CN110914446B - 酮还原酶 - Google Patents
酮还原酶 Download PDFInfo
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- CN110914446B CN110914446B CN201880042466.4A CN201880042466A CN110914446B CN 110914446 B CN110914446 B CN 110914446B CN 201880042466 A CN201880042466 A CN 201880042466A CN 110914446 B CN110914446 B CN 110914446B
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- ketoreductase
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- 101001110310 Lentilactobacillus kefiri NADP-dependent (R)-specific alcohol dehydrogenase Proteins 0.000 title claims abstract description 398
- 102000004190 Enzymes Human genes 0.000 title abstract description 130
- 108090000790 Enzymes Proteins 0.000 title abstract description 130
- 150000002576 ketones Chemical class 0.000 claims abstract description 82
- 150000003333 secondary alcohols Chemical class 0.000 claims abstract description 53
- 239000000758 substrate Substances 0.000 claims description 178
- 150000001299 aldehydes Chemical class 0.000 claims description 76
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 63
- 238000000034 method Methods 0.000 claims description 51
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- 150000003138 primary alcohols Chemical class 0.000 claims description 36
- HPXRVTGHNJAIIH-UHFFFAOYSA-N cyclohexanol Chemical compound OC1CCCCC1 HPXRVTGHNJAIIH-UHFFFAOYSA-N 0.000 claims description 28
- 230000008569 process Effects 0.000 claims description 24
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- 238000007254 oxidation reaction Methods 0.000 abstract description 54
- 230000003647 oxidation Effects 0.000 abstract description 47
- 230000009467 reduction Effects 0.000 abstract description 30
- 230000002255 enzymatic effect Effects 0.000 abstract description 8
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Abstract
本发明涉及酮还原酶及其用途。本发明的酮还原酶特别适用于酶催化酮向手性或非手性仲醇的还原以及手性或非手性仲醇向酮的氧化。
Description
技术领域
本发明涉及酮还原酶及其用途。本发明的酮还原酶特别适用于酶催化酮向手性或非手性仲醇的还原以及手性或非手性仲醇向酮的氧化。
背景技术
根据欧盟第635734号拨款协议,产生本申请的项目已获得欧盟的地平线2020(Horizon 2020)研究与创新计划的资助。
酮还原酶(KRED,也称为“醇脱氢酶”、“ADH”或“羰基还原酶”)催化醛和酮分别向相应的伯醇和仲醇的还原。这些酶还能催化逆反应,即伯醇和仲醇分别向相应的醛和酮的氧化。
酮还原酶是在生命的所有界中发现的普遍存在的酶。已知的商购可得的酮还原酶来自马肝(HLADH)、面包酵母(YADH)和细菌,如布氏热厌氧菌(Thermoanaerobium brockii,TBADH)和开啡尔乳杆菌(Lactobacillus kefir,LKADH)。
基于其序列同一性和生化特性,酮还原酶可分为不同的蛋白家族。SDR-家族(短链-脱氢酶/还原酶)的成员展示短链酶,其不含任何金属离子。相反,MDR-家族(中链-脱氢酶/还原酶)的成员展示中链酶,其取决于Zn2+。酮还原酶的另一组展示取决于Fe2+的长链酶(供查阅:K.Drauz,H.O.May,Enzyme Catalysis in Organic Synthesis,WileyVCH,Weinheim,2012)。某些酮还原酶需要二聚或多聚以提供完全的催化活性。
对于工业应用,将酮立体选择性地还原为仲醇是令人关注的,其中前手性羰基化合物被立体选择性地和/或区域选择性地还原为限定的手性醇。在一些工业应用中,还希望将仲醇立体选择性地转化为酮,以用于所需外消旋化合物的手性拆分,例如,允许分离对映异构体。此外,对于其中存在两个或多个羟基的醇化合物,可能非常感兴趣的是仅将这样的仲醇基团中的一个基团区域选择性地氧化成酮。此外,具有一个或多个酮基的化合物的区域选择性还原可能具有工业相关性。在工业应用中,通常认为伯醇向醛的酶促氧化与醛向伯醇的酶促还原相关性较低,但二者同样被酮还原酶催化。可以通过调节酶反应的化学平衡来影响同一种酮还原酶分别在氧化反应或还原反应中的使用。
可取的是使用具有高比活性(specific activity)、立体选择性和区域选择性的酮还原酶。工业使用酮还原酶的另一个重要标准是长加工稳定性(longprocessstability),其通常与高的高温稳定性(也称为“热稳定性”)和高溶剂稳定性相关。如果底物已经是手性的,则还需要使用具有高立体特异性的酮还原酶。
由酮还原酶催化的还原需要还原型的辅因子(reduced cofactor)作为电子供体。一些酮还原酶使用还原型烟酰胺腺嘌呤二核苷酸(NADH)作为辅因子,其他酮还原酶使用还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH),一些酮还原酶同时接受NADH和NADPH。相应地,由酮还原酶催化的氧化需要氧化型的辅因子(oxidized cofactor)作为电子受体。对于该反应,酮还原酶使用氧化型的烟酰胺腺嘌呤二核苷酸(NAD+)或氧化型的烟酰胺腺嘌呤二核苷酸磷酸(NADP+)或使用NAD+和NADP+两者。
在还原过程中酮还原酶的体外使用需要NAD(P)H的再生系统。常见的再生系统是葡萄糖脱氢酶(GDH)或甲酸脱氢酶,其与酮还原酶联合使用。也可通过共底物(通常是伯醇或仲醇,例如异丙醇氧化为丙酮)的伴随氧化由酮还原酶本身进行辅因子再生。将酮还原酶同时用作还原催化剂和辅因子再生系统需要同时接受酮底物和用于辅因子再生的共底物。特异性共底物的选择可取决于酮还原酶对这种共底物的比活性和在特定辅因子再生条件下的酮还原酶稳定性。
在氧化过程中酮还原酶的体外使用也需要NAD(P)+的再生系统。同样,如果可获得并接受合适的酮或醛,则可以同时将酮还原酶用作氧化催化剂和用于辅因子再生。此外,NAD(P)H-氧化酶(NOX)可用于使用氧气作为共底物将NAD(P)H氧化为NAD(P)+。NOX反应的副产物可能是水或过氧化氢,具体取决于所使用的NOX酶类别。在工业过程中,通常优选形成水的NOX酶。
从现有技术中已知各种酮还原酶/醇脱氢酶。在这方面,可以参考例如以下专利申请和专利:US 6,255,092、US 6,645,746、US 7,393,667、US 7,629,157、US 7,977,078、US8,257,952、US 8,273,554、US 8,288,131、US 8,426,178、US 8,512,973、US 8,617,853、US2002/0064847、US 2010/0035317、US 2010/014399、WO 2009/040080、WO 2011/022548、WO2012/046254、WO 2013/102619、EP 0645453和EP1553170。在这方面,也可以参考例如以下科研论文:Wulfet al.Enzyme Microb Technol.2012Sep 10;51(4):217-24,Paul et al.,ChemCatChem.2013Sep 23,5(12):3875-3881、Mallin et al.Enzyme MicrobTechnol.2013Sep10;53(4):283-7和Schmidt et al.,Angew Chem Int EdEngl.2015Feb23;54(9):2784-7。蛋白质数据库在线,登录号:WP_022887115,涉及Glaciibactersuperstes的短链脱氢酶。选择或改进了现有技术中描述的这些酮还原酶中的大多数,用于在药物应用中的中间体的化学酶促合成的领域中,将特定的酮底物还原转化为仲醇化合物,例如WO 2009/040080中公开的酶。
然而,现有技术的酮还原酶在各方面都不令人满意,需要与常规酮还原酶相比具有优点的酮还原酶。特别地,在醇底物氧化为酮或醛产物以及将酮或醛底物还原为醇产物的过程中需要具有高立体选择性、区域选择性和比活性的酮还原酶,并且在相应的工业生产过程中,需要这种酮还原酶具有高工艺稳定性。在这方面,工业应用中的高工艺稳定性可包括在水环境、非水环境和/或两相体系中,和/或高底物浓度下和/或高温下和/或外加水溶性溶剂和/或4-11的宽pH范围内和/或固体支撑物上(即,当被固定化时)和/或高剪切力下(例如,通过搅拌、抽吸、膜过滤制备)的化学和物理稳定性以及酶活性。其他因素,例如底物选择性、KM、比活性、立体特异性、非对映立体选择性、底物抑制、产物抑制,其他因素(例如粗提取组分、底物污染物或副产物)引起的抑制,以及在适合寄主中的重组可溶性表达能力可起重要作用。当底物为手性底物时,即已经含有一个或多个手性中心和/或其手性轴时,希望所述手性底物中含有的前手性羰基的酶促还原的立体选择性基本上不受所述一个或多个手性中心和/或轴的影响。
发明内容
本发明的目的是提供改进的酮还原酶。
已经通过本专利申请请求保护的主题解决了该问题。
SEQ ID NO:1为本发明的优选的酮还原酶的氨基酸序列。SEQ ID NO:101、SEQ IDNO:113和SEQ ID NO:149与SEQ ID NO:1相同。
SEQ ID NO:2为编码本发明的SEQ ID NO:1的氨基酸序列的DNA序列。
SEQ ID NO:3为从WO 2013/102619获知的比较氨基酸序列,在该文献中它被称为具有识别号10的序列。所述比较序列SEQ ID NO:3与本发明的SEQ ID NO:1的氨基酸序列具有91.1%的同一性。
SEQ ID NO:4为编码如WO 2013/102619中所公开的比较序列SEQ ID NO:3的氨基酸序列的DNA序列,在该文献中它被称为具有识别号9的序列。
SEQ ID NO:5为DNA序列,其是SEQ ID NO:4的密码子优化的变体,编码SEQ ID NO:3的氨基酸序列。
SEQ ID NO:6至SEQ ID NO:100、SEQ ID NO:102至SEQ ID NO:112、SEQ ID NO:114至SEQ ID NO:148、以及SEQ ID NO:150至SEQ ID NO:182是来自SEQ ID NO:1的变体酮还原酶,其具有进一步改善的功能特性,例如在醇底物氧化为酮或醛产物以及酮或醛底物还原为醇产物的过程中的提高的热稳定性、提高的活性,特别是比活性。
具体地,变体酮还原酶在以下一个或多个位置具有以下取代,该位置的编号参考本发明的比较序列SEQ ID NO:1:
当将本发明的SEQ ID NO:1与比较序列SEQ ID NO:3进行比对时,比较序列SEQ IDNO:3的位置1对应于本发明的SEQ ID NO:1的位置9。本发明的SEQ ID NO:1在其N端含有在比较序列SEQ ID NO:3中缺失的8个位置,即M1、T2、S3、S4、S5、S6、P7和S8。因此,在比对中,比较序列SEQ ID NO:3的位置N2、A3、L4、V5、T6、G7、G8、S9、R10、G11、I12、G13、E14和A15分别对应于本发明的SEQ ID NO:1的位置N10、A11、L12、V13、T14、G15、G16、S17、R18、G19、I20、G21、E22和A23。
附图说明
图1显示了将本发明的SEQ ID NO:1的酮还原酶的Tm50值与比较序列SEQ ID NO:3的酮还原酶的Tm50值进行比较的实验结果。
具体实施方式
本发明提供新的酮还原酶,特别是与比较序列SEQ ID NO:3的酮还原酶相比表现出改善的性能的酮还原酶。此外,本发明提供新的酮还原酶,特别是与比较序列SEQ ID NO:1的酮还原酶相比表现出改善的性能的酮还原酶。
令人惊讶地发现,本发明的酮还原酶具有以下优异的性能:
-比活性,特别是还原活性(参见实施例3),和/或氧化活性(参见实施例4),和/或
-氧化过程中的pH特性(“pH相关活性”),特别是在中性pH值下的优异的氧化活性(参见实施例4),和/或
-立体选择性(参见实施例5),和/或
-温度稳定性,即在较高温度下的残余活性(半最高熔解温度,Tm50(40);参见实施例6、图1;Tm50(10);参见实施例12),和/或
-区域选择性;和/或
-对于将酮或醛底物转化为醇产物或相反方向的转化还原活性与氧化活性之比增加;或
-对于将酮或醛底物转化为醇产物或相反方向的转化还原活性与氧化活性之比降低;和/或
-高的辅因子偏好性(cofactorpreference)。
本发明的第一方面涉及一种酮还原酶,其包含与本发明的SEQ ID NO:1的氨基酸序列具有至少92%的同一性的氨基酸序列。
本发明的酮还原酶包含与SEQ ID NO:1的氨基酸序列具有确定同一性的氨基酸序列。这意味着本发明的酮还原酶可包含所述氨基酸序列作为其总氨基酸序列的子序列,或者本发明的酮还原酶可基本上由所述氨基酸序列组成。当本发明的酮还原酶包含所述氨基酸序列作为其总氨基酸序列的子序列时,所述总氨基酸序列可在所述子序列的N端和/或C端延伸,即可包含其他氨基酸残基。这样的延伸可能是有利的,例如,当为了例如纯化目的而将酮还原酶固定在固体支撑物上时。
为了本发明的目的,同源性和同一性被理解为同义词。同一性百分比计算如下:序列同一性[%]=匹配数/L×100,其中L为对齐位置(即相同的和不同的(包括空位,如果有的话))的数目。优选使用BLASTP来计算同一性(参见,例如,Altschul SF et al.(1997)"Gapped BLAST和PSI-BLAST:a new generation ofprotein database search programs",Nucleic Acids Res.25:3389-3402、或Altschul SF(2005)“Protein database searchesusing compositionally adjusted substitution matrices.”FEBS J.272:5101-5109),最好使用以下算法参数:矩阵:BLOSUM62;空位罚分(Gap Cost):存在:11延伸(extension):1,预期阈值:10以及字长:6。筛选结果中覆盖率超过35%的序列。可以在NCBI主页(https://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome)上在线访问BlastP。可以根据需要调整其他程序设置,例如使用以下设置:
-字段“输入查询序列”:查询子范围:无
-字段“选择搜索设置”:数据库:无冗余蛋白质序列(nr);可选参数:无
-字段“程序选择”:算法:blastp(蛋白质-蛋白质BLAST)
-算法参数:字段“一般参数”:最大目标序列:20000;短查询:自动调整短输入序列的参数;期望阈值:10;字长:6;查询范围的最大匹配:0
-算法参数:字段“评分参数”:矩阵:BLOSUM62;空位罚分(Gap Cost):存在:11延伸(Extension):1;组成调整:条件组成得分矩阵调整
-算法参数:字段“滤波器和掩码”:滤波器:无;掩码:无。
优选地,酮还原酶包含与本发明的SEQ ID NO:1的氨基酸序列具有至少92.0%、至少92.1%、至少92.2%、至少92.3%、至少92.4%、至少92.5%、至少92.6%、至少92.7%、至少92.8%、至少92.9%、至少93%、至少93.1%、至少93.2%、至少93.3%、至少93.4%、至少93.5%、至少93.6%、至少93.7%、至少93.8%、至少93.9%、更优选至少94%、至少94.1%、至少94.2%、至少94.3%、至少94.4%、至少94.5%、至少94.6%、至少94.7%、至少94.8%、至少94.9%、还更优选至少95%、至少95.1%、至少95.2%、至少95.3%、至少95.4%、至少95.5%、至少95.6%、至少95.7%、至少95.8%、至少95.9%、更优选至少96%、至少96.1%、至少96.2%、至少96.3%、至少96.4%、至少96.5%、至少96.6%、至少96.7%、至少96.8%、至少96.9%、甚至更优选至少97%、至少97.1%、至少97.2%、至少97.3%、至少97.4%、至少97.5%、至少97.6%、至少97.7%、至少97.8%、至少97.9%、最优选至少98%、至少98.1%、至少98.2%、至少98.3%、至少98.4%、至少98.5%、至少98.6%、至少98.7%、至少98.8%、至少98.9%、以及尤其是至少99%、至少99.1%、至少99.2%、至少99.3%、至少99.4%、至少99.5%、至少99.6%、至少99.7%、至少99.8%、至少99.9%的同一性的氨基酸序列。在一个优选的实施方案中,酮还原酶包含基本上由本发明的SEQ ID NO:1的氨基酸序列组成的氨基酸序列。在另一个优选的实施方案中,酮还原酶由本发明的SEQ ID NO:1的氨基酸序列组成。
在一个优选的实施方案中,本发明的酮还原酶是本发明的SEQ ID NO:1的氨基酸序列与任何其他氨基酸、寡核苷酸或多肽的融合蛋白,其融合到N-末端和/或C端。
在一个优选的实施方案中,本发明的酮还原酶包含本发明的SEQ ID NO:1的氨基酸序列,以及额外的至少10个氨基酸残基、更优选地至少20个氨基酸残基、甚至更优选地至少30个氨基酸残基、最优选至少40个氨基酸残基,氨基酸残基独立地选自Ala、Arg、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Lys、Met、Phe、Pro、Ser、Thr、Trp Tyr和Val。
在一个优选的实施方案中,本发明的具有改善的性质的酮还原酶相对于比较序列SEQ ID NO:3有所不同,即,与比较序列SEQ ID NO:3的酮还原酶有所不同,不同之处在于:至少一个残基改变、优选1-23个残基改变、更优选2-23个残基改变、更优选3-23个残基改变、更优选4-23个残基改变、更优选5-23个残基改变、更优选6-23个残基改变、更优选7-23个残基改变、更优选8-23个残基改变、更优选9-23个残基改变、更优选10-23个残基改变、更优选11-23个残基改变、更优选12-23个残基改变、更优选13-23个残基改变、更优选14-23个残基改变、甚至更优选15-23个残基改变、甚至更优选16-23个残基改变、甚至更优选17-23个残基改变、甚至更优选18-23个残基改变、更优选19-23个残基改变、甚至更优选20-23个残基改变、甚至更优选21-23个残基改变、甚至更优选22-23个残基改变、最优选23个残基改变。
因此,本发明的酮还原酶优选地满足两个要求,一个是关于本发明的SEQ ID NO:1的最小同一性的要求,另一个是关于与比较序列SEQ ID NO:3的某些差异的要求。
关于比较序列SEQ ID NO:3与本发明的SEQ ID NO:1之间的序列变异,差异是指比较序列SEQ ID NO:3的给定位置的一个或多个氨基酸被选自Ala、Arg、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Lys、Met、Phe、Pro、Ser、Thr、Trp、Tyr和Val中的任何其他蛋白氨基酸(proteinogenic amino acid)取代。在一个优选的实施方案中,取代不改变序列长度,即单个氨基酸残基被另一个单个氨基酸残基替换。然而,也可以不替换而删除一个或多个氨基酸残基和/或插入一个或多个氨基酸残基。
原则上,酶的任何位置上的取代可以是保守取代,其中一种氨基酸被具有相当特征的氨基酸取代(例如,用另一种疏水性氨基酸取代一种疏水性氨基酸)。此外,在酶的任何位置上的取代可以是非保守取代,其中一种氨基酸被具有其他特征的氨基酸取代(例如,用亲水性氨基酸取代疏水性氨基酸)。
为了说明的目的,根据氨基酸取代领域中的常规实践描述了本文公开的特定取代,其中选择了格式[未取代氨基酸-序列位置-取代氨基酸],并且该序列位置的左侧或右侧直接与氨基酸字母(单字母代码)相邻。在单个氨基酸要被一个以上的氨基酸取代的情况下,或者一个以上的氨基酸要被单个氨基酸取代的情况下,上述一个以上的氨基酸的位置应按以下顺序排列:从左到右对应于N端到C端。仅出于说明目的,取代“I190V”描述了序列位置190处的氨基酸异亮氨酸(I)被氨基酸缬氨酸(V)取代,即I被V取代。类似地,取代“M1MTS”描述了序列位置1处的氨基酸甲硫氨酸(M)被氨基酸(N-末端)-甲硫氨酸(M)-苏氨酸(T)-丝氨酸(S)-(C-末端)取代,即M被MTS取代。类似地,取代“MTS1M”描述了序列位置1中的氨基酸(N-末端)-甲硫氨酸(M)-苏氨酸(T)-丝氨酸(S)-(C-末端)被氨基酸甲硫氨酸(M)取代,即MTS被M取代。
在下文中,出于关于本发明的第一方面和第二方面的说明的目的,除非另有明确说明,否则位置编号是指从M1、T2、S3、S4、S5、S6、P7和S8开始的比较序列SEQ ID NO:3的位置。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3的酮还原酶的区别在于
-在比较序列SEQ ID NO:3的序列区段A中的至少一个残基改变,优选至少2个、至少3个、至少4个或至少5个残基改变,其中所述序列区段A在比较序列SEQ ID NO:3的位置14至比较序列SEQ ID NO:3的位置28的范围内;优选地,其中所述序列区段A在比较序列SEQID NO:3的位置16至比较序列SEQ ID NO:3的位置26的范围内;优选在选自V16、I18、S22、Q23和G26中的一个或多个或所有位置上的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段B中的至少一个残基改变,其中所述序列区段B在比较序列SEQ ID NO:3的位置29至比较序列SEQ ID NO:3的位置43的范围内;优选地,其中所述序列区段B在比较序列SEQ ID NO:3的位置33至比较序列SEQ ID NO:3的位置35的范围内;优选在位置V34处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段C中的至少一个残基改变,其中所述序列区段C在比较序列SEQ ID NO:3的位置44至比较序列SEQ ID NO:3的位置58的范围内;优选地,其中所述序列区段C在比较序列SEQ ID NO:3的位置45至比较序列SEQ ID NO:3的位置47的范围内;优选在位置V46处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段D中的至少一个残基改变,其中所述序列区段D在比较序列SEQ ID NO:3的位置82到比较序列SEQ ID NO:3的位置96的范围内;优选地,其中所述序列区段D在比较序列SEQ ID NO:3的位置88到比较序列SEQ ID NO:3的位置90的范围内;优选在位置A89处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段E中的至少一个残基改变,其中所述序列区段E在比较序列SEQ ID NO:3的位置123至比较序列SEQ ID NO:3的位置137的范围内;优选地,其中所述序列区段E在比较序列SEQ ID NO:3的位置129至比较序列SEQ ID NO:3的位置131的范围内;优选在位置E130处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段F中的至少一个残基改变,优选至少2个、至少3个、至少4个或至少5个残基改变,其中所述序列区段F在比较序列SEQ ID NO:3的位置185至比较序列SEQ ID NO:3的位置199的范围内;优选地,其中所述序列区段F在比较序列SEQ ID NO:3的位置188至比较序列SEQ ID NO:3的位置195的范围内;优选在选自A189、I190、S191、D192和V194中的一个或多个或所有位置处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段G中的至少一个残基改变,其中所述序列区段G在比较序列SEQ ID NO:3的位置200至比较序列SEQ ID NO:3的位置214的范围内;优选地,其中所述序列区段G在比较序列SEQ ID NO:3的位置203至比较序列SEQ ID NO:3的位置205的范围内;优选在位置S204处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段H中的至少一个残基改变,优选至少2、至少3、至少4个残基改变,其中所述序列区段H在比较序列SEQ ID NO:3的位置221至比较序列SEQ ID NO:3的位置233的范围内;优选地,其中所述序列区段H在比较序列SEQ ID NO:3的位置227至比较序列SEQ ID NO:3的位置232的范围内;优选在选自A228、I229、L230和V231中的一个或多个或所有位置处的残基改变;和/或
-在比较序列SEQ ID NO:3的序列区段I中的至少一个残基改变,优选至少2个、至少3个残基改变,其中所述序列区段I在比较序列SEQ ID NO:3的位置234至比较序列SEQ IDNO:3的位置238的范围内;优选地,其中所述序列区段I在比较序列SEQ ID NO:3的位置236至比较序列SEQ ID NO:3的位置238的范围内;优选在选自F236、S237和T238中的一个或多个或所有位置处的残基改变。
优选地,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶中的残基改变包括比较序列SEQ ID NO:3的以下位置:M1、V16、I18、S22、Q23、G26、V34、V46、A89、E130、A189、I190、S191、D192、V194、S204、A228、I229、L230、V231、F236、S237和T238中的一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个、至少22个或至少23个位置处的残基改变。
在特别优选的实施方案中,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个、至少22个或至少23个位置上不同,以使其包含选自以下的至少一个或多个或所有替换:
-在位置M1处替换为
-M1A、M1R、M1N、M1D、M1C、M1Q、M1E、M1G、M1H、M1I、M1L、M1K、M1F、M1P、M1S、M1T、M1W、M1Y或M1V;优选为M1L、M1P、M1G、M1S、M1Q、M1H、M1K或M1R;更优选为M1K、M1L或M1R;最优选为M1L;或
M1MTSSSSPSM、M1MTSSSSPSA、M1MTSSSSPSR、M1MTSSSSPSN、M1MTSSSSPSD、M1MTSSSSPSC、M1MTSSSSPSQ、M1MTSSSSPSE、M1MTSSSSPSG、M1MTSSSSPSH、M1MTSSSSPSI、M1MTSSSSPSL、M1MTSSSSPSK、M1MTSSSSPSF、M1MTSSSSPSP、M1MTSSSSPSS、M1MTSSSSPST、M1MTSSSSPSW、M1MTSSSSPSY或M1MTSSSSPSV;优选为M1MTSSSSPSL、M1MTSSSSPSP、M1MTSSSSPSG、M1MTSSSSPSS、M1MTSSSSPSQ、M1MTSSSSPSH、M1MTSSSSPSK或M1MTSSSSPSR;更优选地,M1MTSSSSPSK、M1MTSSSSPSL或M1MTSSSSPSR;最优选为M1MTSSSSPSL;和/或
-在位置V16处替换为V16A、V16R、V16N、V16D、V16C、V16Q、V16E、V16G、V16H、V16I、V16L、V16K、V16M、V16F、V16P、V16S、V16T、V16W或V16Y;优选为V16A、V16I、V16L、V16M、V16Y、V16S或V16T;更优选为V16I、V16V、V16M或V16L;最优选为V16I;和/或
-在位置I18处替换为I18A、I18R、I18N、I18D、I18C、I18Q、I18E、I18G、I18H、I18L、I18K、I18M、I18F、I18P、I18S、I18T、I18W、I18Y或I18V;优选为I18A、I18V、I18L、I18M、I18S、I18T、I18Q、I18D、I18E、I18H、I18K或I18R;更优选为I18M、I18R、I18K或I18L;甚至更优选为I18M或I18R;最优选为I18M;和/或
-在位置S22处替换为S22A、S22R、S22N、S22D、S22C、S22Q、S22E、S22G、S22H、S22I、S22L、S22K、S22M、S22F、S22P、S22T、S22W、S22Y或S22V;优选为S22A、S22G、S22T、S22N、S22Q、S22D、S22E、S22K或S22R;更优选地,S22A、S22G、S22T、S22D、S22E、S22H、S22K或S22R;甚至更优选为S22R或S22A;最优选为S22A;和/或
-在位置Q23处替换为Q23A、Q23R、Q23N、Q23D、Q23C、Q23E、Q23G、Q23H、Q23I、Q23L、Q23K、Q23M、Q23F、Q23P、Q23S、Q23T、Q23W、Q23Y或Q23V;优选为Q23A、Q23L、Q23S、Q23C、Q23N、Q23D、Q23E、Q23H、Q23K或Q23R;更优选为Q23N、Q23A、Q23E或Q23D;甚至更优选为Q23N或Q23E;最优选为Q23E;和/或
-在位置G26处替换为G26A、G26R、G26N、G26D、G26C、G26Q、G26E、G26H、G26I、G26L、G26K、G26M、G26F、G26P、G26S、G26T、G26W、G26Y或G26V;优选为G26I、G26F、G26S、G26T、G26N、G26D、G26H、G26K或G26R;更优选为G26T、G26S或G26N;最优选为G26S;和/或
-在位置V34处替换为V34A、V34R、V34N、V34D、V34C、V34Q、V34E、V34G、V34H、V34I、V34L、V34K、V34M、V34F、V34P、V34S、V34T、V34W或V34Y;优选为V34A、V34L、V34I、V34P、V34G、V34S、V34T、V34D、V34E、V34K或V34Y;更优选为V34L、V34A、V34T、V34E或V34Y;甚至更优选为V34L、V34E或V34Y;甚至更优选为V34L或V34Y;最优选为V34L;和/或
-在位置V46处替换为V46A、V46R、V46N、V46D、V46C、V46Q、V46E、V46G、V46H、V46I、V46L、V46K、V46M、V46F、V46P、V46S、V46T、V46W或V46Y;优选为V46A、V46I、V46L、V46G、V46T、V46Q、V46H、V46R或V46K;更优选为V46I、V46A、V46G、V46R或V46T;甚至更优选为V46I或V46R;最优选为V46I;和/或
-在位置A89处替换为A89R、A89N、A89D、A89C、A89Q、A89E、A89G、A89H、A89I、A89L、A89K、A89M、A89F、A89P、A89S、A89T、A89W、A89Y或A89V;优选为A89V、A89I、A89L、A89F、A89W、A89M、A89G、A89S、A89T、A89N、A89D、A89R或A89C;更优选为A89S、A89T、A89V、A89M、A89W、A89I或A89C;甚至更优选为A89V、A89M、A89W、A89M、A89C、A89S或A89T;甚至更优选为A89V、A89M、A89W、A89M、A89C或A89S;最优选为A89S;和/或
-在位置E130处替换为E130R、E130N、E130D、E130C、E130Q、E130G、E130H、E130I、E130L、E130K、E130M、E130F、E130P、E130S、E130T、E130W、E130Y或E130V;优选为E130A、E130L、E130P、E130G、E130S、E130N、E130Q、E130D、E130H、E130K、E130T或E130R;更优选为E130A、E130T、E130S、E130Q、E130N、E130D、E130H、E130K或E130R;甚至更优选为E130A、E130T、E130S、E130D、E130Q或E130N;甚至更优选为E130A、E130T、E130S或E130Q;最优选为E130Q;和/或
-在位置A189处替换为A189R、A189N、A189D、A189C、A189Q、A189E、A189G、A189H、A189I、A189L、A189K、A189M、A189F、A189P、A189S、A189T、A189W、A189Y或A189V;优选为A189P、A189S、A189N、A189Q、A189D、A189E、A189K、A189R或A189H;更优选为A189D、A189E、A189S或A189P;甚至更优选为A189S、A189D或A189E;最优选为A189S;和/或
-在位置I190处替换为I190A、I190R、I190N、I190D、I190C、I190Q、I190E、I190G、I190H、I190L、I190K、I190M、I190F、I190P、I190S、I190T、I190W、I190Y或I190V;优选为I190A、I190L、I190M、I190T、I190N、I190Q或I190D;更优选为I190M、I190N或I190L;最优选为I190M;和/或
-在位置S191处替换为S191A、S191R、S191N、S191D、S191C、S191Q、S191E、S191G、S191H、S191I、S191L、S191K、S191M、S191F、S191P、S191T、S191W、S191Y或S191V;优选为S191A、S191V、S191I、S191L、S191G、S191T、S191N或S191C;更优选为S191C、S191A、S191T或S191G;甚至更优选为S191C、S191A或S191T,甚至更优选为S191C或S191A;最优选为S191A;和/或
-在位置D192处替换为D192A、D192R、D192N、D192C、D192Q、D192E、D192G、D192H、D192I、D192L、D192K、D192M、D192F、D192P、D192S、D192T、D192W、D192Y或D192V;优选为D192A、D192V、D192I、D192L、D192P、D192G、D192S、D192T、D192N、D192Q、D192E、D192K或D192R;更优选为D192V、D192E、D192K、D192R、D192A、D192P、D192S或D192Q;甚至更优选为D192P、D192V、D192E或D192A;甚至更优选为D192P、D192V、D192A或D192E;最优选为D192E;和/或
-在位置V194处替换为V194A、V194R、V194N、V194D、V194C、V194Q、V194E、V194G、V194H、V194I、V194L、V194K、V194M、V194F、V194P、V194S、V194T、V194W或V194Y;优选为V194A、V194I、V194L、V194M、V194F、V194G、V194Y、V194S、V194T、V194N、V194Q或V194D;更优选为V194 L、V194I、V194A、V194M或V194S;甚至更优选为V194L或V194I;最优选为V194I;和/或
-在位置S204处替换为S204A、S204R、S204N、S204D、S204C、S204Q、S204E、S204G、S204H、S204I、S204L、S204K、S204M、S204F、S204P、S204T、S204W、S204Y或S204V;优选为S204A、S204M、S204P、S204G、S204C、S204N、S204Q、S204E、S204D、S204H、S204K、S204R或S204T;更优选为S204N、S204G、S204K、S204P或S204A;甚至更优选为S204A、S204K、S204P、S204N或S204G;甚至更优选为S204A、S204K、S204P或S204N;最优选为S204N;和/或
-在位置A228处替换为A228R、A228N、A228D、A228C、A228Q、A228E、A228G、A228H、A228I、A228L、A228K、A228M、A228F、A228P、A228S、A228T、A228W、A228Y或A228V;优选为A228V、A228I、A228S、A228T、A228N、A228Q、A228D、A228E、A228H或A228R;更优选为A228S、A228Q或A228T;最优选为A228T;和/或
-在位置I229处替换为I229A、I229R、I229N、I229D、I229C、I229Q、I229E、I229G、I229H、I229L、I229K、I229M、I229F、I229P、I229S、I229T、I229W、I229Y或I229V;优选为I229A、I229V、I229L、I229M、I229G、I229S、I229T、I229C、I229N、I229D、I229E或I229H;更优选为I229T、I229S、I229A或I229V;最优选为I229T;和/或
-在位置L230处替换为L230A、L230R、L230N、L230D、L230C、L230Q、L230E、L230G、L230H、L230I、L230K、L230M、L230F、L230P、L230S、L230T、L230W、L230Y或L230V;优选为L230V、L230I、L230M、L230F、L230T、L230S、L230G或L230A;更优选为L230A、L230V、L230I或L230F;甚至更优选为L230F、L230V或L230I;最优选为L230V;和/或
-在位置V231处替换为V231A、V231R、V231N、V231D、V231C、V231Q、V231E、V231G、V231H、V231I、V231L、V231K、V231M、V231F、V231P、V231S、V231T、V231W或V231Y;优选为V231A、V231I、V231L、V231P、V231G、V231S、V231T、V231N、V231Q、V231D、V231E、V231H、V231F、V231K或V231R;更优选为V231F、V231H、V231S或V231T;甚至更优选为V231F、V231S或V231H;甚至更优选为V231F或V231S;最优选为V231S;和/或
-在位置F236处替换为F236A、F236R、F236N、F236D、F236C、F236Q、F236E、F236G、F236H、F236I、F236L、F236K、F236M、F236P、F236S、F236T、F236W、F236Y或F236V;优选为F236A、F236V、F236I、F236L、F236M、F236W、F236G、F236Y、F236S、F236T、F236N、F236Q、F236D、F236E或F236R;更优选为F236Y、F236W或F236M;甚至更优选为F236Y或F236M,最优选为F236Y;和/或
