CN107208077A - 裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法 - Google Patents

裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法 Download PDF

Info

Publication number
CN107208077A
CN107208077A CN201580073275.0A CN201580073275A CN107208077A CN 107208077 A CN107208077 A CN 107208077A CN 201580073275 A CN201580073275 A CN 201580073275A CN 107208077 A CN107208077 A CN 107208077A
Authority
CN
China
Prior art keywords
gly
ala
leu
pro
ser
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201580073275.0A
Other languages
English (en)
Inventor
M·阿莱施克
C·科恩
D·摩尔
E·M·宾德
G·沙兹梅尔
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DSM Austria GmbH
Original Assignee
Erber AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Erber AG filed Critical Erber AG
Publication of CN107208077A publication Critical patent/CN107208077A/zh
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/14—Pretreatment of feeding-stuffs with enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Animal Husbandry (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Birds (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physiology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Fodder In General (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

本发明涉及具有SEQ ID No.46的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体，其中所述多肽变体各自具有在N‑末端处缩短了47个氨基酸的氨基酸序列，并且所述氨基酸序列与SEQ ID No.46的氨基酸序列区段48‑540具有至少70％，优选地80％，特别优选地100％的序列同一性，即SEQ ID No.1；以及编码所述多肽变体的分离的多核苷酸；以及包含至少一种多肽变体和任选地至少一种助剂的裂解镰孢毒素的添加剂；以及所述多肽变体或添加剂用于以水解方式裂解至少一种镰孢毒素的用途，和用于以水解方式裂解至少一种镰孢毒素的方法。

