+E27K+E34N/G/L/D+I39K/R+I58A/C/R/S/T/V/Y+I98V+K106V/T/Q+K112E/R+I223V以及H14R+E27K+E34N/G/L/D/S+I39K/R+I98V+K106V/T/Q+K112E/R+I118F/V/R+I223V，其中，“/”表示“或者”。
5.一种DNA分子，其特征在于，所述DNA分子编码权利要求1至4中任一项所述的脯氨酸羟化酶。
6.一种重组质粒，其特征在于，所述重组质粒连接有权利要求5所述的DNA分子。
7.根据权利要求6所述的重组质粒，其特征在于，所述重组质粒选自如下之一：pET-21b(+)、pET-22b(+)、pET-3a(+)、pET-3d(+)、pET-11a(+)、pET-12a(+)、pET-14b(+)、pET-15b(+)、pET-16b(+)、pET-17b(+)、pET-19b(+)、pET-20b(+)、pET-21a(+)、pET-23a(+)、pET-23b(+)、pET-24a(+)、pET-25b(+)、pET-26b(+)、pET-27b(+)、pET-28a(+)、pET-29a(+)、pET-30a(+)、pET-31b(+)、pET-32a(+)、pET-35b(+)、pET-38b(+)、pET-39b(+)、pET-40b(+)、pET-41a(+)、pET-41b(+)、pET-42a(+)、pET-43a(+)、pET-43b(+)、pET-44a(+)、pET-49b(+)、pQE2、pQE9、pQE30、pQE31、pQE32、pQE40、pQE70、pQE80、pRSET-A、pRSET-B、pRSET-C、pGEX-5X-1、pGEX-6p-1、pGEX-6p-2、pBV220、pBV221、pBV222、pTrc99A、pTwin1、pEZZ18、pKK232-18、pUC-18以及pUC-19。
8.一种宿主细胞，其特征在于，所述宿主细胞含有权利要求6或7所述的重组质粒。
9.根据权利要求8所述的宿主细胞，其特征在于，所述宿主细胞为原核细胞或真核细胞，优选所述真核细胞为酵母细胞。
10.根据权利要求8所述的宿主细胞，其特征在于，所述宿主细胞为感受态细胞，优选所述感受态细胞为大肠杆菌BL21细胞或大肠杆菌W3110。
11.一种生产L-羟脯氨酸衍生物的方法，其特征在于，所述方法包括：以L-脯氨酸衍生物为底物，利用权利要求1至4中任一项所述的脯氨酸羟化酶催化所述底物的羟基化，得到通式(I)所示的L-羟脯氨酸衍生物；
其中，R1选自C1-C5的亚烷基或C2-C5亚烯基；R2选自C0-C4亚烷基或C2-C4亚烯基；R3选自羟基、氨基、C1-C6烷氧基、芳氧基、C1-C6烷硫基或C1-C6芳硫基；R4选自氢、C1-C6烷基、C2-C6烯基或C2-C6炔基。
12.根据权利要求11所述的方法，其特征在于，所述方法包括：以α-酮戊二酸和O2为共同底物，以铁离子为辅因子，利用所述脯氨酸羟化酶催化所述底物的羟基化，得到所述L-羟脯氨酸衍生物。
13.根据权利要求11或12所述的方法，其特征在于，所述L-羟脯氨酸衍生物为顺式-4-羟基-L-脯氨酸或(2S,5S)-5-羟基哌啶-2-羧酸。
14.根据权利要求11或12所述的方法，其特征在于，所述脯氨酸羟化酶在5～45℃的反应温度，优选5～15℃的反应温度下催化所述底物的羟基化，得到所述通式(I)所示的L-羟脯氨酸衍生物。
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