CN114761552A - 新的丝氨酸蛋白酶变体 - Google Patents
新的丝氨酸蛋白酶变体 Download PDFInfo
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- CN114761552A CN114761552A CN202080082417.0A CN202080082417A CN114761552A CN 114761552 A CN114761552 A CN 114761552A CN 202080082417 A CN202080082417 A CN 202080082417A CN 114761552 A CN114761552 A CN 114761552A
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Abstract
本申请涉及新的丝氨酸蛋白酶变体以及包含所述变体的饲料组合物,所述变体具有优于已有丝氨酸蛋白酶的酶活性并且具有高温活性和热抗性。
Description
技术领域
本申请涉及新的丝氨酸蛋白酶变体。
背景技术
蛋白酶参与各种功能,例如生物体内的消化、吸收和防御,并且根据活性位点的结构分为丝氨酸蛋白酶、半胱氨酸蛋白酶、天冬氨酸蛋白酶和金属蛋白酶。丝氨酸蛋白酶(或丝氨酸内肽酶)的特征在于在它们的活性位点中共同地具有切割蛋白质中肽键的活性丝氨酸残基,其中丝氨酸在蛋白酶的活性位点作为亲核氨基酸(Hedstrom,2002.Chem Rev 102:4501-4524)。
丝氨酸蛋白酶已经在多种应用中使用。除了治疗人类疾病(例如,溶解血凝块)的治疗应用之外,丝氨酸蛋白酶不仅用作洗衣除垢剂和隐形眼镜清洁剂的组分,而且还用于乳蛋白的改性、蚕丝脱胶、皮革浸泡、脱毛、寡肽的合成、从肺X射线胶片中回收银、饲料和食品的生产和改进等(韩国专利公开号10-2005-0068750)。目前仍然需要开发具有提高的热稳定性、增加的活性等性质的丝氨酸蛋白酶,以获得更高的工业成本效益和效率。
在这种情况下,本申请发明人进行了大量的努力,并最终开发出源自Thermobifida fusca的丝氨酸蛋白酶的新变体,并且发现该新丝氨酸蛋白酶变体的活性显著大于常规丝氨酸蛋白酶的活性,从而完成了本申请中的各项发明。
实施方案描述
技术问题
本申请的一个目的是提供丝氨酸蛋白酶变体。
本申请的另一个目的是提供编码丝氨酸蛋白酶变体的多核苷酸和包含多核苷酸的载体。
本申请的另一个目的是提供表达丝氨酸蛋白酶变体的微生物。
本申请的另一个目的是提供饲料组合物,所述饲料组合物包含丝氨酸蛋白酶变体和表达该丝氨酸蛋白酶变体的微生物中的至少一种。
本申请有益效果
本申请的丝氨酸蛋白酶变体由于与常规丝氨酸蛋白酶相比具有增强的活性以及高温下活性和耐热性而可有效地用于各种工业领域。
附图简要描述
本申请的丝氨酸蛋白酶变体由于与常规丝氨酸蛋白酶相比具有增强的活性以及高温下活性和耐热性而可有效地用于各种工业领域。
技术方案
下文详细描述本申请的内容。同时,在本申请中公开的每处描述和实施方案可以在本文应用于不同的描述和实施方案。换言之,本申请中公开的各种组分的所有组合都包括在本申请的范围内。此外,本申请的范围不应受下文提供的描述的限制。
此外,本领域普通技术人员可以认识到或能够使用不超过常规实验来确认本申请的具体实施方案的多种等同方案。这种等同方案旨在包含在本申请的范围内。
本申请的一个方面提供了丝氨酸蛋白酶变体。
本文所用术语“丝氨酸蛋白酶”是指属于蛋白酶亚组并具有蛋白水解活性的酶。具体而言,丝氨酸蛋白酶可以是通过水解肽键而降解蛋白质并且基本上在其活性位点具有活性丝氨酸残基的酶,并且更具体而言,是具有组氨酸、天冬氨酸和丝氨酸的氨基酸残基的空间排列(可以被称为催化三联体)的酶。
根据本申请的丝氨酸蛋白酶可以源自Thermobifida属的微生物。特别地,在本申请中,野生型丝氨酸蛋白酶可以是源自Thermobifida fusca的丝氨酸蛋白酶,特别是,具有源自SEQ ID NO:40的氨基酸序列或源自SEQ ID NO:2的氨基酸序列的多肽,更特别是,包括SEQ ID NO:31所示氨基酸序列的多肽,但不限于此。SEQ ID NO:2、31和40的氨基酸序列可以从已知数据库获得,例如国家生物技术信息中心(NCBI)的GenBank数据库。
本申请的丝氨酸蛋白酶可以具有与上述氨基酸序列具有相同活性的任何序列,但不限于此。此外,丝氨酸蛋白酶可以具有SEQ ID NO:31的氨基酸序列或与SEQ ID NO:31的氨基酸序列具有至少80%同源性或同一性的氨基酸序列,但不限于此。具体而言,丝氨酸蛋白酶可以包含SEQ ID NO:31中所示的氨基酸或与SEQ ID NO:31具有至少80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%同源性或同一性的氨基酸。另外,显然,只要氨基酸序列具有上述同源性或同一性以及与丝氨酸蛋白酶等同的效果,在部分序列中具有包括缺失、修饰、取代或添加在内的氨基酸序列的任何蛋白质都在本申请的范围内。
换言之,尽管在本申请中使用的表述是“具有预定的SEQ ID NO:中所示的氨基酸序列的蛋白质或多肽”和“包含预定的SEQ ID NO:中所示的氨基酸序列的蛋白质或多肽”,但是能清楚判断的是,在本申请中也可以使用具有在部分序列中包括缺失、修饰、取代或添加的氨基酸序列的任何蛋白质,只要该蛋白具有与由相应氨基酸序列组成的多肽相同或等同的活性。例如,“包含SEQ ID NO:31的氨基酸序列的多肽”显然属于“由SEQ ID NO:31的氨基酸序列组成的多肽”,只要前者具有与后者相同或等同的活性。
本文所用术语“同源性”或“同一性”是指两个给定氨基酸序列或核苷酸序列之间的相关性程度,并且可以显示为百分比。术语同源性和同一性可以互换使用。
保守多核苷酸或多肽的序列同源性或同一性可以通过标准比对算法确定,并且可以一起使用由要使用的程序建立的默认缺口罚分。基本上,同源或相同的序列通常可以沿着整个序列或整个序列的至少约50%、60%、70%、80%或90%在中等或高度严格条件下彼此杂交。在杂交的多核苷酸中,也考虑含有简并密码子代替密码子的多核苷酸。
任何两个给定多核苷酸或多肽之间的序列同源性、相似性或同一性可以使用已知的计算机算法,如“FASTA”程序,通过使用Pearson et al.(1988)Proc.Natl.Acad.Sci.USA85:2444的默认参数来确定。或者,可以使用Needleman-Wunsch算法(1970,J.Mol.Biol.48:443–453),其在欧洲分子生物学开放软件套件(European Molecular Biology OpenSoftware Suite(EMBOSS))包的Needleman程序中进行(Rice et al.,2000,TrendsGenet.16:276–277)(5.0.0版或其后的版本)(GCG程序包(包括GCG程序包)(Devereux,J.,et al.,Nucleic Acids Research 12:387(1984))、BLASTP、BLASTN、FASTA(Atschul,S.F.,et al.,J MOLEC BIOL 215:403(1990);Guide to Huge Computers,Martin J.Bishop,ed.,Academic Press,San Diego,1994,和CARILLO et al.(1988)SIAM J Applied Math48:1073)。例如,可以使用来自国家生物技术信息中心数据库(National Center forBiotechnology Information database)的BLAST或ClustalW来确定同源性、相似性或同一性。