Description

裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所 述添加剂的用途，以及用于裂解镰孢毒素的方法
本发明涉及裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述多肽变体和/或所述添加剂的用途，以及涉及用于借助于所述多肽变体和/或所述包含所述多肽变体的添加剂来裂解镰孢毒素的方法。
真菌毒素非常频繁地出现在农业植物性产品中，并且依据真菌毒素的种类和浓度而引起严重的经济损害，特别是在从农业产品制备的食物或饲料中，和也在用这样的食物或饲料喂养的动物和人中，其中这样的损害是极其多种多样的。已经开发了许多方法，试图用这些方法来使真菌毒素变得无害，也就是说使真菌毒素解毒或降解，以便在动物和人的营养、动物饲养、饲料和食物加工等的领域中尽可能地阻止由真菌毒素引起的损害。
突出的一组真菌毒素为镰孢毒素(类)，其中术语“镰孢毒素”或“镰孢毒素类”是等价的并且各自涉及由霉菌镰孢属物种(Fusarium sp.)所产生的串珠镰孢菌素以及其衍生物和降解产物中的至少一种或多种或者全部，然而特别地涉及串珠镰孢菌素A1-2(FA1-2)，串珠镰孢菌素B1-4(FB1-4)，串珠镰孢菌素C1、2、4(FC1、FC2、FC4)和HFC1，也涉及经部分水解的串珠镰孢菌素FA1-2、FB1-4、FC1-2、FC4和HFC1。经部分水解的串珠镰孢菌素具有仅一个丙三羧酸残基，而FA1-2、FB1-4、FC1-2、FC4和HFC1具有两个丙三羧酸残基。此外，结构上相似的番茄链格孢(Alternaria alternata lycopersici)(AAL)毒素也被镰孢毒素组所涵盖，其中AAL毒素被再分为AAL-TA1(CAS No 79367-52-5)、AAL-TA2(CAS No 79367-51-4)、AAL-TB1(CAS No 176590-32-2)和AAL-TB2(CAS No 176705-51-4)这些组。FA1-2、FB1-4、FC1-2、FC4和HFC1具有下面的结构式：
FB1是最经常出现的来自镰孢毒素组的毒素，然而已知许多也在人和动物中显示出毒性效应的衍生物和相关分子。将在人或动物中由于摄入真菌毒素而引起的疾病称为真菌毒素中毒，在镰孢毒素的情况下也称为镰孢毒素真菌毒素中毒。因而，已知镰孢毒素通过与神经酰胺合酶的相互作用而阻碍鞘脂代谢。鞘脂不仅是细胞膜的组成部分，而且也在许多基本的细胞过程(例如细胞生长、细胞迁移和细胞连接)中，在炎症过程或细胞内转运程序中，作为信号分子或信使分子发挥重要的作用。由于阻碍鞘脂代谢，镰孢毒素被认为对于在最不同的动物种类和人上的毒性效应负有责任。因而，可以证明，镰孢毒素以免疫抑制方式在啮齿动物中产生致癌效应，并且通过流行病学数据将它们与人中的食管癌和神经管缺陷相联系。在各种不同的动物种类例如猪中，它们被认为对于典型的由中毒引起的肺水肿负有责任。关于镰孢毒素真菌毒素中毒的例子为神经毒性疾病，例如马的脑白质软化或者在猪中的由于串珠镰孢菌素中毒而引起的肺水肿。由于在各种不同的谷类上和特别地在玉米、坚果和蔬菜上镰孢毒素的污染几乎无所不在，因此它们的涉及人和动物的健康的强烈负面的效应是不可忽略的。
在EP-A 1 860 954中已经描述了串珠镰孢菌素的微生物降解，根据该文献使用微生物来使串珠镰孢菌素和串珠镰孢菌素衍生物解毒，在其中向饲料中添加从精确定义的用于使串珠镰孢菌素解毒的菌株中选择的解毒性细菌或酵母。
也已经描述了用于串珠镰孢菌素的生物降解的分解代谢途径和对此负责的基因和酶。因而，例如EP-A 0 988 383描述了使串珠镰孢菌素解毒的组合物和方法，其中首先在转基因植物中产生所使用的降解串珠镰孢菌素的酶，在其中串珠镰孢菌素的解毒借助于氨基氧化酶来实现，所述氨基氧化酶对于其酶促活性来说需要分子氧。
此外，WO 2004/085624描述了氨基转移酶、脱氨酶和氨基变位酶以及用于以酶促方式使胺化毒素(例如串珠镰孢菌素)解毒的方法。在此使用具有脱氨酶活性的多肽用于进行解毒。
上面所述的产品或方法具有缺点，即对于真菌毒素的解毒来说需要分子氧以及任选地辅因子，其中特别地上述氨基氧化酶在无氧的反应条件下不显示出效应。
EP-A 2 326 713涉及添加剂，以及涉及用于制备所述添加剂的方法，用所述添加剂可成功实现在不依赖于氧且无辅因子的酶促反应中降解串珠镰孢菌素。然而，在其中所描述的和对于解毒来说决定性的酶即羧酸酯酶的温度稳定性是低的，以至于添加剂或SEQID No.46的羧酸酯酶不适合于在高的温度下进行使用。
在食品工业和饲料工业中，用于制备经卫生化的产品的热处理(在其中降低了微生物负荷)具有重大意义。在此，特别地，出于众多原因(例如为了改善流动性，为了减少粉尘产生，为了降低微生物负荷，特别是沙门氏菌的微生物负荷)的饲料的粒化是广泛流行的，并且是一种标准化过程。在粒化过程期间，通常借助于热蒸汽来使物品润湿、将其加热并随后在压力下挤压通过模具。使用多肽或酶作为待粒化的食物或饲料的添加剂是一项技术挑战，这是因为酶或多肽通常对于高的温度是敏感的。酶或多肽的热处理可以导致其比活性的下降和/或不可逆的变性。一种对此进行抵抗的可能性是蛋白质的包囊或包衣，例如在WO 92/12645中所描述的那样。由此可成功实现保护蛋白质免于热的影响，然而这种做法带来这样的风险，即蛋白质在口-胃-肠道中不能够足够快地释放，因此其效应要么不能够足够快地发挥要么完全不能够发挥。迄今已知的用于使镰孢毒素解毒的多肽由于其低的温度稳定性而不能够在没有事先包囊或事先包衣的情况下添加给待粒化的饲料或食物。
在其中在高的温度下使镰孢毒素解毒是重要的技术过程为，例如面糊制品和其他玉米产品(例如玉米粥、爆米花、玉米片、玉米面包或玉米粉圆饼)的生产，以及淀粉液化过程、糖化过程或发酵过程(例如制浆过程或发酵过程，特别是在生物乙醇制备中)。在此重要的是确保通过这些过程制备的食物或饲料不包含有害量的镰孢毒素。迄今已知的多肽由于其在各自的工艺温度下具有过低的活性或不存在活性而不能够在这些过程中使用。
因此，对于用于特异性地、安全地和可靠地裂解镰孢毒素(特别地用于使镰孢毒素解毒)的酶或多肽存在需求，其中酶促反应既不需要氧也不需要辅因子，并且所述酶或多肽此外还具有足够的温度稳定性以及足够的温度活性，因此所述酶或多肽可以在处于高的温度下的技术过程中进行使用。
现在，本发明旨在提供SEQ ID No.46的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体以供使用，用所述多肽变体可成功实现在不依赖于氧且无辅因子的情况下将至少一种镰孢毒素裂解为无毒或低毒的产物，其中所述多肽变体具有相比于SEQ ID No.46的镰孢毒素羧酸酯酶而言提高的温度稳定性或提高的温度活性。
为了解决该任务，本发明的特征主要在于，所述多肽变体各自具有在N-末端处缩短了47个氨基酸的氨基酸序列，其中所述氨基酸序列与SEQ ID No.46的氨基酸序列区段48-540具有至少70％，优选地80％，特别优选地100％的序列同一性，即SEQ ID No.1。已经令人惊讶地显示，相比于SEQ ID No.46而言缩短了47个氨基酸的氨基酸序列不仅相比于该序列而言显著地更具有活性，而且具有相比于该序列而言提高的温度稳定性。
如相应于本发明的一个进一步改造方案那样，通过形成SEQ ID No.1的多肽变体，特别是具有与SEQ ID No.1具有至少70％的序列同一性的氨基酸序列并且在至少一个从由下列各项组成的组中选择的位置处具有氨基酸置换的多肽变体：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，获得了相比于具有SEQ IDNo.1的镰孢毒素羧酸酯酶而言提高了至少4％的温度稳定性。通过使用这样的具有裂解镰孢毒素的特性的多肽变体，可成功实现例如在处于高的温度下的技术过程期间使镰孢毒素解毒。这特别地通过除了温度稳定性外同样也提高了的温度活性而成为可能。特别地，由此可以确保，所述多肽变体的酶促活性在高的温度荷载下(如例如在容器中进行运输期间也可以是这种情况)也仍然保持。
名称“羧酸酯酶”涉及所有这样的酶、多肽或多肽变体，其能够借助于水将羧酸酯化合物裂解为相应的醇化合物和羧酸化合物。名称“镰孢毒素羧酸酯酶”涉及所有这样的酶、多肽或多肽变体，其能够水解至少一种镰孢毒素，通过从中以水解方式裂解出至少一个丙三羧酸(丙烷-1,2,3-三羧酸)。在此所描述的“裂解镰孢毒素的特性”涉及以水解方式从至少一种镰孢毒素(特别是FA1-2、FB1-4、FC1-2、FC4、HFC1、AAL-TA1-2和AAL-TB1-2或者其衍生物或降解产物)中裂解出至少一个丙三羧酸残基的能力。
下面所使用的术语取自专业用语并且总是以传统的意义进行使用，除非另外指出。因而，术语“多核苷酸”涉及具有所有长度和序列的每一种类型的遗传物质，例如单链和双链DNA和RNA分子，包括调控元件、结构元件、基因群、质粒、全基因组及其片段。名称“多肽”包括蛋白质，例如酶、抗体，以及具有直至500个氨基酸的多肽，例如肽抑制剂，蛋白质的结构域，还有具有小的序列长度例如少于10个氨基酸的短多肽，例如受体、配体、肽激素、标签等。名称“在多核苷酸或多肽中的‘位置’”涉及在所述多核苷酸或多肽的序列中单个特定的碱基或氨基酸。氨基酸的名称缩写为专业人员熟悉的单字母代码或三字母代码。
名称“单位”或“U”涉及酶、多肽或多肽变体的催化活性的量度，并且被定义为在一分钟内在确定的条件下被转化或裂解的底物(在此为串珠镰孢菌素B1)的微摩尔数(μmol)。例如，如果在15分钟内从60μmol FB1中裂解出至少一个丙三羧酸残基，那么这相应于4个单位的催化活性。对于FB1裂解，如下来定义反应条件：转化反应在具有0.1mg/ml牛血清白蛋白的20mM Tris-HCl缓冲液(pH 8.0)中在30℃的温度下进行30分钟。在反应中的底物浓度为100μΜFB1。
将以“单位/毫升溶液”给出的酶溶液或多肽溶液或多肽变体溶液的酶浓度定义为酶溶液或多肽溶液或多肽变体溶液的“催化活性”、“催化酶活性”或“活性”。
名称“比活性”被定义为每毫克酶、多肽或多肽变体的催化活性，并且从酶溶液的催化活性与在该溶液中酶的质量浓度(质量/体积单位)之比来计算出来。例如，如果酶溶液具有50U/ml的催化活性和1mg/ml的质量浓度，那么比活性为50U/mg。
名称“温度稳定性”涉及酶、多肽或多肽变体在对于高的温度的在时间上受限制的暴露(预温育)之后保持其催化活性的特性。通过测量在相同且确定的条件下在5分钟的热处理后和没有热处理的情况下酶溶液或多肽溶液或多肽变体溶液的活性并进行比较来测定温度稳定性。因此，温度稳定性是关于酶对于在时间上有限制的热作用的抵抗能力的量度。经热处理的酶相比于未经热处理的100％-对照而言的剩余活性为50％时所处的温度为关于温度稳定性的量度，并且缩写为T(50％)。例如，如果没有预温育的酶溶液的活性为50U/ml并且在50℃下预温育5分钟后为25U/ml，那么所述酶的温度稳定性为50℃或者说直至50℃所述酶是温度稳定的。多肽变体相比于SEQ ID No.1的亲本多肽而言的T(50％)的增加被定义为关于提高的温度稳定性的量度，并且可以相对地作为百分比值或者绝对地以摄氏度来给出。
术语“温度活性”定义了这样的温度，在所述温度下所述酶、多肽或多肽变体具有最高的活性，其中在30分钟的时间段内测量催化活性。
术语“多肽变体”或“变体”特别地涉及这样的多肽序列，其相比于SEQ ID No.46而言具有至少一个氨基酸置换，其中仍然保持了酶促功能，即裂解镰孢毒素的特性。此外，“多肽变体”还可以相对于SEQ ID No.46的多肽序列而言具有氨基酸插入或缺失，特别是在C-末端或N-末端处延长或缩短了的序列。如果酶促反应机制保持不变，那么酶促功能基本上得到保持，即镰孢毒素在相同的位点处被水解并且相关于SEQ ID No.46的原始亲本多肽而言所述变体的比活性为至少10％，优选地至少50％，更优选地至少90％，然而特别地大于>100％。
在确定位置处的氨基酸置换用下面的命名法来进行描述：原始氨基酸；位置；新氨基酸。例如，如果在位置134处的脯氨酸被甘氨酸替代，那么这用Pro134Gly或P134G来指明。多重突变用加号或斜线来分开。例如，如果在位置134处的脯氨酸被甘氨酸替代和在位置136处的精氨酸被赖氨酸替代，那么这用Pro134Gly+Arg136Lys或Pro134Gly/Arg136Lys或P134G+R136K或P134G/R136K来指明。如果在一个位置处的氨基酸被两个或更多个备选氨基酸替代，那么备选氨基酸用逗号或斜线来分开。例如，如果在位置134处的脯氨酸不仅被甘氨酸替代而且也被丝氨酸、缬氨酸和甲硫氨酸替代，那么这用Pro134Gly,Ser,Val,Met或Pro134Gly/Ser/Val/Met或P134G,S,V,M或P134G/S/V/M来指明。如果未详细地定义在确定位置处的氨基酸置换或替换，那么这如此地来进行解释，即这些氨基酸可以被所有其他氨基酸替代。例如，如果未详细地定义在位置134处的脯氨酸的突变，那么该脯氨酸可以被下列氨基酸之一替代：A、R、N、D、C、Q、E、G、H、I、L、K、M、F、S、T、W、Y或V。
名称“序列同一性”涉及序列同一性百分比。对于氨基酸序列和核苷酸序列，序列同一性可以目视测定，但是优选地用计算机程序来计算出来。将具有SEQ ID No.1的氨基酸序列定义为参考序列。还在序列区段内进行序列比较，其中参考序列的连续序列被理解为区段。对于核苷酸序列来说，序列区段的长度通常为18至600个核苷酸，优选地45至200个核苷酸，更优选地100至150个核苷酸。对于肽序列来说，序列区段的长度通常为3至200个氨基酸，更优选地15至65个氨基酸，最优选地30至50个氨基酸。存在有许多可购得的或免费可用的生物信息学程序，其可以被召来用于同源性测定并且持续地得到进一步发展。对此的例子为GCG Wisconsin Bestfit软件包(Devereux等人,1984)、BLAST(Altschul等人,1990)或BLAST 2(Tatusova和Madden,1999)。由于这些算法的不同的设置可能性，在相同的输入序列的情况下它们得到不同的结果是可能的。因此，必需定义搜索算法和与此有关的设置。在本情况下，序列同一性借助于程序NCBI BLAST(Basic Local Alignment Search Tool)来进行测定，特别地对于多肽，用BLASTP，和对于多核苷酸，用BLASTN，其提供在“NationalCenter for Biotechnology Information”的主页(NCBI；http://www.ncbi.nlm.nih.gov/)上以供使用。因此，可能的是，根据Altschul等人,1997(NucleicAcids Res.,25:3389-3402)的算法，将两个或更多个序列互相进行比较。在此，使用2014年8月12日的那个版本的程序。作为程序设置，考虑基本设置，特别地，对于氨基酸序列比较：“最大靶序列(max target sequence)”＝100；“期望阈值(expected threshold)”＝10；“字长(word size)”＝3；“矩阵(matrix)”＝BLOSOM62；“缺口成本(gap costs)”＝“存在(Existence)：11；延伸(Extention)：1”；“计算调整(computational adjustment)”＝“条件式组成得分矩阵调整(Conditional compositional score matrix adjustment)”；以及对于核苷酸序列比较：字长：11；期望值(Expect value)：10；缺口成本：存在＝5，延伸＝2；过滤器(Filter)＝低复杂度激活的(low complexity activated)；匹配/错配得分(Match/Mismatch Scores)：2，-3；过滤器字符串(Filter String)：L；m。