-在位置S237处替换为S237A、S237R、S237N、S237D、S237C、S237Q、S237E、S237G、S237H、S237I、S237L、S237K、S237M、S237F、S237P、S237T、S237W、S237Y或S237V;优选为S237A、S237V、S237I、S237L、S237M、S237F、S237G、S237Y、S237T、S237C、S237N、S237Q、S237D、S237E、S237H、S237K或S237R;更优选为S237F、S237A、S237V或S237C;甚至更优选为S237F、S237A或S237C;甚至更优选为S237F或S237C;最优选为S237C;和/或
-在位置T238处替换为T238A、T238R、T238N、T238D、T238C、T238Q、T238E、T238G、T238H、T238I、T238L、T238K、T238M、T238F、T238P、T238S、T238W、T238Y或T238V;优选为T238F、T238A、T238V、T238I、T238L、T238M、T238P、T238G、T238A、T238C、T238N或T238Q;更优选为T238F、T238A、T238V、T238I、T238L、T238M或T238G;甚至更优选为T238F、T238M或T238A;甚至更优选为T238F或T238A;最优选为T238A。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个或至少22个位置处不同,以使其包含选自:V16I、I18M、S22A、Q23E、G26S、V34L、V46I、A89S、E130Q、A189S、I190M、S191A、D192E、V194I、S204N、A228T、I229T、L230V、V231S、F236Y、S237C和T238A中的至少一个或多个或所有替换。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个、至少22个或至少23个位置处不同,以使其包含选自:M1MTSSSSPSL、V16I、I18M、S22A、Q23E、G26S、V34L、V46I、A89S、E130Q、A189S、I190M、S191A、D192E、V194I、S204N、A228T、I229T、L230V、V231S、F236Y、S237C和T238A中的至少一个或多个或所有替换。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个或至少8个位置处不同,以使其包含选自:M1MTSSSSPSL、V16I、S22A、Q23E、V46I、I190M、A228T、I229T中的至少一个或多个或所有替换,并且此外可以与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个或至少15个位置处不同,从而使其进一步包含选自:I18M、G26S、V34L、A89S、E130Q、A189S、S191A、V194I、D192E、S204N、L230V、V231S、F236Y、S237C和T238A中的至少一个或多个或所有替换。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个或至少7个位置处不同,以使其包含选自:M1MTSSSSPSL、V16I、S22A、Q23E、V46I、A228T、I229T中的至少一个或多个或所有替换,并且此外可以与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个、至少15个位置或至少16个位置处不同,以使其进一步包含选自:I18M、G26S、V34L、A89S、E130Q、A189S、I190M、S191A、V194I、D192E、S204N、L230V、V231S、F236Y、S237C,和T238A中的至少一个或多个或所有替换。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个位置或至少9个位置处不同,以使其包含选自:M1MTSSSSPSL、I18M、A89S、E130Q、A189S、D192E、S204N、V231S、S237C中的至少一个或多个或所有替换,并且此外可以与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个或至少14个位置处不同,以使其进一步包含选自:V16I、S22A、Q23E、G26S、V34L、V46I、I190M、S191A、V194I、A228T、I229T、L230V、F236Y,和T238A中的至少一个或多个或所有替换。
优选地,本发明的酮还原酶与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个或至少8个位置处不同,以使其包含选自:M1MTSSSSPSL、I18M、E130Q、A189S、D192E、S204N、V231S、S237C中的至少一个或多个或所有替换,并且此外可以与比较序列SEQ ID NO:3在至少一个或多个,优选至少2个、至少3个、至少4个、至少5个、至少6个、至少7个、至少8个、至少9个、至少10个、至少11个、至少12个、至少13个、至少14个或至少15个位置处不同,以使其进一步包含选自:V16I、S22A、Q23E、G26S、V34L、V46I、A89S、I190M、S191A、V194I、A228T、I229T、L230V、F236Y和T238A中的至少一个或多个或所有替换。
本发明的第二方面涉及酮还原酶,其包含与比较序列SEQ ID NO:3的氨基酸序列具有至少70%的同一性但与比较序列SEQ ID NO:3的区别在于至少有一个残基改变的氨基酸序列。
在本发明的该第二方面的优选实施方案中,酮还原酶包含具有以下特征的氨基酸序列
-与比较序列SEQ ID NO:3的氨基酸序列具有至少70%、或至少71%、或至少72%、或至少73%、或至少74%、或至少75%、或至少76%、或至少77%、或至少78%、或至少79%、或至少80%、或至少81%、或至少82%、或至少83%、或至少84%、或至少85%、或至少86%、或至少87%、或至少88%、或至少89%、或至少90%、或至少91%、或至少92%、或至少93%、或至少94%、或至少95%、或至少96%、或至少97%、或至少98%、或至少99%的同一性;和
-与比较序列SEQ ID NO:3的区别在于1-23个残基改变、或2-23个残基改变、或3-23个残基改变、或4-23个残基改变、或5-23个残基改变、或6-23个残基改变、或7-23个残基改变、或8-23个残基改变、或9-23个残基改变、或10-23个残基改变、或11-23个残基改变、或12-23个残基改变、或13-23个残基改变、或14-23个残基改变、或15-23个残基改变、或16-23个残基改变、或17-23个残基改变、或18-23个残基改变、或19-23个残基改变、或20-23个残基改变、或21-23个残基改变、或22-23个残基改变、或23个残基改变。
因此,本发明的该第二方面的酮还原酶优选地满足两个要求,一个是关于与比较序列SEQ ID NO:3的最小同一性的要求,另一个是关于与比较序列SEQ ID NO:3的某些差异的要求。
上文已经结合本发明的第一方面详细说明了根据本发明第二方面的酮还原酶与比较序列SEQ ID NO:3的优选差异,即,酮还原酶包含与本发明的SEQ ID NO:1的氨基酸序列具有至少具有92%的同一性的氨基酸序列,并且类似地适用于本发明的第二方面,即,酮还原酶包含与比较序列SEQ ID NO:3的氨基酸序列具有至少70%的同一性但与比较序列SEQ ID NO:3的区别在于至少有一个残基改变的氨基酸序列。因此,在下文中不再重复所述优选的差异和特定的替代。
本发明的第三方面涉及一种酮还原酶,其包含与比较序列SEQ ID NO:1的氨基酸序列具有至少70%的同一性的氨基酸序列,其中与比较序列SEQ ID NO:1相比,该氨基酸序列
(i)至少有一个氨基酸残基改变,并且
(ii)表现出改善的热稳定性,和/或
(iii)在醇底物向酮或醛产物的氧化中表现出增加的活性,特别是比活性,和/或
(iv)在酮或醛底物向为醇产物的还原中表现出增加的活性,特别是比活性;和/或
(v)对于将酮或醛底物转化为醇产物或相反方向的转化(vice versa),表现出氧化活性与还原活性之比增加;和/或
(vi)对于将酮或醛底物转化为醇产物或相反方向的转化(vice versa),表现出还原活性与氧化活性之比增加;或
(vii)对于将酮或醛底物转化为醇产物或相反方向的转化(vice versa),表现出还原活性与氧化活性之比降低。
优选地,酮还原酶包含与本发明的SEQ ID NO:1的氨基酸序列具有至少71%、至少72%、至少73%、至少74%、至少75%、至少76%、至少77%、至少78%、至少79%、至少80%、至少81%、至少82%、至少83%、至少84%、至少85%、至少86%、至少87%、至少88%、至少89%、至少90%、更优选至少91.0%、至少91.5%、至少92.0%、至少92.1%、至少92.2%、至少92.3%、至少92.4%、至少92.5%、至少92.6%、至少92.7%、至少92.8%、至少92.9%、至少93%、至少93.1%、至少93.2%、至少93.3%、至少93.4%、至少93.5%、至少93.6%、至少93.7%、至少93.8%、至少93.9%、更优选至少94%、至少94.1%、至少94.2%、至少94.3%、至少94.4%、至少94.5%、至少94.6%、至少94.7%、至少94.8%、至少94.9%、还更优选至少95%、至少95.1%、至少95.2%、至少95.3%、至少95.4%、至少95.5%、至少95.6%、至少95.7%、至少95.8%、至少95.9%、仍然更优选至少96%、至少96.1%、至少96.2%、至少96.3%、至少96.4%、至少96.5%、至少96.6%、至少96.7%、至少96.8%、至少96.9%、甚至更优选至少97%、至少97.1%、至少97.2%、至少97.3%、至少97.4%、至少97.5%、至少97.6%、至少97.7%、至少97.8%、至少97.9%、最优选至少98%、至少98.1%、至少98.2%、至少98.3%、至少98.4%、至少98.5%、至少98.6%、至少98.7%、至少98.8%、至少98.9%、以及尤其是至少99%、至少99.1%、至少99.2%、至少99.3%、至少99.4%、至少99.5%、至少99.6%、至少99.7%、至少99.8%或至少99.9%的同一性的氨基酸序列。
在下文中,出于关于本发明的第三方面的说明的目的,除非另有明确说明,否则位置编号是指本发明的SEQ ID NO:1的位置。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-01中的至少一个残基改变,其中所述序列区段SS-01在本发明的SEQ ID NO:1的位置1至本发明的SEQ ID NO:1的位置10的范围内;优选地,其中所述序列区段SS-01在本发明的SEQ ID NO:1的位置3至本发明的SEQ ID NO:1的位置10的范围内;优选在选自S3、S4、S8、L9和N10中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置S3处的优选突变优选选自S3A、S3R、S3N、S3D、S3C、S3Q、S3E、S3G、S3H、S3I、S3L、S3K、S3M、S3F、S3P、S3T、S3W、S3Y和S3V;更优选为S3I;和/或
本发明的SEQ ID NO:1的位置S4处的优选突变优选选自S4A、S4R、S4N、S4D、S4C、S4Q、S4E、S4G、S4H、S4I、S4L、S4K、S4M、S4F、S4P、S4T、S4W、S4Y和S4V;更优选为S4N;和/或
本发明的SEQ ID NO:1的位置S8处的优选突变优选选自S8A、S8R、S8N、S8D、S8C、S8Q、S8E、S8G、S8H、S8I、S8L、S8K、S8M、S8F、S8P、S8T、S8W、S8Y和S8V;更优选为S8A;和/或
本发明的SEQ ID NO:1的位置L9处的优选突变优选选自L9A、L9R、L9N、L9D、L9C、L9Q、L9E、L9G、L9H、L9I、L9K、L9M、L9F、L9P、L9S、L9T、L9W、L9Y和L9V;更优选选自L9C、L9M、L9R、L9K、L9Q和L9G;和/或
本发明的SEQ ID NO:1的位置N10处的优选突变优选选自N10A、N10R、N10D、N10C、N10Q、N10E、N10G、N10H、N10I、N10L、N10K、N10M、N10F、N10P、N10S、N10T、N10W、N10Y和N10V;更优选为N10G。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-02中的至少一个残基改变,其中所述序列区段SS-02范围在本发明的SEQ ID NO:1的位置11至本发明的SEQ ID NO:1的位置20的范围内;优选地,其中所述序列区段SS-02在本发明的SEQ ID NO:1的位置11至本发明的SEQ IDNO:1的位置16的范围内;优选在选自A11和V13中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置A11处的优选突变优选选自A11R、A11N、A11D、A11C、A11Q、A11E、A11G、A11H、A11I、A11L、A11K、A11M、A11F、A11P、A11S、A11T、A11W、A11Y和A11V;更优选选自A11V和A11C;和/或
本发明的SEQ ID NO:1的位置V13处的优选突变优选选自V13A、V13R、V13N、V13D、V13C、V13Q、V13E、V13G、V13H、V13I、V13L、V13K、V13M、V13F、V13P、V13S、V13T、V13W和V13Y;更优选为V13G。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-03中的至少一个残基改变,其中所述序列区段SS-03在本发明的SEQ ID NO:1的位置21至本发明的SEQ ID NO:1的位置30的范围内;优选地,其中所述序列区段SS-03在本发明的SEQ ID NO:1的位置22至本发明的SEQ ID NO:1的位置30的范围内;优选在选自E22,S25,M26和A30中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置E22处的优选突变优选选自E22A、E22R、E22N、E22D、E22C、E22Q、E22G、E22H、E22I、E22L、E22K、E22M、E22F、E22P、E22S、E22T、E22W、E22Y和E22V;更优选为E22K;和/或
本发明的SEQ ID NO:1的位置S25处的优选突变优选选自S25A、S25R、S25N、S25D、S25C、S25Q、S25E、S25G、S25H、S25I、S25L、S25K、S25M、S25F、S25P、S25T、S25W、S25Y,和S25V;更优选为S25A;和/或
本发明的SEQ ID NO:1的位置M26处的优选突变优选选自M26A、M26R、M26N、M26D、M26C、M26Q、M26E、M26G、M26H、M26I、M26L、M26K、M26F、M26P、M26S、M26T、M26W、M26Y和M26V;更优选为M26R;和/或
本发明的SEQ ID NO:1的位置A30处的优选突变优选选自A30R、A30N、A30D、A30C、A30Q、A30E、A30G、A30H、A30I、A30L、A30K、A30M、A30F、A30P、A30S、A30T、A30W、A30Y和A30V;更优选为A30R。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-04中的至少一个残基改变,其中所述序列区段SS-04在本发明的SEQ ID NO:1的位置31至本发明的SEQ ID NO:1的位置40的范围内;优选地,其中所述序列区段SS-04在本发明的SEQ ID NO:1的位置31至本发明的SEQ ID NO:1的位置39的范围内;优选在选自E31、A38和S39中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置E31处的优选突变优选选自E31A、E31R、E31N、E31D、E31C、E31Q、E31G、E31H、E31I、E31L、E31K、E31M、E31F、E31P、E31S、E31T、E31W、E31Y和E31V;更优选为E31N;和/或
本发明的SEQ ID NO:1的位置A38处的优选突变优选选自A38R、A38N、A38D、A38C、A38Q、A38E、A38G、A38H、A38I、A38L、A38K、A38M、A38F、A38P、A38S、A38T、A38W、A38Y和A38V;更优选为A38M;和/或
本发明的SEQ ID NO:1的位置S39处的优选突变优选选自S39A、S39R、S39N、S39D、S39C、S39Q、S39E、S39G、S39H、S39I、S39L、S39K、S39M、S39F、S39P、S39T、S39W、S39Y和S39V;更优选选自S39G和S39A。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-05中的至少一个残基改变,其中所述序列区段SS-05在本发明的SEQ ID NO:1的位置41至本发明的SEQ ID NO:1的位置50的范围内;优选地,其中所述序列区段SS-05在本发明的SEQ ID NO:1的位置42至本发明的SEQ ID NO:1的位置50的范围内;优选在选自L42、E43、Q44、K49和A50中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置L42处的优选突变优选选自L42A、L42R、L42N、L42D、L42C、L42Q、L42E、L42G、L42H、L42I、L42K、L42M、L42F、L42P、L42S、L42T、L42W、L42Y和L42V;更优选为L42Y;和/或
本发明的SEQ ID NO:1的位置E43处的优选突变优选选自E43A、E43R、E43N、E43D、E43C、E43Q、E43G、E43H、E43I、E43L、E43K、E43M、E43F、E43P、E43S、E43T、E43W、E43Y和E43V;更优选为E43K;和/或
本发明的SEQ ID NO:1的位置Q44处的优选突变优选选自Q44A、Q44R、Q44N、Q44D、Q44C、Q44E、Q44G、Q44H、Q44I、Q44L、Q44K、Q44M、Q44F、Q44P、Q44S、Q44T、Q44W、Q44Y和Q44V;更优选选自Q44T、Q44E和Q44R;和/或
本发明的SEQ ID NO:1的位置K49处的优选突变优选选自K49A、K49R、K49N、K49D、K49C、K49Q、K49E、K49G、K49H、K49I、K49L、K49M、K49F、K49P、K49S、K49T、K49W、K49Y和K49V;更优选为K49R;和/或
本发明的SEQ ID NO:1的位置A50处的优选突变优选选自A50R、A50N、A50D、A50C、A50Q、A50E、A50G、A50H、A50I、A50L、A50K、A50M、A50F、A50P、A50S、A50T、A50W、A50Y和A50V;更优选为A50K。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-06中的至少一个残基改变,其中所述序列区段SS-06在本发明的SEQ ID NO:1的位置51至本发明的SEQ ID NO:1的位置60的范围内;优选地,其中所述序列区段SS-06在本发明的SEQ ID NO:1的位置54至本发明的SEQ ID NO:1的位置60的范围内;优选在选自I54、V55、K56、Q57和T60中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置I54处的优选突变优选选自I54A、I54R、I54N、I54D、I54C、I54Q、I54E、I54G、I54H、I54L、I54K、I54M、I54F、I54P、I54S、I54T、I54W、I54Y和I54V;更优选选自I54R和I54V;和/或
本发明的SEQ ID NO:1的位置V55处的优选突变优选选自V55A、V55R、V55N、V55D、V55C、V55Q、V55E、V55G、V55H、V55I、V55L、V55K、V55M、V55F、V55P、V55S、V55T、V55W和V55Y;更优选地选自V55I和V55C;和/或
本发明的SEQ ID NO:1的位置K56处的优选突变优选选自K56A、K56R、K56N、K56D、K56C、K56Q、K56E、K56G、K56H、K56I、K56L、K56M、K56F、K56P、K56S、K56T、K56W、K56Y和K56V;更优选为K56M;和/或
本发明的SEQ ID NO:1的位置Q57处的优选突变优选选自Q57A、Q57R、Q57N、Q57D、Q57C、Q57E、Q57G、Q57H、Q57I、Q57L、Q57K、Q57M、Q57F、Q57P、Q57S、Q57T、Q57W、Q57Y和Q57V;更优选选自Q57P和Q57A;和/或
本发明的SEQ ID NO:1的位置T60处的优选突变优选选自T60A、T60R、T60N、T60D、T60C、T60Q、T60E、T60G、T60H、T60I、T60L、T60K、T60M、T60F、T60P、T60S、T60W、T60Y和T60V;更优选选自T60C、T60N和T60A。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-07中的至少一个残基改变,其中所述序列区段SS-07在本发明的SEQ ID NO:1的位置61至本发明的SEQ ID NO:1的位置70的范围内;优选地,其中所述序列区段SS-07在本发明的SEQ ID NO:1的位置63至本发明的SEQ ID NO:1的位置70的范围内;优选在选自S69和D70的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置S69处的优选突变优选选自S69A、S69R、S69N、S69D、S69C、S69Q、S69E、S69G、S69H、S69I、S69L、S69K、S69M、S69F、S69P、S69T、S69W、S69Y和S69V;更优选地选自S69V和S69A;和/或
本发明的SEQ ID NO:1的位置D70处的优选突变优选选自D70A、D70R、D70N、D70C、D70Q、D70E、D70G、D70H、D70I、D70L、D70K、D70M、D70F、D70P、D70S、D70T、D70W、D70Y和D70V;更优选选自D70C和D70S。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-08中的至少一个残基改变,其中所述序列区段SS-08在本发明的SEQ ID NO:1的位置71至本发明的SEQ ID NO:1的位置80的范围内;优选地,其中所述序列区段SS-08在本发明的SEQ ID NO:1的位置75至本发明的SEQ ID NO:1的位置77的范围内;优选在选自G75、S76和F77中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置G75处的优选突变优选选自G75A、G75R、G75N、G75D、G75C、G75Q、G75E、G75H、G75I、G75L、G75K、G75M、G75F、G75P、G75S、G75T、G75W、G75Y和G75V;更优选为G75L;和/或
本发明的SEQ ID NO:1的位置S76处的优选突变优选选自S76A、S76R、S76N、S76D、S76C、S76Q、S76E、S76G、S76H、S76I、S76L、S76K、S76M、S76F、S76P、S76T、S76W、S76Y和S76V;更优选选自S76M、S76C、S76L和S76G;和/或
本发明的SEQ ID NO:1的位置F77处的优选突变优选选自F77A、F77R、F77N、F77D、F77C、F77Q、F77E、F77G、F77H、F77I、F77L、F77K、F77M、F77P、F77S、F77T、F77W、F77Y和F77V;更优选为F77L。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-09中的至少一个残基改变,其中所述序列区段SS-09在本发明的SEQ ID NO:1的位置81至本发明的SEQ ID NO:1的位置90的范围内;优选地,其中所述序列区段SS-09在本发明的SEQ ID NO:1的位置82至本发明的SEQ ID NO:1的位置89的范围内。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-10中的至少一个残基改变,其中所述序列区段SS-10在本发明的SEQ ID NO:1的位置91至本发明的SEQ ID NO:1的位置100的范围内;优选地,其中所述序列区段SS-10在本发明的SEQ ID NO:1的位置92至本发明的SEQ ID NO:1的位置100的范围内;优选在选自S92、S97、Q98、F99和S100中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置S92处的优选突变优选选自S92A、S92R、S92N、S92D、S92C、S92Q、S92E、S92G、S92H、S92I、S92L、S92K、S92M、S92F、S92P、S92T、S92W、S92Y和S92V;更优选选自S92M和S92G;和/或
本发明的SEQ ID NO:1的位置S97处的优选突变优选选自S97A、S97R、S97N、S97D、S97C、S97Q、S97E、S97G、S97H、S97I、S97L、S97K、S97M、S97F、S97P、S97T、S97W、S97Y和S97V;更优选选自S97A、S97W、S97V、S97C、S97F和S97M;更优选选自S97V和S97C;最优选地选自S97C;和/或
本发明的SEQ ID NO:1的位置Q98处的优选突变优选选自Q98A、Q98R、Q98N、Q98D、Q98C、Q98E、Q98G、Q98H、Q98I、Q98L、Q98K、Q98M、Q98F、Q98P、Q98S、Q98T、Q98W、Q98Y和Q98V;更优选为Q98E;和/或
本发明的SEQ ID NO:1的位置F99处的优选突变优选选自F99A、F99R、F99N、F99D、F99C、F99Q、F99E、F99G、F99H、F99I、F99L、F99K、F99M、F99P、F99S、F99T、F99W、F99Y和F99V;更优选为F99R;和/或
本发明的SEQ ID NO:1的位置S100处的优选突变优选选自S100A、S100R、S100N、S100D、S100C、S100Q、S100E、S100G、S100H、S100I、S100L、S100K、S100M、S100F、S100P、S100T、S100W、S100Y和S100V;更优选地选自S100E、S100Q、S100R、S100V和S100C;最优选地选自S100R。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-11中的至少一个残基改变,其中所述序列区段SS-11在本发明的SEQ ID NO:1的位置101至本发明的SEQ ID NO:1的位置110的范围内;优选地,其中所述序列区段SS-11在本发明的SEQ ID NO:1的位置101至本发明的SEQ IDNO:1的位置109的范围内;优选在选自P101、A106、A108和D109中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置P101处的优选突变优选选自P101A、P101R、P101N、P101D、P101C、P101Q、P101E、P101G、P101H、P101I、P101L、P101K、P101M、P101F、P101S、P101T、P101W、P101Y和P101V;更优选地选自P101F、P101H和P101W;和/或
本发明的SEQ ID NO:1的位置A106处的优选突变优选选自A106R、A106N、A106D、A106C、A106Q、A106E、A106G、A106H、A106I、A106L、A106K、A106M、A106F、A106P、A106S、A106T、A106W、A106Y和A106V;更优选地选自A106C和A106E;和/或
本发明的SEQ ID NO:1的位置A108处的优选突变优选选自A108R、A108N、A108D、A108C、A108Q、A108E、A108G、A108H、A108I、A108L、A108K、A108M、A108F、A108P、A108S、A108T、A108W、A108Y和A108V;更优选为A108P;和/或
本发明的SEQ ID NO:1的位置D109处的优选突变优选选自D109A、D109R、D109N、D109C、D109Q、D109E、D109G、D109H、D109I、D109L、D109K、D109M、D109F、D109P、D109S、D109T、D109W、D109Y和D109V;更优选选自D109G和D109R。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-12中的至少一个残基改变,其中所述序列区段SS-12在本发明的SEQ ID NO:1的位置111至本发明的SEQ ID NO:1的位置120的范围内;优选地,其中所述序列区段SS-12在本发明的SEQ ID NO:1的位置112至本发明的SEQ IDNO:1的位置120的范围内;优选在选自N112、M113、L114、T119和A120中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置N112处的优选突变优选选自N112A、N112R、N112D、N112C、N112Q、N112E、N112G、N112H、N112I、N112L、N112K、N112M、N112F、N112P、N112S、N112T、N112W、N112Y和N112V;更优选为N112T;和/或
本发明的SEQ ID NO:1的位置M113处的优选突变优选选自M113A、M113R、M113N、M113D、M113C、M113Q、M113E、M113G、M113H、M113I、M113L、M113K、M113F、M113P、M113S、M113T、M113W、M113Y和M113V;更优选为M113V;和/或
本发明的SEQ ID NO:1的位置L114处的优选突变优选选自L114A、L114R、L114N、L114D、L114C、L114Q、L114E、L114G、L114H、L114I、L114K、L114M、L114F、L114P、L114S、L114T、L114W、L114Y和L114V;更优选选自L114V和L114I;和/或
本发明的SEQ ID NO:1的位置T119处的优选突变优选选自T119A、T119R、T119N、T119D、T119C、T119Q、T119E、T119G、T119H、T119I、T119L、T119K、T119M、T119F、T119P、T119S、T119W、T119Y和T119V;更优选为T119L;和/或
本发明的SEQ ID NO:1的位置A120处的优选突变优选选自A120R、A120N、A120D、A120C、A120Q、A120E、A120G、A120H、A120I、A120L、A120K、A120M、A120F、A120P、A120S、A120T、A120W、A120Y和A120V;更优选为A120G。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-13中的至少一个残基改变,其中所述序列区段SS-13在本发明的SEQ ID NO:1的位置121至本发明的SEQ ID NO:1的位置130的范围内;优选地,其中所述序列区段SS-13在本发明的SEQ ID NO:1的位置121至本发明的SEQ IDNO:1的位置127的范围内;优选在选自P121和A127的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置P121处的优选突变优选选自P121A、P121R、P121N、P121D、P121C、P121Q、P121E、P121G、P121H、P121I、P121L、P121K、P121M、P121F、P121S、P121T、P121W、P121Y和P121V;更优选地选自P121V和P121A;最优选地选自P121V;和/或
本发明的SEQ ID NO:1的位置A127处的优选突变优选选自A127R、A127N、A127D、A127C、A127Q、A127E、A127G、A127H、A127I、A127L、A127K、A127M、A127F、A127P、A127S、A127T、A127W、A127Y和A127V;更优选选自A127K、A127C、A127G和A127Q。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-14中的至少一个残基改变,其中所述序列区段SS-14在本发明的SEQ ID NO:1的位置131至本发明的SEQ ID NO:1的位置140的范围内;优选地,其中所述序列区段SS-14在本发明的SEQ ID NO:1的位置132至本发明的SEQ IDNO:1的位置139的范围内;优选在选自I132、S133、P136、Q138和T139中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置I132处的优选突变优选选自I132A、I132R、I132N、I132D、I132C、I132Q、I132E、I132G、I132H、I132L、I132K、I132M、I132F、I132P、I132S、I132T、I132W、I132Y和I132V;更优选地选自I132V、I132W和I132F;和/或
本发明的SEQ ID NO:1的位置S133处的优选突变优选选自S133A、S133R、S133N、S133D、S133C、S133Q、S133E、S133G、S133H、S133I、S133L、S133K、S133M、S133F、S133P、S133T、S133W、S133Y和S133V;更优选选自S133M、S133L、S133A、S133Q和S133V;和/或
本发明的SEQ ID NO:1的位置P136处的优选突变优选选自P136A、P136R、P136N、P136D、P136C、P136Q、P136E、P136G、P136H、P136I、P136L、P136K、P136M、P136F、P136S、P136T、P136W、P136Y和P136V;更优选地选自P136V和P136G;和/或
本发明的SEQ ID NO:1的位置Q138处的优选突变优选选自Q138A、Q138R、Q138N、Q138D、Q138C、Q138E、Q138G、Q138H、Q138I、Q138L、Q138K、Q138M、Q138F、Q138P、Q138S、Q138T、Q138W、Q138Y和Q138V;更优选地选自Q138A、Q138T和Q138S;和/或
本发明的SEQ ID NO:1的位置T139处的优选突变优选选自T139A、T139R、T139N、T139D、T139C、T139Q、T139E、T139G、T139H、T139I、T139L、T139K、T139M、T139F、T139P、T139S、T139W、T139Y和T139V;更优选选自T139V、T139F和T139K。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-15中的至少一个残基改变,其中所述序列区段SS-15在本发明的SEQ ID NO:1的位置141至本发明的SEQ ID NO:1的位置150的范围内;优选地,其中所述序列区段SS-15在本发明的SEQ ID NO:1的位置141至本发明的SEQ IDNO:1的位置149的范围内;优选在选自A141、S147、T148和G149中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置A141处的优选突变优选选自A141R、A141N、A141D、A141C、A141Q、A141E、A141G、A141H、A141I、A141L、A141K、A141M、A141F、A141P、A141S、A141T、A141W、A141Y和A141V;更优选为A141S;和/或
本发明的SEQ ID NO:1的位置S147处的优选突变优选选自S147A、S147R、S147N、S147D、S147C、S147Q、S147E、S147H、S147I、S147L、S147K、S147M、S147F、S147P、S147S、S147T、S147W、S147Y和S147V;更优选为S147C;和/或
本发明的SEQ ID NO:1的位置T148处的优选突变优选选自T148A、T148R、T148N、T148D、T148C、T148Q、T148E、T148G、T148H、T148I、T148L、T148K、T148M、T148F、T148P、T148S、T148W、T148Y和T148V;更优选为T148S;和/或
本发明的SEQ ID NO:1的位置G149处的优选突变优选选自G149A、G149R、G149N、G149D、G149C、G149Q、G149E、G149H、G149I、G149L、G149K、G149M、G149F、G149P、G149S、G149T、G149W、G149Y和G149V;更优选选自G149C、G149V、G149M、G149I和G149S,最优选选自G149C、G149V、G149M和G149I。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-16中的至少一个残基改变,其中所述序列区段SS-16在本发明的SEQ ID NO:1的位置151至本发明的SEQ ID NO:1的位置160的范围内;优选地,其中所述序列区段SS-16在本发明的SEQ ID NO:1的位置151至本发明的SEQ IDNO:1的位置159的范围内;优选在选自S151、K152、A155、M157、V158和G159中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置S151处的优选突变优选选自S151A、S151R、S151N、S151D、S151C、S151Q、S151E、S151G、S151H、S151I、S151L、S151K、S151M、S151F、S151P、S151T、S151W、S151Y和S151V;更优选为S151A;和/或
本发明的SEQ ID NO:1的位置K152处的优选突变优选选自K152A、K152R、K152N、K152D、K152C、K152Q、K152E、K152G、K152H、K152I、K152L、K152M、K152F、K152P、K152S、K152T、K152W、K152Y和K152V;更优选为K152A;和/或
本发明的SEQ ID NO:1的位置A155处的优选突变优选选自A155R、A155N、A155D、A155C、A155Q、A155E、A155G、A155H、A155I、A155L、A155K、A155M、A155F、A155P、A155S、A155T、A155W、A155Y和A155V;更优选为A155W和A155H;和/或
本发明的SEQ ID NO:1的位置M157处的优选突变优选选自M157A、M157R、M157N、M157D、M157C、M157Q、M157E、M157G、M157H、M157I、M157L、M157K、M157F、M157P、M157S、M157T、M157W、M157Y和M157V;更优选选自M157F、M157R、M157G、M157C、M157E、M157D和M157K;和/或
本发明的SEQ ID NO:1的位置V158处的优选突变优选选自V158A、V158R、V158N、V158D、V158C、V158Q、V158E、V158G、V158H、V158I、V158L、V158K、V158M、V158F、V158P、V158S、V158T、V158W和V158Y;更优选为V158M;和/或