多核苷酸或多肽的同源性、相似性或同一性可以通过GAP计算机程序比较序列信息来确定,例如Needleman et al.,(1970),J Mol Biol.48:443,如Smith and Waterman,Adv.Appl.Math(1981)2:482所公开的程序。简言之,GAP程序将相似性定义为通过将相似比对的符号(即核苷酸或氨基酸)的数目除以两个序列中较短序列中符号的总数而获得的值。用于GAP程序的默认参数可以包括:(1)二进制比较矩阵(包含相同的值1和不相同的值0)和Gribskov et al.(1986)Nucl.Acids Res.14:6745的加权比较矩阵,公开于Schwartz andDayhoff,eds.,Atlas Of Protein Sequence And Structure,National BiomedicalResearch Foundation,pp.353–358(1979)(或EDNAFULL(NCBI NUC4.4的EMBOSS版本)取代矩阵);(2)每个缺口的罚分为3.0,并且每个缺口中的每个符号的额外的0.10罚分(或缺口开放罚分为10,缺口延伸罚分为0.5);(3)端部缺口无罚分。
此外,任何两个给定的多核苷酸或多肽之间的序列同源性、相似性或同一性可以通过在限定的严格条件下、通过Southern杂交比较它们的序列来确认,并且限定的适当杂交条件在本技术的范围内并且可以通过本领域普通技术人员熟知的方法来确定。
在具有丝氨酸蛋白酶活性的上述蛋白中,本申请中提供的丝氨酸蛋白酶变体可以指这样的变体:在该变体中,在具体位置上的氨基酸被取代,从而与突变前的蛋白相比具有超过100%的酶活性。
本文所用术语“变体”是指通过保守取代和/或修饰所述序列的至少一个不同氨基酸、同时保留蛋白质的功能或特性而获得的多肽。由于数个氨基酸的取代、缺失或添加,变体具有与所鉴定的序列不同的氨基酸序列。这种变体通常可以通过修饰上述多肽序列之一并评价修饰的多肽的性质来鉴定。也就是说,相对于天然蛋白,变体的能力可以增强、不改变或减弱。
此外,一些变体可以包括其中至少一部分(如N末端前导序列或跨膜结构域)被去除的变体。其它变体可以包括从成熟蛋白的N末端和/或C末端去除一部分或向成熟蛋白的N末端和/或C末端添加一部分的变体。
术语“变体”也可以与其它术语互换使用,例如修饰/突变的蛋白质,修饰,修饰的多肽,突变体,突变蛋白和发散体(divergent),并且也可以没有限制地使用用于指示变异的任何术语。
与天然野生型或未修饰的蛋白质相比,变体可具有增强的活性,但不限于此。
本文所用术语“保守取代”是指用具有相似结构和/或化学性质的不同氨基酸取代一个氨基酸。例如,变体具有至少一个保守取代,同时保留至少一项生物活性。这种氨基酸取代通常可以基于残基的极性、电荷、溶解度、疏水性、亲水性和/或两亲性质的相似性。例如,带正电荷的(碱性)氨基酸包括精氨酸、赖氨酸和组氨酸;带负电的(酸性)氨基酸包括谷氨酸和天冬氨酸;芳族氨基酸包括苯丙氨酸、色氨酸和酪氨酸;疏水氨基酸包括丙氨酸、缬氨酸、异亮氨酸、亮氨酸、蛋氨酸、苯丙氨酸、酪氨酸和色氨酸。另外,氨基酸可以被分类为带有侧链的带电氨基酸和带有侧链的不带电氨基酸。带有侧链的带电氨基酸包括天冬氨酸、谷氨酸、赖氨酸、精氨酸和组氨酸,带有侧链的不带电氨基酸分为非极性氨基酸或极性氨基酸。非极性氨基酸包括甘氨酸、丙氨酸、缬氨酸、亮氨酸、异亮氨酸、蛋氨酸、苯丙氨酸、色氨酸和脯氨酸,极性氨基酸包括丝氨酸、苏氨酸、半胱氨酸、酪氨酸、天冬酰胺和谷氨酰胺。通常,保守取代对产生的多肽的活性几乎没有或没有影响。
变体还可以包括对多肽的性质和二级结构具有最小影响的氨基酸的缺失或添加。例如,以共翻译的方式或翻译后方式,多肽可以与参与蛋白质转移的蛋白质的N末端的信号(或前导)序列缀合。所述多肽也可以与不同的序列或接头缀合,以鉴定、纯化或合成多肽。
本文所用术语“丝氨酸蛋白酶变体”是指包括在具有丝氨酸蛋白酶活性的多肽的氨基酸序列中的至少一个氨基酸的取代的多肽。根据本申请的丝氨酸蛋白酶变体可以包括这样的变体:在所述变体中,对应于SEQ ID NO:31的氨基酸序列自N末端第12位的氨基酸和/或对应于SEQ ID NO:31的氨基酸序列自N末端第116位的氨基酸各自被不同的氨基酸取代。具体而言,变体可以是这样的蛋白质:在所述变体中,SEQ ID NO:31的氨基酸序列自N末端第12位氨基酸或第116位氨基酸或第12位和第116位氨基酸均被不同的氨基酸取代。术语“不同的氨基酸”是指与取代前的氨基酸不同的氨基酸,可以使用与取代前不同的任何氨基酸,但不限于此。
具体而言,根据本申请的丝氨酸蛋白酶变体可以是这样的变体:在所述变体中,SEQ ID NO:31的氨基酸序列的第12位的苯丙氨酸被甘氨酸、丙氨酸、精氨酸、天冬氨酸、半胱氨酸、谷氨酸、天冬酰胺、谷氨酰胺、组氨酸、脯氨酸、丝氨酸、酪氨酸、异亮氨酸、亮氨酸、赖氨酸、色氨酸、缬氨酸、蛋氨酸或苏氨酸取代;和/或在第116位的天冬酰胺被甘氨酸、丙氨酸、精氨酸、天冬氨酸、半胱氨酸、谷氨酸、谷氨酰胺、组氨酸、脯氨酸、丝氨酸、酪氨酸、异亮氨酸、亮氨酸、赖氨酸、苯丙氨酸、色氨酸、缬氨酸、蛋氨酸或苏氨酸取代,但不限于此。
具体而言,变体可以是这样的蛋白质:在所述变体中,对应于SEQ ID NO:31的氨基酸序列的第12位的氨基酸被酪氨酸(Y)、丝氨酸(S)、丙氨酸(A)或精氨酸(R)取代;对应于氨基酸序列第116位的氨基酸被天冬氨酸(D)、丝氨酸(S)、苏氨酸(T)或甘氨酸(G)取代;或者SEQ ID NO:31的氨基酸序列的第12和116位的氨基酸各自被酪氨酸(Y)和天冬氨酸(D),酪氨酸(Y)和丝氨酸(S),丝氨酸(S)和天冬氨酸(D),丝氨酸(S)和苏氨酸(T),或丙氨酸(A)和甘氨酸(G)取代,但不限于此。显然,SEQ ID NO:31的氨基酸序列的第12位和/或第116位氨基酸各自被不同的氨基酸取代的变体包括其中对应于每个位置的氨基酸各自被不同的氨基酸取代的变体。
另外,变体还包括这样的变体:在所述变体中,在对应于SEQ ID NO:31自N末端的第12位和第116位氨基酸的位置上的氨基酸分别被SEQ ID NO:31所示的氨基酸序列或与SEQ ID NO:31的氨基酸序列具有至少80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的同源性或同一性的氨基酸序列中的不同氨基酸取代。
具体而言,在上述变体中,在对应于SEQ ID NO:31的氨基酸序列的第12位和/或第116位氨基酸的位置上的氨基酸各自被不同的氨基酸取代的变体可以由选自SEQ ID NO:32至39的氨基酸序列的任何一个氨基酸序列组成,但不限于此。这些变体可以是具有丝氨酸蛋白酶活性的变体,其中在对应于SEQ ID NO:31的氨基酸序列的第12位和/或第116位的位置上的氨基酸各自被不同的氨基酸取代,并且与SEQ ID NO:31的氨基酸序列具有至少80%、90%、95%、96%、97%、98%或99%且小于100%的同源性。
本申请的丝氨酸蛋白酶变体与修饰前的多肽、天然野生型多肽或未修饰的多肽相比,可具有增强的活性,但不限于此。此外能够清楚理解的是,在部分氨基酸序列具有缺失、修饰、取代和添加的氨基酸序列的任何蛋白质都在本申请的范围内,只要该蛋白质具有上述同源性以及与丝氨酸蛋白酶等同的效果。
此外能够清楚理解的是,除了在第12位和/或第116位氨基酸中提供特定活性的突变之外,在相应的SEQ ID NO的氨基酸序列的正向或反向中添加序列、或具有天然存在的突变、或具有沉默突变的任何变体不排除在本申请的范围之外,只要该变体具有与根据本申请的变体相同或等同的活性。
同时,NCBI参考序列WP_016188200.1(SEQ ID NO:40)的成熟区对应于本申请的SEQ ID NO:31的氨基酸序列,并且SEQ ID NO:40除信号肽之外的序列对应于本申请的SEQID NO:2。
如上所述能够清楚理解的是,本申请的丝氨酸蛋白酶变体可以包括对丝氨酸蛋白酶的性质和二级结构具有很小影响的氨基酸的缺失或添加,其中在对应于SEQ ID NO:31的第12位和/或第116位氨基酸的位置上的氨基酸各自被不同的氨基酸取代。此外,通过本领域公知的序列比对,本领域技术人员能够清楚理解的是,本申请的SEQ ID NO:31自N末端的第12位和第116位对应于SEQ ID NO:40的第193位和第297位以及SEQ ID NO:2的第163位和第267位,并且SEQ ID NO:31包括在SEQ ID NO:40和SEQ ID NO:2中。.