根据本发明的一个进一步改造方案，通过进行下列氨基酸置换而由此获得温度稳定性的特别大的提高：在位置10和456处的氨基酸置换选自Q、E、N、H、K和R，在位置33、107、293和332处的氨基酸置换选自E、Q、D、K、R和N，在位置66、463和478处的氨基酸置换选自D、E、K、N、Q和R，在位置140和490处的氨基酸置换选自P、A、S和N，在位置144和367处的氨基酸置换选自I、L、M和V，在位置149、270、312、329和372处的氨基酸置换选自F、Y、W和H，在位置151和453处的氨基酸置换选自D、E、K和R，在位置157和462处的氨基酸置换选自F、H、W和Y，在位置199、302、365和464处的氨基酸置换选自I、L、M和V，在位置266和455处的氨基酸置换选自A、S和T，在位置267、394和429处的氨基酸置换选自A、N、P和S，在位置272处的氨基酸置换选自H、N、Q和S，在位置275处的氨基酸置换选自A、D、E、G、K、N、Q、R和S，在位置280处的氨基酸置换选自A、D、E、K、N、P、Q、R和S，在位置284处的氨基酸置换选自A、N、P、S、T和V，在位置286处的氨基酸置换选自A、D、E、K、N、P、R和S，在位置360、377、391、419和427处的氨基酸置换选自A、I、L、S、T和V，在位置363、443和457处的氨基酸置换选自A、S、T和V，在位置364处的氨基酸置换选自H、I、L、M、N、Q、S和V，在位置371处的氨基酸置换选自A、I、L、M、S、T和V，在位置389处的氨基酸置换选自I、L、M和V，在位置418、430、447和473处的氨基酸置换选自A、G和S，在位置424处的氨基酸置换选自A、D、E、G、K、R和S，在位置436处的氨基酸置换选自A、G、S和T，在位置440处的氨基酸置换选自A、G、S和T，在位置465处的氨基酸置换选自A、G、H、N、Q、S和T，在位置469处的氨基酸置换选自D、E、K和R，和/或在位置487处的氨基酸置换选自N、D、Q、H和S，其中在上述位点处原来存在的氨基酸无论如何均被替换。
根据本发明的一个优选的进一步改造方案，所述氨基酸置换选自由下列各项组成的组：10Q、33E、66D、107E、140P、144M、149F、151R、157Y、199I、266S、267P、270F、272H、275E、275A、280D、280P、284T、284P、286P、286R、293E、302I、312F、329F、332E、360V、363T、364H、364L、365I、367H、371V、371M、372F、377V、389L、391V、394P、418A、419V、424A、424K、427V、429P、430A、436A、436S、440G、440S、443T、447A、453R、455S、456Q、457T、462Y、463D、464I、465H、465S、465G、469K、473A、478D、487N和490P。通过所述多肽变体具有至少一个这样的置换，可以更加有针对性地提高温度稳定性或温度活性，其中另外地，其他酶参数(例如比活性、pH-稳定性或pH-活性)同样也可以得到改善，然而其至少有着具有SEQ ID No.1的镰孢毒素羧酸酯酶的值。
根据本发明的一个进一步改造方案，所述多肽变体在至少一个从由66、199、302、377、394、424、430和463组成的组中选择的位置处具有氨基酸置换，并且具有相比于具有SEQ ID No.1的镰孢毒素羧酸酯酶而言提高了至少6％的温度稳定性。借助于这样的多肽变体，可能的是，在具有高的温度的技术过程例如面糊制品和其他玉米产品(例如玉米粥、爆米花、玉米片、玉米面包或玉米粉圆饼)的生产，以及淀粉液化过程、糖化过程或发酵过程(例如制浆过程或发酵过程，特别是在生物乙醇制备中)期间裂解镰孢毒素。
根据本发明的一个优选的进一步改造方案，所述氨基酸置换选自由下列各项组成的组：66D、199I、302I、377V、394P、424A、430A和463D。借助于这样的置换，可以使所述多肽变体的温度稳定性以及温度活性相对于SEQ ID No.1的镰孢毒素羧酸酯酶而言提高至少3℃。
根据本发明的一个进一步改造方案，所述多肽变体在该氨基酸序列的至少两个，特别是三个从由下列各项组成的组中选择的位置处具有氨基酸置换：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，并且具有相比于SEQ IDNo.1的镰孢毒素羧酸酯酶而言提高了至少15％的温度稳定性。
根据本发明的一个优选的进一步改造方案，所述氨基酸置换选自由下列各项组成的组：10Q、66D、144M、151R、199I、266S、267P、272H、275E、275A、280D、284T、286P、286R、293E、302I、360V、363T、364H、364L、365I、367H、371V、371M、372F、377V、389L、391V、394P、418A、419V、424A、424K、427V、429P、430A、436A、436S、440G、440S、443T、447A、453R、455S、456Q、457T、462Y、463D、464I、465H、465S、465G、469K、473A、478D、487N和490P。通过多个氨基酸的置换，已经令人惊讶地显示，大致加上了其对于温度稳定性的正面效应，其中通过嵌入至少三个与原来在该序列中存在的氨基酸不同的氨基酸，温度稳定性提高了多于7℃。这样的提高足以例如在生物乙醇制备中在处于大约55℃下的制浆停歇之时或者在处于大约65至70℃的中等温度下的饲料粒化之时使用所述酶。
根据本发明的一个进一步改造方案，所述多肽变体的氨基酸序列包含多个氨基酸置换的组合，其中所述位置组合选自由下列各项组成的组：66/199/302/394/424/430、66/199/302/377/394/424/430、66/199/302/377/394/424/430/463、66/144/199/302/360/372/377/394/424/430/443/463、199/302/377/394/424/430/463、66/199/302/377/394、66/199/302/364/377/394/424/430/463、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/440/463、66/199/302/377/394/424/430/447/463、66/199/302/377/394/418/424/430/463、66/199/302/377/394/424/436/430/463、66/199/302/364/377/394/424/430/463、66/199/302/377/394/424/430/463/490、66/199/302/377/394/424/430/463/469、66/199/302/377/389/394/424/430/463、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/463/464、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/440/463、66/199/302/377/394/424/430/457/463、66/199/302/377/394/424/430/436/463、66/199/302/363/371/377/394/424/430/463、66/199/302/377/394/424/430/447/453/463、66/199/302/377/394/424/430/456/462/463、66/199/302/377/394/419/424/427/430/463、66/199/302/365/377/394/424/430/463/487和66/199/302/371/377/394/424/430/463/487。如此经置换的多肽变体具有相比于SEQ ID No.1的镰孢毒素羧酸酯酶而言提高了至少25％的温度稳定性。通过这样的置换组合，可能的是，不仅提高了所述多肽变体的温度稳定性，而且还提高了所述多肽变体的温度活性。
根据本发明的一个优选的进一步改造方案，所述多肽变体的氨基酸序列选自由SEQ ID No.2至29组成的组。这样的多肽变体具有相比于SEQ ID No.1的酶而言提高了至少11℃，优选地至少13℃和更加优选地至少15℃的温度稳定性，由此在具有高的温度负荷的技术过程(例如在生物乙醇制备中处于65℃下的制浆停歇，或者处于大约75-80℃的温度下的粒化)期间或之后保证各自多肽的活性和因此镰孢毒素的解毒。
根据本发明的一个进一步改造方案，所述多肽变体的每一个氨基酸序列包含多个氨基酸置换的组合，其中所述位置组合选自由下列各项组成的组：66/99/302/364/377/389/394/419/424/427/430/447/463/465/469、66/199/302/377/389/394/419/424/427/430/447/463/465/469、66/199/302/363/364/371/377/389/394/419/424/427/430/447/463/464/465/469、66/199/302/363/371/377/389/394/419/424/427/430/447/463/464/465/469、66/199/302/364/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、66/199/302/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、66/199/302/363/367/371/377/394/424/430/463/490、66/199/302/377/394/418/419/424/427/430/436/440/447/463、66/199/302/377/389/394/424/430/457/463/464/465/469、66/199/302/363/371/377/389/394/419/424/427/430/440/447/457/463/464/469/490、66/199/302/377/394/424/430/463/447/490/469/465、66/199/302/377/394/424/430/463/490/469/465/419/427/371/487、66/199/302/371/377/394/419/424/427/430/447/453/463/465/469/487/490、66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/457/463/464/465/469/490、66/199/302/371/377/389/394/419/424/427/430/436/447/453/456/462/463/465/469/490/487和66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/453/456/457/462/463/464/465/469/487/490。这些多肽变体具有相比于SEQ ID No.1的镰孢毒素羧酸酯酶而言提高了至少40％的温度稳定性以及增高了的温度活性，由此它们在许多需要更高温度的方法中是可使用的。
根据本发明的一个优选的进一步改造方案，所述多肽变体的氨基酸序列选自由SEQ ID No.30至45组成的组。这样的多肽变体具有至少18℃，优选地至少22℃，和更优选地至少27℃的相比于SEQ ID No.1的酶而言提高的温度稳定性，由此在具有高的温度负荷的技术过程(例如处于超过80℃，特别地超过85-90℃的温度下的粒化)期间或之后保证所述多肽的活性和因此镰孢毒素的解毒。特别地，在大约90℃的高的温度下的粒化在家禽工业中具有重大意义，以便保证饲料中沙门氏菌负荷的充分降低。
名称“保守突变”涉及氨基酸由被专业人员视为保守的(即具有类似的特殊特性的)其他氨基酸来进行的置换，或者说该氨基酸的特性仍然是得到保存的，即得以保留的。氨基酸的特殊特性为例如其大小、极性、疏水性、电荷或pK值。可以将氨基酸基于其特性划分为组，并且所述组可以以维恩图(Venn Diagram)来描绘。来自相同组和优选地来自相同亚组的氨基酸可以相互置换。依据疏水性、极性和大小这些特性把氨基酸划分为组和亚组可以从Taylor W.R.(1986)中获知。“保守突变”是指例如将一个酸性氨基酸置换成另一个酸性氨基酸，将一个碱性氨基酸置换成另一个碱性氨基酸，将一个极性氨基酸置换成另一个极性氨基酸，等等。特别地，所述多肽变体另外还可以在至少一个位置处包含至少一个保守氨基酸置换，其中所述保守氨基酸置换选自由下列置换组成的组：G至A，A至G/S，V至I/L/A/T/S，I至V/L/M，L至I/M/V，M至L/I/V，P至A/S/N，F至Y/W/H，Y至F/W/H，W至Y/F/H，R至K/E/D，K至R/E/D，H至Q/N/S，D至N/E/K/R/Q，E至Q/D/K/R/N，S至T/A，T至S/V/A，C至S/T/A，N至D/Q/H/S，和Q至E/N/H/K/R。
如果在根据本发明的多肽变体中在确定位置处的置换导致例如极性氨基酸(例如Asp)被疏水氨基酸(例如Ala)替代，那么“保守突变”也是指所有在所述位置处导致另一个疏水氨基酸(例如甘氨酸、亮氨酸、苯丙氨酸、缬氨酸)的突变。在此也包括这些包含备选的保守突变的进一步的多肽变体。
此外，本发明还旨在提供编码SEQ ID No.1的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体的多核苷酸以供使用，用所述多肽变体可成功实现在不依赖于氧且无辅因子的情况下将至少一种镰孢毒素裂解为无毒或低毒的产物，并且所述多肽变体具有相比于SEQID No.1的镰孢毒素羧酸酯酶而言提高的温度稳定性。
为了解决该任务，本发明的特征在于，所述多核苷酸具有编码具有SEQ ID No.1的氨基酸序列的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体的核苷酸序列，其中所述多肽变体具有与具有SEQ ID No.1的氨基酸序列有着至少70％的序列同一性的氨基酸序列，并且所述多肽变体在至少一个从由下列各项组成的组中选择的位置处具有氨基酸置换：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，然而特别地具有下列置换：10Q、33E、66D、107E、140P、144M、149F、151R、157Y、199I、266S、267P、270F、272H、275E、275A、280D、280P、284T、284P、286P、286R、293E、302I、312F、329F、332E、360V、363T、364H、364L、365I、367H、371V、371M、372F、377V、389L、391V、394P、418A、419V、424A、424K、427V、429P、430A、436A、436S、440G、440S、443T、447A、453R、455S、456Q、457T、462Y、463D、464I、465H、465S、465G、469K、473A、478D、487N和490P或者其组合。用这样的分离的多核苷酸，可以借助于表达载体来制备用于产生根据本发明的多肽变体的转基因宿主细胞。
名称“表达载体”涉及能够在体内或在体外表达基因的DNA构建体。特别地，该名称涵盖这样的DNA构建体，其适合于将编码所述多肽的核苷酸序列转移到宿主细胞中，从而使得所述核苷酸序列整合到基因组中或者游离地存在于染色体外空间中，并且在细胞内表达编码所述多肽的核苷酸序列和任选地从细胞中提取出所述多肽。
名称“宿主细胞”涉及所有这样的细胞，其要么包含待表达的核苷酸序列，要么包含表达载体，并且能够制备根据本发明的酶或多肽。特别地，该名称涵盖原核细胞和/或真核细胞，优选地巴斯德毕赤酵母(Pichia pastoris)，大肠杆菌(E.coli)，枯草芽孢杆菌(Bacillus subtilis)，链霉菌属(Streptomyces)，汉逊酵母属(Hansenula)，木霉属(Trichoderma)，乳杆菌属(Lactobacillus)，曲霉属(Aspergillus)，植物细胞，和/或芽孢杆菌属(Bacillus)、木霉属或曲霉属的孢子。在此所使用的名称“巴斯德毕赤酵母(Pichiapastoris)”是名称“Komagataella pastoris”的同义词，其中巴斯德毕赤酵母(Pichiapastoris)是较老的名称而Komagataella pastoris是系统分类学上较新的名称(Yamada等人,1995)。