本发明的SEQ ID NO:1的位置G159处的优选突变优选选自G159A、G159R、G159N、G159D、G159C、G159Q、G159E、G159H、G159I、G159L、G159K、G159M、G159F、G159P、G159S、G159T、G159W、G159Y和G159V;更优选选自G159S和G159A。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-17中的至少一个残基改变,其中所述序列区段SS-17在本发明的SEQ ID NO:1的位置161至本发明的SEQ ID NO:1的位置170的范围内;优选地,其中所述序列区段SS-17在本发明的SEQ ID NO:1的位置168至本发明的SEQ IDNO:1的位置170的范围内;优选在选自D169中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置D169处的优选突变优选选自D169A、D169R、D169N、D169C、D169Q、D169E、D169G、D169H、D169I、D169L、D169K、D169M、D169F、D169P、D169S、D169T、D169W、D169Y和D169V;更优选为D169E。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-18中的至少一个残基改变,其中所述序列区段SS-18在本发明的SEQ ID NO:1的位置171至本发明的SEQ ID NO:1的位置180的范围内;优选地,其中所述序列区段SS-18在本发明的SEQ ID NO:1的位置172至本发明的SEQ IDNO:1的位置177的范围内;优选在选自M172的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置M172处的优选突变优选选自M172A、M172R、M172N、M172D、M172C、M172Q、M172E、M172G、M172H、M172I、M172L、M172K、M172F、M172P、M172S、M172T、M172W、M172Y和M172V;更优选为M172C。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-19中的至少一个残基改变,其中所述序列区段SS-19在本发明的SEQ ID NO:1的位置181至本发明的SEQ ID NO:1的位置190的范围内;优选地,其中所述序列区段SS-19在本发明的SEQ ID NO:1的位置181至本发明的SEQ IDNO:1的位置185的范围内;优选在选自P181、K182和N185中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置P181处的优选突变优选选自P181A、P181R、P181N、P181D、P181C、P181Q、P181E、P181G、P181H、P181I、P181L、P181K、P181M、P181F、P181S、P181T、P181W、P181Y和P181V;更优选地选自P181V、P181W和P181L;和/或
本发明的SEQ ID NO:1的位置K182处的优选突变优选选自K182A、K182R、K182N、K182D、K182C、K182Q、K182E、K182G、K182H、K182I、K182L、K182M、K182F、K182P、K182S、K182T、K182W、K182Y和K182V;更优选为K182I;和/或
本发明的SEQ ID NO:1的位置N185处的优选突变优选选自N185A、N185R、N185D、N185C、N185Q、N185E、N185G、N185H、N185I、N185L、N185K、N185M、N185F、N185P、N185S、N185T、N185W、N185Y和N185V;更优选为N185R。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-20中的至少一个残基改变,其中所述序列区段SS-20在本发明的SEQ ID NO:1的位置191至本发明的SEQ ID NO:1的位置200的范围内;优选地,其中所述序列区段SS-20在本发明的SEQ ID NO:1的位置194至本发明的SEQ IDNO:1的位置200的范围内;优选在选自T194、A199和E200中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置T194处的优选突变优选选自T194A、T194R、T194N、T194D、T194C、T194Q、T194E、T194G、T194H、T194I、T194L、T194K、T194M、T194F、T194P、T194S、T194W、T194Y和T194V;更优选为T194V;和/或
本发明的SEQ ID NO:1的位置A199处的优选突变优选选自A199R、A199N、A199D、A199C、A199Q、A199E、A199G、A199H、A199I、A199L、A199K、A199M、A199F、A199P、A199S、A199T、A199W、A199Y和A199V;更优选为A199C;和/或
本发明的SEQ ID NO:1的位置E200处的优选突变优选选自E200A、E200R、E200N、E200D、E200C、E200Q、E200G、E200H、E200I、E200L、E200K、E200M、E200F、E200P、E200S、E200T、E200W、E200Y和E200V;更优选选自E200P、E200V和E200A。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-21中的至少一个残基改变,其中所述序列区段SS-21范围在本发明的SEQ ID NO:1的位置201至本发明的SEQ ID NO:1的位置210的范围内;优选地,其中所述序列区段SS-21在本发明的SEQ ID NO:1的位置202至本发明的SEQ IDNO:1的位置210的范围内;优选在选自I202、P204、M206、D208、L209和P210中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置I202处的优选突变优选选自I202A、I202R、I202N、I202D、I202C、I202Q、I202E、I202G、I202H、I202L、I202K、I202M、I202F、I202P、I202S、I202T、I202W、I202Y和I202V;更优选为I202V;和/或
本发明的SEQ ID NO:1的位置P204处的优选突变优选选自P204A、P204R、P204N、P204D、P204C、P204Q、P204E、P204G、P204H、P204I、P204L、P204K、P204M、P204F、P204S、P204T、P204W、P204Y和P204V;更优选为P204L;和/或
本发明的SEQ ID NO:1的位置M206处的优选突变优选选自M206A、M206R、M206N、M206D、M206C、M206Q、M206E、M206G、M206H、M206I、M206L、M206K、M206F、M206P、M206S、M206T、M206W、M206Y和M206V;更优选选自M206T、M206D、M206G和M206A;最优选地选自M206D;和/或
本发明的SEQ ID NO:1的位置D208处的优选突变优选选自D208A、D208R、D208N、D208C、D208Q、D208E、D208G、D208H、D208I、D208L、D208K、D208M、D208F、D208P、D208S、D208T、D208W、D208Y和D208V;更优选地选自D208H、D208C和D208N;和/或
本发明的SEQ ID NO:1的位置L209处的优选突变优选选自L209A、L209R、L209N、L209D、L209C、L209Q、L209E、L209G、L209H、L209I、L209K、L209M、L209F、L209P、L209S、L209T、L209W、L209Y和L209V;更优选为L209C;和/或
本发明的SEQ ID NO:1的位置P210处的优选突变优选选自P210A、P210R、P210N、P210D、P210C、P210Q、P210E、P210G、P210H、P210I、P210L、P210K、P210M、P210F、P210S、P210T、P210W、P210Y和P210V;更优选为P210C。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-22中的至少一个残基改变,其中所述序列区段SS-22在本发明的SEQ ID NO:1的位置211至本发明的SEQ ID NO:1的位置220的范围内;优选地,其中所述序列区段SS-22在本发明的SEQ ID NO:1的位置212至本发明的SEQ IDNO:1的位置220的范围内;优选在选自N212、G213、W214、I215、V217和I220的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置N212处的优选突变优选选自N212A、N212R、N212D、N212C、N212Q、N212E、N212G、N212H、N212I、N212L、N212K、N212M、N212F、N212P、N212S、N212T、N212W、N212Y和N212V;更优选地选自N212K、N212P和N212A;和/或
本发明的SEQ ID NO:1的位置G213处的优选突变优选选自G213A、G213R、G213N、G213D、G213C、G213Q、G213E、G213H、G213I、G213L、G213K、G213M、G213F、G213P、G213S、G213T、G213W、G213Y和G213V;更优选为G213D;和/或
本发明的SEQ ID NO:1的位置W214处的优选突变优选选自W214A、W214R、W214N、W214D、W214C、W214Q、W214E、W214G、W214H、W214I、W214L、W214K、W214M、W214F、W214P、W214S、W214T、W214Y和W214V;更优选为W214G;和/或
本发明的SEQ ID NO:1的位置I215处的优选突变优选选自I215A、I215R、I215N、I215D、I215C、I215Q、I215E、I215G、I215H、I215L、I215K、I215M、I215F、I215P、I215S、I215T、I215W、I215Y和I215V;更优选为I215T;和/或
本发明的SEQ ID NO:1的位置V217处的优选突变优选选自V217A、V217R、V217N、V217D、V217C、V217Q、V217E、V217G、V217H、V217I、V217L、V217K、V217M、V217F、V217P、V217S、V217T、V217W和V217Y;更优选地选自V217N和V217P;和/或
本发明的SEQ ID NO:1的位置I220处的优选突变优选选自I220A、I220R、I220N、I220D、I220C、I220Q、I220E、I220G、I220H、I220L、I220K、I220M、I220F、I220P、I220S、I220T、I220W、I220Y和I220V;更优选为I220V。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-23中的至少一个残基改变,其中所述序列区段SS-23在本发明的SEQ ID NO:1的位置221至本发明的SEQ ID NO:1的位置230的范围内;优选地,其中所述序列区段SS-23在本发明的SEQ ID NO:1的位置225至本发明的SEQ IDNO:1的位置227的范围内;优选在选自T225和L227中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置T225处的优选突变优选选自T225A、T225R、T225N、T225D、T225C、T225Q、T225E、T225G、T225H、T225I、T225L、T225K、T225M、T225F、T225P、T225S、T225W、T225Y和T225V;更优选地选自T225M和T225I;和/或
本发明的SEQ ID NO:1的位置L227处的优选突变优选选自L227A、L227R、L227N、L227D、L227C、L227Q、L227E、L227G、L227H、L227I、L227K、L227M、L227F、L227P、L227S、L227T、L227W、L227Y和L227V;更优选为L227V。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-24中的至少一个残基改变,其中所述序列区段SS-24在本发明的SEQ ID NO:1的位置231至本发明的SEQ ID NO:1的位置240的范围内;优选地,其中所述序列区段SS-24在本发明的SEQ ID NO:1的位置232至本发明的SEQ IDNO:1的位置239的范围内;优选在选自N232、G235、T236、T237、V238和S239中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置N232处的优选突变优选选自N232A、N232R、N232D、N232C、N232Q、N232E、N232G、N232H、N232I、N232L、N232K、N232M、N232F、N232P、N232S、N232T、N232W、N232Y和N232V;更优选地选自N232Y和N232F;和/或
本发明的SEQ ID NO:1的位置G235处的优选突变优选选自G235A、G235R、G235N、G235D、G235C、G235Q、G235E、G235H、G235I、G235L、G235K、G235M、G235F、G235P、G235S、G235T、G235W、G235Y和G235V;更优选为G235F;和/或
本发明的SEQ ID NO:1的位置T236处的优选突变优选选自T236A、T236R、T236N、T236D、T236C、T236Q、T236E、T236G、T236H、T236I、T236L、T236K、T236M、T236F、T236P、T236S、T236W、T236Y和T236V;更优选为T236S;和/或
本发明的SEQ ID NO:1的位置T237处的优选突变优选选自T237A、T237R、T237N、T237D、T237C、T237Q、T237E、T237G、T237H、T237I、T237L、T237K、T237M、T237F、T237P、T237S、T237W、T237Y和T237V;更优选为T237I;和/或
本发明的SEQ ID NO:1的位置V238处的优选突变优选选自V238A、V238R、V238N、V238D、V238C、V238Q、V238E、V238G、V238H、V238I、V238L、V238K、V238M、V238F、V238P、V238S、V238T、V238W和V238Y;更优选为V238F;和/或
本发明的SEQ ID NO:1的位置S239处的优选突变优选选自S239A、S239R、S239N、S239D、S239C、S239Q、S239E、S239G、S239H、S239I、S239L、S239K、S239M、S239F、S239P、S239T、S239W、S239Y和S239V;更优选为S239F。
在优选的实施方案中,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于本发明的SEQ ID NO:1的序列区段SS-25中的至少一个残基改变,其中所述序列区段SS-25在本发明的SEQ ID NO:1的位置241至本发明的SEQ ID NO:1的位置246的范围内;优选地,其中所述序列区段SS-25在本发明的SEQ ID NO:1的位置243至本发明的SEQ IDNO:1的位置246的范围内;优选在选自G243、Y244、C245和A246中的一个或多个或所有位置上的残基改变。
本发明的SEQ ID NO:1的位置G243处的优选突变优选选自G243A、G243R、G243N、G243D、G243C、G243Q、G243E、G243H、G243I、G243L、G243K、G243M、G243F、G243P、G243S、G243T、G243W、G243Y和G243V;更优选为G243V;和/或
本发明的SEQ ID NO:1的位置Y244处的优选突变优选选自Y244A、Y244R、Y244N、Y244D、Y244C、Y244Q、Y244E、Y244G、Y244H、Y244I、Y244L、Y244K、Y244M、Y244F、Y244P、Y244S、Y244T、Y244W和Y244V;更优选为Y244F和Y244L;最优选为Y244L;和/或
本发明的SEQ ID NO:1的位置C245处的优选突变优选选自C245A、C245R、C245N、C245D、C245Q、C245E、C245G、C245H、C245I、C245L、C245K、C245M、C245F、C245P、C245S、C245T、C245W、C245Y和C245V;更优选为C245F;和/或
本发明的SEQ ID NO:1的位置A246处的优选突变优选选自A246R、A246N、A246D、A246C、A246Q、A246E、A246G、A246H、A246I、A246L、A246K、A246M、A246F、A246P、A246S、A246T、A246W、A246Y和A246V;更优选为A246F。
在本发明的第三方面的又一个实施方案中,其也是第三方面的所有先前实施方案的一个实施方案,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于在一个或多个序列区段SS-01至SS-25中的至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变。
在本发明的第三方面的另一个实施方案中,其也是第三方面的所有先前实施方案的一个实施方案,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3、S4、S8、L9、N10、A11、V13、E22、S25、M26、A30、E31、A38、S39、L42、E43、Q44、K49、A50、I54、V55、K56、Q57、T60、S69、D70、G75、S76、F77、S92、S97、Q98、F99、S100、P101、A106、A108、D109、N112、M113、L114、T119、A120、P121、A127、I132、S133、P136、Q138、T139、A141、S147、T148、G149、S151、K152、A155、M157、V158、G159、D169、M172、P181、K182、N185、T194、A199、E200、I202、P204、M206、D208、L209、P210、N212、G213、W214、I215、V217、I220、T225、L227、N232、G235、T236、T237、V238、S239、G243、Y244、C245和A246,优选选自S97、F99、S100、P121、S147、G149、M206、P210和Y244。
在本发明的第三方面的另一个实施方案中,其也是第三方面的所有前述实施方案中的一个实施方案,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,优选选自:S97V、S97C、F99R、S100R、P121V、S147C、G149I、G149M、G149C、G149V、Y244L、M206D和P210C。
在第三方面的一个优选实施方案中,其也是第三方面的所有前述实施方案中的一个实施方案,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于一个或多个序列区段SS-01至SS-25中的至少两个残基改变。
在第三方面的一个优选实施方案中,其也是第三方面的所有前述实施方案中的一个实施方案,至少两个氨基酸位置包括一对两个氨基酸位置,其中该一对两个氨基酸位置选自:S97和F99、S97和S100、S97和P121、S97和S147、S97和G149、S97和M206、S97和P210、S97和Y244、F99和S100、F99和P121、F99和S147、F99和G149、F99和M206、F99和P210、F99和Y244、S100和P121、S100和S147、S100和G149、S100和M206、S100和P210、S100和Y244、P121和S147、P121和G149、P121和M206、P121和P210、P121和Y244、S147和G149、S147和M206、S147和P210、S147和Y244、G149和M206、G149和P210、G149和Y244、M206和P210、M206和Y244、或P210和Y244,更优选为S97和S100、S97和S147、S97和G149、S97和P210、S97和Y244、S100和S147、S100和G149、S100和P210、S100和Y244、S147和G149、S147和P210、S147和Y244、G149和P210、G149和Y244、或P210和Y244,最优选为S97和G149、S97和P210、S97和Y244、G149和P210、G149和Y244、P210和Y244
甚至更优选地,至少两个氨基酸位置包括一对两个氨基酸位置,其中该一对两个氨基酸位置选自:S97V和F99R、S97V和S100R、S97V和P121V、S97V和S147C、S97V和G149I、S97V和G149M、S97V和G149C、S97V和G149V、S97V和Y244L、S97V和M206D、S97V和P210C、S97C和F99R、S97C和S100R、S97C和P121V、S97C和S147C、S97C和G149I、S97C和G149M、S97C和G149C、S97C和G149V、S97C和Y244L、S97C和M206D、S97C和P210C、F99R和S100R、F99R和P121V、F99R和S147C、F99R和G149I、F99R和G149M、F99R和G149C、F99R和G149V、F99R和Y244L、F99R和M206D、F99R和P210C、S100R和P121V、S100R和S147C、S100R和G149I、S100R和G149M、S100R和G149C、S100R和G149V、S100R和Y244L、S100R和M206D、S100R和P210C、P121V和S147C、P121V和G149I、P121V和G149M、P121V和G149C、P121V和G149V、P121V和Y244L、P121V和M206D、P121V和P210C、S147C和G149I、S147C和G149M、S147C和G149C、S147C和G149V、S147C和Y244L、S147C和M206D、S147C和P210C、G149I和Y244L、G149I和M206D、G149I和P210C、G149M和Y244L、G149M和M206D、G149M和P210C、G149C和Y244L、G149C和M206D、G149C和P210C、G149V和Y244L、G149V和M206D、G149V和P210C、Y244L和M206D、Y244L和P210C、M206D和P210C,更优选为S97C和S100R、S97C和S147C、S97C和G149I、S97C和G149C、S97C和G149M、S97C和G149V、S97C和P210C、S97C和Y244L、S100R和S147C、S100R和G149I、S100R和G149C、S100R和G149M、S100R和G149V、S100R和P210C、S100R和Y244L、S147C和G149I、S147C和G149C、S147C和G149M、S147C和G149V、S147C和P210C、S147C和Y244L、G149I和P210C、G149I和Y244L、G149C和P210C、G149C和Y244L、G149M和P210C、G149M和Y244L、G149V和P210C、G149V和Y244L、或P210C和Y244L,最优选为S97C和G149I、S97C和G149C、S97C和G149M、S97C和G149V、S97C和P210C、S97C和Y244L、G149I和P210C、G149I和Y244L、G149C和P210C、G149C和Y244L、G149M和P210C、G149M和Y244L、G149V和P210C、G149V和Y244L、或P210C和Y244L。
在第三方面的一个优选实施方案中,其也是第三方面的所有前述实施方案中的一个实施方案,本发明的酮还原酶与本发明的SEQ ID NO:1的酮还原酶的区别在于一个或多个序列区段SS-01至SS-25中的至少三个残基改变。
在第三方面的一个优选实施方案中,其也是第三方面的所有前述实施方案中的一个实施方案,至少三个氨基酸位置包括三个氨基酸位置的三联体(triplet),其中三个氨基酸位置的三联体选自S97和F99和S100、S97和F99和P121、S97和F99和S147、S97和F99和G149、S97和F99和M206、S97和F99和P210、S97和F99和Y244、S97和S100和P121、S97和S100和S147、S97和S100和G149、S97和S100和M206、S97和S100和P210、S97和S100和Y244、S97和P121和S147、S97和P121和G149、S97和P121和M206、S97和P121和P210、S97和P121和Y244、S97和S147和G149、S97和S147和M206、S97和S147和P210、S97和S147和Y244、S97和G149和M206、S97和G149和P210、S97和G149和Y244、S97和M206和P210、S97和M206和Y244、S97和P210和Y244、F99和S100和P121、F99和S100和S147、F99和S100和G149、F99和S100和M206、F99和S100和P210、F99和S100和Y244、F99和P121和S147、F99和P121和G149、F99和P121和M206、F99和P121和P210、F99和P121和Y244、F99和S147和G149、F99和S147和M206、F99和S147和P210、F99和S147和Y244、F99和G149和M206、F99和G149和P210、F99和G149和Y244、F99和M206和P210、F99和M206和Y244、F99和P210和Y244、S100和P121和S147、S100和P121和G149、S100和P121和M206、S100和P121和P210、S100和P121和Y244、S100和S147和G149、S100和S147和M206、S100和S147和P210、S100和S147和Y244、S100和G149和M206、S100和G149和P210、S100和G149和Y244、S100和M206和P210、S100和M206和Y244、S100和P210和Y244、P121和S147和G149、P121和S147和M206、P121和S147和P210、P121和S147和Y244、P121和G149和M206、P121和G149和P210、P121和G149和Y244、P121和M206和P210、P121和M206和Y244、P121和P210和Y244、S147和G149和M206、S147和G149和P210、S147和G149和Y244、S147和M206和P210、S147和M206和Y244、S147和P210和Y244、G149和M206和P210、G149和M206和Y244、G149和P210和Y244、或M206和P210和Y244,更优选为S97和S100和S147、S97和S100和G149、S97和S100和P210、S97和S100和Y244、S97和S147和G149、S97和S147和P210、S97和S147和Y244、S97和G149和P210、S97和G149和Y244、S97和P210和Y244、S100和S147和G149、S100和S147和P210、S100和S147和Y244、S100和G149和P210、S100和G149和Y244、S100和P210和Y244、S147和G149和P210、S147和G149和Y244、S147和P210和Y244、或G149和P210和Y244,最优选为S97和G149和P210、S97和G149和Y244、S97和P210和Y244、或G149和P210和Y244。
甚至更优选地,至少三个氨基酸位置包括三个氨基酸位置的三联体,其中三个氨基酸位置的三联体选自S97V和F99R和S100R、S97V和F99R和P121V、S97V和F99R和S147C、S97V和F99R和G149I、S97V和F99R和G149M、S97V和F99R和G149C、S97V和F99R和G149V、S97V和F99R和Y244L、S97V和F99R和M206D、S97V和F99R和P210C、S97V和S100R和P121V、S97V和S100R和S147C、S97V和S100R和G149I、S97V和S100R和G149M、S97V和S100R和G149C、S97V和S100R和G149V、S97V和S100R和Y244L、S97V和S100R和M206D、S97V和S100R和P210C、S97V和P121V和S147C、S97V和P121V和G149I、S97V和P121V和G149M、S97V和P121V和G149C、S97V和P121V和G149V、S97V和P121V和Y244L、S97V和P121V和M206D、S97V和P121V和P210C、S97V和S147C和G149I、S97V和S147C和G149M、S97V和S147C和G149C、S97V和S147C和G149V、S97V和S147C和Y244L、S97V和S147C和M206D、S97V和S147C和P210C、S97V和G149I和Y244L、S97V和G149I和M206D、S97V和G149I和P210C、S97V和G149M和Y244L、S97V和G149M和M206D、S97V和G149M和P210C、S97V和G149C和Y244L、S97V和G149C和M206D、S97V和G149C和P210C、S97V和G149V和Y244L、S97V和G149V和M206D、S97V和G149V和P210C、S97V和Y244L和M206D、S97V和Y244L和P210C、S97V和M206D和P210C、S97C和F99R和S100R、S97C和F99R和P121V、S97C和F99R和S147C、S97C和F99R和G149I、S97C和F99R和G149M、S97C和F99R和G149C、S97C和F99R和G149V、S97C和F99R和Y244L、S97C和F99R和M206D、S97C和F99R和P210C、S97C和S100R和P121V、S97C和S100R和S147C、S97C和S100R和G149I、S97C和S100R和G149M、S97C和S100R和G149C、S97C和S100R和G149V、S97C和S100R和Y244L、S97C和S100R和M206D、S97C和S100R和P210C、S97C和P121V和S147C、S97C和P121V和G149I、S97C和P121V和G149M、S97C和P121V和G149C、S97C和P121V和G149V、S97C和P121V和Y244L、S97C和P121V和M206D、S97C和P121V和P210C、S97C和S147C和G149I、S97C和S147C和G149M、S97C和S147C和G149C、S97C和S147C和G149V、S97C和S147C和Y244L、S97C和S147C和M206D、S97C和S147C和P210C、S97C和G149I和Y244L、S97C和G149I和M206D、S97C和G149I和P210C、S97C和G149M和Y244L、S97C和G149M和M206D、S97C和G149M和P210C、S97C和G149C和Y244L、S97C和G149C和M206D、S97C和G149C和P210C、S97C和G149V和Y244L、S97C和G149V和M206D、S97C和G149V和P210C、S97C和Y244L和M206D、S97C和Y244L和P210C、S97C和M206D和P210C、F99R和S100R和P121V、F99R和S100R和S147C、F99R和S100R和G149I、F99R和S100R和G149M、F99R和S100R和G149C、F99R和S100R和G149V、F99R和S100R和Y244L、F99R和S100R和M206D、F99R和S100R和P210C、F99R和P121V和S147C、F99R和P121V和G149I、F99R和P121V和G149M、F99R和P121V和G149C、F99R和P121V和G149V、F99R和P121V和Y244L、F99R和P121V和M206D、F99R和P121V和P210C、F99R和S147C和G149I、F99R和S147C和G149M、F99R和S147C和G149C、F99R和S147C和G149V、F99R和S147C和Y244L、F99R和S147C和M206D、F99R和S147C和P210C、F99R和G149I和Y244L、F99R和G149I和M206D、F99R和G149I和P210C、F99R和G149M和Y244L、F99R和G149M和M206D、F99R和G149M和P210C、F99R和G149C和Y244L、F99R和G149C和M206D、F99R和G149C和P210C、F99R和G149V和Y244L、F99R和G149V和M206D、F99R和G149V和P210C、F99R和Y244L和M206D、F99R和Y244L和P210C、F99R和M206D和P210C、S100R和P121V和S147C、S100R和P121V和G149I、S100R和P121V和G149M、S100R和P121V和G149C、S100R和P121V和G149V、S100R和P121V和Y244L、S100R和P121V和M206D、S100R和P121V和P210C、S100R和S147C和G149I、S100R和S147C和G149M、S100R和S147C和G149C、S100R和S147C和G149V、S100R和S147C和Y244L、S100R和S147C和M206D、S100R和S147C和P210C、S100R和G149I和Y244L、S100R和G149I和M206D、S100R和G149I和P210C、S100R和G149M和Y244L、S100R和G149M和M206D、S100R和G149M和P210C、S100R和G149C和Y244L、S100R和G149C和M206D、S100R和G149C和P210C、S100R和G149V和Y244L、S100R和G149V和M206D、S100R和G149V和P210C、S100R和Y244L和M206D、S100R和Y244L和P210C、S100R和M206D和P210C、P121V和S147C和G149I、P121V和S147C和G149M、P121V和S147C和G149C、P121V和S147C和G149V、P121V和S147C和Y244L、P121V和S147C和M206D、P121V和S147C和P210C、P121V和G149I和Y244L、P121V和G149I和M206D、P121V和G149I和P210C、P121V和G149M和Y244L、P121V和G149M和M206D、P121V和G149M和P210C、P121V和G149C和Y244L、P121V和G149C和M206D、P121V和G149C和P210C、P121V和G149V和Y244L、P121V和G149V和M206D、P121V和G149V和P210C、P121V和Y244L和M206D、P121V和Y244L和P210C、P121V和M206D和P210C、S147C和G149I和Y244L、S147C和G149I和M206D、S147C和G149I和P210C、S147C和G149M和Y244L、S147C和G149M和M206D、S147C和G149M和P210C、S147C和G149C和Y244L、S147C和G149C和M206D、S147C和G149C和P210C、S147C和G149V和Y244L、S147C和G149V和M206D、S147C和G149V和P210C、S147C和Y244L和M206D、S147C和Y244L和P210C、S147C和M206D和P210C、G149I和Y244L和M206D、G149I和Y244L和P210C、G149I和M206D和P210C、G149M和Y244L和M206D、G149M和Y244L和P210C、G149M和M206D和P210C、G149C和Y244L和M206D、G149C和Y244L和P210C、G149C和M206D和P210C、G149V和Y244L和M206D、G149V和Y244L和P210C、G149V和M206D和P210C、或Y244L和M206D和P210C、更优选为S97C和S100R和S147C、S97C和S100R和G149I、S97C和S100R和G149C、S97C和S100R和G149M、S97C和S100R和G149V、S97C和S100R和P210C、S97C和S100R和Y244L、S97C和S147C和G149I、S97C和S147C和G149C、S97C和S147C和G149M、S97C和S147C和G149V、S97C和S147C和P210C、S97C和S147C和Y244L、S97C和G149I和P210C、S97C和G149I和Y244L、S97C和G149C和P210C、S97C和G149C和Y244L、S97C和G149M和P210C、S97C和G149M和Y244L、S97C和G149V和P210C、S97C和G149V和Y244L、S97C和P210C和Y244L、S100R和S147C和G149I、S100R和S147C和G149C、S100R和S147C和G149M、S100R和S147C和G149V、S100R和S147C和P210C、S100R和S147C和Y244L、S100R和G149I和P210C、S100R和G149I和Y244L、S100R和G149C和P210C、S100R和G149C和Y244L、S100R和G149M和P210C、S100R和G149M和Y244L、S100R和G149V和P210C、S100R和G149V和Y244L、S100R和P210C和Y244L、S147C和G149I和P210C、S147C和G149I和Y244L、S147C和G149C和P210C、S147C和G149C和Y244L、S147C和G149M和P210C、S147C和G149M和Y244L、S147C和G149V和P210C、S147C和G149V和Y244L、S147C和P210C和Y244L、G149I和P210C和Y244L、G149C和P210C和Y244L、G149M和P210C和Y244L、或G149V和P210C和Y244L、最优选为S97C和G149I和P210C、S97C和G149I和Y244L、S97C和G149C和G149M、S97C和G149C和P210C、S97C和G149C和Y244L、S97C和G149M和P210C、S97C和G149M和Y244L、S97C和G149V和P210C、S97C和G149V和Y244L、S97C和P210C和Y244L、G149I和P210C和Y244L、G149C和P210C和Y244L、G149M和P210C和Y244L、或G149V和P210C和Y244L。