因此,关于各自包括SEQ ID NO:31的氨基酸序列的SEQ ID NO:2和40的氨基酸序列,本申请的丝氨酸蛋白酶变体包括这样的变体:在所述变体中,对应于SEQ ID NO:31的第12位和第116位氨基酸(SEQ ID NO:2中的第163位和/或第267位氨基酸以及SEQ ID NO:40中的第193位和/或第297位氨基酸)的位置处的氨基酸各自被取代。另外,上文关于SEQ IDNO:31及第12位和第116位氨基酸给出的描述也适用于SEQ ID NO:2及其第163位和第267位氨基酸和SEQ ID NO:40及其第193位和第297位氨基酸。
根据一个实施方案,本申请的丝氨酸蛋白酶变体可以是这样的变体:在所述变体具有的氨基酸序列中,对应于SEQ ID NO:31的第12位和/或第116位的位置的氨基酸各自被不同的氨基酸取代,并且所述变体与SEQ ID NO:2的氨基酸序列具有至少80%且小于100%的同源性。根据另一个实施方案,本申请的丝氨酸蛋白酶变体可以是这样的变体:在所述变体中,SEQ ID NO:2的第163位和/或第267位氨基酸各自被不同的氨基酸取代,并且所述变体与SEQ ID NO:2的氨基酸序列具有至少80%且小于100%的同源性,并且所述变体与选自SEQ ID NO:3至10的氨基酸序列的任何一个氨基酸序列具有至少80%的同源性,但不限于此。
本申请的另一方面提供了编码丝氨酸蛋白酶变体的多核苷酸。
本文所用术语“多核苷酸”是指核苷酸的聚合物,其中核苷酸单体通过共价键以长链形状彼此连接,并且通常是指具有特定最小长度的DNA或RNA链,更具体而言,是指编码本申请变体的多核苷酸片段。
编码本申请的丝氨酸蛋白酶变体的多核苷酸可以具有编码根据本申请的具有增强的活性的丝氨酸蛋白酶变体的任何核苷酸序列,但不限于此。在本申请中,编码野生型丝氨酸蛋白酶的基因可以源自Thermobifida属的微生物,特别是Thermobifida fusca,但不限于此。
在不改变氨基酸序列的范围内,本申请的多核苷酸可以包括在多肽的氨基酸序列中的编码区中进行的各种修饰,这是由于密码子简并性或考虑了多肽将表达于的活生物体的偏好密码子。具体而言,在对应于SEQ ID NO:31的自N末端的第12位和/或第116位氨基酸的位置上的氨基酸分别被不同的氨基酸取代的编码变体的任何核苷酸序列都可以包括在其中,但不限于此。
例如,本申请的多核苷酸可以是编码本申请的变体的多核苷酸序列,并且具体而言,编码由SEQ ID NO:32至39中所示的氨基酸序列中的任一个组成的多肽或与其具有同源性的多肽,但不限于此。更具体而言,多核苷酸可以是由选自SEQ ID NO:41至48的任一SEQID NO中所示的多核苷酸序列组成的多核苷酸,但不限于此。此外,如上所述,由于本申请的丝氨酸蛋白酶变体包括其中对应于SEQ ID NO:31的第12位和/或第116位氨基酸的氨基酸(SEQ ID NO:2中的第163位和第267位氨基酸以及SEQ ID NO:40中的第193位和/或第297位氨基酸)各自被取代的那些变体,其中SEQ ID NO:2和40的氨基酸序列包括SEQ ID NO:31的氨基酸序列,编码丝氨酸蛋白酶变体的多核苷酸序列也在本申请的范围内。例如,编码丝氨酸蛋白酶变体的多核苷酸序列可以是编码选自SEQ ID NO:3至10的任一SEQ ID NO中所示的氨基酸序列的多核苷酸序列,具体而言,可以是由选自SEQ ID NO:23至30的任一SEQ IDNO中所示的多核苷酸序列组成的多核苷酸序列,但不限于此。
另外,可以包括编码具有变体活性的蛋白质的任何序列,其中在严格条件下,通过与可以由已知基因序列(例如,与核苷酸序列的全部或部分互补的序列)制备的探针杂交,在所述变体中对应于SEQ ID NO:31的自N末端的第12位和/或第116位氨基酸的位置处的氨基酸各自被不同的氨基酸取代,但不限于此。
术语“严格条件”是指能够在多核苷酸之间进行特异性杂交的条件。这样的条件具体公开于文献中。例如,严格条件可以包括如下所述的条件,具有高同源性的基因(至少40%、特别是至少90%、更特别是至少95%、还更特别是至少97%、并且甚至还更特别是至少99%的同源性的基因)实现杂交,但具有比上述同源性更低的同源性的基因不实现杂交的条件;或者用于Southern杂交的典型洗涤条件,即,洗涤在对应于60℃、1×SSC和0.1%SDS,特别是60℃、0.1×SSC和0.1%SDS,更特别是68℃、0.1×SSC和0.1%SDS的盐浓度和温度下进行一次,特别是两次或三次。然而,严格条件不限于此,而是可以由本领域技术人员根据其目的适当地调节。
杂交需要两个核苷酸具有互补序列,但是根据杂交严格性,碱基之间的错配是可能的。术语“互补”用于描述可彼此杂交的核苷酸碱基之间的关系。例如,在DNA中,腺嘌呤与胸腺嘧啶互补,胞嘧啶与鸟嘌呤互补。因此,本申请不仅可以包括基本上相似的核苷酸序列,而且可以包括与整个序列互补的分离的核酸片段。
具体而言,具有同源性的多核苷酸可以在55℃的Tm值下通过使用包括杂交步骤的杂交条件和使用上述条件来检测。此外,Tm值可以是60℃、63℃或65℃,但不限于此,并且可以由本领域技术人员根据目的适当地控制。
多核苷酸杂交的适当严格程度可以取决于多核苷酸的长度和互补程度,其参数是本领域熟知的。
本申请的又一方面提供了包含编码本申请的丝氨酸蛋白酶变体的多核苷酸的载体。
本文所用术语“载体”是指含有编码目标蛋白的多核苷酸的核苷酸序列的DNA构建体,所述编码目标蛋白的多核苷酸与适当的控制序列可操作地连接,以在适当的宿主中表达目标多核苷酸。控制序列可以包括能够启动转录的启动子,用于控制转录的任何操纵子序列,用于编码适当的mRNA核糖体结合位点的序列,和用于控制转录和翻译终止的序列,在转化入适当的宿主后,载体可以独立于宿主的基因组复制或起作用,或者可以整合到基因组本身中。
本文所用术语“可操作地连接”是指编码本申请的目标蛋白的多核苷酸序列与启动和介导多核苷酸转录的启动子序列功能性连接。可操作的连接可以通过本领域已知的遗传重组技术制备,位点特异性DNA切割和连接可以使用本领域已知的限制酶、连接酶等制备,但不限于此。
本申请中使用的载体没有特别限制,可以使用本领域已知的任何载体。常用载体的实例可包括天然或重组形式的质粒、粘粒、病毒和噬菌体。例如,pWE15、M13、MBL3、MBL4、IXII、ASHII、APII、t10、t11、Charon4A、Charon21A等可以用作噬菌体载体或粘粒载体,并且基于pBR、基于pUC、基于pBluescriptII、基于pGEM、基于pTZ、基于pCL、基于pET、基于pUB110的载体可以用作质粒载体。具体而言,可以使用pDZ、pACYC177、pACYC184、pCL、pECCG117、pUC19、pBR322、pMW118、pCC1BAC和pSM704等载体。可用于本申请的载体没有特别限制,而是可以使用任何已知的表达载体。
在一个实施方案中,使用用于细胞中染色体插入的载体,可以用突变的多核苷酸取代编码染色体中的变体目标的多核苷酸。将多核苷酸插入染色体可以通过本领域已知的任何方法进行,例如同源重组,但不限于此。载体可以进一步包括选择标记,以确认染色体插入。选择标记用于选择被载体转化的细胞,即,确认是否插入目标核酸分子。选择标记的实例可以包括提供选择性表型的标记,例如耐药性,营养缺陷型,对细胞毒性剂的抗性,或表面突变多肽的表达。在被选择性试剂处理的环境中,只有表达选择标记的细胞可以存活或显示不同的表型,因此可以选择被转化的细胞。
本申请的又一方面提供了表达本申请的丝氨酸蛋白酶变体的微生物。
本文所用术语“待表达/所表达/表达”蛋白质是指将目标蛋白引入微生物或在微生物中表达的状态。考虑到本申请的目的,“目标蛋白”可以是上述丝氨酸蛋白酶变体。
具体而言,术语“引入蛋白质”是指在最初不具有具体蛋白质的微生物中表现出该具体蛋白质的活性,或者与该蛋白质的内源性活性或修饰前的活性相比,表现出该蛋白质的增强活性。例如,蛋白质的引入可以指将编码具体蛋白质的多核苷酸引入微生物的染色体,或将包含编码具体蛋白质的多核苷酸的载体引入微生物,从而显示蛋白质的活性。
表达丝氨酸蛋白酶变体的微生物可以包括本申请的丝氨酸蛋白酶变体、编码所述变体的多核苷酸和包含所述多核苷酸的载体中的至少一种。
具体而言,包含丝氨酸蛋白酶变体、编码变体的多核苷酸和包含多核苷酸的载体中的至少一种的微生物可以是通过用包含编码变体的多核苷酸的载体转化而制备的微生物,但不限于此。
微生物可以是重组微生物,重组可以通过遗传修饰(如转化)来实现。
本文所用术语“转化”是指将含有编码目标蛋白的多核苷酸的载体引入宿主细胞,以允许所述多核苷酸编码的蛋白在宿主细胞中表达的过程。无论被转化的多核苷酸是插入宿主细胞的染色体中的形式还是位于染色体外的形式,两种形式的被转化的多核苷酸都属于本申请的范围内,只要蛋白在宿主细胞中表达。此外,多核苷酸包括编码目标蛋白的DNA和RNA。多核苷酸可以以任何形式引入宿主细胞,只要多核苷酸被引入宿主细胞中并在其中表达。例如,多核苷酸可以以表达盒的形式引入宿主细胞,表达盒是含有自身复制所需的所有必需元件的基因构建体。表达盒通常可以包括与多核苷酸可操作地连接的启动子,转录终止信号,核糖体结合位点和翻译终止信号。表达盒可以是能够自我复制的表达载体。另外,多核苷酸可以以其原始形式被引入到宿主细胞中,并且可操作地连接到在宿主细胞中表达所需的序列,而不限于此。用于转化的方法包括用于将多核苷酸引入细胞的任何方法,并且可以通过本领域已知的合适的标准技术进行。例如,转化方法包括电穿孔法、磷酸钙(Ca(H2PO4)2、CaHPO4或Ca3(PO4)2)沉淀法,氯化钙(CaCl2)沉淀法,显微注射法,聚乙二醇(PEG)法,DEAE-葡聚糖法,阳离子脂质体法,天然感受(例如,参见Perry and Kuramitsu,1981,Infect.Immun.32:1295-1297]),和乙酸锂-DMSO方法,但不限于此。
重组微生物可以是本申请的丝氨酸蛋白酶的活性得到增强的微生物。
“活性增强”可以指微生物的具体蛋白质的活性与内源性活性或修饰前的活性相比得到增强。术语“内源活性”可以指当微生物由天然或人工因素引起的遗传突变转化时,转化前微生物的亲本菌株所具有的具体蛋白质的活性。
具体而言,本申请中的蛋白质变体的活性的增强可以通过以下至少一种方法实现:增加编码蛋白质变体的基因的细胞内拷贝数的方法,将突变引入编码蛋白质变体的基因的表达控制序列的方法,用具有更强活性的序列替换编码蛋白质变体的基因的表达控制序列的方法,用编码蛋白变体的基因替换编码具有丝氨酸蛋白酶活性的野生型蛋白的染色体基因的方法,和进一步将突变引入编码蛋白变体的基因中,以增强蛋白变体的活性的方法,但不限于此。
接下来,通过缺失、插入、非保守取代、保守取代或上述方式的组合,在核酸序列中引入突变,以进一步增强表达控制序列的活性,或者通过用具有更强活性的核酸序列替换核酸序列,可以进行增加多核苷酸表达的表达控制序列的修饰,但不限于此。表达控制序列可以包括启动子、操纵子序列、核糖体结合位点编码序列、用于调节转录和翻译的序列等,但不限于此。
用于替换内源启动子的强启动子可以连接在多核苷酸表达单元的上游,但本申请不限于此。强启动子的实例可包括CJ1至CJ7启动子(韩国专利号10-0620092),lac启动子,trp启动子,trc启动子,tac启动子,λ噬菌体PR启动子,PL启动子,tet启动子,gapA启动子,SPL7启动子,SPL13(sm3)启动子(韩国专利号10-1783170),O2启动子(韩国专利号10-1632642),tkt启动子,yccA启动子等,但不限于此。
此外,染色体上多核苷酸序列的修饰可以通过缺失、插入、非保守取代、保守取代或以上方式的组合在表达控制序列中引入突变,以进一步增强多核苷酸序列的活性,或者通过用修饰为具有更强活性的多核苷酸序列替换该序列而进行,但不限于此。
通常,蛋白质活性的引入和增强可以将蛋白质的活性或浓度从野生型或未修饰的微生物株中的活性或浓度增加1%、10%、25%、50%、75%、100%、150%、200%、300%、400%或500%至最大1000%或2000%,但不限于此。
根据本申请的宿主细胞或微生物可以是通过包含本申请的多核苷酸或本申请的载体而表达丝氨酸蛋白酶变体的任何微生物。具体而言,宿主细胞或微生物的实例可包括属于埃希氏菌属(Escherichia),沙雷氏菌属(Serratia),欧文氏菌属(Erwinia),肠细菌属(Enterobacteria),普罗维登斯菌属(Providencia),沙门氏菌属(Salmonella),链霉菌属(Streptomyces),假单胞菌属(Pseudomonas),短杆菌属(Brevibacterium),棒状杆菌属(Corynebacterium)或芽孢杆菌属(Bacillus等的微生物菌株,具体而言,可以是枯草芽孢杆菌(Bacillus subtilis)、地衣芽孢杆菌(Bacillus licheniformis),解淀粉芽孢杆菌(Bacillus amyloliquefaciens),韦利茨芽孢杆菌(Bacillus velezensis),大肠杆菌(Escherichia coli),谷氨酸棒杆菌(Corynebacterium glutamicum)或米曲霉(Aspergillus oryzae)的菌株,更具体而言,可以是枯草芽孢杆菌(Bacillus subtilis),但不限于此。
本申请的又一方面提供了包含本申请的丝氨酸蛋白酶变体和表达丝氨酸蛋白酶变体的微生物中的至少一种的饲料组合物。
包含在本申请变体的饲料组合物中的丝氨酸蛋白酶变体可以以这样的方式包含在饲料组合物中:表达丝氨酸蛋白酶变体的微生物包含在饲料组合物中,或者可以是从表达丝氨酸蛋白酶变体的微生物中分离并纯化的形式,但是本申请不限于此。
本文所用术语“饲料组合物”是指任何天然或人造食物、膳食等的任何制备物,或用于动物食用、摄入和消化或适合的膳食组分,并且饲料可以以本领域已知的各种形式制备。
饲料组合物可以是饲料添加剂。
饲料的类型没有特别限制,并且可以使用本领域中常用的饲料。饲料的非限制性实例可以包括:蔬菜饲料,例如谷物、根/果实、食品加工副产物、藻类、纤维、药物副产物、油和脂肪、淀粉、瓜类、以及谷物副产物;和动物饲料,例如蛋白质,无机材料,油和脂肪,矿物质,单细胞蛋白质,动物浮游生物或食品。这些饲料可以单独使用或至少两种组合使用。
本申请的饲料组合物可以进一步包括选自以下的至少一种:有机酸,例如柠檬酸、富马酸、己二酸、乳酸和苹果酸;磷酸盐,例如磷酸钠,磷酸钾,酸式焦磷酸盐和多磷酸盐(聚合磷酸盐);以及天然抗氧化剂,例如多酚,儿茶素,α-生育酚,迷迭香提取物,维生素C,绿茶提取物,甘草提取物,壳聚糖,鞣酸和植酸。