此外，本发明还旨在提供裂解镰孢毒素的添加剂以供使用，所述添加剂包含至少一种具有SEQ ID No.46的氨基酸序列的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体，其中各自多肽变体在不依赖于氧且无辅因子的情况下将至少一种镰孢毒素裂解为无毒或低毒的产物并且具有相比于SEQ ID No.46的镰孢毒素羧酸酯酶而言提高的温度稳定性。
为了解决该任务，本发明的特征在于，所述裂解镰孢毒素的添加剂包含至少一种根据本发明的镰孢毒素羧酸酯酶的多肽变体，和任选地，至少一种助剂。通过将这样的添加剂添加至被镰孢毒素污染的饲料，可以成功实现使镰孢毒素解毒，这可以显著地通过在用所述添加剂饲喂的受试者的血浆和/或肾脏和/或肺和/或肝脏中二氢鞘氨醇与鞘氨醇的比例的降低来进行测量。
在动物的不同器官中以及在动物的血浆中二氢鞘氨醇与鞘氨醇的比例是关于镰孢毒素(特别是FB1)的毒性效应的普遍接受且灵敏的生物标志物。由镰孢毒素引起的鞘脂代谢紊乱尤其与马的脑病或猪中的肺水肿有关。作为生物标志物的二氢鞘氨醇与鞘氨醇的比例的重要性以及其分析测量描述在Grenier等人(Biochem.Pharmaceuticals,第83卷,(2012),第1465-1473页)中以及描述在EFSA Journal(2014,12(5):3667)中。
根据本发明的一个进一步改造方案，如此地来形成所述添加剂，从而使得所述助剂选自由下列各项组成的组：惰性载体、维生素、矿物质、植物源性物质、酶和其他用于使真菌毒素解毒的组分，例如真菌毒素降解酶，特别是黄曲霉毒素氧化酶、麦角胺水解酶、麦角胺酰胺酶、玉米赤霉烯酮酯酶、玉米赤霉烯酮内酯酶、玉米赤霉烯酮水解酶、赭曲毒素酰胺酶、串珠镰孢菌素氨基转移酶、氨基多元醇氨基氧化酶、脱氧瓜萎镰菌醇环氧化物水解酶、脱氧瓜萎镰菌醇脱氢酶、脱氧瓜萎镰菌醇氧化酶、单端孢霉烯脱氢酶、单端孢霉烯氧化酶；降解真菌毒素的微生物；和结合真菌毒素的物质，例如微生物细胞壁或无机材料例如膨润土。通过使用这样的添加剂，可以例如在饲料或食物中确保，所任选地包含的镰孢毒素的量安全可靠地被裂解特别是解毒至如此程度，即对于摄入了这些饲料或食物的受试者的机体的有害效应不存在。
本发明的另外的使用领域为这样的添加剂，其除了包含至少一种根据本发明的多肽变体外，还包含至少一种酶，所述酶例如参与蛋白质的降解，例如蛋白酶，或者所述酶参与淀粉或纤维或脂肪或糖原的代谢，例如淀粉酶、纤维素酶或葡聚糖酶，以及例如水解酶、脂解酶、甘露糖苷酶、氧化酶、氧化还原酶、植酸酶或木聚糖酶。
此外，本发明还旨在根据本发明的添加剂用于裂解在食物或饲料中的至少一种镰孢毒素的用途，所述食物或饲料特别地是经粒化的，所述饲料特别地为用于猪、家禽、牛、马、鱼类或水产养殖的饲料。作为食物或饲料，可以使用所有适合于人或动物营养(特别地也适合于家畜、绵羊或山羊)的食物或饲料，特别地也使用干酒槽。
此外，本发明还旨在根据本发明的添加剂用于在用于制备或加工食物或饲料的过程中裂解至少一种镰孢毒素的用途，所述过程特别地在至少50℃的温度下进行。通过所述添加剂的这样的用途，保证了镰孢毒素的解毒，例如在其中在高的温度下的处理是重要的食品技术过程期间，例如在玉米或谷物的加工中，在淀粉液化过程中，在糖化过程中，或在发酵过程(例如制浆过程或发酵过程，特别是在生物乙醇制备中)中。由此可以确保，在来自于此类方法的产品(例如饲料颗粒、面糊制品、玉米粥、爆米花、玉米片、玉米面包、玉米粉圆饼、干酒糟或淀粉)中没有重大的(特别地有损于健康的)量的镰孢毒素保持完整。
此外，本发明还旨在至少一种根据本发明的多肽变体或者至少一种根据本发明的添加剂在制备用于预防和/或治疗镰孢毒素真菌毒素中毒的制剂中的用途。通过多肽变体或添加剂的这样的用途，在预防的情况下可能的是，尽管经口摄入了镰孢毒素，但是将人或动物的健康状况基本上保持在这样的水平，该水平相应于没有经口摄入镰孢毒素或经口摄入了减少的镰孢毒素的水平。在治疗镰孢毒素真菌毒素中毒的情况下，可能的是，缓解此类疾病的症状和特别地显著改善在器官和/或血浆中二氢鞘氨醇与鞘氨醇的比例。此外，通过这样的用途，还可能的是，改善有用动物的生产能力，特别是饲料利用率和体重增加，并且降低死亡率。
此外，本发明还旨在提供用于以酶促方式裂解至少一种镰孢毒素的方法以供使用，采用所述方法可以用多肽将至少一种镰孢毒素不依赖于氧地、特异性地、安全地和可靠地以水解方式裂解为无毒或低毒的产物，其中所述以水解方式的裂解要么在热处理期间要么在热处理之后进行。
为了解决该任务，如此地来施行所述方法，从而通过根据本发明的多肽变体或根据本发明的添加剂从镰孢毒素中以水解方式裂解出至少一个丙三羧酸。在此，将所述至少一种镰孢毒素与至少一种根据本发明的多肽变体或至少一种根据本发明的添加剂相混合，通过至少一种多肽变体从至少一种镰孢毒素中以水解方式裂解出至少一个丙三羧酸，由此使所述镰孢毒素解毒，其中使由各自多肽变体和镰孢毒素组成的混合物经历至少50℃，优选地至少70℃的热处理，并且所述以水解方式的裂解要么在所述热处理期间要么在所述热处理之后进行。
在本发明的一个优选的进一步改造方案中，如此地来施行所述方法，从而将所述多肽变体或所述添加剂与被至少一种镰孢毒素污染的饲料或食物相混合，并且进行热处理(任选地通过粒化过程)。由此确保，一旦由所述多肽变体和所述镰孢毒素组成的混合物与湿气相接触，在被污染的和任选地经粒化的饲料或食物中所包含的镰孢毒素就被裂解。在湿润的饲料或食物例如麦芽浆或糊浆的情况下，镰孢毒素的水解在经口摄入之前在该湿润的饲料或食物中发生。由此达到，最大程度地消除或至少降低镰孢毒素对于人和动物的有害效应。在此，湿气是指水或含水液体的存在，其中例如唾液或其他在消化道中存在的液体也属于该范畴。消化道被定义为口腔、咽(咽喉)、食管和胃肠道或者其等价物，其中在动物中可以存在不同的名称或者个别成分可以不存在于动物的消化道中。
在本发明的一个优选的进一步改造方案中，如此地来施行所述方法，以5U至500U，优选地10U至300U，和更优选地15U至100U/千克饲料或食物的浓度范围使用所述多肽变体。通过添加这些量的多肽变体，可成功实现，取决于镰孢毒素的浓度，将在食物或饲料中(特别地在干酒糟中)的镰孢毒素裂解并因此使其解毒至如此程度，即至少70％，优选地至少80％，特别地至少90％的至少一种镰孢毒素被裂解。
如果没有明确地另外说明，那么单数名称例如“ein”、“eine”、“das”、“die”或“der”被理解为示例性的并且包括许多可能性。例如，如果涉及“基因”、“酶”或“细胞”，那么总是意图包括复数。
下面将依据实施例更详细地阐明本发明。
多肽变体
实施例1：编码裂解镰孢毒素的多肽的多核苷酸的修饰、克隆和表达
按照说明书，使用“Quick-change Site-directed Mutagenesis Kits”(Stratagene)，借助于PCR，通过核苷酸序列的突变来进行氨基酸置换、插入或缺失。备选地，为此还合成制备了完整的核苷酸序列(GeneArt)。借助于标准方法，将借助于PCR诱变而产生的或从GeneArt购进的核苷酸序列整合到用于在大肠杆菌(E.coli)或巴斯德毕赤酵母(P.pastoris)中进行表达的表达载体之中，转化到大肠杆菌或巴斯德毕赤酵母中，以及在大肠杆菌或巴斯德毕赤酵母中进行表达(J.M.Cregg,Pichia Protocols,第二版,ISBN-10:1588294293、2007；J.Sambrook等人,2012,Molecular Cloning,A Laboratory Manual,第4版,Cold Spring Harbor)，其中也可以为该任务考虑任何其他合适的宿主细胞。
名称“表达载体”涉及能够在体内或在体外表达基因的DNA构建体。特别地，该名称涵盖这样的DNA构建体，其适合于将编码所述多肽的核苷酸序列转移到宿主细胞中，以便在那里整合到基因组中或者游离地存在于染色体外空间中，并且在细胞内表达编码所述多肽的核苷酸序列和任选地从细胞中提取出所述多肽。名称“宿主细胞”涉及所有这样的细胞，其要么包含待表达的核苷酸序列，要么包含表达载体，并且能够制备根据本发明的酶或多肽。特别地，该名称涵盖原核细胞和/或真核细胞，优选地巴斯德毕赤酵母(P.pastoris)，大肠杆菌(E.coli)，枯草芽孢杆菌(Bacillus subtilis)，链霉菌属(Streptomyces)，汉逊酵母属(Hansenula)，木霉属(Trichoderma)，乳杆菌属(Lactobacillus)，曲霉属(Aspergillus)，植物细胞，和/或芽孢杆菌属(Bacillus)、木霉属或曲霉属的孢子。考虑将可溶的细胞裂解物(在大肠杆菌的情况下)或者培养物上清液(在巴斯德毕赤酵母的情况下)用于测定所述多肽变体的催化特性。
实施例2：降解镰孢毒素的多肽的催化活性和比活性的测定
借助于标准方法将编码降解镰孢毒素的多肽变体的相应基因克隆到大肠杆菌中，在细胞内进行表达，并随后借助于超声波进行裂解并进行离心。将清澈的上清液用具有0.1mg/ml牛血清白蛋白的20mM Tris-HCl缓冲液(pH 8.0)进行稀释(大约10-3至10-5)，并且如此地用于FB1降解批料中，从而通过该多肽将10％至90％的在降解批料中所包含的FB1量降解。
为了测定催化酶活性，进行用于以水解方式裂解串珠镰孢菌素B1(FB1)的试验，其中使试验批料在具有0.1mg/ml牛血清白蛋白的20mM Tris-HCl缓冲液(pH 8.0)中和在30℃的温度下进行30分钟。此外，所述批料还包含100μΜFB1的底物浓度(BiopureReferenzsubstanzen GmbH Tulln,Austria,BRM 001007)和待测试的多肽。在30分钟的温育后，将所述批料在99℃下热灭活5分钟以终止反应。
为了测定饲料样品的酶促活性，必须在所述试验之前从饲料样品中提取出转化镰孢毒素的多肽变体。为此将10g饲料溶解在100ml的具有0.1mg/ml牛血清白蛋白的20mMTris-HCl缓冲液(pH 8.0)中，并在20℃和150rpm下摇动1小时。然后，将样品在4000g下离心15分钟，并且将清澈的上清液根据需要进行稀释(10-2至10-3)并用于FB1批料中。
按照Heinl等人(J.of Biotechnology,2010,145,第120-129页,2.6.3.“Liquidchromatography-mass spectrometry”)的方法，借助于LC-MS(液相色谱法-质谱法)来进行FB1的定量。为此，考虑用FB1标准物进行校准，所述FB1标准物还包含完整的经13C标记的内部FB1标准物(Biopure Referenzsubstanzen GmbH Tulln,Austria)。与Heinl等人(2010)不同，仅测量了FB1的降解，以便测定所使用的多肽溶液的催化酶活性。所使用的多肽溶液的催化酶活性以“单位/ml”来给出，其中将一个“单位”定义为在该试验中在上述反应条件下1μmol FB1/分钟的减少。
为了测定比活性，借助于定量Western印迹或ELISA来测定酶浓度。通过将活性(单位)与所使用的酶量相关联来计算比酶活性，并且以“单位/mg”来给出。
实施例3：降解镰孢毒素的多肽的温度稳定性
如在实施例1和2中所描述的那样，进行降解镰孢毒素的多肽的表达和定量。在活性测定之前，将细胞裂解物的量分为多个部分(每个大约60μL)。使2至10个部分在商用PCR-循环仪(例如EppendorfMastercycler Gradient)中经历热处理5分钟，其中将每个部分在不同的温度下进行温育。同时，将细胞裂解物的另一部分，100％-对照，在冰上进行温育。在热处理之后，将所有样品/试验批料在10℃下温育一分钟以使温度相称。如在实施例2中所描述的那样，测定经热处理的样品以及100％-对照的酶促活性。在热处理后的残留活性被称为剩余活性。将相比于未经热处理的100％-对照而言的剩余活性为50％时所处的温度缩写为T(50％)，并且其是关于多肽的温度稳定性的量度。
相比于具有SEQ ID No.46或SEQ ID No.1的多肽而言多肽变体的T(50％)(其以摄氏度给出)的增加是关于提高的温度稳定性的量度。T(50％)-值的增加可以以℃，但也可以以相对于亲本多肽的T(50％)-值而言的百分比值来给出。下面的例子将会用于进行阐明：如果亲本酶在冰上温育5分钟后具有50U/ml的催化活性并且在48℃下温育5分钟后具有25U/ml的催化活性，那么T(50％)为48℃。如果多肽变体具有51℃的T(50％)，那么温度稳定性(T(50％))的相对提高为6.25。这从所述两个T(50％)-值的3℃的差除以亲本起始酶的48℃的T(50％)-值再乘以100而得出。
为了测定温度稳定性，也可以考虑比活性来代替催化活性。
温度稳定性的测定也可以借助于备选的酶测定法来进行，甚至无需测定FB1转化的催化活性。在此重要的是，在批料中无论如何一定要使用相同量的经热处理的多肽和100％-对照，这例如通过使用相同体积的细胞裂解物来提供。
为了测定温度稳定性，也可以考虑酶促降解试验的测量信号(例如MS信号、消光等)来代替催化活性。如果所述测量信号与酶促活性(例如，经转化的FB1的峰面积)成正比，那么多肽的T(50％)-值为这样的温度，在所述温度下，经热处理的多肽的测量信号值具有该多肽的100％-对照的测量信号值的50％。
测得具有SEQ ID No.46的多肽的温度稳定性(T(50％))为42℃，具有SEQ ID No.1的多肽的温度稳定性为45℃。因此，通过缩短N-末端序列可以达到的温度稳定性的相对提高为大约7％。此外，还可以查明具有SEQ ID No.1的多肽相比于具有SEQ ID No.46的亲本多肽而言的提高的酶促活性。
具有SEQ ID No.1的多肽的温度稳定性可以通过单个氨基酸的有针对性的置换而进一步得到提高。这些多肽变体相比于具有SEQ ID No.1的亲本多肽而言的温度稳定性的相对提高呈现在表1中。
表1：多肽变体的修饰以及其相比于具有SEQ ID No.1的亲本酶而言的以百分比表示的温度稳定性的相对提高
实施例4：降解镰孢毒素的多肽的依赖于温度的活性(温度活性)
在试验进行之前纯化待就其依赖于温度的活性进行测试的、降解镰孢毒素的多肽变体。为此，借助于阴离子交换色谱法和随后的大小排阻色谱法以两步骤过程从发酵上清液中纯化出多肽变体。将多肽变体调整至1mg/ml的浓度，并且在1ml的反应体积中以10-5-10-6的稀释度用于反应批料中。如在实施例3中所描述的试验那样，通过FB1水解和随后的借助于LC-MS的FB1的定量来进行活性测定，其中所述试验在各种不同的温度下进行。用两个加热板(Eppendorf,ThermoMixer)在温度10℃、20℃、30℃、35℃、40℃、45℃、50℃、55℃、60℃、65℃和70℃下进行温育。在热作用开始后30分钟，各取出100μl的反应批料并在99℃下热灭活5分钟。在此，在加热板中于30℃下进行的试验用作100％-对照。在表2中列举了示例性结果。表2：降解镰孢毒素的多肽的依赖于温度的活性。<LOQ的值低于检测极限(定量水平(Level of Quantification)：<0.15U/l，在试验批料中)
实施例5：降解镰孢毒素的多肽的粒化稳定性的测定
借助于标准方法，将所选择的多肽变体克隆到巴斯德毕赤酵母中，在受控的有氧条件下在生物反应器中进行表达，并分泌到细胞外。将清澈的上清液与生物质分开，向其掺入载体物质(麦芽糖糊精)，并且借助于喷雾干燥设备将其加工成可粒化的粉末。将以粉末形式存在的降解镰孢毒素的多肽变体各自以100U/kg的相同浓度与仔猪饲养饲料相混合，并且在受控的过程中加工成饲料颗粒。在颗粒化过程期间，将饲料以单个批次在精确确定的温度(75-95℃，以5℃的间距)下借助于热蒸汽进行润湿并加热。在该准备阶段后，进行真正的粒化过程。如在实施例2中所描述的那样，测定在所述颗粒中所包含的降解镰孢毒素的多肽变体的剩余活性，其中未经粒化的具有各自的降解镰孢毒素的多肽变体的饲料用作100％-对照。因此，在颗粒化过程后残留的酶活性被定义为剩余活性。所述值在表3中给出。