在本发明的第三方面的一个优选实施方案中,其也是所有前述实施方案中的一个实施方案,所述酮还原酶包含选自以下的氨基酸序列或由选自以下的氨基酸序列组成:SEQID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ IDNO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ IDNO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ IDNO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ IDNO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ IDNO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ IDNO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47、SEQ IDNO:48、SEQ ID NO:49、SEQ ID NO:50、SEQ ID NO:51、SEQ ID NO:52、SEQ ID NO:53、SEQ IDNO:54、SEQ ID NO:55、SEQ ID NO:56、SEQ ID NO:57、SEQ ID NO:58、SEQ ID NO:59、SEQ IDNO:60、SEQ ID NO:61、SEQ ID NO:62、SEQ ID NO:63、SEQ ID NO:64、SEQ ID NO:65、SEQ IDNO:66、SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69、SEQ ID NO:70、SEQ ID NO:71、SEQ IDNO:72、SEQ ID NO:73、SEQ ID NO:74、SEQ ID NO:75、SEQ ID NO:76、SEQ ID NO:77、SEQ IDNO:78、SEQ ID NO:79、SEQ ID NO:80、SEQ ID NO:81、SEQ ID NO:82、SEQ ID NO:83、SEQ IDNO:84、SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87、SEQ ID NO:88、SEQ ID NO:89、SEQ IDNO:90、SEQ ID NO:91、SEQ ID NO:92、SEQ ID NO:93、SEQ ID NO:94、SEQ ID NO:95、SEQ IDNO:96、SEQ ID NO:97、SEQ ID NO:98、SEQ ID NO:99、SEQ ID NO:100、SEQ ID NO:102、SEQID NO:103、SEQ ID NO:104、SEQ ID NO:105、SEQ ID NO:106、SEQ ID NO:107、SEQ ID NO:108、SEQ ID NO:109、SEQ ID NO:110、SEQ ID NO:111、SEQ ID NO:112、SEQ ID NO:114、SEQID NO:115、SEQ ID NO:116、SEQ ID NO:117、SEQ ID NO:118、SEQ ID NO:119、SEQ ID NO:120、SEQ ID NO:121、SEQ ID NO:122、SEQ ID NO:123、SEQ ID NO:124、SEQ ID NO:125、SEQID NO:126、SEQ ID NO:127、SEQ ID NO:128、SEQ ID NO:129、SEQ ID NO:130、SEQ ID NO:131、SEQ ID NO:132、SEQ ID NO:133、SEQ ID NO:134、SEQ ID NO:135、SEQ ID NO:136、SEQID NO:137、SEQ ID NO:138、SEQ ID NO:139、SEQ ID NO:140、SEQ ID NO:141、SEQ ID NO:142、SEQ ID NO:143、SEQ ID NO:144、SEQ ID NO:145、SEQ ID NO:146、SEQ ID NO:147、SEQID NO:148、SEQ ID NO:150、SEQ ID NO:151、SEQ ID NO:152、SEQ ID NO:153、SEQ ID NO:154、SEQ ID NO:155、SEQ ID NO:156、SEQ ID NO:157、SEQ ID NO:158、SEQ ID NO:159、SEQID NO:160、SEQ ID NO:161、SEQ ID NO:162、SEQ ID NO:163、SEQ ID NO:164、SEQ ID NO:165、SEQ ID NO:166、SEQ ID NO:167、SEQ ID NO:168、SEQ ID NO:169、SEQ ID NO:170、SEQID NO:171、SEQ ID NO:172、SEQ ID NO:173、SEQ ID NO:174、SEQ ID NO:175、SEQ ID NO:176、SEQ ID NO:177、SEQ ID NO:178、SEQ ID NO:179、SEQ ID NO:180、SEQ ID NO:181、和/或SEQ ID NO:182。
在本发明的第三方面的其他优选实施方案中,本发明的酮还原酶为选自SEQ IDNO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ IDNO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ IDNO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ IDNO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ IDNO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ IDNO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47、SEQ IDNO:48、SEQ ID NO:49、SEQ ID NO:50、SEQ ID NO:51、SEQ ID NO:52、SEQ ID NO:53、SEQ IDNO:54、SEQ ID NO:55、SEQ ID NO:56、SEQ ID NO:57、SEQ ID NO:58、SEQ ID NO:59、SEQ IDNO:60、SEQ ID NO:61、SEQ ID NO:62、SEQ ID NO:63、SEQ ID NO:64、SEQ ID NO:65、SEQ IDNO:66、SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69、SEQ ID NO:70、SEQ ID NO:71、SEQ IDNO:72、SEQ ID NO:73、SEQ ID NO:74、SEQ ID NO:75、SEQ ID NO:76、SEQ ID NO:77、SEQ IDNO:78、SEQ ID NO:79、SEQ ID NO:80、SEQ ID NO:81、SEQ ID NO:82、SEQ ID NO:83、SEQ IDNO:84、SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87、SEQ ID NO:88、SEQ ID NO:89、SEQ IDNO:90、SEQ ID NO:91、SEQ ID NO:92、SEQ ID NO:93、SEQ ID NO:94、SEQ ID NO:95、SEQ IDNO:96、SEQ ID NO:97、SEQ ID NO:98、SEQ ID NO:99、SEQ ID NO:100、SEQ ID NO:102、SEQID NO:103、SEQ ID NO:104、SEQ ID NO:105、SEQ ID NO:106、SEQ ID NO:107、SEQ ID NO:108、SEQ ID NO:109、SEQ ID NO:110、SEQ ID NO:111、SEQ ID NO:112、SEQ ID NO:114、SEQID NO:115、SEQ ID NO:116、SEQ ID NO:117、SEQ ID NO:118、SEQ ID NO:119、SEQ ID NO:120、SEQ ID NO:121、SEQ ID NO:122、SEQ ID NO:123、SEQ ID NO:124、SEQ ID NO:125、SEQID NO:126、SEQ ID NO:127、SEQ ID NO:128、SEQ ID NO:129、SEQ ID NO:130、SEQ ID NO:131、SEQ ID NO:132、SEQ ID NO:133、SEQ ID NO:134、SEQ ID NO:135、SEQ ID NO:136、SEQID NO:137、SEQ ID NO:138、SEQ ID NO:139、SEQ ID NO:140、SEQ ID NO:141、SEQ ID NO:142、SEQ ID NO:143、SEQ ID NO:144、SEQ ID NO:145、SEQ ID NO:146、SEQ ID NO:147、SEQID NO:148、SEQ ID NO:150、SEQ ID NO:151、SEQ ID NO:152、SEQ ID NO:153、SEQ ID NO:154、SEQ ID NO:155、SEQ ID NO:156、SEQ ID NO:157、SEQ ID NO:158、SEQ ID NO:159、SEQID NO:160、SEQ ID NO:161、SEQ ID NO:162、SEQ ID NO:163、SEQ ID NO:164、SEQ ID NO:165、SEQ ID NO:166、SEQ ID NO:167、SEQ ID NO:168、SEQ ID NO:169、SEQ ID NO:170、SEQID NO:171、SEQ ID NO:172、SEQ ID NO:173、SEQ ID NO:174、SEQ ID NO:175、SEQ ID NO:176、SEQ ID NO:177、SEQ ID NO:178、SEQ ID NO:179、SEQ ID NO:180、SEQ ID NO:181、和/或SEQ ID NO:182中的一种或多种酮还原酶的至少72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、优选至少92.1%、至少92.2%、至少92.3%、至少92.4%、至少92.5%、至少92.6%、至少92.7%、至少92.8%、至少92.9%、至少93%、至少93.1%、至少93.2%、至少93.3%、至少93.4%、至少93.5%、至少93.6%、至少93.7%、至少93.8%、至少93.9%、更优选至少94%、至少94.1%、至少94.2%、至少94.3%、至少94.4%、至少94.5%、至少94.6%、至少94.7%、至少94.8%、至少94.9%、仍然更优选至少95%、至少95.1%、至少95.2%、至少95.3%、至少95.4%、至少95.5%、至少95.6%、至少95.7%、至少95.8%、至少95.9%、更优选至少96%、至少96.1%、至少96.2%、至少96.3%、至少96.4%、至少96.5%、至少96.6%、至少96.7%、至少96.8%、至少96.9%、甚至更优选至少97%、至少97.1%、至少97.2%、至少97.3%、至少97.4%、至少97.5%、至少97.6%、至少97.7%、至少97.8%、至少97.9%、最优选至少98%、至少98.1%、至少98.2%、至少98.3%、至少98.4%、至少98.5%、至少98.6%、至少98.7%、至少98.8%、至少98.9%、尤其是至少99%、至少99.1%、至少99.2%、至少99.3%、至少99.4%、至少99.5%、至少99.6%、至少99.7%、至少99.8%、或至少99.9%。
为了本发明的目的,关于两个氨基酸序列的术语“区别在于至少一个残基改变”或“区别在于至少一个氨基酸残基改变”是指该两个序列在1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个或更多个氨基酸残基上有所不同。
为了本发明的第三方面的目的,在醇底物向酮或醛产物的氧化中的改善的活性,特别是比活性,是指与SEQ ID NO:1的酮还原酶相比,某种酮还原酶变体的以U/ml溶解产物给出的体积活性(volumetric activity)的活性提高率,或以U/mg冻干的溶解产物或U/mg纯化或部分纯化的蛋白质给出的活性的活性提高率。
可以用本领域已知的任何醇底物,并且优选地通过使用底物乙酰乙酸乙酯(EAA)或环己醇,来检测不同于本发明的SEQ ID NO:1的酮还原酶的酮还原酶变体的活性的提高。与本发明的SEQ ID NO:1的酮还原酶不同的改进的酮还原酶变体的活性与SEQ ID NO:1相比增加,与SEQ ID NO:1相比,活性为
-优选至少101%至2000%、至少101%至1900%、至少101%至1800%、至少101%至1700%、至少101%至1600%、至少101%至1500%、至少102%至1500%、至少103%至1500%、至少104%至1500%、至少105%至1500%、至少106%至1500%、至少107%至1500%、至少108%至1500%、至少109%至1500%、至少110%至1500%、至少111%至1500%、至少112%至1500%、至少113%至1500%、至少114%至1500%、至少115%至1500%、至少116%至1500%、至少117%至1500%、至少118%至1500%、至少119%至1500%、至少120%至1500%、至少121%至1500%、至少122%至1500%、至少123%至1500%、至少124%至1500%、至少125%至1500%、至少126%至1500%、至少127%至1500%、至少128%至1500%、至少129%至1500%、至少130%至1500%、至少131%至1500%、至少132%至1500%、至少133%至1500%、至少134%至1500%、至少135%至1500%、至少136%至1500%、至少137%至1500%、至少138%至1500%、至少139%至1500%、至少140%至1500%、至少141%至1500%、至少142%至1500%、至少143%至1500%、至少144%至1500%、至少145%至1500%、至少146%至1500%、至少147%至1500%、至少148%至1500%、至少149%至1500%、至少150%至1500%、至少151%至1500%、至少152%至1500%、至少153%至1500%、至少154%至1500%、至少155%至1500%、至少156%至1500%、至少157%至1500%、至少158%至1500%、至少159%至1500%、至少160%至1500%、至少161%至1500%、至少162%至1500%、至少163%至1500%、至少164%至1500%、至少165%至1500%、至少166%至1500%、至少167%至1500%、至少168%至1500%、至少169%至1500%、至少170%至1500%、至少171%至1500%、至少172%至1500%、至少173%至1500%、至少174%至1500%、至少175%至1500%、至少176%至1500%、至少177%至1500%、至少178%至1500%、至少179%至1500%、至少180%至1500%、至少181%至1500%、至少182%至1500%、至少183%至1500%、至少184%至1500%、至少185%至1500%、至少186%至1500%、至少187%至1500%、至少188%至1500%、至少189%至1500%、至少190%至1500%、至少191%至1500%、至少192%至1500%、至少193%至1500%、至少194%至1500%、至少195%至1500%、至少196%至1500%、至少197%至1500%、至少198%至1500%、至少199%至1500%、至少200%至1500%、至少101%至1200%、至少102%至1200%、至少103%至1200%、优选至少104%至1200%、至少105%至1200%、至少106%至1200%、至少107%至1200%、至少108%至1200%、至少109%至1200%、至少110%至1200%、至少111%至1200%、至少112%至1200%、至少113%至1200%、至少114%至1200%、至少115%至1200%、至少116%至1200%、至少117%至1200%、至少118%至1200%、至少119%至1200%、至少120%至1200%、至少121%至1200%、至少122%至1200%、至少123%至1200%、至少124%至1200%、至少125%至1200%、至少126%至1200%、至少127%至1200%、至少128%至1200%、至少129%至1200%、至少130%至1200%、至少131%至1200%、至少132%至1200%、至少133%至1200%、至少134%至1200%、至少135%至1200%、至少136%至1200%、至少137%至1200%、至少138%至1200%、至少139%至1200%、至少140%至1200%、至少141%至1200%、至少142%至1200%、至少143%至1200%、至少144%至1200%、至少145%至1200%、至少146%至1200%、至少147%至1200%、至少148%至1200%、至少149%至1200%、至少150%至1200%、至少151%至1200%、至少152%至1200%、至少153%至1200%、至少154%至1200%、至少155%至1200%、至少156%至1200%、至少157%至1200%、至少158%至1200%、至少159%至1200%、至少160%至1200%、至少161%至1200%、至少162%至1200%、至少163%至1200%、至少164%至1200%、至少165%至1200%、至少166%至1200%、至少167%至1200%、至少168%至1200%、至少169%至1200%、至少170%至1200%、至少171%至1200%、至少172%至1200%、至少173%至1200%、至少174%至1200%、至少175%至1200%、至少176%至1200%、至少177%至1200%、至少178%至1200%、至少179%至1200%、至少180%至1200%、至少181%至1200%、至少182%至1200%、至少183%至1200%、至少184%至1200%、至少185%至1200%、至少186%至1200%、至少187%至1200%、至少188%至1200%、至少189%至1200%、至少190%至1200%、至少191%至1200%、至少192%至1200%、至少193%至1200%、至少194%至1200%、至少195%至1200%、至少196%至1200%、至少197%至1200%、至少198%至1200%、至少199%至1200%、至少200%至1200%。
可以使用本领域已知的任何酮底物,并且优选地通过使用环己酮来检测与本发明的SEQ ID NO:1的酮还原酶不同的酮还原酶变体的活性的提高。与本发明的SEQ ID NO:1的酮还原酶不同的改进的酮还原酶变体的活性与SEQ ID NO:1相比增加,与SEQ ID NO:1相比,活性为
-至少101%至2000%、至少101%至1900%、至少101%至1800%、至少101%至1700%、至少101%至1600%、至少101%至1500%、至少102%至1500%、至少103%至1500%、至少104%至1500%、至少105%至1500%、至少101%至1000%、至少101%至1000%、至少102%至1000%、至少103%至1000%、至少104%至1000%、至少105%至1000%、至少101%至500%、至少101%至500%、至少102%至500%、至少103%至500%、至少104%至500%、至少105%至500%、以及至少105%至479%。
可以使用本领域已知的任何醇底物,并且优选通过使用底物乙酰乙酸乙酯(EAA)或环己醇中的任一种来检测与发明的SEQ ID NO:1的酮还原酶不同的酮还原酶变体的活性的提高。与SEQ ID NO:1相比,与本发明的SEQ ID NO:1的酮还原酶不同的改进的酮还原酶变体的活性增加,并且以百分比表示,其活性增加了:
-优选至少1%至300%、至少2%至300%、至少3%至300%、至少4%至300%、至少5%至300%、至少6%至300%、至少7%至300%、至少8%至300%、至少9%至300%、至少10%至300%、至少11%至300%、至少12%至300%、至少13%至300%、至少14%至300%、至少15%至300%、至少16%至300%、至少17%至300%、至少18%至300%、至少19%至300%、至少20%至300%、至少21%至300%、至少22%至300%、至少23%至300%、至少24%至300%、至少25%至300%、至少26%至300%、至少27%至300%、至少28%至300%、至少29%至300%、至少30%至300%、至少31%至300%、至少32%至300%、至少33%至300%、至少34%至300%、至少35%至300%、至少36%至300%、至少37%至300%、至少38%至300%、至少39%至300%、至少40%至300%、至少41%至300%、至少42%至300%、至少43%至300%、至少44%至300%、至少45%至300%、至少46%至300%、至少47%至300%、至少48%至300%、至少49%至300%、至少50%至300%、至少51%至300%、至少52%至300%、至少53%至300%、至少54%至300%、至少55%至300%、至少56%至300%、至少57%至300%、至少58%至300%、至少59%至300%、至少60%至300%、至少61%至300%、至少62%至300%、至少63%至300%、至少64%至300%、至少65%至300%、至少66%至300%、至少67%至300%、至少68%至300%、至少69%至300%、至少70%至300%、至少71%至300%、至少72%至300%、至少73%至300%、至少74%至300%、至少75%至300%、至少76%至300%、至少77%至300%、至少78%至300%、至少79%至300%;
-或者优选至少1%至200%、至少2%至200%、至少3%至200%、至少4%至200%、至少5%至200%、至少6%至200%、至少7%至200%、至少8%至200%、至少9%至200%、至少10%至200%、至少11%至200%、至少12%至200%、至少13%至200%、至少14%至200%、至少15%至200%、至少16%至200%、至少17%至200%、至少18%至200%、至少19%至200%、至少20%至200%、至少21%至200%、至少22%至200%、至少23%至200%、至少24%至200%、至少25%至200%、至少26%至200%、至少27%至200%、至少28%至200%、至少29%至200%、至少30%至200%、至少31%至200%、至少32%至200%、至少33%至200%、至少34%至200%、至少35%至200%、至少36%至200%、至少37%至200%、至少38%至200%、至少39%至200%、至少40%至200%、至少41%至200%、至少42%至200%、至少43%至200%、至少44%至200%、至少45%至200%、至少46%至200%、至少47%至200%、至少48%至200%、至少49%至200%、至少50%至200%、至少51%至200%、至少52%至200%、至少53%至200%、至少54%至200%、至少55%至200%、至少56%至200%、至少57%至200%、至少58%至200%、至少59%至200%、至少60%至200%、至少61%至200%、至少62%至200%、至少63%至200%、至少64%至200%、至少65%至200%、至少66%至200%、至少67%至200%、至少68%至200%、至少69%至200%、至少70%至200%、至少71%至200%、至少72%至200%、至少73%至200%、至少74%至200%、至少75%至200%、至少76%至200%、至少77%至200%、至少78%至200%、至少79%至200%;
-或优选至少1%至100%、至少2%至100%、至少3%至100%、至少4%至100%、至少5%至100%、至少6%至100%、至少7%至100%、至少8%至100%、至少9%至100%、至少10%至100%、至少11%至100%、至少12%至100%、至少13%至100%、至少14%至100%、至少15%至100%、至少16%至100%、至少17%至100%、至少18%至100%、至少19%至100%、至少20%至100%、至少21%至100%、至少22%至100%、至少23%至100%、至少24%至100%、至少25%至100%、至少26%至100%、至少27%至100%、至少28%至100%、至少29%至100%、至少30%至100%、至少31%至100%、至少32%至100%、至少33%至100%、至少34%至100%、至少35%至100%、至少36%至100%、至少37%至100%、至少38%至100%、至少39%至100%、至少40%至100%、至少41%至100%、至少42%至100%、至少43%至100%、至少44%至100%、至少45%至100%、至少46%至100%、至少47%至100%、至少48%至100%、至少49%至100%、至少50%至100%、至少51%至100%、至少52%至100%、至少53%至100%、至少54%至100%、至少55%至100%、至少56%至100%、至少57%至100%、至少58%至100%、至少59%至100%、至少60%至100%、至少61%至100%、至少62%至100%、至少63%至100%、至少64%至100%、至少65%至100%、至少66%至100%、至少67%至100%、至少68%至100%、至少69%至100%、至少70%至100%、至少71%至100%、至少72%至100%、至少73%至100%、至少74%至100%、至少75%至100%、至少76%至100%、至少77%至100%、至少78%至100%、至少79%至100%;
-或优选至少1%至70%、至少2%至70%、至少3%至70%、至少4%至70%、至少5%至70%、至少6%至70%、至少7%至70%、至少8%至70%、至少9%至70%、至少10%至70%、至少11%至70%、至少12%至70%、至少13%至70%、至少14%至70%、至少15%至70%、至少16%至70%、至少17%至70%、至少18%至70%、至少19%至70%、至少20%至70%、至少21%至70%、至少22%至70%、至少23%至70%、至少24%至70%、至少25%至70%、至少26%至70%、至少27%至70%、至少28%至70%、至少29%至70%、至少30%至70%、至少31%至70%、至少32%至70%、至少33%至70%、至少34%至70%、至少35%至70%、至少36%至70%、至少37%至70%、至少38%至70%、至少39%至70%、至少40%至70%、至少41%至70%、至少42%至70%、至少43%至70%、至少44%至70%、至少45%至70%、至少46%至70%、至少47%至70%、至少48%至70%、至少49%至70%、至少50%至70%、至少51%至70%、至少52%至70%、至少53%至70%、至少54%至70%、至少55%至70%。
或者,在醇底物向酮或醛产物的氧化中或在酮或醛底物向醇产物的还原中的改善的活性,特别是比活性,可以表示为改进的变体的相对于SEQ ID NO:1的更高的活性值,其中体积活性以U/ml培养物、U/ml溶解产物给出,或比活性以U/mg冻干的溶解产物,或U/mg纯化或部分纯化的蛋白质给出,
-其中与SEQ ID NO:1相比,改进的变体的环己醇至环己酮的氧化活性将提高至少1.1至12倍、至少1.2至12倍、至少1.3至12倍、至少1.4至12倍、至少1.5至12倍、至少1.6至12倍、至少1.7至12倍、至少1.8至12倍、至少1.9至12倍、至少2至12倍、至少2.5至12倍、至少3至12倍、至少3.5至12倍、至少4至12倍、至少4.5至12倍、至少5至12倍、至少5.5至12倍、至少5至10倍;和/或优选将提高至少1.1至15倍、至少1.2至15倍、至少1.3至15倍、至少1.4至15倍、至少1.5至15倍、至少1.6至15倍、至少1.7至15倍、至少1.8至15倍、至少1.9至15倍、至少2至15倍、至少2.5至15倍、至少3至15倍、至少3.5至15倍、至少4至15倍、至少4.5至15倍、至少5至15倍、至少5.5至15倍、至少5至12倍;和/或
-其中与SEQ ID NO:1相比,改进的变体的环己酮至环己醇的还原活性将提高至少20倍至200倍、至少20倍至190倍、至少20倍至180倍、至少20倍至170倍、至少20倍至160倍、至少20倍至150倍、至少22倍至150倍、更优选至少30倍至150倍、至少40倍至150倍、至少40倍至140倍、至少40倍至130倍、至少40倍至120倍、至少40倍至110倍、至少40倍至100倍;和/或优选将提高至少1.1倍至10倍、至少1.1倍至9倍、至少1.1倍至8倍、至少1.1倍至7倍、至少1.5倍至7倍、更优选至少2倍至7倍、至少2倍至6.5倍、至少2倍至6倍、至少2倍至5.5倍、至少2倍至5倍,优选至少1.1倍至4.8倍;和/或
-其中与SEQ ID NO:1相比,改进的变体的环己酮/环己醇的还原活性与氧化活性之比将增加至少20倍至200倍、至少20倍至190倍、至少20倍至180倍、至少20倍至170倍、至少20倍至160倍、至少20倍至150倍、至少30倍至150倍、至少40倍至150倍、至少45倍至150倍、至少50倍至150倍,优选至少70倍至120倍,和/或优选将增加至少1.1至10倍、至少1.1至9倍、至少1.1至8倍、至少1.4至7倍、至少1.6至7倍,优选至少1.4至5倍;或
-其中与SEQ ID NO:1相比,改进的变体的环己酮/环己醇的还原活性与氧化活性之比将降低至少0.9至0.05倍、至少0.9至0.06倍、至少0.9至0.07倍、至少0.9至0.08倍、至少0.9至0.09倍、至少0.8至0.09倍、0.7至0.09倍、0.6至0.09倍、0.8至0.1倍,优选至少0.65至0.13倍。
或者,在醇底物向酮或醛产物的氧化中或在酮或醛底物向醇产物的还原中的改善的活性,特别是比活性,可以表示为改进的变体的相对于SEQ ID NO:1的更高的活性值,其中体积活性以U/ml培养物、U/ml溶解产物给出,或比活性以U/mg冻干的溶解产物,或U/mg纯化或部分纯化的蛋白质给出,
-其中与SEQ ID NO:1相比,改进的变体的环己醇至环己酮的氧化活性比SEQ IDNO:1高至少1.1至15倍、至少1.2至15倍、至少1.3至15倍、至少1.4至15倍、至少1.5至15倍、至少1.6至15倍、至少1.7至15倍、至少1.8至15倍、至少1.9至15倍、至少2至15倍、至少2.5至15倍、至少3至15倍、至少3.5至15倍、至少4至15倍、至少4.5至15倍、至少5至15倍、至少5.5至15倍;和/或优选高至少1.1至12倍、至少1.2至12倍、至少1.3至12倍、至少1.4至12倍、至少1.5至12倍、至少1.6至12倍、至少1.7至12倍、至少1.8至12倍、至少1.9至12倍、至少2至12倍、至少2.5至12倍、至少3至12倍、至少3.5至12倍、至少4至12倍、至少4.5至12倍、至少5至12倍、至少5.5至12倍、至少5至12倍;和/或
-其中与SEQ ID NO:1相比,改进的变体的环己酮至环己醇的还原活性比SEQ IDNO:1高至少1.1倍至10倍、至少1.1倍至9倍、至少1.1倍至8倍、至少1.1倍至7倍、至少1.5倍至7倍,更优选至少2倍至7倍、至少2倍至6.5倍、至少2倍至6倍、至少2倍至5.5倍、至少2倍至5倍,优选至少1.1倍至4.8倍;和/或
-其中与SEQ ID NO:1相比,改进的变体的环己酮/环己醇的还原活性与氧化活性之比比SEQ ID NO:1高至少1.1至10倍、至少1.1至9倍、至少1.1至8倍、至少1.4至7倍、至少1.6至7倍,优选至少1.4至5倍;或
-其中与SEQ ID NO:1相比,改进的变体的环己酮/环己醇的还原活性与氧化活性之比比SEQ ID NO:1低至少0.9至0.05倍、至少0.9至0.06倍、至少0.9至0.07倍、至少0.9至0.08倍、至少0.9至0.09倍、至少0.8至0.09倍、0.7至0.09倍、0.6至0.09倍、0.8至0.1倍,优选至少0.65至0.13倍。
在本发明的一个优选的实施方案中,与SEQ ID NO:1相比显示出改进的环己酮至环己醇的还原活性的SEQ ID NO:1的改进的变体,与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少一个残基改变、至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,并且优选选自:G149M、G149C、G149V、P210C、Y244L,并且最优选选自:G149V和Y244L。
在本发明的一个优选的实施方案中,与SEQ ID NO:1相比显示出改进的环己醇至环己酮的氧化活性的SEQ ID NO:1的改进的变体,与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少一个残基改变、至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,并且优选选自:S97V、S147C、G149M、G149V、G149I、M206D、P210C、Y244L,并且最优选选自:S147C、G149M、P210C。
在本发明的一个优选的实施方案中,与SEQ ID NO:1相比显示出提高的环己酮/环己醇的还原活性与氧化活性之比的SEQ ID NO:1的改进的变体,与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少一个残基改变、至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,并且优选选自:S100R、P121V、M206D和P210C,并且最优选选自:S100R和P210C。
在本发明的一个优选的实施方案中,与SEQ ID NO:1相比显示出降低的环己酮/环己醇的还原活性与氧化活性之比的SEQ ID NO:1的改进的变体,与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少一个残基改变、至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,并且优选选自:S97V、S97C、S147C、G149I、G149M、G149C、G149V、Y244L,并且最优选选自:S147C、G149I和G149M。
为了本发明的第三方面的目的,本发明的酶的改进的热稳定性是这种酶在升高的温度下温育(incubation)给定时间后保留酶活性的特性。由此可以使用任何测定条件来测定酮还原酶的活性。为了本发明的目的,通过测量以下一种或多种特性来确定某些酮还原酶的热稳定性的提高:
Tm50(40)值:为了本发明的目的,Tm50(40)值是在将40mg/ml的溶液温育15分钟后,酮还原酶具有其初始活性的50%的温度。初始活性是未经温度处理即在室温下温育15分钟后相应的酶的活性。酶活性原则上可以通过使用任何活性检测法来测定。在本发明中,使用了如实施例中规定的用于测定氧化活性或还原活性的检测法,这些检测法成为说明书的一部分。
Tm50(10)值:为了本发明的目的,Tm50(10)值是在将对应于OD600为10的粗提物温育15分钟后,酮还原酶具有其初始活性的50%的温度。初始活性是未经温度处理即在室温下温育15分钟后相应的酶的活性。酶活性原则上可以通过使用任何活性检测法来测定。在本发明中,使用了如实施例中规定的用于测定氧化活性或还原活性的检测法,这些检测法成为说明书的一部分。
残余活性:为了本发明的目的,15分钟/42℃后的残余活性定义为:与未在42℃下进行温育的活性相比,在42℃下温育15分钟后的酮还原酶的活性。残余活性的计算方法为:在42℃下温育15分钟后的活性除以未经42℃下温育的活性,然后将该值乘以100,以百分比形式得出残余活性值。酶活性原则上可以通过使用任何活性检测法来测定。为了本发明的目的,使用了实施例中所述的检测法,这些检测法成为说明书的一部分。
相对于SEQ ID NO:1而言,SEQ ID NO:1的改进的变体表现出热稳定性,其表示为在42℃下温育15分钟后的至少20%至100%、至少21%至100%、至少22%至100%、至少23%至100%、至少24%至100%、至少25%至100%、至少26%至100%的残余活性,并且优选表现出下述的热稳定性,该稳定性表示为在42℃下温育15分钟后的至少27%至100%、至少28%至100%、至少29%至100%、至少30%至100%、至少31%至100%、至少32%至100%、至少33%至100%、至少34%至100%、至少35%至100%、至少36%至100%、至少37%至100%、至少38%至100%、至少39%至100%、至少40%至100%、至少41%至100%、至少42%至100%、至少43%至100%、至少44%至100%、至少45%至100%、至少46%至100%、至少47%至100%、至少48%至100%、至少49%至100%、至少50%至100%、至少51%至100%、至少52%至100%、至少53%至100%、至少54%至100%、至少55%至100%、至少56%至100%、至少57%至100%、至少58%至100%、至少59%至100%、60%至100%、至少61%至100%、至少62%至100%、至少63%至100%、至少64%至100%、优选至少40%至100%、至少41%至100%、至少42%至100%、至少43%至100%、至少44%至100%、至少45%至100%、至少46%至100%、至少47%至100%、至少48%至100%、至少49%至100%、至少50%至100%、至少51%至100%、至少52%至100%、至少53%至100%、至少54%至100%、至少55%至100%、至少56%至100%、至少57%至100%、至少58%至100%、至少59%至100%、60%至100%、至少61%至100%、至少62%至100%、至少63%至100%、至少64%至100%、更优选至少50%至100%、至少51%至100%、至少52%至100%、至少53%至100%、至少54%至100%、至少55%至100%、至少56%至100%、至少57%至100%、至少58%至100%、至少59%至100%、60%至100%、至少61%至100%、至少62%至100%、至少63%至100%、至少64%至100%、最优选至少65%至100%、至少66%至100%、至少67%至100%、至少68%至100%、至少69%至100%、至少70%至100%、至少71%至100%、至少72%至100%、至少73%至100%、至少74%至100%、至少75%至100%、至少76%至100%、至少77%至100%、至少78%至100%、至少79%至100%、或80%至100%的残余活性。
相对于SEQ ID NO:1而言,SEQ ID NO:1的改进的变体以Tm50(40)值表示的热稳定性为至少47.0℃至90℃、至少47.0℃至85℃、至少47.0℃至80℃、至少47.0℃至75℃、至少47.0℃至70℃、至少47.5℃至70℃、至少48.0℃至70℃、至少48.5℃至70℃、至少49.0℃至70℃、至少49.5℃至70℃、至少50.0℃至70℃、至少50.5℃至70℃、至少51.0℃至70℃、至少51.5℃至70℃、至少52.0℃至70℃、至少52.5℃至70℃、至少53.0℃至70℃、至少53.5℃至70℃、至少54.0℃至70℃、至少54.5℃至70℃、至少55.0℃至70℃、至少55.5℃至70℃、至少56.0℃至70℃、至少56.5℃至70℃、至少57.0℃至70℃、至少57.5℃至70℃、至少58.0℃至70℃、至少58.0℃至70℃、至少58.0℃至69℃、至少58.0℃至68℃、至少58.0℃至67℃、至少58.0℃至66℃、至少58.0℃至65℃、至少58.0℃至64℃、至少58.0℃至63℃、和至少58.0℃至62℃,并且优选至少47.0℃至90℃、至少47.0℃至87.5℃、至少47.0℃至85℃、至少47.0℃至82.5℃、至少47.0℃至80℃、至少47.5℃至80℃、至少48.0℃至80℃、至少48.5℃至80℃、至少49.0℃至80℃、至少49.5℃至80℃、至少50.0℃至80℃、至少50.5℃至80℃、至少51.0℃至80℃、至少51.5℃至80℃、至少52.0℃至80℃、至少52.5℃至80℃、至少53.0℃至80℃、至少53.5℃至80℃、至少54.0℃至80℃、至少54.5℃至80℃、至少55.0℃至80℃、至少55.5℃至80℃、至少56.0℃至80℃、至少56.5℃至80℃、至少57.0℃至80℃、至少57.5℃至80℃、至少58.0℃至80℃、至少58.0℃至80℃、至少58.0℃至79℃、至少58.0℃至78℃、至少58.0℃至77℃、至少58.0℃至76℃、至少58.0℃至75℃、至少58.0℃至74℃、至少58.0℃至73℃、和至少58.0℃至72℃。
相对于SEQ ID NO:1而言,SEQ ID NO:1的改进的变体以Tm50(10)值表示的热稳定性为至少42.0℃至90℃、至少42.0℃至85℃、至少42.0℃至80℃、至少42.0℃至75℃、至少42.5℃至75℃、至少43.0℃至75℃、至少44℃至75℃、至少44.5℃至75℃、至少45℃至75℃、至少45.5℃至75℃、至少46℃至75℃、至少46.5℃至75℃、至少47℃至75℃、至少47.5℃至75℃、至少48℃至75℃、至少48.5℃至75℃、至少49℃至75℃、至少49.5℃至75℃、至少50℃至75℃、至少50.5℃至75℃、至少51℃至75℃、至少51.5℃至75℃、至少52℃至75℃、至少52.5℃至75℃、至少53℃至75℃、至少53.0℃至74℃、至少53.0℃至73℃、至少53.0℃至72℃、至少53.0℃至71℃、至少53.0℃至70℃、至少53.0℃至69℃、至少53.0℃至68℃、至少53.0℃至67℃、至少53.0℃至66℃和至少53.0℃至66.5℃。
在本发明的一个优选的实施方案中,与SEQ ID NO:1相比显示出改进的热稳定性的SEQ ID NO:1的改进的变体与本发明的SEQ ID NO:1的酮还原酶的区别在于选自以下序列位置的至少一个残基改变、至少两个残基改变、至少三个残基改变、至少四个残基改变或至少六个残基改变:S3I、S4N、S8A、L9G、L9M、L9C、L9R、L9K、L9Q、N10G、A11V、A11C、V13G、E22K、S25A、M26R、A30R、E31N、A38M、S39G、S39A、L42Y、E43K、Q44T、Q44E、Q44R、K49R、A50K、I54R、I54V、V55I、V55C、K56M、Q57P、Q57A、T60C、T60N、T60A、S69V、S69A、D70C、D70S、G75L、S76M、S76C、S76L、S76G、F77L、S92M、S92G、S97A、S97W、S97V、S97C、S97M、S97F、Q98E、F99R、S100E、S100Q、S100R、S100V、S100C、P101F、P101H、P101W、A106C、A106E、A108P、D109G、D109R、N112T、M113V、L114V、L114I、T119L、A120G、P121V、P121A、A127K、A127C、A127G、A127Q、I132V、I132W、I132F、S133L、S133A、S133Q、S133M、S133V、P136V、P136G、Q138A、Q138T、Q138S、T139V、T139F、T139K、A141S、S147C、T148S、G149C、G149V、G149S、G149M、G149I、S151A、K152A、A155W、A155H、M157F、M157R、M157G、M157C、M157E、M157K、M157D、V158M、G159S、G159A、D169E、M172C、P181V、P181W、P181L、K182I、N185R、T194V、A199C、E200P、E200V、E200A、I202V、P204L、M206T、M206D、M206G、M206A、D208H、D208C、D208N、L209C、P210C、N212K、N212P、N212A、G213D、W214G、I215T、V217N、V217P、I220V、T225I、T225M、L227V、N232Y、N232F、G235F、T236S、T237I、V238F、S239F、G243V、Y244F、Y244L、C245F和A246F,并且优选选自:S97V、S97C、F99R、P121V、G149C、G149M、Y244L,并且最优选选自:S97C、G149C和G149M。
说明书的以下部分适用于所有方面,即本发明的第一方面、本发明的第二方面以及本发明的第三方面。
优选地,本发明的酮还原酶能够将酮底物立体选择性和/或区域选择性地还原为仲醇和/或能够将仲醇立体选择性和/或区域选择性地氧化为酮产物。
优选地,本发明的酮还原酶能够将伯醇氧化成醛和/或能够将醛还原成伯醇。
在一些实施方案中,本发明的酮还原酶能够将醛化合物氧化为羧酸。
在一个优选的实施方案中,本发明的酮还原酶能够优选地立体选择性和/或区域选择性地将通式(I)的酮底物还原为仲醇
或优选地区域选择性地将通式(I')的醛底物还原为伯醇
或优选地立体选择性和/或区域选择性地将通式(II)的仲醇氧化为酮
或优选地区域选择性地将通式(II')的伯醇氧化为醛
其中X和Y各自独立地选自
-饱和或不饱和的、未取代或单取代或多取代的、支链或直链的C1-C12脂肪族残基;
-饱和或不饱和的、未取代或单取代或多取代的C3-C12脂环族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
-饱和或不饱和的、未取代或单取代或多取代的C1-C12杂脂环族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
-未取代或单取代或多取代的C6-C10芳族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;或
-未取代或单取代或多取代的C1-C12杂芳族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
或其中X和Y与它们所连接的碳原子一起可以形成饱和或不饱和的、未取代或单取代或多取代的C3-C12脂环族残基,或饱和或不饱和的、未取代或单取代或多取代的C1-C12杂脂环族残基;
其中单取代或多取代是指独立地被选自以下官能团中的一个或多个官能团取代:-F、-Cl、-Br、-C1-12-烷基、-C1-12-烷基-F、-C1-12-烷基-Cl、-C1-12-烷基-Br、-OH、=O、-OC1-12-烷基、-OC6-10-芳基、-O-杂芳基、-OCOC1-12-烷基、-OCOC6-10-芳基、-OCO-杂芳基、-SH、-SC1-12-烷基、-SC6-10-芳基、-S-杂芳基、-S(=O)1-2OH、-NO、-NO2、-N3、-NH2、-NH(C1-12-烷基)、-N(C1-12-烷基)2、-NH(C6-10-芳基)、-N(C6-10-芳基)2、-NH(杂芳基)、-N(杂芳基)2、-CN、-CHO、-CO2H、CO-C1-2-烷基、-CO-C6-10-芳基和-CO-杂芳基;
其中杂脂环族或杂芳族是指具有至少两种不同元素的原子作为其环的成员的环状结构,而杂原子优选选自元素S、O、N和B的原子。
在优选的实施方案中,X和Y与它们连接的碳原子一起形成选自:环丙基、环丁基、环戊基、环己基和环庚基中的环,在每种情况下均可为饱和或不饱和的、未取代或单取代或多取代的。
在另一个优选的实施方案中,X和Y与它们所连接的碳原子一起形成选自:环氧乙烷、氧杂环丁烷、氧戊环(oxolane)、氧杂茂(oxole)、氮杂环丁烷、甲氢氮杂茂(azolidine)、吡咯烷、哌啶、哌嗪和吗啉中的环,在每种情况下均可为饱和的或不饱和的、未取代或单取代或多取代的。优选地,所述环被=O取代,更优选在环氧原子或环氮原子的α位取代,使得各个环可以分别被认为是内酯和内酰胺,任选地进一步被取代,优选被一个或多个羟基取代。优选地,所述环被-OH取代,更优选在环氧原子或环氮原子的α位取代,使得各个环分别可以被认为是缩醛、缩酮和缩醛胺,任选地进一步被取代,优选被一个或多个羟基取代。优选的实例包括吡喃醛糖(aldopyranose)、吡喃酮糖(ketopyranose)和糖醇。
在另一个优选的实施方案中,X和Y与它们所连接的碳原子一起形成杂脂环族环,而一个或多个杂原子优选地选自元素S、O、N和B的原子,更优选选自元素O和N的原子。优选地,至少一个所述杂原子与通式(I)或(I')的羰基直接相邻,使得各个环可以被视为内酯或内酰胺,和/或所述杂原子中的至少一个与通式(II)或(II')的羟基直接相邻,使得各个环可分别被认为是环状半缩醛、半缩酮和相应的缩醛胺。
在一个优选的实施方案中,通式(I)的酮底物为酮糖,其可以以开链形式或环状形式(例如呋喃糖形式或吡喃糖形式)存在。优选的酮糖包括但不限于酮戊糖和酮己糖,尤其是阿洛酮糖、果糖、山梨糖和塔格糖。酮糖也可以为二糖或多糖的一部分。
在优选的实施方案中,通式(I')的醛底物是醛糖,其可以以开链形式或环状形式(例如呋喃糖形式或吡喃糖形式)存在。优选的醛糖包括但不限于戊醛糖和己醛糖,尤其是核糖、阿拉伯糖、木糖、来苏糖、阿洛糖、阿卓糖、葡萄糖、甘露糖、古洛糖、艾杜糖、半乳糖、塔罗糖和前述任何一种的脱氧衍生物,例如鼠李糖。醛糖也可以为二糖或多糖的一部分。
在一个优选的实施方案中,通式(II)的仲醇或通式(II')的伯醇为酮糖,其可以以开链形式或环状形式存在(例如呋喃糖形式或吡喃糖形式);或为醛糖,其可以以开链形式或环状形式存在(例如呋喃糖形式或吡喃糖形式);或为酮糖或醛糖的糖醇,或另外的多元醇(例如甘油)。酮糖也可以为二糖或多糖的一部分。醛糖也可以为二糖或多糖的一部分。
为了说明的目的,饱和或不饱和的、未取代或单取代或多取代的、支链或直链的C1-C12脂肪族残基包括但不限于烷基残基、烯基残基和炔基残基,例如-CH3、-CH2CH3、-CH2CH2CH3、-CH(CH3)2、-CH2CH2CH2CH3、-CH(CH3)CH2CH3、-CH2CH(CH3)2、-C(CH3)3、-CH=CH2、-CH=CHCH=CH2、-C≡CH和-CH=CHC≡CH。
出于说明的目的,饱和或不饱和的、未取代或单取代或多取代的C3-C12脂环族残基包括但不限于C3-12-环烷基,例如环戊基和环己基。
为了说明的目的,饱和或不饱和的、未取代或单取代或多取代的C1-C12杂脂环族残基包括但不限于C3-12-环烷基,其中1或2个环碳原子被选自N、O和S的杂原子取代(C1-12-杂环烷基),例如环氧乙烷、氧杂环丁烷、氧戊环、氧杂茂、氮杂环丁烷、甲氢氮杂茂、吡咯烷、哌啶、哌嗪和吗啉。
为了说明的目的,未取代或单取代或多取代的C6-C10芳族残基包括但不限于苯基和萘基。
为了说明的目的,未取代或单取代或多取代的C1-C12杂芳香族残基包括但不限于单环环系、二环环系和三环环系。单环杂芳基的实例包括但不限于氮杂环丁烷基、氮杂环庚烷基、吖丙啶基、二吖庚因基、1,3-二氧戊环基、二氧六环基、二噻烷基、呋喃基、咪唑基、咪唑啉基、咪唑烷基、异噻唑基、异噻唑啉基、异噻唑烷基、异噁唑基、异噁唑啉基、异噁唑烷基、吗啉基、噁二唑基、噁二唑啉基、噁二唑烷基、噁唑基、噁唑啉基、噁唑烷基、哌嗪基、哌啶基、吡喃基、吡嗪基、吡唑基、吡唑啉基、吡唑烷基、吡啶基、嘧啶基、哒嗪基、吡咯基、吡咯啉基、吡咯烷基、四氢呋喃基、四氢噻吩基、四嗪基、四唑基、噻二唑基、噻二唑啉基、噻二唑烷基、噻唑基、噻唑啉基、噻唑烷基、噻吩基、硫代吗啉基、1,1-二氧代硫代吗啉基(硫代吗啉砜)、噻喃基、三嗪基、三唑基和三噻烷基。二环杂芳基的实例包括但不限于苯并咪唑基、苯并噻二唑基、苯并噻唑基,苯并噻吩基、苯并三唑基、苯并噁唑基、苯并呋喃基、苯并吡喃基、苯并噻喃基、噌啉基、吲唑基、吲哚基、2,3-二氢吲哚基、吲嗪基、萘啶基、异苯并呋喃基、异苯并噻吩基、异吲哚基、异喹啉基、酞嗪基、4H-吡啶并(1,2-a)嘧啶-4-酮、吡喃并吡啶基、喹啉基、喹嗪基、喹喔啉基、喹唑啉基、四氢异喹啉基、四氢喹啉基和噻喃并吡啶基。三环杂芳基的实例包括但不限于吖啶基、咔唑基、咔啉基、二苯并(b,d)呋喃基、二苯并(b,d)噻吩基、萘并(2,3-b)呋喃、萘并(2,3-b)噻吩基、吩嗪基、吩噻嗪基、吩噁嗪基、噻蒽基、噻吨基和呫吨基。
为了说明的目的,单取代或多取代,优选地独立地是指核心结构(core)的氢被选自下组中的一种或多种官能团取代:-卤素(优选为-F、-Cl、-Br、-I)、-OH、=O、-OC1-12-烷基、-OC6-10-芳基、-O-杂芳基、-OCOC1-12-烷基、-OCOC6-10-芳基、-OCO-杂芳基、-SH、-SC1-12-烷基、-SC6-10-芳基、-S-杂芳基、-S(=O)1-2OH、-NO、-NO2、-N3、-NH2、-NH(C1-12-烷基)、-N(C1-12-烷基)2、-NH(C6-10-芳基)、-N(C6-10-芳基)2、-NH(杂芳基)、-N(杂芳基)2、-CN、-CHO、-CO2H、CO-C1-2-烷基、-CO-C6-10-芳基和-CO-杂芳基。
为了说明的目的,包含单糖、双糖或寡糖的糖残基或包含单糖、双糖或寡糖的脱氧糖残基是指通式(I)的酮底物或通式(I')的醛底物可以为多羟基羰基化合物,任选地通过缩醛键和/或缩酮键连接至其他多羟基羰基化合物。例如,当X为被-OH单取代的C1烷基和Y为被-OH多取代的C2烷基时,其中每个碳原子具有单一-OH取代基,通式(I)的酮底物为包含两种对映异构体的丁酮糖,即D-赤藓酮糖以及L-赤藓酮糖。类似地,通式的(I)酮底物可以为戊酮糖或己酮糖,其反过来可以连接至其他糖残基从而形成双糖或寡糖。相应地,当X为被-OH多取代的C2烷基时,其中每个碳原子具有单一-OH取代基,通式(I')的醛底物为包含两种对映异构体的丙醛糖,即D-甘油醛以及L-甘油醛。类似地,通式(I')的醛底物可以为丁醛糖、戊醛糖或己醛糖,其反过来可以连接至其他糖残基从而形成双糖或寡糖。
为了说明的目的,能够优选地立体选择性地和/或区域选择性地还原酮底物是指在合适的条件下(优选在pH 7.0和37℃的水中)在存在合适的辅因子的情况下,酮还原酶对至少一种酮底物表现出至少一些活性,从而产生仲醇。
为了说明的目的,能够优选地立体选择性地氧化醇底物是指在合适的条件下(优选在pH 7.0和37℃的水中)存在合适的辅因子的情况下,酮还原酶对至少一种伯醇底物或仲醇底物表现出至少一些活性,从而产生醛或酮。
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地将通式(III)的苯乙酮-衍生物还原为相应的仲醇
其中
R12、R13、R14、R15和R16各自独立地选自下组:-H;-卤素(优选为氯、溴和碘);未取代或单取代或多取代的-C1-12-烷基;未取代或单取代或多取代的-C6-10-芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;未取代或单取代或多取代的-杂芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;以及-OR18,其中R18为-H,未取代或单取代或多取代的-C1-12-烷基,或者未取代或单取代或多取代的-C6-10-芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;
优选地,R12、R13、R14、R15和R16独立地为-H、-卤素或OR18;更优选为-H、-Cl或OCH3;
R17为-H;-卤素(优选为氯、溴和碘);未取代或单取代或多取代的-C1-12-烷基;未取代或单取代或多取代的-C6-10-芳基,任选地通过饱和或不饱和、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;未取代或单取代或多取代的-杂芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;-OR19,-NH2,-NHR19或-NR19R20,其中R19和R20各自独立地选自未取代或单取代或多取代的-C1-12-烷基;未取代或单取代或多取代的-C6-10-芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;或者未取代或单取代或多取代的-杂芳基,任选地通过饱和或不饱和的、未取代或单取代或多取代的脂肪族C1-12-烃残基桥连;
优选地,R17为-H;
其中该类型的特别优选的酮底物为1-(4-氯苯基)乙酮(1-(4-chlorophenyl)ethanone,4-CAP),
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地催化逆反应,即,将相应的仲醇氧化为根据通式(III)的苯乙酮衍生物。
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性和/或区域选择性地将通式(IV)的开环二酮(secodione)-衍生物还原为相应的仲醇
其中
R23和R24各自独立地选自-H和-C1-12-烷基;且
优选地,R23为-CH2CH3,R24为-C1-12-烷基;更优选地,R23为-CH2CH3,R24为-CH3;
其中该类型的特别优选的酮底物为乙基开环二酮(乙基-3-甲氧基-8,14-开环-甾烷-1,3,5(10),9(11)-四烯-14,17-二酮)
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地催化逆反应,即,将相应的仲醇氧化为根据通式(IV)的开环二酮-衍生物。
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地将乙基-4-氯-3-氧代-丁酸酯(COBE)还原为相应的仲醇。在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地催化逆反应,即将相应的仲醇氧化为乙基-4-氯-3-氧代-丁酸酯(COBE)。
在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性和/或区域选择性地将乙基-3-氧代-3-苯基-丙酸酯还原为相应的仲醇。在本发明的一些实施方案中,本发明的酮还原酶能够优选地立体选择性地和/或区域选择性地催化逆反应,即将相应的仲醇氧化为乙基-3-氧代-3-苯基-丙酸酯。
在本发明的一些实施方案中,本发明的酮还原酶能够优选区域选择性地将选自脂肪醇的酮类(例如2-丁醛(与异丁醛同义)或优选地2-庚醛)的醛底物还原为相应的伯醇。在本发明的一些实施方案中,本发明的酮还原酶能够优选地区域选择性催化逆反应,即,将相应的伯醇氧化为脂肪醇(例如2-丁醛或2-庚醛)的酮。
在一些实施方案中,本发明的酮还原酶能够将醛化合物氧化为羧酸。
在一些实施方案中,本发明的酮还原酶能够将醛底物氧化为羧酸,其中醛底物优选选自脂肪族醛底物、芳香族醛底物和杂芳族醛底物。
令人惊讶地发现,本发明的酮还原酶表现出
-关于氧化和还原二者的高活性;和/或
-在不同的pH值下的高氧化活性(“pH相关活性”);和/或
-广泛的底物耐受性;和/或
-高的辅因子偏好性;和/或
-高立体选择性;和/或
-高立体特异性;和/或
-高温下的高残余活性;和/或
-高区域选择性。
在本发明的酮还原酶的优选实施方案中,
(i)酮还原酶的比活性高于比较氨基酸序列SEQ ID NO:1和/或SEQ ID NO:3的比活性;和/或
(ii)酮还原酶的热稳定性高于比较氨基酸序列SEQ ID NO:1和/或SEQ ID NO:3的热稳定性;和/或
(iii)酮还原酶的立体选择性高于比较氨基酸序列SEQ ID NO:1和/或SEQ ID NO:3的立体选择性;和/或
(iv)酮还原酶的在pH 7对pH 9的比活性方面的pH相关活性比高于比较氨基酸序列SEQ ID NO:1和/或SEQ ID NO:3的立体选择性;和/或
(v)通过酮还原酶将伯醇转化为醛和/或仲醇转化为酮的氧化相关活性高于SEQID NO:1和/或SEQ ID NO:3的氨基酸序列的相应的氧化相关活性。
在本发明的酮还原酶的其他优选实施方案中,SEQ ID NO:1的酮还原酶变体
(i)表现出比对比序列SEQ ID NO:1的酮还原酶更高的热稳定性,其表示为Tm50(40)-值;和/或
(ii)表现出比对比序列SEQ ID NO:1的酮还原酶更高的热稳定性,其表示为Tm50(10)值;和/或
(iii)表现出在42℃下保持15分钟后相对于比较序列SEQ ID NO:1的改善的热稳定性,其表示为残余活性,和/或
(iv)相对于比较序列SEQ ID NO:1,在醇底物向酮产物或醛产物的氧化中表现出增加的活性,特别是比活性;和/或
(v)相对于比较序列SEQ ID NO:1,在酮底物或醛底物向醇产物的还原中表现出增加的活性,特别是比活性;和/或
(vi)对于将酮或醛底物转化为醇产物或相反反向的转化,表现出氧化活性与还原活性之比增加;和/或
(vi)与比较序列SEQ ID NO:1相比,对于将酮或醛底物转化为醇产物或相反方向的转化,表现出还原活性与氧化活性之比增加;或
(vii)与比较序列SEQ ID NO:1相比,对于将酮或醛底物转化为醇产物或相反方向的转化,表现出还原活性与氧化活性之比降低。
本发明的酮还原酶还可以在任选包含共底物(例如异丙醇)的各种溶剂中表现出高立体特异性和/或高活性。
为了说明的目的,优选地在标准条件下用标准底物测量本发明的酮还原酶的功能特性,并且任选地在相同条件下与其他酮还原酶(例如比较序列SEQ ID NO:3)的各个特性进行比较。优选地,除非另有明确说明,否则所述标准底物选自乙酰乙酸乙酯/3-羟基丁酸乙酯、环己酮/环己醇、丙酮/异丙醇、乙基-4-氯-3-羟基-丁酸酯(COBE)以及1-(4-氯-苯基)乙醇(4-CAP);所述标准条件如实验部分所述。
为了说明的目的,立体选择性是下述的化学反应的特性:在非立体特异性生成新立体中心期间或在预先存在的立体中心的非立体特异性转化期间,单一反应物被转化为立体异构体不均等的混合物。选择性通常源于在产生不同产物的机理途径(mechanisticpathway)中空间效应和电子效应的差异。优选地,在本发明的酮还原酶的催化下,底物向手性产物的转化以至少50%ee、更优选至少75%ee、还更优选至少90%ee、还更优选至少95%ee、甚至更优选至少96%ee、甚至更优选至少97%ee、最优选至少98%ee、特别是至少99%ee的对映体过量提供所需手性产物。高立体选择性描述了酶将仲醇的(R)-和(S)-对映异构体混合物中的一种对映异构体选择性氧化为前手性酮而不会使另一种对映异构体明显转化的能力,或选择性地将前手性酮还原成仅一种对映异构体((R)-或(S))的仲醇产物而不会大量产生另一种对映异构体的能力。立体选择性可以通过使用分析手性色谱法(例如使用具有手性固定相的能够分离不同的对映异构体的合适色谱柱的气相色谱法(GC)或高效液相色谱法(HPLC))分析反应混合物来测定。
为了说明的目的,立体特异性是下述的反应机理特性:其由不同的立体异构体反应物产生不同的立体异构体反应产物,或仅对立体异构体中的一种(或一个子集(subset))起作用。优选地,在本发明的酮还原酶的催化下,手性底物向另一种手性产物的转化以至少50%ee、更优选至少75%ee、还更优选至少90%ee。90%ee、更优选至少95%ee、甚至更优选至少96%ee、甚至更优选至少97%ee、最优选至少98%ee、特别是至少99%ee的非对映体过量提供所需手性产物。
为了说明的目的,区域选择性是从一组相同类型的各种替代功能中特定地转换特定功能而不同时转换其他功能的特性。具有多于一个可转换官能度(functionality)的底物的实例包括但不限于鼠李糖-1,5-内酯、甲羟戊酸内酯、葡糖醇内酯(glucanolactone)、糖类等。因此,出于说明的目的,高区域选择性描述了酶选择性地仅将两个或多个醇基中的一个氧化为相应的羰基产物或选择性地仅将两个或多个羰基中的一个还原为相应的醇产物而不显著氧化或还原其他基团的能力。区域选择性可以使用分析色谱法(例如采用合适的色谱柱和分离方案来分离不同的可能产物的气相色谱法(GC)或高效液相色谱法(HPLC))来测定。通过与分析标准进行比较,可以确定生成的产物。可选择性地,质谱或核磁共振(NMR)谱也可以用于产物鉴定。
为了说明的目的,底物转化为产物的活性是指在某些反应条件下酶将底物转化为产物的转化率,优选初始转化率。优选地,该活性为比活性,其中酶活性与第二个通常是定量参数的参数有关,表明该酶相对于该第二个参数的有效性。通常使用的第二参数可以为(冻干的)酶的重量(每克酶)、纯化的(冻干的)酶的重量(每克纯化的酶)或在酶的生产和纯化中使用的每细胞培养物体积(每培养物体积)。酮还原酶冻干物可例如通过如实施例2(参见下文)所述的破坏细胞并随后冻干粗提取物来获得。
为了说明的目的,酶的pH相关活性描述了在不同pH值下酶的相对活性,优选相对比活性。优选地,酶的pH特性(pHprofile)提供了在各种pH值下的宽的活性范围。不同pH值下的相对活性可以表示为“pH相关活性比”,其可以描述为在pH值(1)和pH值(2)下的活性的pH相关活性比(百分比)。最佳pH值是酶表现出最高比活性的pH值。理想的是,酶在宽的pH范围内表现出高的pH相关活性。具体而言,许多酮还原酶在弱碱性pH值下比在中性或弱酸性pH值下对氧化反应表现出更高的比活性值,在弱酸性/中性pH值下比在弱碱性pH值下对还原反应表现出更高的比活性值。优选地,酮还原酶在弱酸性和中性pH值下对氧化反应表现出高pH相关活性。本发明的“pH相关活性比”分别涉及较高的pH7对pH9的比活性之比(以百分比表示)或较高的pH7.5对pH9的比活性之比(以百分比表示),其中某些酮还原酶在pH 9下对底物的比活性定义为100%值,相同酮还原酶在pH 7或pH 7.5下对相同底物的比活性的百分比分别描述了pH相关的活性比。
为了说明的目的,辅因子偏好性描述了在辅因子NAD+/NADH或NADP+/NADPH的存在下将给定底物转化为产物时酶,特别是酮还原酶的相对活性,优选相对比活性。对于某些应用,不需要/强制性地要求相对于另一辅因子NADH或NADPH的偏好。然而,对于某些反应,需要使用特定的辅因子的偏好。因此,有利的是提供具有高的辅因子偏好性的酮还原酶,其可以为对NAD+/NADH的偏好或对NADP+/NADPH的偏好。给定反应中的辅因子偏好可以表示为辅因子(1)相对于辅因子(2)的比活性的百分比。优选地,本发明的SEQ ID NO:1的酮还原酶显示出37.3的相对于乙酰乙酸乙酯对NADPH/NADP的辅因子偏好比,3.3的对COBE的辅因子偏好比。
优选地,本发明的酮还原酶能够以高比活性来还原或氧化底物,优选以下比活性:0.001-100U/mg,更优选0.01-50U/mg、0.01-25U/mg、0.01-10U/mg、0.01-7.5U/mg、0.01-6U/mg、0.01-5.5U/mg、0.01-5U/mg、0.01-4.5U/mg、0.01-4.4U/mg、0.01-4.3U/mg、0.01-4.2U/mg、0.02-4.2U/mg、0.03-4.2U/mg、0.03-4.1U/mg,甚至更优选0.1-50U/mg、0.1-25U/mg、0.1-10U/mg、0.1-7.5U/mg、0.1-6U/mg、0.1-5.5U/mg、0.1-5U/mg、0.1-4.5U/mg、0.1-4.4U/mg、0.1-4.3U/mg、0.1-4.2U/mg、0.1-4.2U/mg、0.1-4.2U/mg、0.1-4.1U/mg,甚至更优选0.3-50U/mg、0.3-25U/mg、0.3-10U/mg、0.3-7.5U/mg、0.3-6U/mg、0.3-5.5U/mg、0.3-5U/mg、0.3-4.5U/mg、0.3-4.4U/mg、0.3-4.3U/mg、0.3-4.2U/mg、0.3-4.2U/mg、0.3-4.2U/mg、0.3-4.1U/mg,还更优选0.5-50U/mg、0.5-25U/mg、0.5-10U/mg、0.5-7.5U/mg、0.5-6U/mg、0.5-5.5U/mg、0.5-5U/mg、0.5-4.5U/mg、0.5-4.4U/mg、0.5-4.3U/mg、0.5-4.2U/mg、0.5-4.2U/mg、0.5-4.2U/mg、0.5-4.1U/mg酮还原酶的冻干物。在本发明的意义上的底物为通过本发明的酮还原酶而被转化的伯醇或仲醇、醛或酮。
优选地,本发明的酮还原酶能够在NADPH的存在下以高比活性还原或氧化底物,优选以下比活性:0.01-100U/mg、0.01-50U/mg、0.01-25U/mg、0.01-10U/mg、0.01-7.5U/mg、0.01-6U/mg、0.01-5.5U/mg、0.01-5U/mg,更优选0.1-100U/mg、0.1-50U/mg、0.1-25U/mg、0.1-10U/mg、0.1-7.5U/mg、0.1-6U/mg、0.1-5.5U/mg、0.1-5U/mg,更优选1-100U/mg、1-50U/mg、1-25U/mg、1-10U/mg、1-7.5U/mg、1-6U/mg、1-5.5U/mg、1-5U/mg,甚至更优选2-100U/mg、2-50U/mg、2-25U/mg、2-10U/mg、2-7.5U/mg、2-6U/mg、2-5.5U/mg、2-5U/mg,最优选2.4-4.1U/mg。在本发明的意义上的底物为通过本发明的酮还原酶而被转化的伯醇或仲醇、醛或酮。具体地,本发明的酮还原酶能够在NADPH的存在下以4.1U/mg酮还原酶冻干物的比活性还原乙酰乙酸乙酯,在NADPH的存在下以3.2U/mg酮还原酶冻干物的比活性还原环己酮,以及在NADPH的存在下以2.4U/mg酮还原酶冻干物的比活性还原4-CAP。
优选地,本发明的酮还原酶能够以高比活性还原乙酰乙酸乙酯,优选以下比活性:0.01-100U/mg,更优选0.01-50U/mg、0.01-25U/mg、0.01-10U/mg、0.01-7.5U/mg、0.01-6U/mg、0.01-5.5U/mg、0.01-5U/mg,更优选0.1-100U/mg、0.1-50U/mg、0.1-25U/mg、0.1-10U/mg、0.1-7.5U/mg、0.1-6U/mg、0.1-5.5U/mg、0.1-5U/mg,甚至更优选0.2-100U/mg、0.2-50U/mg、0.2-25U/mg、0.2-10U/mg、0.2-7.5U/mg、0.2-6U/mg、0.2-5.5U/mg、0.2-5U/mg,还更优选0.3-100U/mg、0.3-50U/mg、0.3-25U/mg、0.3-10U/mg、0.3-7.5U/mg、0.3-6U/mg、0.3-5.5U/mg、0.3-5U/mg,更优选0.1-4.5U/mg,更优选0.11-4.5U/mg,更优选0.15-4.5U/mg,更优选0.2-4.5U/mg,更优选0.1-4.3U/mg,更优选0.15-4.3U/mg,更优选0.2-4.3U/mg,更优选0.1-4.2U/mg,更优选0.1-4.2U/mg,更优选0.1-4.1U/mg,更优选0.11-4.2U/mg、最优选0.11-4.1U/mg酮还原酶冻干物。具体地,本发明的酮还原酶能够在NADPH的存在下以4.1U/mg的酮还原酶冻干物的比活性以及在NADH的存在下以0.11U/mg的酮还原酶冻干物的比活性还原乙酰乙酸乙酯。
优选地,本发明的酮还原酶能够以高比活性还原4-CAP,优选以下比活性:0.001-100U/mg,更优选0.01-50U/mg,更优选0.01-25U/mg,更优选0.01-10U/mg,更优选0.01-7.5U/mg,更优选0.01-5U/mg,更优选0.01-4U/mg,更优选0.01-3U/mg,更优选0.05-3U/mg,更优选0.1-3U/mg,更优选0.2-2.9U/mg,更优选0.25-2.8U/mg,更优选0.5-2.7U/mg,更优选1-2.6U/mg,更优选1-2.5U/mg,更优选1-2.4U/mg,更优选2-2.4U/mg,最优选2.2-2.4U/mg酮还原酶冻干物。具体地,本发明的酮还原酶能够在NADPH的存在下以2.4U/mg的酮还原酶冻干物的比活性还原4-CAP。
优选地,本发明的酮还原酶能够以高比活性还原环己酮,优选以下比活性:0.01-100U/mg,优选0.1-100U/mg、0.5-100U/mg,更优选0.5-50U/mg,更优选0.5-25U/mg,更优选0.5-10U/mg,更优选0.5-7.5U/mg,更优选0.5-5U/mg,更优选1-5U/mg,更优选1.5-5U/mg,更优选2-5U/mg,更优选2-4U/mg,更优选2-3.9U/mg,更优选2.5-3.8U/mg,更优选3-3.7U/mg,更优选3-3.6U/mg,更优选3-3.5U/mg,更优选3-3.4U/mg,更优选3-3.3U/mg,最优选3.2U/mg酮还原酶冻干物。具体地,本发明的酮还原酶能够在NADPH的存在下以3.2U/mg的酮还原酶冻干物的比活性还原环己酮。
优选地,本发明的酮还原酶能够以高比活性还原COBE,优选以下比活性:0.01-100U/mg,更优选0.06-50U/mg、0.08-50U/mg,更优选0.08-25U/mg,更优选0.8-10U/mg,更优选0.08-5U/mg,更优选0.08-2.5U/mg,更优选0.08-1U/mg,更优选0.08-0.9U/mg,更优选0.08-0.8U/mg,更优选0.08-0.7U/mg,更优选0.08-0.6U/mg,更优选0.08-0.5U/mg,更优选0.08-0.4U/mg,更优选0.09-0.4U/mg,最优选0.09-0.3U/mg酮还原酶冻干物。具体地,本发明的酮还原酶能够在NADPH的存在下以0.3U/mg的酮还原酶冻干物的比活性以及在NADH的存在下以0.09U/mg酮还原酶冻干物的比活性还原COBE。
优选地,本发明的酮还原酶能够以高比活性氧化3-羟基丁酸乙酯,优选以下比活性:0.03-100U/mg,更优选0.05-50U/mg,更优选0.05-25U/mg,更优选0.05-10U/mg,更优选0.06-5U/mg,更优选0.06-2.5U/mg,更优选0.07-1U/mg,更优选0.07-0.9U/mg,更优选0.07-0.8U/mg,更优选0.07-0.7U/mg,更优选0.07-0.6U/mg,更优选0.07-0.5U/mg,更优选0.07-0.4U/mg,更优选0.07-0.3U/mg,更优选0.08-0.25U/mg,最优选0.09-0.22U/mg酮还原酶冻干物。具体而言,本发明的酮还原酶能够在pH 7下以0.07-0.1U/mg,最优选0.09U/mg的酮还原酶冻干物的高比活性,以及在pH 9下以0.07-0.25U/mg,更优选0.1-0.25U/mg、0.15-0.25U/mg、0.2-0.25U/mg,最优选0.22U/mg的酮还原酶冻干物的高比活性氧化3-羟基丁酸乙酯。
优选地,本发明的酮还原酶能够以高的比活性氧化环己醇,优选以下比活性:0.25-100U/mg,更优选0.26-50U/mg、0.26-25U/mg、0.26-10U/mg、0.27-5U/mg、0.28-2.5U/mg、0.29-2.5U/mg、0.29-1U/mg、0.29-0.9U/mg、0.29-0.8U/mg,更优选0.29-0.7U/mg,更优选0.3-0.6U/mg,最优选0.31-0.5U/mg酮还原酶冻干物。具体而言,本发明的酮还原酶能够在pH7.5下以0.3-0.5U/mg,最优选以0.31U/mg的酮还原酶冻干物的高比活性氧化环己醇,并且能够在pH 9下以0.3-1U/mg,更优选0.3-0.8U/mg、0.3-0.7U/mg、0.3-0.6U/mg,最优选以0.5U/mg酮还原酶冻干物的高比活性氧化环己醇。
优选地,本发明的酮还原酶能够在pH 7下以高比活性氧化异丙醇,优选以下比活性:0.02-10U/mg、0.02-5U/mg、0.02-2U/mg、0.02-1U/mg、0.02-0.5U/mg、0.02-0.25U/mg、0.02-0.1U/mg、0.02-0.09U/mg、0.02-0.08U/mg、0.02-0.07U/mg、0.02-0.06U/mg、0.03-0.05U/mg,最优选0.03U/mg酮还原酶冻干物。
优选地,本发明的酮还原酶能够在pH 9下以高的比活性氧化异丙醇,优选以下比活性:0.04-10U/mg、0.04-5U/mg、0.04-2U/mg、0.04-1U/mg、0.04-0.5U/mg、0.04-0.25U/mg、0.04-0.1U/mg、0.04-0.09U/mg、0.04-0.08U/mg、0.04-0.07U/mg、0.04-0.06U/mg,最优选0.05U/mg酮还原酶冻干物。
为了测定给定底物的氧化活性,优选将本发明的酮还原酶在30℃下在含有底物(50-100mM或20%(v/v))和辅因子(例如0.25mM的NAD(P)+)的缓冲液中温育。通过测量由NAD(P)+变化导致的340nm处的吸光度变化来测定氧化活性。
为了测定给定底物的还原活性,优选将本发明的酮还原酶在30℃下在含有底物(50-100mM或20%(v/v))和辅因子(例如0.25mM的NAD(P)H)的缓冲液中温育。通过测量由NAD(P)H变化导致的340nm处的吸光度变化来测定还原活性。
用于酮还原酶反应中辅因子再生的合适的共底物可以根据它们的比活性来选择,优选选自:异丙醇、2-丁醇、2-戊醇、2-己醇和2-庚醇。
用于在酮还原酶中辅因子再生的其他合适方法可以是与本发明的酮还原酶偶联的酶系统。特别地,NAD(P)H-氧化酶(NOX)可用于使用氧气作为共底物将NAD(P)H氧化为NAD(P)+。对于涉及NADPH作为辅因子的方法,NOX系统特别受关注,因为某些NOX系统的特征是相对于NAD+对NADP+的辅因子偏好。
优选地,本发明的酮还原酶在共底物中,优选在异丙醇、2-丁醇、2-戊醇、2-己醇和/或2-庚醇中表现出高稳定性。优选地,于30℃下在50%水性共底物(优选为异丙醇)中预温育酮还原酶6小时之后,酮还原酶相对于其预温育之前的活性表现出至少1%,更优选为至少10%或至少20%,还更优选为至少30%或至少40%,还更优选为至少50%或至少60%,甚至更优选为至少70%或至少80%,最优选为至少85%或至少90%,特别是至少95%或至少99%的残余活性。在这方面,本发明意义中的残余活性描述了与不使用共底物的预温育之后的酶的活性相比,在使用共底物的预温育之后的酮还原酶的剩余活性。为了测定在共底物(包括例如异丙醇)中的稳定性,将本发明的酮还原酶在缓冲液温育(优选于30℃下在pH为7或pH为9,含有50%共底物例如异丙醇)中预温育6小时,将酮还原酶的酶活性与不使用共底物的预温育后的酶进行比较。根据本发明,当两种酶活性相同时,即当与不使用共底物的预温育后的活性相比没有活性损失时,酮还原酶的残余活性为100%。
共底物通过酮还原酶的有效转化以及酮还原酶在共底物中的稳定性与建立用于将酮或醛底物还原为仲醇或伯醇的有效工业方法特别相关。本发明的酮还原酶在共底物中的稳定性及其转化本发明的共底物的能力也与将伯醇和仲醇逆向转化为醛和酮底物相关。
为了说明的目的,高的热稳定性描述了酶在给定的时间段内暴露于指定的高温后抵抗不可逆失活的能力。将在高温下温育一定时间的酶的残余活性计算为相对于尚未在高温下温育的酶样品的酶活性的相对值。酶活性原则上可以通过使用任何活性检测法来测定。为了本发明的目的,通过测量和描述以下一种或多种特性来确定热稳定性:
-Tm50(40)值:为了本发明的目的,Tm50(40)值是在Tris/HCl缓冲液(50mM,pH 7,2mMMgCl2)中将40mg/ml冻干酶溶液温育15分钟后,酮还原酶具有其初始活性的50%的温度。初始活性是未经温度处理即在室温下温育15分钟后相应的酶的活性。酶活性原则上可以通过使用任何活性检测法来测定。在本发明中,使用了如实施例中规定的用于测定氧化活性或还原活性的检测法,这些检测法成为说明书的一部分。
-Tm50(10)值:为了本发明的目的,Tm50(10)值是在对应于OD600为10的粗提物温育15分钟后,酮还原酶具有其初始活性的50%的温度。初始活性是未经温度处理即在室温下温育15分钟后相应的酶的活性。酶活性原则上可以通过使用任何活性检测法来测定。在本发明中,使用了如实施例中规定的用于测定氧化活性或还原活性的检测法,这些检测法成为说明书的一部分。
-残余活性:为了本发明的目的,15分钟/42℃后的残余活性定义为:与未在42℃下进行温育的活性相比,在42℃下温育15分钟后的酮还原酶的活性。残余活性的计算方法为:在42℃下温育15分钟后的活性除以未经42℃下温育的活性,然后将该值乘以100,以百分比形式得出残余活性值。酶活性原则上可以通过使用任何活性检测法来测定。为了本发明的目的,使用了实施例中所述的检测法,这些检测法成为说明书的一部分。
在一些实施方案中,本发明的酮还原酶表现出优异的氧化和还原活性,优选优异的比氧化和还原活性、优异的热稳定性和/或立体选择性,这起因于与比较序列SEQ ID NO:3的序列差异。
为了本说明书的比较目的,将比较序列SEQ ID NO:3称为“比较”,是指超出了WO2013/102619的公开范围,在该文献中其被称为具有识别号10的序列,没有对比较序列SEQID NO:3进行任何序列位置的进一步改变。