本申请的饲料组合物可以进一步包括选自以下的至少一种:辅助物质,例如氨基酸、矿物质、维生素、抗生素、抗菌物质、抗氧化剂、抗真菌剂和不同益生菌形式的微生物制剂;谷物,(例如,粉末或粉碎的小麦、燕麦、大麦、玉米和大米);植物蛋白饲料,包括油菜、菜豆、向日葵为主要成分;动物蛋白饲料,例如血粉,肉粉,骨粉和鱼粉;糖和乳制品(例如,由各种类型的干奶粉和乳清粉形成的干组分;脂质(例如,活性成分,诸如通过加热液化的任何动物脂肪和植物油);以及添加剂,例如营养补充剂,消化和吸收促进剂,生长促进剂和预防剂。
本申请的饲料组合物可以是干燥或液体制剂的形式,并且可以进一步包括用于饲料的赋形剂。用于饲料的赋形剂可以是,例如,沸石,玉米粉,米糠等,但不限于此。
除了丝氨酸蛋白酶变体之外,本申请的饲料组合物还可以包括酶制剂。例如,饲料组合物可进一步包括至少一种选自以下的酶:脂质降解酶,如脂肪酶,将植酸降解成磷酸盐和磷酸肌醇的植酸酶,催化淀粉、糖原等中含有的α-1,4-糖苷键水解的淀粉酶,催化有机磷酸酯水解的磷酸酶,催化麦芽糖生成两个葡萄糖分子的麦芽酶,以及催化蔗糖水解为葡萄糖-果糖混合物的转化酶。然而,本申请不限于此。
本申请的饲料组合物可以单独施用于动物,或与包含在可食用载体中的其它饲料添加剂组合施用于动物。此外,饲料组合物可以作为饲料添加剂或追加饲料容易地施用,直接施用到家畜饲料中或与饲料分开施用,或在单独的口服制剂中施用或与其它成分组合施用。此外,每日剂量可通过如本申请所属领域中通常已知的每日一次剂量或每日多次剂量来使用。
施用本申请的饲料组合物的动物的实例可以包括家畜,例如肉牛、奶牛、小牛、猪、仔猪、绵羊、山羊、马、兔、狗和猫;和家禽,例如小鸡、母鸡、家鸡、公鸡、鸭、鹅、火鸡、鹌鹑和小禽,但不限于此。
包括在本申请的饲料组合物中的丝氨酸蛋白酶变体的量没有特别限制,并且可以根据目的适当地调节。在一个实施方案中,可以以适当的量包含丝氨酸蛋白酶变体,用于降解蛋白质源材料,同时在家畜的消化道中长期存有,如本申请所属领域中通常已知的,但本申请不限于此。
本申请的又一方面提供了包含本申请的丝氨酸蛋白酶变体和表达丝氨酸蛋白酶变体的微生物中的至少一种的食物组合物。丝氨酸蛋白酶变体可用于液体或固体食物组合物中。此外,食品可以是粉剂、丸剂、饮料、茶或普通食品的添加剂。
在一个实施方案中,食物可以是一组需要蛋白酶的食物,例如乳制品,用于改善肠运动和体重减轻的健康功能食物,和用于预防高血压的健康功能食物。
在另一个实施方案中,丝氨酸蛋白酶变体可以作为食品增溶剂、食品软化剂和肉质改良剂包含在各种食品组合物中。在各种其它实施方案中,丝氨酸蛋白酶变体可以在麸质网络分解的步骤中加入到焙烤混合物中。或者,丝氨酸蛋白酶变体可用于催化食物蛋白(例如乳蛋白)的水解。或者,为了提供、制备调味剂,降低苦味,改变乳化性,产生生物活性肽,降低蛋白质中引起变态反应的抗原,丝氨酸蛋白酶变体可以包含在各种食品组合物中。然而,这仅仅是说明性的实施方案,丝氨酸蛋白酶变体的用途不限于此。
本申请的食物组合物中包含的丝氨酸蛋白酶变体的量可以由本领域普通技术人员根据目的适当地调节。
本申请的又一方面提供包含本申请的丝氨酸蛋白酶变体和表达丝氨酸蛋白酶变体的微生物中的至少一种的除垢剂组合物。
本申请的除垢剂组合物可以是第一和第二水性除垢剂组合物、非水性液体除垢剂组合物、流延固体、粗粒、颗粒、压缩片剂、凝胶、糊剂或浆料的形式。除垢剂组合物可用于除去顽固的食物污渍、食物残渣膜和其它少量的食物组合物。
根据本申请的除垢剂组合物可以以用于清洁硬表面的除垢剂组合物,用于清洁织物的除垢剂组合物,用于洗碗的除垢剂组合物,用于口腔清洁的除垢剂组合物,用于清洁义齿的除垢剂或隐形眼镜清洁溶液的形式提供。然而,本申请不限于此。
本申请的又一方面提供包含本申请的丝氨酸蛋白酶变体和表达丝氨酸蛋白酶变体的微生物中的至少一种的药物组合物。
本申请的药物组合物可用作消化酶的药物组合物,以改善消化疾病、消化障碍和消化道手术后异常,直接施用于血凝块以溶解纤维蛋白的溶栓或抗血栓组合物,用作体内防御系统以去除炎性物质或坏死组织的抗炎药物,或用于减轻手术或伤口后的水肿的抗炎药物。
根据使用的方法或目的,药物组合物可以进一步包括药学上可接受的载体、赋形剂、稀释剂或辅助成分。载体、赋形剂或稀释剂可包括选自乳糖、右旋糖、蔗糖、山梨糖醇、甘露糖醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、阿拉伯胶、藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁、矿物油、糊精、碳酸钙、丙二醇、液体石蜡和盐水中的至少一种,但不限于此。
除了上述目的之外,本申请的丝氨酸蛋白酶变体或表达丝氨酸蛋白酶变体的微生物还可以用于生产化妆品、加工皮革、制备药物、制备诊断剂和管理废物、制备用于学术研究的化学品的目的。然而,示例性地描述了目的,并且丝氨酸蛋白酶变体也可以用于本领域已知的变性、降解或去除蛋白质材料的任何其它目的。
实施例
在下文中,将参考以下实施例和实验例更详细地描述本申请。然而,这些实施例和实验例仅用于说明目的,而不旨在限制本申请的范围。
实施例1.源自Thermobifida fusca的丝氨酸蛋白酶变体的筛选
实施例1-1:源自Thermobifida fusca的丝氨酸蛋白酶文库的制备
通过易错PCR将随机突变引入编码对应于源自Thermobifida fusca的丝氨酸蛋白酶的成熟区的氨基酸(SEQ ID NO:31)的基因中。使用DiversifyTM PCR RandomMutagenesis Kit(Clontech,货号630703)实施易错PCR,并且使用的PCR条件如下表1中所述。证实突变以6.2个突变/kb的频率被引入。
表1
使用下表2中所示的引物扩增,使用In-FusionR HD克隆试剂盒(Clontech),将在上述过程中获得的PCR片段各自连接到载体上,并转化到DH5α细胞中以获得菌落。纯化所得菌落中的质粒,以获得大小为约5×104的文库。
表2
模板DNA(pBE-S-TAP) | SEQ ID NO:1 |
正向引物 | SEQ ID NO:13 |
反向引物 | SEQ ID NO:14 |
实施例1-2:源自Thermobifida fusca的丝氨酸蛋白酶文库的筛选
用实施例1-1中制备的蛋白酶文库转化容易释放蛋白质的枯草芽孢杆菌LB700菌株并进行筛选。筛选通过两阶段方法进行。在第一阶段,将用文库转化的枯草芽孢杆菌菌株接种在2%脱脂奶平板上,并根据晕圈大小选择所希望的菌落。根据Groningen方法进行枯草芽孢杆菌的转化,筛选中使用的脱脂奶的组成如下表3所示。
表3
(每1L)
第二阶段涉及通过偶氮酪蛋白显色来重新选择在第一阶段中选择的菌落的方法。将含有卡那霉素抗生素的脑心灌注(BHI,bd,货号53286)液体培养基添加到96深孔板中,并且将在第一阶段中选择的菌落接种到其中,随后在37℃下孵育20至24小时。孵育后,通过离心获得包含酶的上清液,并将上清液与等量的作为底物的2%(w/v)偶氮酪蛋白混合,然后在37℃反应1小时。通过向酶反应溶液中加入3倍体积的10%三氯乙酸(TCA)终止反应,并通过离心除去凝结的蛋白质。通过将产物与等量的NaOH混合进行显色反应,然后在440nm处测量吸光度,以比较显色程度。通过该方法,选择与野生型丝氨酸蛋白酶相比吸光度增加150%或更多的菌落。
实施例2.选择的变体的制备和活性评价
实施例2-1:变体制备
作为分析选自筛选的变体序列的结果,证实SEQ ID NO:31的第12位氨基酸(苯丙氨酸,Phe)和第116位氨基酸(天冬酰胺,Asn)分别被酪氨酸(Tyr)和天冬氨酸(Asp)取代。在图1中,显示了突变的位置。通过定点诱变将两个选择的突变(F12和N116)以单一突变的形式重新引入到pBE-S-TAP质粒中,将具有双突变和单一突变的菌株的活性分别与野生型菌株的活性进行比较。用于制备变体的引物如下表4所示。
表4
TAP_F12Y_F | SEQ ID NO:15 |
TAP_F12Y_R | SEQ ID NO:16 |
TAP_N116D_F | SEQ ID NO:17 |
TAP_N116D_R | SEQ ID NO:18 |
实施例2-2:活性评价
在用制备的质粒转化枯草芽孢杆菌LB700菌株后,使用N-琥珀酰-ALA-ALA-PRO-PHE-p-硝基苯胺(Sigma,货号S7388,下文称为SUC-AAPF-pNA)肽作为底物进行转化株的活性评价。将转化的枯草芽孢杆菌菌株接种到含有卡那霉素抗生素的脑心灌注(BHI,bd,货号53286)液体培养基中,并在37℃下培养20至24小时,将除细胞外的一部分培养液与25mMTris-HCl(pH7.5)缓冲液和1mM Suc-AAPF-pNA混合,然后在37℃下反应30分钟。在410nm测量反应溶液的吸光度。由文献中已知的酶产生的对硝基苯胺的消光系数在410nm处为8800M-1cm-1,并基于此计算酶的单位(Barrett,A.J.,Cathepsin G.Methods Enzymol.,80,Pt.C,561-565,(1981))。测量的活性示于下表5中。
表5
酶活性(单位/ml) | |
野生型 | 16.3 |
F12Y | 34.0 |
F12YN116D | 63.8 |
作为测量的结果,证实F12Y和F12YN116D变体的活性在pH 7.5和37℃的条件下分别增加约2.1倍和3.9倍。
实施例2-3:热稳定性的评价
由于热稳定性是酶的非常重要的性质,因此进行下面描述的实验以证实引入突变对热稳定性的影响。
具体而言,在将实施例2-2中使用的样品分别置于室温、70℃、80℃和90℃下5分钟之后,测量样品的酶活性,用于活性评价。测量的活性示于下表6中。
表6
作为测量的结果证实,即使在80℃下,F12Y和F12YN116D突变显示出比野生型菌株高约2倍和4倍的酶活性。由于证实本申请的丝氨酸蛋白酶变体在高温下也保持高活性,因此丝氨酸蛋白酶变体可以有效地用于工业中。
实施例3.饱和诱变文库的制备和筛选
实施例3-1.F12和N116残基的饱和诱变文库制备
为了鉴定用酪氨酸和天冬氨酸以外的残基取代F12和N116的每个残基(即,先前选择的突变)对它们的活性的影响,制备了关于这两个残基的饱和诱变文库。
分别使用pBE-S-TAP质粒作为模板和引物对SEQ ID NO:11和12以及SEQ ID NO:13和14获得两个PCR片段。使用In-Fusion HD克隆试剂盒将各片段连接,然后转化到DH5α细胞中,从而获得菌落。纯化所得菌落中的质粒以获得大小为约4×103的文库。
表7
模板DNA(pBE-S-TAP) | SEQ ID NO:1 |
饱和诱变_F_1 | SEQ ID NO:19 |
饱和诱变_R_1 | SEQ ID NO:20 |
饱和诱变_F_2 | SEQ ID NO:21 |
饱和诱变_R_2 | SEQ ID NO:22 |
实施例3-2:饱和诱变文库的筛选和活性评价
以与实施例1-2相同的方式对实施例3-1中制备的饱和诱变文库进行筛选。通过筛选选择与F12YN116D变体相比具有相同或增加的活性的变体,并通过序列分析、使用Suc-AAPF-pNA作为底物对这些变体进行活性评价。
表8
酶活性(单位/mL) | |
野生型 | 23.52 |
F12YN116D | 65 |
F12YN116S | 77.35 |
F12SN116D | 61.75 |
F12SN116T | 117.65 |
F12AN116G | 94.9 |
F12A | 94.25 |
F12R | 58.5 |
结果发现,F12和N116变体增加它们的活性,即使它们的残基各自被除了实施例2中证实的酪氨酸和天冬氨酸以外的不同氨基酸(例如,F12S、F12A、F12R、N116S、N116T、N116G等)取代。
根据上述结果,证实了F12和N116残基是展现丝氨酸蛋白酶活性的重要残基,并且丝氨酸蛋白酶的酶活性可以通过用不同的氨基酸取代这些残基中的每一个来提高。因此,本申请的具有增加的酶活性的丝氨酸蛋白酶变体可有效地用于工业中。
综上所述,本申请所属领域的技术人员将能够理解,本申请可以以其它具体形式来实现,而不改变本申请的技术构思或本质特征。在这方面,本文公开的示例性实施方案仅用于说明目的,而不应被解释为限制本申请的范围。相反,本申请旨在不仅涵盖示例性实施方案,而且涵盖可包括在如所附权利要求书所定义的本申请的实质和范围内的各种替代、修改、等同方案和其它实施方案。
序列表
<110> CJ 第一制糖株式会社(CJ CheilJedang Corporation)
<120> 新的丝氨酸蛋白酶变体
<130> OPA20086
<150> KR 10-2019-0157400
<151> 2019-11-29
<160> 48
<170> KoPatentIn 3.0
<210> 1
<211> 6922
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> pBE-S-TFP
<400> 1
actagtgttc ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga 60
tgatatacct aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc 120
catctattac aataaattca cagaatagtc ttttaagtaa gtctactctg aacttaagca 180
aaaggagagg gacgcgtgtg agaagcaaaa aattgtggat cagcttgttg tttgcgttaa 240
cgttaatctt tacgatggcg ttcagcaaca tgtctgcgca ggctgcggcc ggtgcacata 300
tgcaagagct ggcgttgaaa cgggacctcg gcctctctga cgcagaagta gccgaactcc 360
gggctgctga ggcggaagcg gtcgagctcg aggaggagct ccgcgattca ttagggtcag 420
acttcggcgg tgtatatctg gatgctgaca ccaccgaaat tacggtcgcg gtaaccgacc 480
cggcagcggt aagtcgtgtc gacgcggatg atgtcacagt tgatgttgtc gatttcgggg 540
aaacagcttt gaatgatttt gtggcttcat taaatgccat tgccgacacg gcagacccta 600