表3：降解镰孢毒素的多肽的粒化温度和剩余活性。<LOQ的值低于检测极限(定量水平：<0.15U/l，在试验批料中)
序列表
<110> Erber Aktiengesellschaft
<120> 裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法
<130> P05368PCT
<160> 46
<170> PatentIn version 3.5
<210> 1
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> 从SEQ ID No. 46进行修饰的
<400> 1
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asn Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Leu Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Leu Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Ala Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Ala Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Glu Gly Gly Ala Gly Ala Ser Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Glu Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 2
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 2
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Ala Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Glu Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 3
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 3
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Glu Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 4
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 4
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 5
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 5
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Met
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Val Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Phe Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Thr Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 6
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 6
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asn Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 7
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 7
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Glu Gly Gly Ala Gly Ala Ser Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Glu Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 8
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 8
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser His Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 9
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 9
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
His Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 10
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 10
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Gly Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 11
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 11
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 12
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 12
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 13
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 13
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 14
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 14
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Leu Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 15
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 15
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 16
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 16
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 17
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 17
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 18
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 18
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 19
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 19
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Ile
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 20
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 20
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Gly Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 21
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 21
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ser Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 22
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 22
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Thr His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 23
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 23
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ser Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 24
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 24
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Thr Gln Phe Asn Asn Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 25
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 25
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Arg Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 26
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 26
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Gln Ser His Trp Pro Arg Tyr Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 27
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 27
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 28
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 28
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Ile Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Gln Pro Ser Lys
485 490
<210> 29
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 29
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Met Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Gln Pro Ser Lys
485 490
<210> 30
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 30
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Leu Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 31
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 31
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 32
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 32
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Thr Leu Phe Asn Asn Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 33
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 33
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Thr Gln Phe Asn Asn Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 34
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 34
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Leu Phe Asn His Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 35
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 35
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn His Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 36
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 36
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Thr Gln Phe Asn His Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 37
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 37
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Arg Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 38
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 38
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Thr His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