本发明的优异的比活性涉及高于比较序列SEQ ID NO:3的酮还原酶的比活性的本发明的酮还原酶的比活性。优选地,本发明的酮还原酶的比活性比比较序列SEQ ID NO:3的酮还原酶的比活性高至少10%、至少50%、至少100%、至少1.4倍、至少1.5倍、至少2倍、至少2.5倍、至少3倍、至少3.5倍、至少4倍、至少4.5倍、至少5倍、至少5.5倍、至少6倍、至少6.5倍、至少7倍、至少7.5倍、至少8倍、至少8.5倍、至少9倍、至少9.5倍、至少10倍、至少14倍、至少20倍、至少46倍、至少50倍、至少100倍、至少200倍、至少300倍、至少500倍、至少700倍、至少250倍、至少10000倍、或至少100000倍。改善的比活性还可以意味着本发明的酮还原酶确实对期望的底物表现出一定的活性,而比较序列SEQ ID NO:3的酮还原酶对所述底物没有明显的活性。
本发明的优异的“pH相关活性比”涉及高于比较序列SEQ ID NO:3的pH相关活性比的本发明的酮还原酶的pH相关活性比。优选地,本发明的酮还原酶的pH相关活性比的值为至少35%、至少36%、至少37%、至少38%、至少39%、至少40%、至少41%、至少42%、至少43%、至少44%、至少45%、至少46%、至少47%、至少48%、至少49%、至少50%、至少51%、至少52%、至少53%、至少54%、至少55%、至少56%、至少57%、至少58%、至少59%、至少60%、至少61%、至少62%、至少63%、至少64%、至少65%、至少66%、至少67%、至少68%、至少69%、至少70%、至少75%、至少80%或至少90%。具体地,本发明的酮还原酶对底物-3-羟基丁酸乙酯的pH相关活性比的值优选为35%-70%,更优选为37%-60%,最优选为41%,本发明的酮还原酶对底物环己醇的pH相关活性比的值优选为35%-80%,更优选为45%-75%,最优选为62%,并且本发明的酮还原酶对底物异丙醇的pH相关活性比的值优选为40%-80%,更优选为50%-75%,最优选为60%。
与比较序列SEQ ID NO:3的酮还原酶相比,本发明的优异的热稳定性涉及在给定温度下一定时间段后,例如在46℃下温育至少15分钟后,本发明的酮还原酶具有更高的残余比活性。或者,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的改进的热稳定性可以涉及本发明的酮还原酶具有相同的残余比活性,但是其是在较高的温度下温育相同的时间,或在相同的温度下温育更长的时间。优选地,本发明的酮还原酶提供的热稳定性对应于至少40℃、至少41℃、至少42℃、至少43℃、至少44℃、至少45℃、至少46℃、至少47℃、至少48℃、至少49℃、至少50℃、至少55℃、至少60℃、至少65℃、至少70℃或至少72℃的Tm50(40)值。根据本发明还优选的是,本发明的酮还原酶表现出对应于下述Tm50(40)值的热稳定性:与比较序列SEQ ID NO:3的Tm50(40)值相比,该Tm50(40)值增加了至少1℃、至少1℃-2℃、至少1℃-3℃、至少1℃-4℃、至少1℃-5℃、至少1℃-6℃、至少1℃-7℃、至少1℃-8℃、至少1℃-9℃、至少1℃-10℃、至少1℃-15℃、至少1℃-20℃、至少1℃-25℃、至少1℃-30℃、至少1℃-35℃、至少1℃-40℃、至少1℃-45℃、至少1℃-50℃、或至少1℃-55℃、优选至少2℃-3℃、至少2℃-4℃、至少2℃-5℃、至少2℃-6℃、至少2℃-7℃、至少2℃-8℃、至少2℃-9℃、至少2℃-10℃、至少2℃-15℃、至少2℃-20℃、至少2℃-25℃、至少2℃-30℃、至少2℃-35℃、至少2℃-40℃、至少2℃-45℃或至少2℃-50℃,更优选至少3℃-4℃、至少3℃-5℃、至少3℃-6℃、至少3℃-7℃、至少3℃-8℃、至少3℃-9℃、至少3℃-10℃、至少3℃-15℃、至少3℃-20℃、至少3℃-25℃、至少3℃-30℃、至少3℃-35℃、至少3℃-40℃、至少3℃-45℃或至少3℃-50℃,甚至更优选4℃-5℃、至少4℃-6℃、至少4℃-7℃、至少4℃-8℃、至少4℃-9℃、至少4℃-10℃、至少4℃-15℃、至少4℃-20℃、至少4℃-25℃、至少4℃-30℃、至少4℃-35℃、至少4℃-40℃、至少4℃-45℃,甚至更优选5℃-6℃、至少5℃-7℃、至少5℃-8℃、至少5℃-9℃或至少5℃-10℃、至少5℃-15℃、至少5℃-20℃、至少5℃-25℃、至少5℃-30℃、至少5℃-35℃、至少5℃-40℃,更优选6℃-7℃、至少6℃-8℃、至少6℃-9℃或至少6℃-10℃、至少6℃-15℃、至少6℃-20℃、至少6℃-25℃、至少6℃-30℃、至少6℃-35℃,最优选30℃。
在某些实施方案中,本发明的酮还原酶表现出与下述Tm50(40)值相对应的热稳定性:与比较序列SEQ ID NO:3的Tm50(40)值相比,该Tm50(40)值增加了至少1℃、至少2℃、至少3℃、至少4℃、至少5℃、至少6℃、至少7℃、至少8℃、至少9℃、至少10℃、至少15℃、至少20℃、至少25℃或至少30℃。
本发明的优异的立体选择性涉及通过本发明的酮还原酶提供的产物的对映体过量高于通过比较序列SEQ ID NO:3的酮还原酶提供的产物的对映体过量。优选地,通过本发明的SEQ ID NO:1的本发明的酮还原酶提供的对映体过量为至少20%ee、至少30%ee、至少40%ee、至少50%ee、至少60%ee、至少70%ee、至少80%ee、至少90%ee、至少91%ee、至少92%ee、至少93%ee、至少94%ee、至少95%ee、至少96%ee、至少97%ee、至少98%ee或至少99%ee。根据本发明还优选的是,与比较序列SEQ ID NO:3相比,本发明的SEQ ID NO:1的酮还原酶提供的对映体过量增加至少0.1%ee、至少0.5%ee、至少1%ee、至少1.5%ee、至少2%ee、至少2.5%ee、至少3%ee、至少3.5%ee、至少4%ee或至少4.5%ee。
优选在标准反应条件下,典型地于30℃下在含有底物、辅因子、选择性增补物和酮还原酶的缓冲水溶液中,测定与比较序列SEQ ID NO 3的酮还原酶相比的本发明的酮还原酶的优异的比活性。缓冲液优选地选自含有2mM MgCl2、pH为7-9的10-200mM Tris/HCl,pH为6-8的10-200mM磷酸钠/NaOH,或pH为7-9的10-200mM三乙醇胺/HCl。优选加入用于还原反应的辅因子NAD(P)H或用于氧化反应的辅因子NAD(P)至最终浓度范围为0.05-10mM。可选的其他增补物优选地选自1-5%TritonTMX-100(v/v)和0.5-10%DMSO(v/v)。酮还原酶优选地作为粗提取物上清液加入。通过破碎含有酮还原酶的表达宿主,随后离心以分离细胞碎片和含有酮还原酶的粗提取物上清液来获得粗提取物。优选地通过测量产物形成、还原型辅因子NADPH或NADH(优选NADPH)的消耗和/或底物消耗来测定比活性。在通过测量产物形成或底物消耗来测定比活性的情况中,辅因子再生体系(异丙醇、GDH/葡萄糖或NOX)可应用于标准反应中;在通过测量还原型辅因子NADPH或NADH(优选NADPH)的消耗来测定比活性的情况中,不应用辅因子再生体系。在应用辅因子再生体系的情况中,还原型辅因子NADPH或NADH(优选NADPH)可用被辅因子再生体系还原的氧化型辅因子NAD+或NADP+(优选NADP+)替代。
优选地通过在PCR循环仪中于给定温度下(优选地在比较序列SEQ ID NO:3的酮还原酶表现出10%的残余活性的温度下)温育含有酮还原酶的粗提取物15分钟来测定与比较序列SEQ ID NO:3的酮还原酶相比的本发明的酮还原酶的优异的热稳定性。然后将粗提取物在冰上温育30分钟。通过离心分离不溶蛋白质,在标准酮还原酶试验中分析上清液的酮还原酶的剩余活性。在该标准测定中,适用于酮还原酶的底物通过酮还原酶转化,并伴随NADPH氧化为NADP+。通过在标准光度计中测量340nm处的吸收来监测NADPH的增加。在标准反应条件下即典型地于30℃下在含有底物(例如乙酰乙酸乙酯)、辅因子和酮还原酶的缓冲水溶液中进行试验。酮还原酶优选地作为粗提取物上清液加入。通过破碎含有酮还原酶的表达宿主,随后离心以分离细胞碎片和含有酮还原酶的粗提取物上清液来获得粗提取物。缓冲液优选地选自pH为7-9、含有2mM MgCl2的10-200mM Tris/HCl,pH为6-8的10-200mM磷酸钠/NaOH,或pH为7-9的10-200mM三乙醇胺/HCl。优选地加入辅因子NADP+至最终浓度范围为0.05-10mM。优选地酮还原酶作为粗提取物上清液加入。通过破碎含有酮还原酶的表达宿主,随后离心以分离细胞碎片和含有酮还原酶的粗提取物上清液来获得粗提取物。
与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶的优异的立体选择性优选在如上所述的用于测定改善的比活性的标准反应条件下测定。手性分析用于分析反应中形成的产物。
优选地,本发明的酮还原酶能够还原任何醛底物或能够立体选择性和/或区域选择性地还原任何酮底物。
优选地,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶除了改善的比活性、热稳定性和/或立体选择性外,还表现出高的共底物活性和/或共底物稳定性,特别是对于异丙醇、2-丁醇、2-戊醇、2-己醇或2-庚醇的高稳定性和/或活性。
优选地,本发明的酮还原酶表现出的热稳定性高于比较序列SEQ ID NO:3的酮还原酶的热稳定性。优选地,本发明的酮还原酶表现出提高的Tm50(40)-值,其与比较序列SEQID NO:3的酮还原酶相比提高1℃-40℃,优选地1℃-30℃,更优选地1℃-20℃,甚至更优选3℃-15℃,最优选7℃,更优选表现出提高的Tm50(40)-值,其与比较序列SEQ ID NO:3的酮还原酶相比提高1℃-60℃,优选1℃-50℃,更优选1℃-40℃,甚至更优选3℃-35℃,最优选30℃。
酮还原酶优选作为粗提取物上清液添加。通过破碎包含酮还原酶的表达宿主并随后离心以分离细胞碎片和含有酮提取酶的粗提物的上清液来获得粗提取物。
本发明的另一方面涉及酮底物向仲醇的立体选择性还原或手性醇的一种对映异构体向前手性酮的立体选择性氧化,包括使酮底物与适合的辅因子在本发明的酮还原酶的存在下反应的步骤。优选地,用于还原反应的所述适合的辅因子为NADH或NADPH或用于氧化反应的所述适合的辅因子为NAD+或NADP+。如上限定的关于本发明酮还原酶的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
本发明的另一方面涉及将醛还原为伯醇的方法,包括使醛与适合的辅因子在本发明酮还原酶的存在下反应。优选地,所述适合的辅因子为NADH或NADPH。如上限定的关于本发明酮还原酶的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
优选地,该方法包括通过相应的酮还原酶以高比活性伴随转化用于辅因子再生的共底物,优选地根据其比活性从下组选择这样的共底物:异丙醇、2-丁醇、2-戊醇、2-己醇和2-庚醇,最优选地以高比活性将异丙醇转化为丙酮。在本发明方法的优选实施方案中,通过使用NADPH:NADH氧化酶(NOX),例如来自桑树杆菌(Lactobacillus sanfranciscensis)(Lountos et al.,Acta Cryst.(2004),D60,2044-2047),以及适合的共底物(例如异丙醇)再生氧化型辅因子NAD(P)+。
本发明的另一方面涉及将仲醇立体选择性地氧化为酮产物的方法,包括使仲醇与适合的辅因子在本发明的酮还原酶的存在下反应的步骤。优选地,所述适合的辅因子为NAD+或NADP+。如上限定的关于本发明酮还原酶的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
本发明的另一方面涉及将伯醇氧化为醛的方法,包括使伯醇与适合的辅因子在本发明的酮还原酶的存在下反应的步骤。优选地,所述适合的辅因子为NAD+或NADP+。如上限定的关于本发明酮还原酶的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
优选地,该方法包括通过相应的酮还原酶以高比活性伴随转化用于辅因子再生的共底物,优选地根据其比活性从下组选择这样的共底物:异丙醇、2-丁醇、2-戊醇、2-己醇和2-庚醇,最优选地以高比活性将异丙醇转化为丙酮。在本发明方法的优选实施方案中,通过使用NADPH:NADH氧化酶(NOX),例如来自桑树杆菌(Lountos et al.,Acta Cryst.(2004),D60,2044-2047),以及适合的共底物(例如丙酮)再生还原型辅因子NAD(P)H。
优选地,在用于将酮底物立体选择性地还原为仲醇和用于将醛底物还原为伯醇的本发明的方法中,醛底物和酮底物分别可以为任意醛底物或任意酮底物,优选地为通式(I')的醛底物或通式(I)的酮底物。
如上限定的关于本发明的酮底物和醛底物的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
本发明的另一方面涉及将醛底物氧化为羧酸的方法,包括使醛底物与适合的辅因子在本发明的酮还原酶的存在下反应的步骤。优选地,所述适合的辅因子为NAD+或NADP+。如上限定的关于本发明酮还原酶的所有优选的实施方案也适用于本发明的方法,因此在下文中不再重复。
优选地,醛底物具有通式(V)
其中Z选自
-饱和或不饱和的、未取代或单取代或多取代的、支链或直链的C1-C12脂肪族残基;
-饱和或不饱和的、未取代或单取代或多取代的C3-C12脂环族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
-饱和或不饱和的、未取代或单取代或多取代的C1-C12杂脂环族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
-未取代或单取代或多取代的C6-C10芳族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;或
-未取代或单取代或多取代的C1-C12杂芳族残基,任选地通过饱和或不饱和的、未取代或单取代或多取代的C1-C12脂肪族残基桥接至CO-部分;
其中单取代或多取代是指独立地被选自:-F、-Cl、-Br、-C1-12-烷基、-C1-12-烷基-F、-C1-12-烷基-Cl、-C1-12-烷基-Br、-OH、=O、-OC1-12-烷基、-OC6-10-芳基、-O-杂芳基、-OCOC1-12-烷基、-OCOC6-10-芳基、-OCO-杂芳基、-SH、-SC1-12-烷基、-SC6-10-芳基、-S-杂芳基、-S(=O)1-2OH、-NO、-NO2、-N3、-NH2、-NH(C1-12-烷基)、-N(C1-12-烷基)2、-NH(C6-10-芳基)、-N(C6-10-芳基)2、-NH(杂芳基)、-N(杂芳基)2、-CN、-CHO、-CO2H、CO-C1-2-烷基、-CO-C6-10-芳基和-CO-杂芳基中的一个或多个官能团取代;
其中杂脂环族或杂芳族是指具有至少两种不同元素的原子作为其环成员的环状结构,而杂原子优选选自元素S、O、N和B的原子。
优选地,该方法包括通过相应的酮还原酶以高比活性辅助转化用于辅因子再生的共底物,优选地根据其比活性从下组选择这样的共底物:异丙醇、2-丁醇、2-戊醇、2-己醇和2-庚醇,最优选地以高比活性将异丙醇转化为丙酮。在本发明方法的优选实施方案中,通过使用NADPH:NADH氧化酶(NOX),例如来自桑树杆菌(Lountos et al.,Acta Cryst.(2004),D60,2044-2047),以及适合的共底物(例如丙酮)再生还原型辅因子NAD(P)H。
因此,本发明还涉及用于以下的方法:
-仲醇的制备,涉及将酮底物优选地立体选择性地还原为所述仲醇;
-伯醇的制备,涉及将醛底物还原为所述伯醇;
-酮产物的制备,涉及将仲醇底物氧化为所述酮产物;
-醛的制备,涉及将伯醇底物氧化为所述醛;和/或
-羧酸的制备,涉及将醛底物氧化为所述羧酸;
其中该方法包括使底物与适合的辅因子在本发明的酮还原酶的存在下反应的步骤。对于由酮还原酶催化的还原反应(酮底物还原为相应的仲醇,醛还原为相应的伯醇),需要还原形式的适合的辅因子作为电子供体。对于由酮还原酶催化的氧化反应(仲醇氧化为相应的酮,伯醇氧化为相应的醛,醛氧化为相应的羧酸),需要氧化形式的适合的辅因子作为电子受体。
当底物为手性时,该反应可以立体选择性和/或区域选择性地进行,因此可用于例如,动力学外消旋拆分。
在以制备规模设置过程例如酮的还原时,考虑到选自以下参数的一个或多个参数,可在考虑具体的其他反应条件下进行本发明的方法:可以例如使用用于辅因子再生的异丙醇或葡萄糖/GDH进行制备规模还原。可以在水性环境中、非水性环境中或两相系统中实施本发明的方法。可以在pH值为4-11,和/或5℃-90℃的温度下和/或底物浓度为1-800g/L下实施本发明的方法。可以用游离酶或用固定化酶实施本发明的方法。可以以间歇方法或通过连续移除产物实施本发明的方法。可以以高底物进料,和/或通过将产物结合(例如通过吸附)至反应容器中的固相,和/或加入水溶性溶剂和/或在高剪切力下实施本发明的方法。
本发明的另一方面涉及制备产物(优选为手性产物)的方法,其包括:
·根据本发明将酮底物立体选择性地还原为仲醇的方法;
·根据本发明将醛还原为伯醇的方法;
·根据本发明将仲醇立体选择性地氧化为酮产物的方法;
·根据本发明将伯醇氧化为醛的方法;和/或
·根据本发明将醛底物氧化为羧酸的方法。
如上限定的关于本发明各种方法的所有优选实施方案,包括本发明的酮还原酶,本发明的酮底物,本发明的仲醇底物,本发明的醛底物,本发明的伯醇底物,也适用于本发明的方法,因此在下文中不再重复。
本发明的另一方面涉及本发明的酮还原酶在将酮底物立体选择性地还原为仲醇中的用途。
本发明的另一方面涉及本发明的酮还原酶在将醛还原为伯醇中的用途。
本发明的另一方面涉及本发明的酮还原酶在将仲醇氧化成酮产物中的用途。
本发明的另一方面涉及本发明的酮还原酶在将伯醇氧化为醛产物中的用途。
本发明的另一方面涉及本发明的酮还原酶在将醛氧化为羧酸中的用途。
优选地,在本发明的分别用于将酮底物立体选择性还原为仲醇以及用于将醛底物还原为伯醇的用途中,醛底物和酮底物可以为任意醛底物或任意酮底物,优选地为通式(I')的醛底物或通式(I)的酮底物。
如上限定的关于本发明的酮底物和醛底物的所有优选实施方案也适用于本发明的方法,因此在下文中不再重复。
以下实施例进一步说明本发明,但不应解释为限制其范围。
实施例1检测对应于本发明SEQ ID NO:1的新的酮还原酶基因
在为了获得来自木兰假丝酵母(Candida magnoliae)DSMZ 70638的新的酮还原酶的、使用简并PCR引物的基于PCR的基因组筛选中鉴定出具有SEQ ID NO:2的新的酮还原酶的基因。令人惊讶地,发现并克隆了对应于本发明的SEQ ID NO:1的新的酮还原酶的基因区段,其表示本发明的未知的新酮还原酶。通过使用精制的PCR引物,鉴定了SEQ ID NO:2所示的酮还原酶基因的全开放阅读框,其编码新鉴定的本发明的SEQ ID NO:1的酮还原酶。
实施例2SEQ ID NO:2和SEQ ID NO:5的新酮还原酶基因的表达
新发现的编码本发明的酮还原酶(本发明的SEQ ID NO:1)的酮还原酶基因SEQ IDNO 2,以及编码对应于比较序列SEQ ID NO:3的酮还原酶的酮还原酶基因SEQ ID NO:5,被克隆到表达载体pLE1A17(pRSF-1b的衍生物,Novagen)中。所得到的质粒用于大肠杆菌(E.coli)BL21(DE3)细胞的转化。SEQ ID NO:5为SEQ ID NO:4的密码子优化的DNA变体。SEQID NO:4和SEQ ID NO:5这两个DNA序列编码相同的酮还原酶序列SEQ ID NO:3。
为了表达酮还原酶基因SEQ ID NO:2和SEQ ID NO:5,将细胞在37℃下在补充有卡那霉素(50mg/l)的ZYM505培养基(F.William Studier、Protein Expression andPurification 41(2005)207–234)中培养。通过IPTG(0.1mM)在对数期诱导基因的表达,并在30℃进行16-18小时。
通过离心(3220×g,20分钟,4℃)收获细胞,并用细胞裂解缓冲液(50mMTris-HClpH 7.0;2mM MgCl2,1×CelLytic B(Sigma);DNA核酸酶0.02U,溶菌酶0.5mg/ml)破碎细胞。通过离心(3220×g 30分钟,4℃)从细胞碎片中分离出粗提取物。将上清液在0.2μm的膜滤器上无菌过滤并冷冻干燥。获得了褐色粉末。
通过变性SDS-PAGE研究粗提取物上清液和不溶性细胞碎片的酮还原酶含量的水平。在粗提取物上清液中发现了约75%的本发明的SEQ ID NO:1的过表达蛋白,而在不溶部分中发现了25%。从比较序列SEQ ID NO:3酮还原酶中,在可溶性部分中仅发现了50%的过表达蛋白质,而在不溶性细胞碎片中发现了50%。
实施例3乙酰乙酸乙酯、环己酮、COBE、2-庚酮、4-CAP的还原活性:
通过监测由NADH或NADPH氧化导致的在340nm处的吸光度的降低,来光度测定地测量本发明的SEQ ID NO:1和比较序列SEQ ID NO:3的酮还原酶还原乙酰乙酸乙酯、环己酮、COBE、2-庚酮、4-CAP的初始活性。一个单位对应于每分钟1μmolNADH或NADPH的氧化。
为了测量还原活性,在具有0.5mM NADH或NADPH的Tris-HCl缓冲液(50mM,pH 7.0,2mM MgCl2)中使用100mM乙酰乙酸乙酯或50mM COBE或4-CAP或2-庚酮或环己酮。分别由对应于本发明的SEQ ID NO:1和比较序列SEQ ID NO:3的每种被测试的酮还原酶的冻干物制备20mg/ml的储备溶液,并根据各自对不同底物的活性将其稀释用于活性测定(通常为1:2.5-1:100)。实验结果示于下表。
从比较数据清楚可见,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)具有显著改善的还原活性(单位/mg冻干物)。
本发明的SEQ ID NO:1的酮还原酶相对于NADH表现出对NADPH的强的辅因子偏好(例如,辅因子偏好比:对于乙酰乙酸乙酯:37.3;对于COBE:3.3)
实验结果汇总如下:
*不可检测,活性低于检测极限
实施例4对3-羟基丁酸乙酯、环己醇、异丙醇的氧化活性;pH特性:
对于氧化活性的测量,通过测量由NADP+的氧化导致的在340nm处的吸光度的增加,来光度测定地测量氧化3-羟基丁酸乙酯、环己醇和异丙醇的初始活性。一个单位对应于每分钟1μmolNADH的氧化。
在磷酸钾缓冲液(50mM,pH 7.0)或Tris/HCl缓冲液(50mM,pH 9.0,MgCl2)中使用100mM 3-羟基丁酸乙酯或50mM环己醇或20%(v/v)异丙醇。分别由对应于本发明的SEQ IDNO:1和比较序列SEQ ID NO:3的每种被测试的酮还原酶的冻干物制备20mg/ml的储备溶液,并根据各自对不同底物的活性将其稀释用于活性测定(通常为1:1-1:10)。
从比较数据清楚可见,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)具有显著改善的氧化活性。此外,本发明的SEQ ID NO:1的酮还原酶相对于比较序列SEQ ID NO:3显示出在较低的pH值(分别为pH 7.0或7.5)下比在较高的pH(pH 9.0)下更优的pH特性(profile),这反映在如下的上部表格中所示的、与在pH 9.0下相比,在pH 7.0下的更高的“改善因子”,其中“改善因子”描述了在各个pH值的测定条件下SEQ ID NO:1相对于比较序列SEQ ID NO:3的活性比。作为表达本发明的SEQ ID NO:1的优异pH特性的另一种方式,在如下的下部表格中显示了相应酶在pH 7(或分别在pH7.5)下的氧化活性与其在pH 9下的活性相比的“活性百分比”。虽然比较序列SEQ ID NO:3的比较酮还原酶显示对于环己醇和异丙醇的活性百分比为27%和33%,但是本发明的SEQ ID NO:1的酮还原酶对于相同的底物显示为60%或62%的活性百分比。这表明本发明的酮还原酶(本发明的SEQ ID NO:1)显著改善了在中性至弱酸性条件下的氧化。
实验结果汇总如下:
*不可检测,活性低于检测极限
如以上两表所示的实验结果表明,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)在不同的pH值下具有显著改善的氧化活性(单位/mg冻干物),与在pH 9下的初始活性相比,在中性pH(分别为pH 7和pH 7.5)下具有的较低的初始活性降低(较高的“活性百分比”)。
实施例5立体选择性:
用4mg/ml的本发明的SEQ ID NO:1和比较序列SEQ ID NO:3的酮还原酶进行分析性生物转化。为了辅因子再循环,使用了葡萄糖脱氢酶(GDH)系统,该系统包含0.1mg/mlGDH-03(来自德国c-LEcta GmbH的商品)和150mM葡萄糖,以将NADP+恒定还原为NADPH。反应在含1mM NADP+和50mM 2-庚酮或4-CAP或COBE的磷酸钾缓冲液(50mM,pH 7.0,2mM MgCl2)中进行。将反应混合物在200rpm和30℃下温育24小时。使用五体积的MTBE(2-甲氧基-2-甲基丙烷)萃取反应混合物。有机相用于手性GC分析。在配备FID检测器的Shimadzu GC2010中使用Ivadex-3 GC色谱柱(IVA-Analysentechnik,手性相:二乙酰基叔丁基甲硅烷基β-环糊精,长度25m,ID 0.25mm,膜0.25μm)。通过与分析标准底物比较鉴定对映异构体。由对映体的峰面积计算对映体过量。
实验结果汇总如下:
从以上比较数据清楚地看出,对于所有测试底物而言,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)具有显著更高的立体选择性。
实施例6Tm值-Tm50(40)
在不同温度下将本发明的SEQ ID NO:1和比较序列SEQ ID NO:3的酮还原酶各自的40mg/ml溶液温育15分钟后,测量各酶的热稳定性。Tm50(40)值定义为在将40mg/ml溶液温育15分钟后相比于在25℃下储存的样品仍保留50%残余活性的温度。如实施例1中所述,使用乙酰乙酸乙酯用光度测定法检测活性。
实验结果如图1所示。
从比较数据清楚可见,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)具有+7℃的显著改善的半最大熔解温度(Tm50(40))。半最大熔解温度是各个酮还原酶相对于其在25℃时的活性仍具有50%残余活性的温度。因此,与比较序列SEQ ID NO:3的酮还原酶相比,本发明的酮还原酶(本发明的SEQ ID NO:1)在较高温度下具有显著改善的活性。
实施例7SEQ ID NO:1和变体对乙酰乙酸乙酯的还原活性:
分子生物学方法:SEQ ID NO:1的变体通过本领域已知的标准定点诱变技术产生。
通过监测由NADPH的氧化引起的在340nm处的吸光度的降低,来光度测定地测量酮还原酶还原乙酰乙酸乙酯的初始活性。一个单位对应于每分钟1μmolNADPH的氧化。
为了测量还原活性,在30℃下在具有2mM NADPH的Tris-HCl缓冲液(50mM,pH 7.0,2mM MgCl2)中使用100mM乙酰乙酸乙酯。取决于各自活性和筛选形式,将粗细胞提取物稀释以用于活性测定。在比色皿中测量所选酶的特征。在96孔微量滴定板中进行高通量筛选。
实施例8SEQ ID NO:1和变体对环己酮的还原活性:
通过监测由NADPH的氧化引起的在340nm处的吸光度的降低,来测量酮还原酶还原环己酮的初始活性。一个单位对应于每分钟1μmol NADPH的氧化。取决于各自的活性,将粗细胞提取物稀释以用于活性测定。
为了测量还原活性,在30℃下在具有0.5mMNADPH的磷酸钾缓冲液(50mM,pH 7.0)中使用50mM环己酮。
实验结果汇总如下:
与SEQ ID NO:1相比,一些变体表现出较低的环己酮活性,但是无论其如何,提供了如实施例10中所示的改善的还原活性/氧化活性(red/ox)比。
实施例9SEQ ID NO:1及其变体对环己醇的氧化活性:
通过监测由NADP+的还原引起的在340nm处吸光度的增加来测量酮还原酶氧化环己醇的初始活性。一个单位对应于每分钟1μmolNADP+的氧化。取决于各自的活性,将粗细胞提取物稀释以用于活性测定。
为了测量氧化活性,在30℃下在具有1mM NADP+的磷酸钾缓冲液(50mM,pH 7.0)中使用50mM环己酮。
在96孔微量滴定板中进行高通量筛选。在高通量筛选中,无法准确测量到与SEQID NO:1相比改善5倍以上的值,并以≥5给出。
高通量筛选的实验结果汇总如下:
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在比色皿中测量所选酶的详细活性表征。相应的实验结果汇总如下:
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与SEQ ID NO:1相比,一些变体表现出较低的环己醇活性,但是无论其如何,提供了如实施例10中所示的改善的还原活性/氧化活性比。
实施例10确定SEQ ID NO:1变体的还原/氧化比:
酮还原酶通常显示出对特定底物的氧化形式的还原活性远高于对该底物的还原形式的氧化活性。因此,对于某些应用非常需要具有改善的氧化特性的酮还原酶。
由对环己酮的初始还原活性(参见实施例8)除以对环己醇的氧化活性(参见实施例9)计算还原活性和氧化活性之间的比值。计算出的比值越低,相应变体的氧化活性越优选。
实验结果汇总如下:
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还原活性与氧化活性之间的比值表明了酶的偏好性,以用于根据需要氧化或还原的应用来选择酶。
实施例11在高通量筛选中筛选提高的热稳定性:
生物催化过程非常需要提高热稳定性。更稳定的酶可以耐受更高的温度而不会劣化。此外,更稳定的酶通常在中等温度下可在更长的时间内保持活性,对有机助溶剂或较高的底物或产物浓度更耐受。这通常使稳定的酶变体比不稳定的酶具有更高的加工性能。
为了在高通量筛选期间鉴定更稳定的酶变体,使用如实施例5所述的对乙酰乙酸乙酯的还原检测法,在42℃下将稀释至OD为10的粗提取物温育15分钟后,测量酶变体的残余活性,并与未经热处理的样品的活性进行比较。残余活性表示为热处理后相对于初始活性的残余的活性,以百分比表示。SEQ ID NO:1的野生型酶通常显示出10-25%之间的残余活性。如果在指定温度下温育后显示出更高的残余活性,则可以鉴定为更稳定的酶。
实验结果汇总如下:
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实施例12-测量Tm值Tm50(10)
在不同温度下将OD600=10的粗提取物温育15分钟后,测量所选的酮还原酶变体的热稳定性。Tm50(10)值定义为与储存在冰上的样品相比,在将对应于OD600为10的粗提物温育15分钟后,仍保留50%残余活性的温度。如实施例5所述测量活性。
实验结果汇总如下:
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实施例13乙酰乙酸乙酯的活性
如实施例3中所述,使用NADPH作为辅因子,测试了几种酮还原酶变体的还原乙酰乙酸乙酯的初始活性。
实验结果汇总如下:
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序列表
<110> C-乐克塔股份有限公司
<120> 酮还原酶
<130> IV0017 (EP)
<160> 182
<170> BiSSAP 1.3.6
<210> 1
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 人工的,本发明
<400> 1
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
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Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
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Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
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Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
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Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 2
<211> 741
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<223> 编码SEQ ID NO:1
<400> 2
atgacgtcct cctcttctcc ctccttgaac gctctggtca cgggcggcag ccgcggcatt 60
ggcgaggcca tttccatgca gctggccgcc gagggctaca gcgtcacaat tgcgtctcgc 120
gggctcgagc agctcgaggc cgtgaaggca aagctgccca ttgtgaagca gggccagacg 180
caccacgtgt ggcagctcga tctcagtgat gtcgaggccg ccggctcctt caagggtgcg 240
ccgctgccgg ccagcagcta cgacgtgttt gtgagcaatg ccggcatttc tcagttctcg 300
ccgattgcgg agcacgccga cgcggattgg cagaacatgc tgaccgtgaa cctgacggcg 360
ccgattgcgc tcacgaaggc cgtcgtgaag gcaatcagcg acaagccgcg ccagacgcct 420
gcgcacatca tcttcatctc gacgggcctg tcgaagcgcg gcgccccaat ggttggcgtc 480
tacagcgcct cgaaggccgg catcgacggg ttcatgcgct cgctggcccg cgagctcggc 540
ccgaagggca tcaacgtgaa ctgcgtgagc cccggcgtga caaggacgag catggccgag 600
ggcatcgacc ccagcatgtt cgaccttcct atcaacgggt ggattgaggt cgacgcgatc 660
gccgacgcgg tcacgtacct cgtgaagtcg aaaaacgtca cgggcacgac cgtttccgtc 720
gacaacggct actgcgccta a 741
<210> 3
<211> 238
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 人工的,比较
<400> 3
Met Asn Ala Leu Val Thr Gly Gly Ser Arg Gly Ile Gly Glu Ala Val
1 5 10 15
Ser Ile Gln Leu Ala Ser Gln Gly Tyr Gly Val Thr Ile Ala Ser Arg
20 25 30
Gly Val Glu Gln Leu Glu Ala Val Lys Ala Lys Leu Pro Val Val Lys
35 40 45
Gln Gly Gln Thr His His Val Trp Gln Leu Asp Leu Ser Asp Val Glu
50 55 60
Ala Ala Gly Ser Phe Lys Gly Ala Pro Leu Pro Ala Ser Ser Tyr Asp
65 70 75 80
Val Phe Val Ser Asn Ala Gly Ile Ala Gln Phe Ser Pro Ile Ala Glu
85 90 95
His Ala Asp Ala Asp Trp Gln Asn Met Leu Thr Val Asn Leu Thr Ala
100 105 110
Pro Ile Ala Leu Thr Lys Ala Val Val Lys Ala Ile Ser Asp Lys Pro
115 120 125
Arg Glu Thr Pro Ala His Ile Ile Phe Ile Ser Thr Gly Leu Ser Lys
130 135 140
Arg Gly Ala Pro Met Val Gly Val Tyr Ser Ala Ser Lys Ala Gly Ile
145 150 155 160
Asp Gly Phe Met Arg Ser Leu Ala Arg Glu Leu Gly Pro Lys Gly Ile
165 170 175
Asn Val Asn Cys Val Ser Pro Gly Val Thr Arg Thr Ala Ile Ser Asp
180 185 190
Gly Val Asp Pro Ser Met Phe Asp Leu Pro Ile Ser Gly Trp Ile Glu
195 200 205
Val Asp Ala Ile Ala Asp Ala Val Thr Tyr Leu Val Lys Ser Lys Asn
210 215 220
Val Thr Gly Ala Ile Leu Val Val Asp Asn Gly Phe Ser Thr
225 230 235
<210> 4
<211> 717
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<223> 编码SEQ ID NO:3
<400> 4
atgaacgctc tagtgacggg tggcagtcgt ggcattggcg aggccgtttc cattcaattg 60
gcttcgcagg gctacggcgt cacaattgca tctcgcggcg ttgagcagct cgaggccgta 120
aaggcgaagc tgcccgttgt gaagcagggc cagacgcacc acgtgtggca gctcgacttg 180
agcgacgtcg aggcggccgg ctccttcaag ggtgcgcccc tgccggccag cagctacgac 240
gtgtttgtga gcaatgccgg cattgctcag ttctcgccga ttgcggaaca cgccgacgcg 300
gactggcaaa acatgctgac cgtgaacctg acggcgccga tcgccctcac gaaggcggtc 360
gtgaaggcca tcagcgacaa gccgcgcgag acgccggccc atatcatttt catctcaacg 420
ggcctatcga agcgcggcgc tccgatggtc ggtgtctaca gcgcttcaaa ggccggtatc 480
gacggattca tgcgctcgct ggcccgcgag ctcggcccaa agggcatcaa cgtgaactgc 540
gtcagccccg gagtgacaag gacggccatt tccgatggtg tggaccccag catgttcgac 600
ctgcctatca gcgggtggat cgaggtcgac gccattgccg acgccgttac gtacctcgtg 660
aagtcgaaga acgtcacggg cgcgatcctc gtagtcgaca acggcttctc gacgtag 717
<210> 5
<211> 717
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<223> 编码SEQ ID NO:3, 密码子优化的
<400> 5
atgaatgcgc tggttacggg cggatctcgc ggtattggcg aagctgtgag catccagctg 60
gcatcacagg ggtatggcgt tacgattgcc tctcgtggcg tggaacaact ggaagcggta 120
aaagccaaac ttcccgtagt caaacagggg caaacacacc atgtgtggca actggacttg 180
tcggatgtag aagcggctgg aagcttcaaa ggtgcaccct taccagcgag ttcgtatgac 240
gtgtttgtgt ccaatgctgg gattgcccag ttttcgccga ttgcggaaca tgctgatgca 300
gactggcaga acatgctgac cgtcaatctg accgcaccga ttgcgctcac caaggcagtc 360
gttaaagcca tctccgacaa accacgggaa actcctgctc acatcatctt cattagcacc 420
ggactgtcaa aacgcggtgc accgatggtt ggtgtctact cagccagcaa agccggcatc 480
gatggcttca tgcgctctct ggcccgtgag ttaggcccga aaggcatcaa cgtgaattgc 540
gttagcccgg gtgtcactcg taccgcgatt agcgatggtg tggatccgag tatgtttgat 600
ctccctatca gtggttggat tgaggtagac gcgattgccg atgcggttac ctacttggtg 660
aagtcgaaga acgttacagg tgcgattctt gtggtggata acggcttttc cacgtaa 717
<210> 6
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 6
<400> 6
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Val Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 7
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 7
<400> 7
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Phe Cys Ala
245
<210> 8
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 8
<400> 8
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Leu Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 9
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 9
<400> 9
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe His
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 10
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 10
<400> 10
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Asp Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 11
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 11
<400> 11
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Cys
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 12
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 12
<400> 12
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Val Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 13
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 13
<400> 13
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Cys Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 14
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 14
<400> 14
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Asn Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 15
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 15
<400> 15
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Pro Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 16
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 16
<400> 16
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Val Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 17
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 17
<400> 17
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Thr Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 18
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 18
<400> 18
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 19
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 19
<400> 19
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Gly Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Met Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 20
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 20
<400> 20
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Ala Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 21
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 21
<400> 21
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Val Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 22
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 22
<400> 22
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Ala Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 23
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 23
<400> 23
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Pro Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 24
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 24
<400> 24
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Ile Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 25
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 25
<400> 25
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Val Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 26
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 26
<400> 26
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Tyr Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 27
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 27
<400> 27
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Phe Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 28
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 28
<400> 28
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Ser Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 29
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 29
<400> 29
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Glu Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Thr
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 30
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 30
<400> 30
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Gln Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 31
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 31
<400> 31
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Gly Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 32
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 32
<400> 32
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Arg Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 33
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 33
<400> 33
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Phe Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 34
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 34
<400> 34
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser His Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 35
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 35
<400> 35
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Trp Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 36
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 36
<400> 36
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Arg Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 37
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 37
<400> 37
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Val Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 38
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 38
<400> 38
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Cys Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 39
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 39
<400> 39
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Val Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 40
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 40
<400> 40
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Ser Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 41
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 41
<400> 41
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Trp Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 42
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 42
<400> 42
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Val Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 43
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 43
<400> 43
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Cys Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 44
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 44
<400> 44
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Arg Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 45
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 45
<400> 45
Met Thr Ser Ser Ser Ser Pro Ser Gly Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 46
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 46
<400> 46
Met Thr Ser Ser Ser Ser Pro Ser Met Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 47
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 47
<400> 47
Met Thr Ser Ser Ser Ser Pro Ser Leu Gly Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 48
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 48
<400> 48
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Cys
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 49
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 49
<400> 49
Met Thr Ser Ser Ser Ser Pro Ser Arg Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 50
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 50
<400> 50
Met Thr Ser Ser Ser Ser Pro Ser Lys Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 51
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 51
<400> 51
Met Thr Ser Ser Ser Ser Pro Ser Gln Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 52
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 52
<400> 52
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ala Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 53
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 53
<400> 53
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Lys Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 54
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 54
<400> 54
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Arg Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 55
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 55
<400> 55
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Arg Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 56
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 56
<400> 56
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Asn Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 57
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 57
<400> 57
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Met Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 58
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 58
<400> 58
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Gly Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 59
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 59
<400> 59
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Tyr Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 60
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 60
<400> 60
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ala Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 61
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 61
<400> 61
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Arg Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 62
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 62
<400> 62
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Val Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 63
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 63
<400> 63
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Ile Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 64
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 64
<400> 64
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Met Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 65
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 65
<400> 65
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Gly Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 66
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 66
<400> 66
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Phe Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 67
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 67
<400> 67
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Val Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 68
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 68
<400> 68
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Glu Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 69
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 69
<400> 69
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Leu Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 70
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 70
<400> 70
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Ala Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 71
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 71
<400> 71
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Ala Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Met Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 72
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 72
<400> 72
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Phe Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 73
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 73
<400> 73
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Arg Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 74
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 74
<400> 74
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Gly Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 75
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 75
<400> 75
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ala Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 76
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 76
<400> 76
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Gly Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Thr Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Val Tyr Phe Phe
245
<210> 77
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 77
<400> 77
Met Thr Ser Ser Ser Ser Pro Ser Gly Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 78
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 78
<400> 78
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Gly Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 79
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 79
<400> 79
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ala Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asn
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 80
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 80
<400> 80
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Arg Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Phe Thr Ile Phe Phe Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 81
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 81
<400> 81
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Cys Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 82
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 82
<400> 82
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Glu Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 83
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 83
<400> 83
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Lys Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 84
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 84
<400> 84
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Ser Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 85
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 85
<400> 85
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Cys Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 86
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 86
<400> 86
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Glu Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 87
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 87
<400> 87
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Pro Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 88
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 88
<400> 88
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Ser Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 89
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 89
<400> 89
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ala Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 90
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 90
<400> 90
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Trp Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 91
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 91
<400> 91
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Val Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 92
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 92
<400> 92
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 93
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 93
<400> 93
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Met Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 94
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 94
<400> 94
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Cys Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 95
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 95
<400> 95
Met Thr Ser Asn Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 96
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 96
<400> 96
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Lys Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 97
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 97
<400> 97
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Val Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 98
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 98
<400> 98
Met Thr Ile Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 99
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 99
<400> 99
Met Thr Ser Ser Ser Ser Pro Ala Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 100
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 100
<400> 100
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Cys Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 101
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 101
<400> 101
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 102
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 102
<400> 102
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Thr Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 103
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 103
<400> 103
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Arg Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 104
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 104
<400> 104
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Glu Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 105
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 105
<400> 105
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Lys Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 106
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 106
<400> 106
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Arg Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 107
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 107
<400> 107
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Met Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 108
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 108
<400> 108
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Pro Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 109
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 109
<400> 109
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Ala Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 110
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 110
<400> 110
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Cys His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 111
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 111
<400> 111
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Asn His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 112
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 112
<400> 112
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Ala His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 113
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 113
<400> 113
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 114
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 114
<400> 114
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Val Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 115
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 115
<400> 115
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Cys Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 116
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 116
<400> 116
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Ser Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 117
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 117
<400> 117
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ala Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 118
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 118
<400> 118
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Met Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 119
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 119
<400> 119
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Cys Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 120
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 120
<400> 120
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Leu Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 121
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 121
<400> 121
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Gly Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 122
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 122
<400> 122
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Leu Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 123
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 123
<400> 123
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Leu Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 124
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 124
<400> 124
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Cys Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 125
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 125
<400> 125
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Lys Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 126
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 126
<400> 126
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Pro Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 127
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 127
<400> 127
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Ala Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 128
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 128
<400> 128
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Lys Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 129
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 129
<400> 129
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Cys Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 130
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 130
<400> 130
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Gly Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 131
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 131
<400> 131
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Val Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 132
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 132
<400> 132
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Gln Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 133
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 133
<400> 133
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Leu Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 134
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 134
<400> 134
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ala Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 135
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 135
<400> 135
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Gln Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 136
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 136
<400> 136
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Met Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 137
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 137
<400> 137
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Val Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 138
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 138
<400> 138
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Val Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 139
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 139
<400> 139
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Ala Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 140
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 140
<400> 140
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Thr Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 141
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 141
<400> 141
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Gly Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 142
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 142
<400> 142
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Ser Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 143
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 143
<400> 143
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Trp Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 144
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 144
<400> 144
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Met Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 145
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 145
<400> 145
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Ala Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 146
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 146
<400> 146
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Ala Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 147
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 147
<400> 147
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Glu Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 148
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 148
<400> 148
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Cys Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 149
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 149
<400> 149
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 150
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 150
<400> 150
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Val Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 151
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 151
<400> 151
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Trp Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 152
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 152
<400> 152
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Leu Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 153
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 153
<400> 153
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Ile Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 154
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 154
<400> 154
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Val Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 155
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 155
<400> 155
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Ile Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 156
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 156
<400> 156
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Gly Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 157
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 157
<400> 157
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Val Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 158
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 158
<400> 158
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Phe Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 159
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 159
<400> 159
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Lys Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 160
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 160
<400> 160
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ser His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 161
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> SEQ ID NO: 161
<400> 161
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Arg Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 162
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 162
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 163
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 163
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Leu Cys Ala
245
<210> 164
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 164
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Ile Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 165
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 165
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 166
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 166
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 167
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 167
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Cys Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 168
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 168
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Ala Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 169
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 169
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Arg Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 170
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 170
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 171
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 171
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Arg Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Ala Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Phe Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 172
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 172
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Arg Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Val Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Asp Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 173
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 173
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Ser Gln Phe Arg Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Val Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 174
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 174
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 175
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 175
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Ala Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 176
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 176
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Val Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Asp Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Phe Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 177
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 177
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Met Leu Ser Lys Arg Gly Ala Pro Asp Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Asp Phe Asp
195 200 205
Leu Pro Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Phe Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 178
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 178
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Thr Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Leu Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 179
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 179
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Lys Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Ser Gly Leu Ser Lys Arg Gly Ala Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Val Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Val Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 180
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 180
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Ser Gly Leu Ser Lys Arg Gly His Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Val Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Tyr Cys Ala
245
<210> 181
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 181
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Glu Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Ser Gly Leu Ser Lys Arg Gly His Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Val Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Phe Cys Ala
245
<210> 182
<211> 246
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<223> 变体
<400> 182
Met Thr Ser Ser Ser Ser Pro Ser Leu Asn Ala Leu Val Thr Gly Gly
1 5 10 15
Ser Arg Gly Ile Gly Glu Ala Ile Ser Met Gln Leu Ala Ala Glu Gly
20 25 30
Tyr Ser Val Thr Ile Ala Ser Arg Gly Leu Lys Gln Leu Glu Ala Val
35 40 45
Lys Ala Lys Leu Pro Ile Val Lys Gln Gly Gln Thr His His Val Trp
50 55 60
Gln Leu Asp Leu Ser Asp Val Glu Ala Ala Gly Ser Phe Lys Gly Ala
65 70 75 80
Pro Leu Pro Ala Ser Ser Tyr Asp Val Phe Val Ser Asn Ala Gly Ile
85 90 95
Cys Gln Phe Ser Pro Ile Ala Glu His Ala Asp Ala Asp Trp Gln Asn
100 105 110
Met Leu Thr Val Asn Leu Thr Ala Pro Ile Ala Leu Thr Lys Ala Val
115 120 125
Val Lys Ala Ile Ser Asp Lys Pro Arg Gln Thr Pro Ala His Ile Ile
130 135 140
Phe Ile Ser Ser Gly Leu Ser Lys Arg Gly His Pro Met Val Gly Val
145 150 155 160
Tyr Ser Ala Ser Lys Ala Gly Ile Asp Gly Phe Met Arg Ser Leu Ala
165 170 175
Arg Glu Leu Gly Pro Lys Gly Ile Asn Val Asn Cys Val Ser Pro Gly
180 185 190
Val Thr Arg Thr Ser Met Ala Glu Gly Ile Asp Pro Ser Met Phe Asp
195 200 205
Leu Cys Ile Asn Gly Trp Ile Glu Val Asp Ala Ile Ala Asp Ala Val
210 215 220
Thr Tyr Val Val Lys Ser Lys Asn Val Thr Gly Thr Thr Val Ser Val
225 230 235 240
Asp Asn Gly Phe Cys Ala
245
Claims (6)
1.一种酮还原酶,其氨基酸序列选自SEQ ID NO:1、7、8、9、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、32、35、36、38、39、40、41、44、46、47、48、49、50、52、55、56、58、60、63、64、65、67、68、69、70、71、72、73、74、75、76、78、79、80、81、82、83、84、88、89、90、91、92、93、96、104、106、110、115、116、123、124、125、126、127、131、133、135、136、137、143、144、145、146、147、159、161、162、163、164、165、166、167、168、169、170、171、172、173、174、175、176、177、178、179、180、181和182。
2.根据权利要求1所述的酮还原酶,其中所述酮还原酶能够转化仲醇底物3-羟基丁酸乙酯和/或环己醇。
3.用于以下的方法:
-将仲醇底物氧化为酮产物;和/或
-将伯醇底物氧化为醛;
其中所述方法包括在根据权利要求1至2中任一项所述的酮还原酶的存在下使底物与适合的辅因子反应的步骤,其中所述仲醇底物如权利要求2中所定义,所述伯醇底物为异丙醇。
4.根据权利要求3所述的方法,其中所述仲醇底物是环己醇。
5.根据权利要求1至2中任一项所述的酮还原酶用于以下的用途:
-将仲醇底物氧化为酮产物;和/或
-将伯醇底物氧化为醛;
其中所述仲醇底物如权利要求2中所定义,所述伯醇为异丙醇。
6.根据权利要求5所述的用途,其中所述仲醇底物是环己醇。
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EP18156201.8 | 2018-02-11 | ||
EP18159099 | 2018-02-28 | ||
EP18159099.3 | 2018-02-28 | ||
PCT/EP2018/069058 WO2019012095A1 (en) | 2017-07-14 | 2018-07-13 | TCO-reductase |
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WO2021100848A1 (ja) * | 2019-11-22 | 2021-05-27 | 株式会社エーピーアイ コーポレーション | カルボニル還元酵素、これをコードする核酸、及びこれらを利用した光学活性化合物の製造方法 |
CN110938609B (zh) * | 2019-12-27 | 2021-07-23 | 南京朗恩生物科技有限公司 | 一种活性增强的酮还原酶突变体及其应用 |
CN110923277B (zh) * | 2019-12-27 | 2021-05-11 | 南京朗恩生物科技有限公司 | 一种生物催化制备s-3-二甲胺基-1-(2-噻吩基)-1-丙醇的方法 |
CN111690696B (zh) * | 2020-07-07 | 2021-12-31 | 南京朗恩生物科技有限公司 | 一种生物催化制备达芦那韦中间体的方法 |
CN112680425B (zh) * | 2021-01-13 | 2022-06-10 | 江南大学 | 一种醇脱氢酶突变体及其应用 |
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