aagtcactgg atggtatacc gatctcgaaa gtgatgccgt agtcattacg accttgcgtg 660
gcgggactcc tgctgccgag gaacttgctg agagagcggg tctcgacgaa agagccgttc 720
ggattgtgga agaagatgaa gaaccacaga gcttggctgc aattattggt ggaaacccgt 780
actatttcgg aaattacaga tgcagtatcg ggtttagtgt ccgtcagggc tctcaaacgg 840
gattcgcgac cgcaggccac tgcggatcca cggggacgcg tgtgtcttct ccttcaggaa 900
cagttgcagg aagttatttc ccgggtcgcg atatgggctg ggtgcggatt acatcagcag 960
atactgtaac accactcgta aatcggtata atgggggaac tgttacggtc actgggtcac 1020
aagaagctgc caccggatcc tccgtttgtc gctctggagc aacaacgggc tggcgctgcg 1080
gaactatcca atcaaaaaac caaacggttc gctatgcaga agggactgtt actggtttaa 1140
caagaactac agcctgtgct gaaggtgggg attctggagg gccatggctc acaggtagcc 1200
aggcgcaagg ggttacaagc ggcggaacag gcgattgcag aagtggaggg attacctttt 1260
tccaaccaat caatccattg cttagctatt tcggccttca attagtgacc ggctgaaagc 1320
ttgtcgacct gcagtctaga catcaccatc atcaccacta atgcggtagt ttatcacagt 1380
taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc 1440
tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc 1500
tatttcactt tttgcattct acaaactgca taactattat gtaaatcgct cctttttagg 1560
tggcacaaat gtgaggcatt ttcgctcttt ccggcaacca cttccaagta aagtataaca 1620
cactatactt tatattcata aagtgtgtgc tctgcgaggc tgtcggcagt gccgaccaaa 1680
accataaaac ctttaagacc tttctttttt ttacgagaaa aaagaaacaa aaaaacctgc 1740
cctctgccac ctcagcaaag gggggttttg ctctcgtgct cgtttaaaaa tcagcaaggg 1800
acaggtagta ttttttgaga agatcactca aaaaatctcc acctttaaac ccttgccaat 1860
ttttattttg tccgttttgt ctagcttacc gaaagccaga ctcagcaaga ataaaatttt 1920
tattgtcttt cggttttcta gtgtaacgga caaaaccact caaaataaaa aagatacaag 1980
agaggtctct cgtatctttt attcagcaat cgcgcccgat tgctgaacag attaataata 2040
gattttagct ttttatttgt tgaaaaaagc taatcaaatt gttgtcggga tcaattactg 2100
caaagtctcg ttcatcccac cactgatctt ttaatgatgt attggggtgc aaaatgccca 2160
aaggcttaat atgttgatat aattcatcaa ttccctctac ttcaatgcgg caactagcag 2220
taccagcaat aaacgactcc gcacctgtac aaaccggtga atcattacta cgagagcgcc 2280
agccttcatc acttgcctcc catagatgaa tccgaacctc attacacatt agaactgcga 2340
atccatcttc atggtgaacc aaagtgaaac ctagtttatc gcaataaaaa cctatactct 2400
ttttaatatc cccgactggc aatgccggga tagactgtaa cattctcacg cataaaatcc 2460
cctttcattt tctaatgtaa atctattacc ttattattaa ttcaattcgc tcataattaa 2520
tcctttttct tattacgcaa aatggcccga tttaagcaca ccctttattc cgttaatgcg 2580
ccatgacagc catgataatt actaatacta ggagaagtta ataaatacga gcaaaaggcc 2640
agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 2700
cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 2760
tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 2820
tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 2880
gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 2940
acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 3000
acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 3060
cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 3120
gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 3180
gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 3240
agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 3300
ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 3360
ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 3420
atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 3480
tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 3540
gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 3600
ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 3660
caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 3720
cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 3780
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 3840
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 3900
agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 3960
tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 4020
agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 4080
atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 4140
ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 4200
cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 4260
caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 4320
attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 4380
agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct 4440
aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc 4500
gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca gtaaccaaca tgattaacaa 4560
ttattagagg tcatcgttca aaatggtatg cgttttgaca catccactat atatccgtgt 4620
cgttctgtcc actcctgaat cccattccag aaattctcta gcgattccag aagtttctca 4680
gagtcggaaa gttgaccaga cattacgaac tggcacagat ggtcataacc tgaaggaaga 4740
tctgattgct taactgcttc agttaagacc gaagcgctcg tcgtataaca gatgcgatga 4800
tgcagaccaa tcaacatggc acctgccatt gctacctgta cagtcaagga tggtagaaat 4860
gttgtcggtc cttgcacacg aatattacgc catttgcctg catattcaaa cagctcttct 4920
acgataaggg cacaaatcgc atcgtggaac gtttgggctt ctaccgattt agcagtttga 4980
tacactttct ctaagtatcc acctgaatca taaatcggca aaatagagaa aaattgacca 5040
tgtgtaagcg gccaatctga ttccacctga gatgcataat ctagtagaat ctcttcgcta 5100
tcaaaattca cttccacctt ccactcaccg gttgtccatt catggctgaa ctctgcttcc 5160
tctgttgaca tgacacacat catctcaata tccgaatagg gcccatcagt ctgacgacca 5220
agagagccat aaacaccaat agccttaaca tcatccccat atttatccaa tattcgttcc 5280
ttaatttcat gaacaatctt cattctttct tctctagtca ttattattgg tccattcact 5340
attctcattc ccttttcaga taattttaga tttgcttttc taaataagaa tatttggaga 5400
gcaccgttct tattcagcta ttaataactc gtcttcctaa gcatccttca atccttttaa 5460
taacaattat agcatctaat cttcaacaaa ctggcccgtt tgttgaacta ctctttaata 5520
aaataatttt tccgttccca attccacatt gcaataatag aaaatccatc ttcatcggct 5580
ttttcgtcat catctgtatg aatcaaatcg ccttcttctg tgtcatcaag gtttaatttt 5640
ttatgtattt cttttaacaa accaccatag gagattaacc ttttacggtg taaaccttcc 5700
tccaaatcag acaaacgttt caaattcttt tcttcatcat cggtcataaa atccgtatcc 5760
tttacaggat attttgcagt ttcgtcaatt gccgattgta tatccgattt atatttattt 5820
ttcggtcgaa tcatttgaac ttttacattt ggatcatagt ctaatttcat tgcctttttc 5880
caaaattgaa tccattgttt ttgattcacg tagttttctg tattcttaaa ataagttggt 5940
tccacacata ccaatacatg catgtgctga ttataagaat tatctttatt atttattgtc 6000
acttccgttg cacgcataaa accaacaaga tttttattaa tttttttata ttgcatcatt 6060
cggcgaaatc cttgagccat atctgacaaa ctcttattta attcttcgcc atcataaaca 6120
tttttaactg ttaatgtgag aaacaaccaa cgaactgttg gcttttgttt aataacttca 6180
gcaacaacct tttgtgactg aatgccatgt ttcattgctc tcctccagtt gcacattgga 6240
caaagcctgg atttacaaaa ccacactcga tacaactttc tttcgcctgt ttcacgattt 6300
tgtttatact ctaatatttc agcacaatct tttactcttt cagccttttt aaattcaaga 6360
atatgcagaa gttcaaagta atcaacatta gcgattttct tttctctcca tggtctcact 6420
tttccacttt ttgtcttgtc cactaaaacc cttgattttt catctgaata aatgctacta 6480
ttaggacaca taatattaaa agaaaccccc atctatttag ttatttgttt agtcacttat 6540
aactttaaca gatggggttt ttctgtgcaa ccaattttaa gggttttcaa tactttaaaa 6600
cacatacata ccaacacttc aacgcacctt tcagcaacta aaataaaaat gacgttattt 6660
ctatatgtat caagataaga aagaacaagt tcaaaaccat caaaaaaaga caccttttca 6720
ggtgcttttt ttattttata aactcattcc ctgatctcga cttcgttctt tttttacctc 6780
tcggttatga gttagttcaa attcgttctt tttaggttct aaatcgtgtt tttcttggaa 6840
ttgtgctgtt ttatccttta ccttgtctac aaacccctta aaaacgtttt taaaggcttt 6900
taagccgtct gtacgttcct aa 6922
<210> 2
<211> 338
<212> PRT
<213> 未知(Unknown)
<220>
<221>
<222>
<223> Thermobifida fusca
<400> 2
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Phe Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asn Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 3
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163Y)
<400> 3
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Tyr Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asn Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 4
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163Y_N267D)
<400> 4
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Tyr Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asp Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 5
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163Y_N267S)
<400> 5
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Tyr Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Ser Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 6
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163S_N267D)
<400> 6
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Ser Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asp Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 7
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163S_N267T)
<400> 7
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Ser Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Thr Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 8
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163A_N267G)
<400> 8
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Ala Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Gly Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 9
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163A)
<400> 9
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Ala Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asn Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 10
<211> 338
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163R)
<400> 10
Met Gln Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu
1 5 10 15
Val Ala Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu
20 25 30
Glu Leu Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp
35 40 45
Ala Asp Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val
50 55 60
Ser Arg Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly
65 70 75 80
Glu Thr Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp
85 90 95
Thr Ala Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp
100 105 110
Ala Val Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu
115 120 125
Leu Ala Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu
130 135 140
Glu Asp Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro
145 150 155 160
Tyr Tyr Arg Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln
165 170 175
Gly Ser Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly
180 185 190
Thr Arg Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro
195 200 205
Gly Arg Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr
210 215 220
Pro Leu Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser
225 230 235 240
Gln Glu Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr
245 250 255
Gly Trp Arg Cys Gly Thr Ile Gln Ser Lys Asn Gln Thr Val Arg Tyr
260 265 270
Ala Glu Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu
275 280 285
Gly Gly Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly
290 295 300
Val Thr Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe
305 310 315 320
Phe Gln Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val
325 330 335
Thr Gly
<210> 11
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_成熟区_F
<400> 11
aagaagatga agaaccacag agcttggctg caattattgg tggaaac 47
<210> 12
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_成熟区_R
<400> 12
gactgcaggt cgacaagctt tcagccggtc actaattgaa ggcc 44
<210> 13
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> pBE-S-vector-F
<400> 13
aagcttgtcg acctgcagtc tag 23
<210> 14
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> pBE-S-vector-R
<400> 14
ctgtggttct tcatcttctt ccac 24
<210> 15
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_F12Y_F
<400> 15
gtggaaaccc gtactattac ggaaattaca gatgcag 37
<210> 16
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_F12Y_R
<400> 16
ctgcatctgt aatttccgta atagtacggg tttccac 37
<210> 17
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_N116D_F
<400> 17
ggaactatcc aatcaaaaga ccaaacggtt cgctatgc 38
<210> 18
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> TAP_N116D_R
<400> 18
gcatagcgaa ccgtttggtc ttttgattgg atagttcc 38
<210> 19
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> 饱和诱变_F_1
<400> 19
gtggaaaccc gtactatnnn ggaaattaca gatgcag 37
<210> 20
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> 饱和诱变_R_1
<400> 20
gcatagcgaa ccgtttgnnn ttttgattgg atagttcc 38
<210> 21
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> 饱和诱变_F_2
<400> 21
ggaactatcc aatcaaaann ncaaacggtt cgctatgc 38
<210> 22
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> 饱和诱变_R_2
<400> 22
ctgcatctgt aatttccnnn atagtacggg tttccac 37
<210> 23
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F12Y)
<400> 23
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactattacg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaaa ccaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 24
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163Y_N267D)
<400> 24
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactattacg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaga ccaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 25
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163Y_N267S)
<400> 25
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactattacg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaag tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 26
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163S_N267D)
<400> 26
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactatagtg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaga tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 27
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163S_N267T)
<400> 27
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactatagtg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaac tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 28
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163A_N267G)
<400> 28
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactatgctg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaagg tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 29
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163A)
<400> 29
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactatgctg gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaaa tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 30
<211> 1017
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca (F163R)
<400> 30
atgcaagagc tggcgttgaa acgggacctc ggcctctctg acgcagaagt agccgaactc 60
cgggctgctg aggcggaagc ggtcgagctc gaggaggagc tccgcgattc attagggtca 120
gacttcggcg gtgtatatct ggatgctgac accaccgaaa ttacggtcgc ggtaaccgac 180
ccggcagcgg taagtcgtgt cgacgcggat gatgtcacag ttgatgttgt cgatttcggg 240
gaaacagctt tgaatgattt tgtggcttca ttaaatgcca ttgccgacac ggcagaccct 300
aaagtcactg gatggtatac cgatctcgaa agtgatgccg tagtcattac gaccttgcgt 360
ggcgggactc ctgctgccga ggaacttgct gagagagcgg gtctcgacga aagagccgtt 420
cggattgtgg aagaagatga agaaccacag agcttggctg caattattgg tggaaacccg 480
tactatagag gaaattacag atgcagtatc gggtttagtg tccgtcaggg ctctcaaacg 540
ggattcgcga ccgcaggcca ctgcggatcc acggggacgc gtgtgtcttc tccttcagga 600
acagttgcag gaagttattt cccgggtcgc gatatgggct gggtgcggat tacatcagca 660
gatactgtaa caccactcgt aaatcggtat aatgggggaa ctgttacggt cactgggtca 720
caagaagctg ccaccggatc ctccgtttgt cgctctggag caacaacggg ctggcgctgc 780
ggaactatcc aatcaaaaaa tcaaacggtt cgctatgcag aagggactgt tactggttta 840
acaagaacta cagcctgtgc tgaaggtggg gattctggag ggccatggct cacaggtagc 900
caggcgcaag gggttacaag cggcggaaca ggcgattgca gaagtggagg gattaccttt 960
ttccaaccaa tcaatccatt gcttagctat ttcggccttc aattagtgac cggctga 1017
<210> 31
<211> 187
<212> PRT
<213> 未知(Unknown)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区
<400> 31
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Phe Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asn Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 32
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y)
<400> 32
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Tyr Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asn Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 33
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y_N116D)
<400> 33
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Tyr Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asp Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 34
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y_N116S)
<400> 34
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Tyr Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Ser Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 35
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12S_N116D)
<400> 35
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Ser Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asp Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 36
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12S_N116T)
<400> 36
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Ser Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Thr Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 37
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12A_N116G)
<400> 37
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Ala Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Gly Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 38
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12A)
<400> 38
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Ala Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asn Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 39
<211> 187
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12R)
<400> 39
Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr Arg Gly Asn Tyr Arg
1 5 10 15
Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser Gln Thr Gly Phe Ala
20 25 30
Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg Val Ser Ser Pro Ser
35 40 45
Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg Asp Met Gly Trp Val
50 55 60
Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu Val Asn Arg Tyr Asn
65 70 75 80
Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu Ala Ala Thr Gly Ser
85 90 95
Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp Arg Cys Gly Thr Ile
100 105 110
Gln Ser Lys Asn Gln Thr Val Arg Tyr Ala Glu Gly Thr Val Thr Gly
115 120 125
Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly Asp Ser Gly Gly Pro
130 135 140
Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr Ser Gly Gly Thr Gly
145 150 155 160
Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln Pro Ile Asn Pro Leu
165 170 175
Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
180 185
<210> 40
<211> 369
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca
<400> 40
Met Asn His Ser Ser Arg Arg Thr Thr Ser Leu Leu Phe Thr Ala Ala
1 5 10 15
Leu Ala Ala Thr Ala Leu Val Ala Ala Thr Thr Pro Ala Ser Ala Gln
20 25 30
Glu Leu Ala Leu Lys Arg Asp Leu Gly Leu Ser Asp Ala Glu Val Ala
35 40 45
Glu Leu Arg Ala Ala Glu Ala Glu Ala Val Glu Leu Glu Glu Glu Leu
50 55 60
Arg Asp Ser Leu Gly Ser Asp Phe Gly Gly Val Tyr Leu Asp Ala Asp
65 70 75 80
Thr Thr Glu Ile Thr Val Ala Val Thr Asp Pro Ala Ala Val Ser Arg
85 90 95
Val Asp Ala Asp Asp Val Thr Val Asp Val Val Asp Phe Gly Glu Thr
100 105 110
Ala Leu Asn Asp Phe Val Ala Ser Leu Asn Ala Ile Ala Asp Thr Ala
115 120 125
Asp Pro Lys Val Thr Gly Trp Tyr Thr Asp Leu Glu Ser Asp Ala Val
130 135 140
Val Ile Thr Thr Leu Arg Gly Gly Thr Pro Ala Ala Glu Glu Leu Ala
145 150 155 160
Glu Arg Ala Gly Leu Asp Glu Arg Ala Val Arg Ile Val Glu Glu Asp
165 170 175
Glu Glu Pro Gln Ser Leu Ala Ala Ile Ile Gly Gly Asn Pro Tyr Tyr
180 185 190
Phe Gly Asn Tyr Arg Cys Ser Ile Gly Phe Ser Val Arg Gln Gly Ser
195 200 205
Gln Thr Gly Phe Ala Thr Ala Gly His Cys Gly Ser Thr Gly Thr Arg
210 215 220
Val Ser Ser Pro Ser Gly Thr Val Ala Gly Ser Tyr Phe Pro Gly Arg
225 230 235 240
Asp Met Gly Trp Val Arg Ile Thr Ser Ala Asp Thr Val Thr Pro Leu
245 250 255
Val Asn Arg Tyr Asn Gly Gly Thr Val Thr Val Thr Gly Ser Gln Glu
260 265 270
Ala Ala Thr Gly Ser Ser Val Cys Arg Ser Gly Ala Thr Thr Gly Trp
275 280 285
Arg Cys Gly Thr Ile Gln Ser Lys Asn Gln Thr Val Arg Tyr Ala Glu
290 295 300
Gly Thr Val Thr Gly Leu Thr Arg Thr Thr Ala Cys Ala Glu Gly Gly
305 310 315 320
Asp Ser Gly Gly Pro Trp Leu Thr Gly Ser Gln Ala Gln Gly Val Thr
325 330 335
Ser Gly Gly Thr Gly Asp Cys Arg Ser Gly Gly Ile Thr Phe Phe Gln
340 345 350
Pro Ile Asn Pro Leu Leu Ser Tyr Phe Gly Leu Gln Leu Val Thr Gly
355 360 365
Ala
<210> 41
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y)
<400> 41
ttggctgcaa ttattggtgg aaacccgtac tattacggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaaacca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 42
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y_N116D)
<400> 42
ttggctgcaa ttattggtgg aaacccgtac tattacggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaagacca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 43
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12Y_N116S)
<400> 43
ttggctgcaa ttattggtgg aaacccgtac tattacggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaagtca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 44
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12S_N116D)
<400> 44
ttggctgcaa ttattggtgg aaacccgtac tatagtggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaagatca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 45
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12S_N116T)
<400> 45
ttggctgcaa ttattggtgg aaacccgtac tatagtggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaactca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 46
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12A_N116G)
<400> 46
ttggctgcaa ttattggtgg aaacccgtac tatgctggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaggtca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 47
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12A)
<400> 47
ttggctgcaa ttattggtgg aaacccgtac tatgctggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaaatca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
<210> 48
<211> 564
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221>
<222>
<223> Thermobifida fusca_成熟区 (F12R)
<400> 48
ttggctgcaa ttattggtgg aaacccgtac tatagaggaa attacagatg cagtatcggg 60
tttagtgtcc gtcagggctc tcaaacggga ttcgcgaccg caggccactg cggatccacg 120
gggacgcgtg tgtcttctcc ttcaggaaca gttgcaggaa gttatttccc gggtcgcgat 180
atgggctggg tgcggattac atcagcagat actgtaacac cactcgtaaa tcggtataat 240
gggggaactg ttacggtcac tgggtcacaa gaagctgcca ccggatcctc cgtttgtcgc 300
tctggagcaa caacgggctg gcgctgcgga actatccaat caaaaaatca aacggttcgc 360
tatgcagaag ggactgttac tggtttaaca agaactacag cctgtgctga aggtggggat 420
tctggagggc catggctcac aggtagccag gcgcaagggg ttacaagcgg cggaacaggc 480
gattgcagaa gtggagggat tacctttttc caaccaatca atccattgct tagctatttc 540
ggccttcaat tagtgaccgg ctga 564
Claims (11)
1.丝氨酸蛋白酶变体,所述丝氨酸蛋白酶变体与SEQ ID NO:31的氨基酸序列具有至少80%且小于100%的序列同源性,其中对应于SEQ ID NO:31的氨基酸序列的第12位的氨基酸和对应于SEQ ID NO:31的氨基酸序列的第116位的氨基酸中的至少一个氨基酸被不同的氨基酸取代。
2.如权利要求1所述的丝氨酸蛋白酶变体,其中对应于第12位的氨基酸被酪氨酸(Y)、丙氨酸(A)、丝氨酸(S)或精氨酸(R)取代。
3.如权利要求1所述的丝氨酸蛋白酶变体,其中对应于第116位的氨基酸被天冬氨酸(D)、丝氨酸(S)、苏氨酸(T)或甘氨酸(G)取代。
4.丝氨酸蛋白酶变体,所述丝氨酸蛋白酶变体包含权利要求1所述的丝氨酸蛋白酶变体的氨基酸序列,并且与SEQ ID NO:2具有至少80%且小于100%的同源性。
5.如权利要求1所述的丝氨酸蛋白酶变体,其中所述丝氨酸蛋白酶变体包含选自SEQID NO:3至10和SEQ ID NO:32至39的任一项中所示的氨基酸序列。
6.编码权利要求1-5中任一项所述的丝氨酸蛋白酶变体的多核苷酸。
7.包含权利要求6所述的多核苷酸的载体。
8.表达权利要求1-5中任一项所述的丝氨酸蛋白酶变体的微生物。
9.如权利要求8所述的微生物,其中所述微生物属于芽孢杆菌(Bacillus)属。
10.如权利要求9所述的微生物,其中所述微生物是枯草芽孢杆菌(Bacillussubtilis)。
11.饲料组合物,所述饲料组合物包含权利要求1所述的丝氨酸蛋白酶变体和表达所述丝氨酸蛋白酶变体的微生物中的至少一种。
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Publication number | Priority date | Publication date | Assignee | Title |
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CN109563497A (zh) * | 2016-06-17 | 2019-04-02 | 丹尼斯科美国公司 | 蛋白酶变体及其用途 |
CN109642225A (zh) * | 2016-07-21 | 2019-04-16 | 诺维信公司 | 丝氨酸蛋白酶变体以及对其进行编码的多核苷酸 |
CN109661465A (zh) * | 2016-07-21 | 2019-04-19 | 诺维信公司 | 丝氨酸蛋白酶变体以及对其进行编码的多核苷酸 |
Family Cites Families (15)
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US7985569B2 (en) * | 2003-11-19 | 2011-07-26 | Danisco Us Inc. | Cellulomonas 69B4 serine protease variants |
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KR20090129425A (ko) * | 2007-03-12 | 2009-12-16 | 다니스코 유에스 인크. | 개질된 프로테아제 |
CN103396904B (zh) * | 2008-11-11 | 2015-12-23 | 宝洁公司 | 包含丝氨酸蛋白酶变体的组合物和方法 |
AU2014214032A1 (en) * | 2013-02-06 | 2015-07-09 | Novozymes A/S | Use of polypeptides having protease activity in animal feed |
EP3013967B1 (en) * | 2013-06-24 | 2021-10-20 | Novozymes A/S | Processes for recovering oil from fermentation product processes and processes for producing fermentation products |
CN107002111A (zh) * | 2014-12-01 | 2017-08-01 | 诺维信公司 | 用于生产蛋白质水解产物的方法 |
KR101632642B1 (ko) | 2015-01-29 | 2016-06-22 | 씨제이제일제당 주식회사 | 신규한 프로모터 및 그의 용도 |
CN108350444A (zh) * | 2015-09-25 | 2018-07-31 | 诺维信公司 | 丝氨酸蛋白酶用于改进乙醇产量的用途 |
WO2018043856A1 (ko) | 2016-08-31 | 2018-03-08 | 씨제이제일제당 (주) | 신규 프로모터 및 이의 용도 |
MX2019006425A (es) * | 2016-12-01 | 2019-08-14 | Basf Se | Estabilizacion de enzimas en composiciones. |
US20190330577A1 (en) * | 2016-12-21 | 2019-10-31 | Dupont Nutrition Biosciences Aps | Methods of using thermostable serine proteases |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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CN109642225A (zh) * | 2016-07-21 | 2019-04-16 | 诺维信公司 | 丝氨酸蛋白酶变体以及对其进行编码的多核苷酸 |
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