485 490
<210> 39
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 39
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Thr Gln Phe Asn Asn Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Thr His Trp Pro Arg Phe Asp Ile
450 455 460
Arg Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 40
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 40
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Leu Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Met Phe Gly Ser Leu Ala Gly Gly Ala Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 41
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 41
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Met Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Pro Pro Ser Lys
485 490
<210> 42
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 42
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Met Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Thr Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Arg Gly Thr Lys Ser His Trp Pro Arg Phe Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Pro Pro Ser Lys
485 490
<210> 43
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 43
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn His Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Val Phe Gly Ser Leu Ala Gly Gly Val Gly Pro Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Gln Gly Thr Lys Thr His Trp Pro Arg Phe Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Lys Ala Cys Pro Pro Ser Lys
485 490
<210> 44
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 44
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly
355 360 365
Ile Glu Met Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ser Val Phe Gly Ser Leu Ala Gly Gly Val Gly Ala Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ala Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Arg Gly Thr Gln Ser His Trp Pro Arg Tyr Asp Arg
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Pro Pro Ser Lys
485 490
<210> 45
<211> 493
<212> PRT
<213> 人工序列
<220>
<223> SEQ ID No. 1的多肽变体
<400> 45
Gln Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val
1 5 10 15
Glu Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala
20 25 30
Ala Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg
35 40 45
Ala Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile
50 55 60
Gly Asp Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu
65 70 75 80
Asp Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly
85 90 95
Gly Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly
100 105 110
Ser Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly
115 120 125
Val Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu
130 135 140
Ala His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn
145 150 155 160
Tyr Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn
165 170 175
Ile Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu
180 185 190
Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser
195 200 205
Glu Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg
210 215 220
Pro Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly
225 230 235 240
Ala Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys
245 250 255
Ile Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg
260 265 270
Pro Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val
275 280 285
Asp Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Ile Val Gly
290 295 300
Gly Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys
305 310 315 320
Thr Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu
325 330 335
Ala Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val
340 345 350
Pro Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn His Gly
355 360 365
Ile Glu Val Leu Ser Ala Ala Phe Val Lys Trp Arg Thr Pro Leu Trp
370 375 380
Arg Tyr Arg Phe Leu Gly Ile Pro Gly Pro Gly Arg Arg Pro Ala Thr
385 390 395 400
His Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser
405 410 415
Val Ala Val Phe Gly Ser Leu Ala Gly Gly Val Gly Pro Ala Asp Ile
420 425 430
Lys Leu Ala Ala Glu Met Ser Ser Ala Trp Val Ser Phe Ala Ala His
435 440 445
Gly Val Pro Asp Arg Gly Thr Gln Thr His Trp Pro Arg Tyr Asp Ile
450 455 460
Ser Gly Glu Ile Lys Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly
465 470 475 480
Leu Gly Val Ser Pro Ser Asn Ala Cys Pro Pro Ser Lys
485 490
<210> 46
<211> 540
<212> PRT
<213> Sphingopyxis sp.
<400> 46
Val Lys Glu His Gln Cys Arg Gly Gly Arg Ala Ser Pro Ala Ala Pro
1 5 10 15
Ala Thr Trp Leu Ala Arg Ile Ser Val Ser Arg Gly Ala Ser Ala Ile
20 25 30
Ala Trp Thr Phe Met Leu Gly Ala Thr Ala Ile Pro Val Ala Ala Gln
35 40 45
Thr Asp Asp Pro Lys Leu Val Arg His Thr Gln Ser Gly Ala Val Glu
50 55 60
Gly Val Glu Gly Asp Val Glu Thr Phe Leu Gly Ile Pro Phe Ala Ala
65 70 75 80
Pro Pro Val Gly Asp Leu Arg Trp Arg Pro Pro Ala Pro Pro Arg Ala
85 90 95
Trp Ala Gly Thr Arg Asp Gly Arg Arg Phe Ala Pro Asp Cys Ile Gly
100 105 110
Asn Glu Arg Leu Arg Glu Gly Ser Arg Ala Ala Gly Thr Ser Glu Asp
115 120 125
Cys Leu Tyr Leu Asn Ile Trp Ser Pro Lys Gln Val Gly Lys Gly Gly
130 135 140
Leu Pro Val Met Ile Trp Val Tyr Gly Gly Gly Phe Ser Gly Gly Ser
145 150 155 160
Gly Ala Val Pro Tyr Tyr Asp Gly Ser Ala Leu Ala Gln Lys Gly Val
165 170 175
Val Val Val Thr Phe Asn Tyr Arg Ala Gly Ile Leu Gly Phe Leu Ala
180 185 190
His Pro Ala Leu Ser Lys Glu Ser Pro Asn Gly Val Ser Gly Asn Tyr
195 200 205
Gly Leu Leu Asp Met Leu Ala Ala Phe Lys Trp Val Gln Asn Asn Ile
210 215 220
Arg Glu Phe Gly Gly Asp Pro Asn Arg Val Thr Val Phe Gly Glu Ser
225 230 235 240
Ala Gly Ala Ser Ala Leu Gly Leu Leu Leu Thr Ser Pro Leu Ser Glu
245 250 255
Ser Ala Phe Asn Gln Ala Ile Leu Gln Ser Pro Gly Leu Ala Arg Pro
260 265 270
Leu Ala Thr Leu Ser Glu Ser Glu Ala Asn Gly Leu Glu Leu Gly Ala
275 280 285
Asp Ile Ser Ala Leu Arg Arg Ala Asp Ala Gly Glu Leu Thr Lys Ile
290 295 300
Ala Gln Ser Arg Ile Pro Met Ser Arg Gln Phe Thr Lys Pro Arg Pro
305 310 315 320
Met Gly Pro Ile Leu Asp Gly Tyr Val Leu Arg Thr Leu Asp Val Asp
325 330 335
Ala Phe Ala Lys Gly Ala Phe Arg Lys Ile Pro Val Leu Val Gly Gly
340 345 350
Asn Ala Asp Glu Gly Arg Ala Phe Thr Asp Arg Leu Pro Val Lys Thr
355 360 365
Val Leu Glu Tyr Arg Ala Tyr Leu Thr Glu Gln Phe Gly Asp Glu Ala
370 375 380
Asp Ala Trp Glu Arg Cys Tyr Pro Ala Asn Ser Asp Ala Asp Val Pro
385 390 395 400
Ala Ala Val Ala Arg Leu Phe Gly Asp Ser Gln Phe Asn Asn Gly Ile
405 410 415
Glu Leu Leu Ser Ala Ala Phe Ala Lys Trp Arg Thr Pro Leu Trp Arg
420 425 430
Tyr Arg Phe Thr Gly Ile Pro Gly Ala Gly Arg Arg Pro Ala Thr His
435 440 445
Gly Asp Glu Ile Pro Tyr Val Phe Ala Asn Leu Gly Pro Ser Ser Val
450 455 460
Ser Met Phe Gly Ser Leu Glu Gly Gly Ala Gly Ala Ser Asp Ile Lys
465 470 475 480
Leu Ala Thr Glu Met Ser Ala Ala Trp Val Ser Phe Ala Val His Gly
485 490 495
Val Pro Asp Gln Gly Thr Lys Ser His Trp Pro Arg Phe Glu Arg Arg
500 505 510
Gly Glu Ile Met Thr Phe Gly Ser Gln Val Gly Ser Gly Glu Gly Leu
515 520 525
Gly Val Ser Pro Ser Lys Ala Cys Gln Pro Ser Lys
530 535 540

Claims (21)

1.具有SEQ ID No.46的镰孢毒素羧酸酯酶的裂解镰孢毒素的多肽变体，其特征在于，所述多肽变体各自具有在N-末端处缩短了47个氨基酸的氨基酸序列，其中所述氨基酸序列与SEQ ID No.46的氨基酸序列区段48-540具有至少70％，优选地80％，特别优选地100％的序列同一性，即SEQ ID No.1。
2.根据权利要求1的多肽变体，其特征在于，所述多肽变体在至少一个从由下列各项组成的组中选择的位置处具有氨基酸置换：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490。
3.根据权利要求2的多肽变体，其特征在于，在位置10和456处的氨基酸置换选自Q、E、N、H、K和R，在位置33、107、293和332处的氨基酸置换选自E、Q、D、K、R和N，在位置66、463和478处的氨基酸置换选自D、E、K、N、Q和R，在位置140和490处的氨基酸置换选自P、A、S和N，在位置144和367处的氨基酸置换选自I、L、M和V，在位置149、270、312、329和372处的氨基酸置换选自F、Y、W和H，在位置151和453处的氨基酸置换选自D、E、K和R，在位置157和462处的氨基酸置换选自F、H、W和Y，在位置199、302、365和464处的氨基酸置换选自I、L、M和V，在位置266和455处的氨基酸置换选自A、S和T，在位置267、394和429处的氨基酸置换选自A、N、P和S，在位置272处的氨基酸置换选自H、N、Q和S，在位置275处的氨基酸置换选自A、D、E、G、K、N、Q、R和S，在位置280处的氨基酸置换选自A、D、E、K、N、P、Q、R和S，在位置284处的氨基酸置换选自A、N、P、S、T和V，在位置286处的氨基酸置换选自A、D、E、K、N、P、R和S，在位置360、377、391、419和427处的氨基酸置换选自A、I、L、S、T和V，在位置363、443和457处的氨基酸置换选自A、S、T和V，在位置364处的氨基酸置换选自H、I、L、M、N、Q、S和V，在位置371处的氨基酸置换选自A、I、L、M、S、T和V，在位置389处的氨基酸置换选自I、L、M和V，在位置418、430、447和473处的氨基酸置换选自A、G和S，在位置424处的氨基酸置换选自A、D、E、G、K、R和S，在位置436处的氨基酸置换选自A、G、S和T，在位置440处的氨基酸置换选自A、G、S和T，在位置465处的氨基酸置换选自A、G、H、N、Q、S和T，在位置469处的氨基酸置换选自D、E、K和R，和/或在位置487处的氨基酸置换选自N、D、Q、H和S。
4.根据权利要求2或3之一的多肽变体，其特征在于，所述氨基酸置换选自由下列各项组成的组：10Q、33E、66D、107E、140P、144M、149F、151R、157Y、199I、266S、267P、270F、272H、275E、275A、280D、280P、284T、284P、286P、286R、293E、302I、312F、329F、332E、360V、363T、364H、364L、365I、367H、371V、371M、372F、377V、389L、391V、394P、418A、419V、424A、424K、427V、429P、430A、436A、436S、440G、440S、443T、447A、453R、455S、456Q、457T、462Y、463D、464I、465H、465S、465G、469K、473A、478D、487N和490P。
5.根据权利要求2的多肽变体，其特征在于，它们在至少一个从由下列各项组成的组中选择的位置处具有氨基酸置换：66、199、302、377、394、424、430和463。
6.根据权利要求5的多肽变体，其特征在于，所述氨基酸置换选自由下列各项组成的组：66D、199I、302I、377V、394P、424A、430A和463D。
7.根据权利要求2至6之一的多肽变体，其特征在于，所述多肽变体在该氨基酸序列的至少两个，特别是三个从由下列各项组成的组中选择的位置处具有氨基酸置换：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490。
8.根据权利要求7的多肽变体，其特征在于，所述氨基酸置换选自由下列各项组成的组：10Q、66D、144M、151R、199I、266S、267P、272H、275E、275A、280D、284T、286P、286R、293E、302I、360V、363T、364H、364L、365I、367H、371V、371M、372F、377V、389L、391V、394P、418A、419V、424A、424K、427V、429P、430A、436A、436S、440G、440S、443T、447A、453R、455S、456Q、457T、462Y、463D、464I、465H、465S、465G、469K、473A、478D、487N和490P。
9.根据权利要求2、3或4之一的多肽变体，其特征在于，所述多肽变体的氨基酸序列包含多个氨基酸置换的组合，其中所述位置组合选自由下列各项组成的组：66/199/302/394/424/430、
66/199/302/377/394/424/430、66/199/302/377/394/424/430/463、
66/144/199/302/360/372/377/394/424/430/443/463、
199/302/377/394/424/430/463、66/199/302/377/394、
66/199/302/364/377/394/424/430/463、
66/199/302/377/394/424/430/463/465、
66/199/302/377/394/424/430/440/463、
66/199/302/377/394/424/430/447/463、
66/199/302/377/394/418/424/430/463、
66/199/302/377/394/424/436/430/463、
66/199/302/364/377/394/424/430/463、
66/199/302/377/394/424/430/463/490、
66/199/302/377/394/424/430/463/469、
66/199/302/377/389/394/424/430/463、
66/199/302/377/394/424/430/463/465、
66/199/302/377/394/424/430/463/464、
66/199/302/377/394/424/430/463/465、
66/199/302/377/394/424/430/440/463、
66/199/302/377/394/424/430/457/463、
66/199/302/377/394/424/430/436/463、
66/199/302/363/371/377/394/424/430/463、
66/199/302/377/394/424/430/447/453/463、
66/199/302/377/394/424/430/456/462/463、
66/199/302/377/394/419/424/427/430/463、
66/199/302/365/377/394/424/430/463/487和
66/199/302/371/377/394/424/430/463/487。
10.根据权利要求9的多肽变体，其特征在于，所述多肽变体的氨基酸序列选自由SEQID No.2至29组成的组。
11.根据权利要求2、3或4之一的多肽变体，其特征在于，所述多肽变体的氨基酸序列包含多个氨基酸置换的组合，其中所述位置组合选自由下列各项组成的组：
66/99/302/364/377/389/394/419/424/427/430/447/463/465/469、
66/199/302/377/389/394/419/424/427/430/447/463/465/469、
66/199/302/363/364/371/377/389/394/419/424/427/430/447/463/464/465/469、
66/199/302/363/371/377/389/394/419/424/427/430/447/463/464/465/469、
66/199/302/364/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、
66/199/302/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、66/199/302/363/367/371/377/394/424/430/463/490、
66/199/302/377/394/418/419/424/427/430/436/440/447/463、
66/199/302/377/389/394/424/430/457/463/464/465/469、
66/199/302/363/371/377/389/394/419/424/427/430/440/447/457/463/464/469/490、66/199/302/377/394/424/430/463/447/490/469/465、
66/199/302/377/394/424/430/463/490/469/465/419/427/371/487、
66/199/302/371/377/394/419/424/427/430/447/453/463/465/469/487/490、
66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/457/463/464/465/469/490、
66/199/302/371/377/389/394/419/424/427/430/436/447/453/456/462/463/465/469/490/487和
66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/453/456/457/462/463/464/465/469/487/490。
12.根据权利要求11的多肽变体，其特征在于，所述多肽变体的氨基酸序列选自由SEQID No.30至45组成的组。
13.分离的多核苷酸，其特征在于，所述多核苷酸具有编码根据权利要求1至12之一的多肽变体的核苷酸序列。
14.裂解镰孢毒素的添加剂，其特征在于，所述添加剂包含至少一种根据权利要求1至12之一的多肽变体，和任选地包含至少一种助剂。
15.根据权利要求14的添加剂，其特征在于，所述助剂选自由下列各项组成的组：惰性载体、维生素、矿物质、植物源性物质、酶和其他用于使真菌毒素解毒的组分，例如真菌毒素降解酶，特别是黄曲霉毒素氧化酶、麦角胺水解酶、麦角胺酰胺酶、玉米赤霉烯酮酯酶、玉米赤霉烯酮内酯酶、玉米赤霉烯酮水解酶、赭曲毒素酰胺酶、串珠镰孢菌素氨基转移酶、氨基多元醇氨基氧化酶、脱氧瓜萎镰菌醇环氧化物水解酶、脱氧瓜萎镰菌醇脱氢酶、脱氧瓜萎镰菌醇氧化酶、单端孢霉烯脱氢酶、单端孢霉烯氧化酶；和降解真菌毒素的微生物；和结合真菌毒素的物质，例如微生物细胞壁或无机材料例如膨润土。
16.根据权利要求14或15之一的添加剂用于裂解在食物或饲料中的至少一种镰孢毒素的用途，所述食物或饲料特别地是经粒化的，所述饲料特别地为用于猪、家禽、牛、马、鱼类或水产养殖的饲料。
17.根据权利要求14或15之一的添加剂用于在用于制备或加工食物或饲料的方法中裂解至少一种镰孢毒素的用途，所述方法特别地在至少50℃，优选地至少70℃的温度下进行。
18.至少一种根据权利要求1至12之一的多肽变体或者至少一种根据权利要求14或15之一的添加剂在制备用于预防和/或治疗镰孢毒素真菌毒素中毒的制剂中的用途。
19.用于以水解方式裂解至少一种镰孢毒素的方法，其特征在于，使至少一种镰孢毒素与至少一种根据权利要求1至12之一的多肽变体或者至少一种根据权利要求14或15之一的添加剂相接触，并且使由所述多肽变体和所述镰孢毒素组成的混合物经历在至少50℃下，优选地在至少70℃下的热处理，并且要么在该热处理期间要么在该热处理之后使所述混合物与对于以水解方式的裂解来说足够的量的湿气相接触。
20.根据权利要求19的方法，其特征在于，使所述多肽变体或所述添加剂与被至少一种镰孢毒素污染的饲料或食物相混合，和任选地在粒化过程期间进行所述热处理。
21.根据权利要求19或20之一的方法，其特征在于，以5U至500U，优选地10U至300U，和更优选地15U至100U/千克饲料或食物的浓度范围添加所述多肽变体。
CN201580073275.0A 2015-02-24 2015-02-24 裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法 Pending CN107208077A (zh)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/AT2015/000032 WO2016134387A1 (de) 2015-02-24 2015-02-24 Fusariumtoxine spaltende polypeptidvarianten, zusatzstoff enthaltend dieselben und verwendung derselben sowie verfahren zur spaltung von fusariumtoxinen

Publications (1)

Publication Number Publication Date
CN107208077A true CN107208077A (zh) 2017-09-26

Family

ID=52828929

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201580073275.0A Pending CN107208077A (zh) 2015-02-24 2015-02-24 裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法

Country Status (22)

Country Link
US (1) US10264807B2 (zh)
EP (1) EP3262163B1 (zh)
JP (1) JP6506857B2 (zh)
CN (1) CN107208077A (zh)
AR (1) AR103755A1 (zh)
AU (1) AU2015384258C1 (zh)
BR (1) BR112017016184A2 (zh)
CA (1) CA2976514C (zh)
CO (1) CO2017008520A2 (zh)
CU (1) CU24505B1 (zh)
DK (1) DK3262163T3 (zh)
ES (1) ES2745685T3 (zh)
HU (1) HUE046772T2 (zh)
MX (1) MX2017010880A (zh)
PH (1) PH12017501520A1 (zh)
PL (1) PL3262163T3 (zh)
PT (1) PT3262163T (zh)
RU (1) RU2720517C2 (zh)
SG (1) SG11201706705YA (zh)
UA (1) UA119809C2 (zh)
UY (1) UY36565A (zh)
WO (1) WO2016134387A1 (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108251399A (zh) * 2016-12-29 2018-07-06 中粮营养健康研究院有限公司 伏马毒素降解酶、编码基因、重组载体、细胞、添加剂及其应用
CN111467476A (zh) * 2020-03-05 2020-07-31 山东农之安生物科技有限公司 解毒药物组合物及其用途
CN111471659A (zh) * 2020-03-05 2020-07-31 山东农之安生物科技有限公司 具有去环氧基催化活性的多肽及其编码核酸和用途
CN112239755A (zh) * 2020-09-22 2021-01-19 天津科技大学 伏马毒素降解酶FumDSS及其基因和应用

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109825484B (zh) * 2017-11-23 2022-06-28 吉林中粮生化有限公司 玉米赤霉烯酮水解酶zhd101突变体及利用该突变体水解玉米赤霉烯酮的方法
US20220233496A1 (en) 2019-07-03 2022-07-28 Intervet Inc. Conjugated deoxynivalenol to protect against mycotoxicosis
MX2023003959A (es) 2020-10-08 2023-05-25 DSM Austria GmbH Variantes de hidrolasa alfa/beta tetramerica con estabilidad a la temperatura incrementada y metodos de uso y produccion de las mismas.
US20240067950A1 (en) 2020-12-18 2024-02-29 DSM Austria GmbH Means and methods for detoxifying ochratoxin a
WO2023025938A1 (en) 2021-08-27 2023-03-02 DSM Austria GmbH Means and methods for detoxifying ochratoxin a
WO2023144753A2 (en) * 2022-01-26 2023-08-03 Danstar Ferment Ag Recombinant expression of fumonisin esterase, compositions and uses thereof
KR102699553B1 (ko) * 2022-12-29 2024-08-28 씨제이제일제당 주식회사 신규한 푸모니신 분해 효소 및 그의 용도
KR20240108806A (ko) * 2022-12-29 2024-07-10 씨제이제일제당 (주) 신규 푸모니신 분해 효소 및 그의 용도
WO2024180045A1 (en) 2023-02-28 2024-09-06 Dsm Ip Assets B.V. Means and methods for modifying fumonisins

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110189755A1 (en) * 2008-09-18 2011-08-04 Wulf-Dieter Moll Method for the production of an additive for the enzymatic decomposition of mycotoxins, additive, and use thereof

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK13491D0 (da) 1991-01-25 1991-01-25 Novo Nordisk As Anvendelse af et enzymholdigt granulat og fremgangsmaade til fremstilling af et forderstof i tabletform
EP0988383B1 (en) 1997-07-07 2002-10-16 Pioneer Hi-Bred International, Inc. Fumonisin detoxification compositions and methods
US6538177B1 (en) * 1998-07-15 2003-03-25 Pioneer Hi-Bred International, Inc. Compositions and methods for fumonisin detoxification
WO2004085624A2 (en) 2003-03-24 2004-10-07 Diversa Corporation Transaminases, deaminases and aminomutases and compositions and methods for enzymatic detoxification
AT501359B1 (de) 2004-11-16 2007-10-15 Erber Ag Verfahren und mikroorganismus zur entgiftung von fumonisinen sowie verwendung und futtermittelzusatz
EP2326714B8 (de) * 2008-09-18 2016-01-06 Erber Aktiengesellschaft Verfahren zur herstellung eines futtermittelzusatzes zum sauerstoffunabhängigen, enzymatischen abbau von mykotoxinen sowie futtermittelzusatz und verwendung desselben

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110189755A1 (en) * 2008-09-18 2011-08-04 Wulf-Dieter Moll Method for the production of an additive for the enzymatic decomposition of mycotoxins, additive, and use thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HEINL,S.等: "RecName: Full=Fumonisin B1 esterase [Sphingopyxis macrogoltabida]Sequence ID: D2D3B6.1", 《NCBI》 *
HEINL等: "Degradation of fumonisin B1 by te consecutive action of two bacterial enzymes", 《J.BIOTECHNOL. 》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108251399A (zh) * 2016-12-29 2018-07-06 中粮营养健康研究院有限公司 伏马毒素降解酶、编码基因、重组载体、细胞、添加剂及其应用
CN111467476A (zh) * 2020-03-05 2020-07-31 山东农之安生物科技有限公司 解毒药物组合物及其用途
CN111471659A (zh) * 2020-03-05 2020-07-31 山东农之安生物科技有限公司 具有去环氧基催化活性的多肽及其编码核酸和用途
CN112239755A (zh) * 2020-09-22 2021-01-19 天津科技大学 伏马毒素降解酶FumDSS及其基因和应用

Also Published As

Publication number Publication date
PH12017501520A1 (en) 2018-02-05
CA2976514C (en) 2022-08-16
PL3262163T3 (pl) 2019-12-31
SG11201706705YA (en) 2017-09-28
AR103755A1 (es) 2017-05-31
UA119809C2 (uk) 2019-08-12
WO2016134387A1 (de) 2016-09-01
HUE046772T2 (hu) 2020-03-30
CA2976514A1 (en) 2016-09-01
EP3262163B1 (de) 2019-06-19
US20180235255A1 (en) 2018-08-23
JP2018506993A (ja) 2018-03-15
RU2017129899A3 (zh) 2019-03-25
MX2017010880A (es) 2019-05-16
ES2745685T3 (es) 2020-03-03
EP3262163A1 (de) 2018-01-03
BR112017016184A2 (pt) 2018-04-17
JP6506857B2 (ja) 2019-04-24
DK3262163T3 (da) 2019-09-23
CU20170112A7 (es) 2018-05-08
RU2017129899A (ru) 2019-03-25
RU2720517C2 (ru) 2020-04-30
CO2017008520A2 (es) 2017-11-10
CU24505B1 (es) 2021-04-07
AU2015384258A1 (en) 2017-08-10
AU2015384258C1 (en) 2020-12-17
UY36565A (es) 2016-09-30
PT3262163T (pt) 2019-10-08
US10264807B2 (en) 2019-04-23
AU2015384258B2 (en) 2020-07-02

Similar Documents

Publication Publication Date Title
CN107208077A (zh) 裂解镰孢毒素的多肽变体、包含所述多肽变体的添加剂和所述添加剂的用途，以及用于裂解镰孢毒素的方法
US11998027B2 (en) Polypeptide for hydrolytic cleavage of zearalenone and/or zearalenone derivatives, isolated polynucleotide thereof as well as a polypeptide containing an additive, use of same as well as a process
WO2012113827A1 (en) Feed processing enzymes
Graminho et al. Purification, biochemical characterization, and genetic cloning of the phytase produced by Burkholderia sp. strain a13
NZ717021B2 (en) Polypeptide for hydrolytic cleavage of zearalenone and/or zearalenone derivatives, isolated polynucleotide thereof as well as a polypeptide containing an additive, use of same as well as a process

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination