CN110831618B - 酮还原酶多肽及多核苷酸 - Google Patents

酮还原酶多肽及多核苷酸 Download PDF

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CN110831618B
CN110831618B CN201880043854.4A CN201880043854A CN110831618B CN 110831618 B CN110831618 B CN 110831618B CN 201880043854 A CN201880043854 A CN 201880043854A CN 110831618 B CN110831618 B CN 110831618B
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杰弗里·C·穆尔
杰克·梁
乔纳森·彭菲尔德
约瓦娜·纳佐尔
尼基·德拉斯
韦丝娜·米切尔
段达
伊曼·法拉萨特
阿古斯蒂纳·罗德里格斯-格拉尼利奥
格兰特·墨菲
尼古拉斯·马歇尔
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Abstract

本发明提供了与天然存在的野生型酮还原酶和亚磷酸脱氢酶相比具有改进的特性的工程化酮还原酶和亚磷酸脱氢酶、以及编码工程化酮还原酶和亚磷酸脱氢酶的多核苷酸、能够表达工程化酮还原酶和亚磷酸脱氢酶的宿主细胞、以及使用工程化酮还原酶和亚磷酸脱氢酶来合成用于抗病毒化合物诸如核苷抑制剂的合成的手性催化剂的方法。本发明还提供了使用工程化酶在一锅多酶系统中使手性醇去外消旋化的方法。

Description

酮还原酶多肽及多核苷酸
本申请要求2017年4月27日提交的美国临时专利申请序列号62/491,161的优先权，该美国临时专利申请在此通过引用以其整体并入以用于所有目的。
对序列表、表格或计算机程序的引用
根据37C.F.R.§1.821，以计算机可读形式(CRF)通过EFS-Web以文件名CX2-166USP1_ST25.txt与本申请同时提交的序列表通过引用并入本文。序列表的电子副本创建于2017年4月25日，具有544千字节的文件大小。
发明领域
本发明提供了与天然存在的野生型酮还原酶和亚磷酸脱氢酶(phosphitedehydrogenase)相比具有改进的特性的工程化酮还原酶和亚磷酸脱氢酶、以及编码工程化酮还原酶和亚磷酸脱氢酶的多核苷酸、能够表达工程化酮还原酶和亚磷酸脱氢酶的宿主细胞、以及使用工程化酮还原酶和亚磷酸脱氢酶来合成用于抗病毒化合物诸如核苷抑制剂的合成的手性催化剂的方法。本发明还提供了使用工程化酶在一锅多酶系统(one-pot,multi-enzyme system)中使手性醇去外消旋化(deracemize)的方法。
背景
属于酮还原酶(KRED)或羰基还原酶类(EC1.1.1.184)的酶可用于从对应的前手性酮底物并通过对应的外消旋醛底物的立体选择性还原合成光学活性醇。KRED通常将酮底物和醛底物转化为对应的醇产物，但是也可以催化逆反应，将醇底物氧化为对应的酮/醛产物。酶诸如KRED对酮和醛的还原以及对醇的氧化需要辅因子，最常见的是还原型烟酰胺腺嘌呤二核苷酸(NADH)或还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)，以及用于氧化反应的烟酰胺腺嘌呤二核苷酸(NAD)或烟酰胺腺嘌呤二核苷酸磷酸(NADP)。NADH和NADPH充当电子供体，而NAD和NADP充当电子受体。经常观察到酮还原酶和醇脱氢酶接受磷酸化或非磷酸化的辅因子(处于其氧化态和还原态)，但最通常不是两者都接受。
为了绕过用于产生关键化合物的许多化学合成步骤，酮还原酶越来越多地被用于将不同的酮底物和醛底物酶促转化为手性醇产物。这些应用可以利用表达酮还原酶的全细胞生物催化酮和醛的还原或生物催化醇的氧化，或者在全细胞中多种酮还原酶的存在将不利地影响期望产物的立体纯度(stereopurity)和产率的那些情况下使用纯化的酶。对于体外应用，可以将辅因子(NADH或NADPH)再生酶诸如葡萄糖脱氢酶(GDH)、甲酸脱氢酶、亚磷酸脱氢酶等与酮还原酶一起使用。鉴定可以用于进行多种酮底物向对应的手性醇产物的转化或多种醇底物向对应的酮产物的转化的其他酮还原酶是令人期望的。
发明概述
本发明提供了与天然存在的野生型酮还原酶和亚磷酸脱氢酶相比具有改进的特性的工程化酮还原酶和亚磷酸脱氢酶、以及编码工程化酮还原酶和亚磷酸脱氢酶的多核苷酸、能够表达工程化酮还原酶和亚磷酸脱氢酶的宿主细胞、以及使用工程化酮还原酶和亚磷酸脱氢酶来合成用于抗病毒化合物诸如核苷抑制剂的合成的手性催化剂的方法。本发明还提供了使用工程化酶在一锅多酶系统中使手性醇去外消旋化的方法。
此外，本发明提供了与天然存在的野生型亚磷酸脱氢酶相比具有改进的特性的工程化亚磷酸脱氢酶、以及编码工程化亚磷酸脱氢酶的多核苷酸、能够表达工程化亚磷酸脱氢酶的宿主细胞、以及使用工程化亚磷酸脱氢酶在一锅多酶系统中使手性醇去外消旋化的方法。
本发明提供了工程化酮还原酶(“KRED”)，其能够在一锅多酶系统中立体选择性地使外消旋醇底物去外消旋化为光学纯的醇产物，并且当与天然存在的从近平滑假丝酵母(Candida parapsilosis)获得的野生型KRED酶(SEQ ID NO:2)、从鲑色锁掷酵母(Sporidiobolus salmonicolor)获得的野生型KRED酶(SEQ ID NO:112)相比时，或者当与其他工程化酮还原酶相比时，具有改进的特性。此外，本发明提供了能够在同一一锅多酶系统中优先使NADPH再循环的工程化亚磷酸脱氢酶(“PDH”)。
在一些另外的实施方案中，工程化酶除了改变的酶活性以外还具有一种或更多种改进的特性。例如，在一些实施方案中，与野生型酮还原酶相比，工程化酮还原酶多肽具有增加的立体选择性，以用于将底物还原为产物和/或优先氧化(S)对映异构体。酶特性的改进包括但不限于热稳定性、溶剂稳定性的增加和/或减少的产物抑制。
本发明提供了工程化酮还原酶变体，所述工程化酮还原酶变体与SEQID NO:2、112、124和/或138具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性。
本发明还提供了工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:2具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：37、37/211、37/211/229、37/229、45、52、52/57/110/272/296、52/57/272、52/57/272/274/279/296、52/57/272/279/296、55/57/276、56、57、57/104/114、57/104/114/229、57/286、79/83/275/276、83、83/275/276、83/276、104、110、114、138/146/258/289、211、211/229、228、229、263、268、272、274、275/276、276、279和309，其中所述位置根据SEQ ID NO:2来编号。在一些另外的实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：37R、37R/211R、37R/211R/229R、37R/229R、45R、52D、52D/57L/272H、52S、52S/57L/110T/272H/296F、52S/57L/272H/279H/296F、52S/57L/272H/274V/279H/296F、55F/57A/276M、56L、57I、57I/104G/114H、57L、57L/104G/114H/229R、57X/286X、79T/83S/275N/276M、83I、83S/275N/276M、83S/276M、104G、110T、114H/K/M、138V/146S/258V/289S、211R、211R/229R、228S、229R、263H/Y、268M/W、272H/I/L/P/Q/S/T/V/W、274I/V、275N/276M、276F/M、279H/Q/R和309F，其中所述位置根据SEQID NO:2来编号。在一些另外的实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：K37R、K37R/K211R、K37R/K211R/G229R、K37R/G229R、H45R、Y52D、Y52D/C57L/G272H、Y52S、Y52S/C57L/K110T/G272H/L296F、Y52S/C57L/G272H/I279H/L296F、Y52S/C57L/G272H/L274V/I279H/L296F、L55F/C57A/L276M、D56L、C57I、C57I/A104G/G114H、C57L、C57L/A104G/G114H/G229R、C57X/W286X、I79T/V83S/A275N/L276M、V83I、V83S/A275N/L276M、V83S/L276M、A104G、K110T、G114H/K/M、S138V/A146S/M258V/T289S、K211R、K211R/G229R、P228S、G229R、G263H/Y、S268M/W、G272H/I/L/P/Q/S/T/V/W、L274I/V、A275N/L276M、L276F/M、I279H/Q/R和R309F，其中所述位置根据SEQ ID NO:2来编号。
本发明还提供了工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:112具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：24/106/136/220/258/260/314/315、24/106/214/250/258/260/314/315、24/220/314/315、122/159/316/318、135、139/207、159/251/272/277/316/318/330和207，其中所述位置根据SEQ IDNO:112来编号。在一些实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：24I/106P/136A/220G/258V/260A/314R/315A、24I/106P/214L/250V/258V/260A/314R/315A、24I/220G/314R/315A、122E/159V/316E/318L、135F、139V/207S、159V/251Q/272F/277P/316E/318L/330L和207G，其中所述位置根据SEQ IDNO:112来编号。在一些另外的实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：V24I/T106P/S136A/S220G/L258V/C260A/P314R/S315A、V24I/T106P/F214L/A250V/L258V/C260A/P314R/S315A、V24I/S220G/P314R/S315A、T122E/I159V/L316E/I318L、V135F、I139V/N207S、I159V/V251Q/Y272F/T277P/L316E/I318L/I330L和N207G，其中所述位置根据SEQ ID NO:112来编号。
本发明还提供了工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:124具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且具有选自以下位置的至少一个取代集：2/101/179/182/228/238/282、3/95、3/95/228/314、24/95/228、95、95/135/139/207和159/228/309/330，其中所述位置根据SEQID NO:124来编号。在一些实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：2T/101P/179L/182M/228R/238L/282E、3Y/95T、3Y/95T/228T/314R、24I/95T/228T、95T、95T/135F/139V/207N和159V/228L/309Q/330L，其中所述位置根据SEQ ID NO:124来编号。在一些另外的实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：A2T/Y101P/A179L/T182M/M228R/A238L/T282E、K3Y/V95T、K3Y/V95T/M228T/P314R、V24I/V95T/M228T、V95T、V95T/V135F/I139V/G207N和I159V/M228L/K309Q/I330L，其中所述位置根据SEQ ID NO:124来编号。
本发明还提供了工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:138具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：19、24/43/47/49/67/68/70/91/220、24/68/91/218/220、67、72、74/75/78/108、75/78/99/108/215/224、78/107、95、96和114，其中所述位置根据SEQ ID NO:138来编号。在一些实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：19S、24I/43V/47E/49N/67V/68E/70P/91V/220G、24I/68E/91V/218N/220G、67W、72Q、74A/75E/78F/108V、75E/78F/99P/108V/215S/224A、78F/107G、95C、96G和114V，其中所述位置根据SEQ ID NO:138来编号。在一些另外的实施方案中，工程化酮还原酶变体包含选自以下的至少一个取代或取代集：G19S、V24I/A43V/S47E/L49N/A67V/V68E/E70P/I91V/S220G、V24I/V68E/I91V/T218N/S220G、A67W、M72Q、K74A/Q75E/Y78F/A108V、Q75E/Y78F/N99P/A108V/D215S/S224A、Y78F/P107G、T95C、S96G和N114V，其中所述位置根据SEQ ID NO:138来编号。
本发明还提供了工程化酮还原酶变体，所述工程化酮还原酶变体包含含有与SEQID NO:2、112、124和/或138具有至少90％序列同一性的序列的多肽序列。在一些实施方案中，工程化酮还原酶变体包含含有与SEQID NO:2、112、124和/或138具有至少95％序列同一性的序列的多肽序列。在一些另外的实施方案中，工程化酮还原酶变体包含SEQ ID NO:2、112、124或138中列出的多肽序列。在一些另外的实施方案中，工程化酮还原酶变体包含编码表5.1、表6.1、表7.1和/或表8.1中提供的变体的多肽序列。在一些另外的实施方案中，工程化酮还原酶变体包含选自SEQ ID NO:4至SEQ ID NO:170中列出的偶数编号的序列的多肽序列。
本发明还提供了编码本文提供的工程化酮还原酶变体的工程化多核苷酸序列。在一些实施方案中，工程化多核苷酸序列包含与选自SEQ IDNO:3至SEQ ID NO:169中列出的奇数编号的序列的序列至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多相同的多核苷酸序列。本发明还提供了包含编码本文提供的工程化酮还原酶变体的工程化多核苷酸序列的载体。在一些实施方案中，载体还包含至少一种控制序列。
本发明还提供了包含载体的宿主细胞，所述载体包含编码本文提供的工程化酮还原酶变体的多核苷酸。
本发明还提供了产生本文提供的工程化酮还原酶变体的方法，所述方法包括在本文提供的宿主细胞藉以产生工程化酮还原酶变体的条件下培养所述宿主细胞。在一些实施方案中，该方法还包括回收由宿主细胞产生的工程化酮还原酶变体的步骤。
本发明还提供了固定化的工程化酮还原酶变体。
本发明还提供了包含至少一种本文提供的工程化酮还原酶变体的组合物。在一些实施方案中，组合物包含至少一种本文提供的固定化的工程化酮还原酶变体。
本发明还提供了工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQID NO:172和/或SEQ ID NO:208具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性。
本发明还提供了工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQID NO:172具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：10/73/78/137/323/325、10/73/78/233/323、10/73/137、13/41/63/132/193/195、18/44/119/124/132/137/145/158/175/177/293/317/323、18/44/119/124/132/137/145/158/177/293/323、18/44/119/124/132/137/145/293/323/334/336、32/44/132/137/145/186/233/293/323/336、41/44/88/193/195、44/69/120/132/137/145/175/195/293/323、44/113/132/145、44/119/132/137/145/158/175/177/293/317/323、44/132/135/136/137/145/293、44/132/136/137/145/293、44/132/137/145/233/308/323、44/132/137/145/293/323、44/132/145、44/132/145/195/293/323、137/233/303/323和266，其中所述位置根据SEQ ID NO:172来编号。在一些实施方案中，工程化亚磷酸脱氢酶变体包含选自以下的至少一个取代或取代集：10K/73A/78Y/137Q/323D/325A、10K/73A/78Y/233I/323D、10K/73A/137Q、13D/41A/63A/132Q/193S/195E、18M/44A/119F/124E/132Q/137I/145G/158K/175S/177T/293L/317R/323D、18M/44A/119F/124E/132Q/137I/145G/158K/177T/293L/323D、18M/44A/119F/124E/132Q/137I/145G/293L/323D/334K/336R、32V/44A/132Q/137I/145G/186T/233I/293L/323D/336S、41A/44A/88R/193S/195E、44A/69K/120V/132Q/137I/145G/175T/195E/293L/323D、44A/113S/132Q/145G、44A/119F/132Q/137I/145G/158K/175S/177T/293L/317R/323D、44A/132Q/135A/136D/137I/145G/293L、44A/132Q/136D/137Q/145G/293L、44A/132Q/137I/145G/233I/308V/323D、44A/132Q/137I/145G/293L/323D、44A/132Q/145G、44A/132Q/145G/195E/293L/323D、137Q/233I/303A/323D和266S/V/W，其中所述位置根据SEQ ID NO:172来编号。在一些另外的实施方案中，工程化亚磷酸脱氢酶变体包含选自以下的至少一个取代或取代集：R10K/C73A/F78Y/R137Q/N323D/V325A、R10K/C73A/F78Y/V233I/N323D、R10K/C73A/R137Q、E13D/R41A/Q63A/R132Q/A193S/S195E、L18M/R44A/L119F/A124E/R132Q/R137I/N145G/L158K/A175S/K177T/I293L/A317R/N323D、L18M/R44A/L119F/A124E/R132Q/R137I/N145G/L158K/K177T/I293L/N323D、L18M/R44A/L119F/A124E/R132Q/R137I/N145G/I293L/N323D/A334K/C336R、S32V/R44A/R132Q/R137I/N145G/R186T/V233I/I293L/N323D/C336S、R41A/R44A/A88R/A193S/S195E、R44A/R69K/R120V/R132Q/R137I/N145G/A175T/S195E/I293L/N323D、R44A/V113S/R132Q/N145G、R44A/L119F/R132Q/R137I/N145G/L158K/A175S/K177T/I293L/A317R/N323D、R44A/R132Q/Q135A/P136D/R137I/N145G/I293L、R44A/R132Q/P136D/R137Q/N145G/I293L、R44A/R132Q/R137I/N145G/V233I/A308V/N323D、R44A/R132Q/R137I/N145G/I293L/N323D、R44A/R132Q/N145G、R44A/R132Q/N145G/S195E/I293L/N323D、R137Q/V233I/E303A/N323D和E266S/V/W，其中所述位置根据SEQ IDNO:172来编号。
本发明还提供了工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQID NO:208具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：32/59/124/177/191/327、78/150/198/327/328、83/266、95/211/213/322、104、178/194/211/213/322、206、211/213/322、215、262、266和323，其中所述位置根据SEQ ID NO:208来编号。在一些实施方案中，工程化亚磷酸脱氢酶变体包含选自以下的至少一个取代或取代集：32V/59M/124E/177S/191H/327D、78Y/150I/198L/327S/328P、83A/266A、95I/211A/213Q/322M、104F/L、178P/194L/211A/213Q/322Q、206N、211A/213Q/322Q、215P、262D/P、266S和323N，其中所述位置根据SEQ ID NO:208来编号。在一些另外的实施方案中，工程化亚磷酸脱氢酶变体包含选自以下的至少一个取代或取代集：S32V/A59M/A124E/T177S/Q191H/R327D、F78Y/F150I/F198L/R327S/L328P、V83A/E266A、F95I/N211A/D213Q/I322M、T104F/L、A178P/C194L/N211A/D213Q/I322Q、L206N、N211A/D213Q/I322Q、L215P、V262D/P、E266S和D323N，其中所述位置根据SEQ ID NO:208来编号。
本发明还提供了工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体包含含有与SEQ ID NO:172和/或SEQ ID NO:208具有至少90％序列同一性的序列的多肽序列。在一些实施方案中，工程化亚磷酸脱氢酶变体包含含有与SEQ ID NO:172和/或SEQ ID NO:208具有至少95％序列同一性的序列的多肽序列。在一些另外的实施方案中，工程化亚磷酸脱氢酶变体包含SEQ ID NO:172或SEQ ID NO:208中列出的多肽序列。在一些另外的实施方案中，工程化亚磷酸脱氢酶变体包含编码表9.1、表10.1和/或表11.1中提供的变体的多肽序列。在仍一些另外的实施方案中，工程化亚磷酸脱氢酶变体包含选自SEQ ID NO:174至SEQ ID NO:260中列出的偶数编号的序列的多肽序列。
本发明还提供了固定化的工程化亚磷酸脱氢酶变体。在一些实施方案中，本发明提供了至少一种本文提供的固定化的工程化酮还原酶变体和至少一种本文提供的工程化亚磷酸脱氢酶变体的混合物。
本发明还提供了包含至少一种本文提供的亚磷酸脱氢酶变体的组合物。在一些实施方案中，本发明还提供了包含至少一种本文提供的工程化酮还原酶变体和至少一种本文提供的工程化亚磷酸脱氢酶的混合物的组合物。
本发明还提供了编码本文提供的工程化亚磷酸脱氢酶变体的工程化多核苷酸序列。在一些实施方案中，工程化多核苷酸序列包含与选自SEQID NO:171至SEQ ID NO:259中列出的奇数编号的序列的序列至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多相同的多核苷酸序列。
本发明还提供了包含编码本文提供的工程化亚磷酸脱氢酶变体的工程化多核苷酸序列的载体。在一些实施方案中，载体还包含至少一种控制序列。在仍一些另外的实施方案中，载体包含编码本文提供的工程化亚磷酸脱氢酶变体的至少一种工程化多核苷酸序列和编码本文提供的工程化酮还原酶变体的至少一种工程化多核苷酸序列。本发明还提供了包含本文提供的载体的宿主细胞。
本发明还提供了用于产生本文提供的工程化亚磷酸脱氢酶变体的方法，所述方法包括在包含含有编码至少一种本发明的工程化亚磷酸脱氢酶的至少一种工程化多核苷酸序列的载体的宿主细胞藉以产生工程化亚磷酸脱氢酶变体的条件下培养所述宿主细胞。在一些实施方案中，宿主细胞包含载体，所述载体包含多核苷酸序列，所述多核苷酸序列包含本文提供的至少一种工程化酮还原酶和至少一种工程化亚磷酸脱氢酶。在一些另外的实施方案中，宿主细胞包含至少一种本文未提供的酮还原酶，但包含至少一种本文提供的工程化亚磷酸脱氢酶变体。在一些另外的实施方案中，宿主细胞包含至少一种本文未提供的亚磷酸脱氢酶，但包含至少一种本文提供的工程化酮还原酶变体。在一些实施方案中，该方法还包括回收由宿主细胞产生的工程化亚磷酸脱氢酶变体的步骤。在宿主细胞产生至少一种酮还原酶和至少一种亚磷酸脱氢酶的实施方案中，一些方法还包括回收由宿主细胞产生的酮还原酶和/或亚磷酸脱氢酶的步骤。
本发明还提供了使手性醇去外消旋化的方法，所述方法包括，在使得手性醇被去外消旋化的条件下，提供至少一种本文提供的工程化酮还原酶变体，提供至少一种本文提供的工程化亚磷酸脱氢酶变体、至少一种手性醇和至少一种辅因子。在一些实施方案中，该方法以一锅反应来进行，而在一些可选择的实施方案中，使用多个反应容器。
附图简述
图1提供了本发明提出的反应方案。
图2提供了底物异构体和产物异构体的结构。
图3提供了一锅多酶反应方案。
图4和图5提供了辅因子竞争测定方案。
图6提供了一锅多酶反应中获得的产物的HPLC色谱图。
发明描述
本发明提供了与天然存在的野生型酮还原酶和亚磷酸脱氢酶相比具有改进的特性的工程化酮还原酶和工程化亚磷酸脱氢酶、以及编码工程化酮还原酶和工程化亚磷酸脱氢酶的多核苷酸、能够表达工程化酮还原酶和工程化亚磷酸脱氢酶的宿主细胞、以及使用工程化酮还原酶和工程化亚磷酸脱氢酶在一锅多酶系统中使外消旋醇去外消旋化的方法。
定义
关于本发明，本文描述中使用的技术和科学术语将具有本领域普通技术人员通常理解的含义，除非另有具体定义。因此，意图以下术语具有以下含义。本文提及的所有专利和出版物，包括这些专利和出版物中公开的所有序列，都通过引用明确并入。除非另外说明，否则本发明的实践涉及本领域技术人员已知的分子生物学、发酵、微生物学和相关领域中通常使用的常规技术。除非本文另外定义，否则本文使用的所有技术和科学术语具有与本发明所属领域的普通技术人员通常理解的相同的含义。尽管与本文描述的那些相似或等效的任何方法和材料都可以用于实践或测试本发明，但描述了优选的方法和材料。实际上，意图是本发明不受限于本文描述的特定方法、方案和试剂，因为这些可以根据使用它们的情况而变化。本文提供的标题不是对本发明的各方面或实施方案的限制。
尽管如此，为了便于理解本发明，以下定义了许多术语。数值范围包括限定该范围的数值。因此，本文公开的每个数值范围意图包括落在此较宽数值范围内的每个较窄数值范围，如同这些较窄数值范围被全部明确写入本文。还意图本文公开的每个最大(或最小)的数值限制包括每个较低(或较高)的数值限制，如同这些较低(或较高)的数值限制被明确写入本文。
如本文使用的，术语“包括/包含/含有(comprising)”及其同源词以其包含性含义来使用(即，等同于术语“包括/包含/含有(including)”及其对应的同源词)。
如本文和所附权利要求中使用的，单数形式“一(a)”、“一(an)”和“该(the)”包括复数指代物，除非上下文另外清楚指明。因此，例如，对“宿主细胞”的提及包括多于一个这样的宿主细胞。
除非另外指示，否则，分别地，核酸以5'至3'方向从左至右书写；并且氨基酸序列以氨基至羧基方向从左至右书写。
本文提供的标题不是对通过参考本说明书整体可以获得的本发明的各方面或实施方案的限制。因此，以下定义的术语通过参考本说明书整体而被更充分地定义。
“酮还原酶”和“KRED”在本文中可互换使用，以指具有将羰基基团还原为其对应醇的酶促能力的多肽。更具体地，如方案1中示出的(参见图1)，本发明的酮还原酶多肽能够在一锅多酶系统中将式(I)的醇立体选择性地去外消旋化为式(II)的对应产物。
亚磷酸脱氢酶和“PDH”在本文中可互换使用，以指具有使NADPH辅因子再生的酶促能力的多肽。
如本文使用的，术语“一锅反应”是指在一个反应容器中使用多种酶(即KRED和PDH)从原材料产生产物。
如本文使用的，术语“蛋白”、“多肽”和“肽”在本文中可互换使用，以表示通过酰胺键共价连接的至少两个氨基酸的聚合物，而不论长度或翻译后修饰(例如，糖基化、磷酸化、脂质化、豆蔻酰化、泛素化等)。这个定义中包括D-氨基酸和L-氨基酸以及D-氨基酸和L-氨基酸的混合物。
如本文使用的，“多核苷酸”和“核酸”是指共价连接在一起的两个或更多个核苷。多核苷酸可以完全包含核糖核苷(即RNA)、完全包含2'脱氧核糖核苷酸(即DNA)或核糖核苷和2'脱氧核糖核苷的混合物。虽然核苷通常将通过标准磷酸二酯键连接在一起，但多核苷酸可以包括一个或更多个非标准连接。多核苷酸可以是单链或双链的，或者可以包括单链区和双链区两者。此外，虽然多核苷酸通常将变化天然存在的编码核苷碱基(即，腺嘌呤、鸟嘌呤、尿嘧啶、胸腺嘧啶和胞嘧啶)，但它也可以包含一种或更多种修饰和/或合成的核苷碱基(例如，肌苷、黄嘌呤、次黄嘌呤等)。优选地，这些修饰的或合成的核苷碱基将是编码核苷碱基。
如本文使用的，“编码序列”是指编码蛋白的氨基酸序列的那部分核酸(例如，基因)。
如本文使用的，“天然存在的”或“野生型”是指在自然界中存在的形式。例如，天然存在的或野生型多肽或多核苷酸序列是存在于可以从自然界的来源分离的生物体中并且未被通过人工操作有意修饰的序列。
如本文使用的，当在本发明中关于(例如，细胞、核酸或多肽)使用时，“非天然存在的”或“工程化”或“重组”是指如下材料或与该材料的天然或自然形式对应的材料：已经以自然界本来不存在的方式被修饰或与其相同但由合成材料产生或衍生和/或通过使用重组技术操作产生。非限制性实例包括，除其他以外，表达在天然(非重组)形式的细胞中不存在的基因或表达本来以不同水平表达的天然基因的重组细胞。
如本文使用的，“序列同一性百分比”、“同一性百分比”和“相同百分比”是指多核苷酸序列或多肽序列之间的比较，并通过在比较窗上比较两个最佳比对的序列来确定，其中与用于两个序列的最佳比对的参考序列相比，比较窗中的多核苷酸或多肽序列的部分可以包括添加或缺失(即，空位)。百分比如下计算：通过确定两个序列中出现相同的核酸碱基或核酸碱基或氨基酸残基中与空位对齐的位置的数目，以产生匹配位置的数目，将匹配位置的数目除以比较窗中位置的总数，并将结果乘以100以产生序列同一性的百分比。对最佳比对和序列同一性百分比的确定使用BLAST和BLAST 2.0算法来进行(例如参见，Altschul等人，J.Mol.Biol.215:403-410[1990]；和Altschul等人，Nucleic Acids Res.3389-3402[1977])。用于进行BLAST分析的软件可通过国家生物技术信息中心(National Center forBiotechnology Information)网站公开获得。
简言之，BLAST分析包括首先通过识别查询序列中长度W的短字(short words)来识别高评分序列对(HSP)，所述长度W的短字在与数据库序列中的相同长度的字比对时，匹配或满足一定的正值的阈值评分T。T被称为相邻字评分阈值(Altschul等人，同上)。这些初始的相邻字击中(word hit)充当种子，用于启始搜索以发现包含它们的更长的HSP。然后，字击中沿每个序列在两个方向上延伸，直至累积比对评分不能增加。对于核苷酸序列，累积评分使用参数M(对于匹配残基对的奖励评分；永远＞0)和N(对于错配残基的惩罚评分；永远＜0)来计算。对于氨基酸序列，使用评分矩阵来计算累积评分。当以下情况时，字击中在每个方向上的延伸停止：累积比对评分从其达到的最大值下降了量X；由于累积一个或更多个负评分的残基比对，累积评分达到零或以下；或到达任一序列的末端。BLAST算法参数W、T和X决定比对的灵敏度和速度。BLASTN程序(对于核苷酸序列)使用以下作为缺省值：字长(W)为11、期望值(E)为10、M＝5、N＝-4、以及双链比较。对于氨基酸序列，BLASTP程序使用以下作为缺省值：字长(W)为3、期望值(E)为10、以及BLOSUM62评分矩阵(参见例如，Henikoff和Henikoff,Proc Natl Acad Sci USA 89:10915[1989])。
许多其他算法是可获得及本领域已知的，这些算法在提供两个序列的同一性百分比方面与BLAST起相似作用。用于比较的序列的最佳比对可以使用本领域已知的任何合适的方法进行(例如，通过Smith和Waterman,Adv.Appl.Math.2:482[1981]的局部同源性算法；通过Needleman和Wunsch,J.Mol.Biol.48:443[1970]的同源性比对算法；通过Pearson和Lipman,Proc.Natl.Acad.Sci.USA 85:2444[1988]的搜索相似性的方法；和/或通过这些算法的计算机化实现[GCG Wisconsin软件包中的GAP、BESTFIT、FASTA和TFASTA])，或通过使用本领域通常已知的方法进行目视检查。此外，序列比对和序列同一性百分比的确定可以使用所提供的缺省参数，利用GCG Wisconsin软件包(Accelrys,Madison WI)中的BESTFIT或GAP程序。
如本文中使用的，“参考序列”指另一序列被与其比较的限定序列。参考序列可以是更大的序列的子集；例如，全长基因或多肽序列的区段。通常，参考序列为至少20个核苷酸或氨基酸残基的长度、至少25个残基的长度、至少50个残基的长度，或者核酸或多肽的全长。由于两个多核苷酸或多肽可以各自(1)包含两个序列之间相似的序列(即，完整序列的一部分)，和(2)还可以包含两个序列之间不同的序列，因此两个(或更多个)多核苷酸或多肽之间的序列比较通常通过比较两个多核苷酸在“比较窗”上的序列来鉴定和比较序列相似性的局部区域来进行。术语“参考序列”不意图受限于野生型序列，并且可以包括工程化序列或改变的序列。例如，在一些实施方案中，“参考序列”可以是先前工程化或改变的氨基酸序列。
如本文使用的，“比较窗”是指至少约20个连续核苷酸位置或氨基酸残基的概念性区段，其中序列可以与至少20个连续核苷酸或氨基酸的参考序列比较，并且其中与参考序列(其不包含添加或缺失)相比，序列在比较窗中的部分可以包含20％或更少的添加或缺失(即，空位)，以获得两个序列的最佳比对。比较窗可以比20个连续残基更长，并且任选地包括30个、40个、50个、100个或更长的窗口。
当如本文使用的，当在特定氨基酸或多核苷酸序列的编号的上下文中使用时，“对应于”、“关于”或“相对于”是指，当将特定氨基酸或多核苷酸序列与指定参考序列相比时，该参考序列的残基的编号。换言之，特定聚合物的残基编号或残基位置相对于参考序列来指定，而不是由残基在特定氨基酸或多核苷酸序列内的实际数值位置指定。例如，可以通过引入空位将特定氨基酸序列诸如工程化酮还原酶的氨基酸序列与参考序列比对，以优化两个序列之间的残基匹配。在这些情况下，尽管存在空位，但是特定氨基酸或多核苷酸序列中残基的编号相对于与其比对的参考序列来进行。如本文使用的，对残基位置的指代，诸如以下进一步描述的“Xn”，应被理解为是指“对应于……的残基”，除非另外明确说明。因此，例如，“X94”是指多肽序列中位置94处的任何氨基酸。
如本文使用的，“立体选择性”指一种立体异构体相对于另一种立体异构体或另一组立体异构体在化学或酶促反应中的优先形成。立体选择性可以是部分的，此时一种立体异构体的形成优于另一种，或者立体选择性可以是完全的，此时仅形成一种立体异构体。当立体异构体是对映异构体时，立体选择性被称为对映选择性，即两种对映异构体的总和中一种对映异构体的分数(通常报道为百分比)。本领域中通常可选择地将其报道(通常为百分比)为根据下式从中计算的对映异构体过量(e.e.)：[主要对映异构体-次要对映异构体]/[主要对映异构体+次要对映异构体]。当立体异构体是非对映异构体时，立体选择性被称为非对映选择性，即两种非对映异构体的混合物中一种非对映异构体的分数(通常报道为百分比)，通常可选择地报告为非对映异构体过量(d.e.)。对映异构体过量和非对映异构体过量是立体异构体过量的类型。还应理解，立体选择性不限于单一立体异构体，并且可以描述立体异构体的组。
如本文使用的，“高立体选择性”是指能够以至少约75％的立体异构体过量将底物转化为其对应的手性醇产物的化学或酶促反应。
如本文使用的，“增加的酶促活性”和“增加的活性”是指工程化酶的改进的特性，其可以通过与参考酶相比，比活性(例如，产生的产物/时间/重量蛋白)的增加或底物向产物的转化百分比(例如，使用指定量的酮还原酶，在指定时间段内，起始量的底物向产物的转化百分比)的增加来表示。确定酶活性的示例性方法在实施例中提供。与酶活性相关的任何特性都可以被影响，包括经典的酶特性Km、Vmax或kcat，其改变可以导致酶促活性的增加。酮还原酶活性可以通过用于测量酮还原酶的标准测定中的任何一种来测量，诸如底物或产物浓度的变化，或辅因子浓度的变化(在辅因子再生系统的不存在下)。酶活性的比较使用限定的酶制剂、设定条件下的限定的测定和一种或更多种限定的底物来进行，如本文进一步详细描述的。通常，当比较细胞裂解物中的酶时，确定细胞的数目和测定的蛋白的量，并使用相同表达系统和相同宿主细胞以使由宿主细胞产生并存在于裂解物中的酶的量的变化最小化。
如本文使用的，“转化”是指将底物向对应产物的酶促转化。
如本文使用的，“转化百分比”是指在指定条件下在一定时间段内转化为产物的底物的百分比。因此，例如，酮还原酶多肽的“酶促活性”或“活性”可以表示为底物向产物的“转化百分比”。
如本文使用的，“热稳定的(thermostable)”或“热稳定的(thermal stable)”可互换使用以指这样的多肽，其与未处理的酶相比，当暴露于一组温度条件(例如，40℃-80℃)一定时间段(例如，0.5hr-24hr)时耐受失活，因此在暴露于升高的温度后，保持一定水平的残余活性(诸如例如，多于60％至80％)。
如本文使用的，“溶剂稳定的”是指与未处理的酶相比，多肽在暴露于不同浓度(例如，5％-99％)的溶剂后，维持相似活性(例如，多于例如60％至80％)的能力。
如本文使用的，“氨基酸差异”或“残基差异”是指多肽序列的一个位置处的氨基酸残基相对于参考序列中对应位置处的氨基酸残基的差异。本文中氨基酸差异的位置通常被称为“Xn”，其中n是指残基差异所基于的参考序列中的对应位置。例如，“与SEQ ID NO:2相比在位置X40处的残基差异”是指在对应于SEQ ID NO:2的位置40的多肽位置处的氨基酸残基的差异。因此，如果SEQ ID NO:2的参考多肽在位置40处具有组氨酸，那么“与SEQ ID NO:2相比在位置X40处的残基差异”是指在对应于SEQ ID NO:2的位置40的多肽位置处除组氨酸以外的任何残基的氨基酸取代。在本文的大多数情况下，在一个位置处的特定氨基酸残基差异指示为“XnY”，其中“Xn”指定如上文描述的对应位置，并且“Y”是在工程化多肽中发现的氨基酸的单字母标识符(即，与参考多肽中的不同的残基)。在一些情况下，本发明还提供由常规符号“AnB”表示的特定氨基酸差异，其中A是参考序列中的残基的单字母标识符，“n”是参考序列中的残基位置的编号，并且B是工程化多肽序列中残基取代的单字母标识符。在一些情况下，本发明的多肽可以包含相对于参考序列的一个或更多个氨基酸残基差异，所述氨基酸残基差异由相对于参考序列存在残基差异的指定位置的列表指示。在一些实施方案中，当多于一个氨基酸可以在多肽的特定残基位置中使用时，可以使用的不同氨基酸残基由“/”隔开(例如，X192A/G)。本发明包括包含一个或更多个氨基酸差异的工程化多肽序列，所述一个或更多个氨基酸差异包括保守氨基酸取代和非保守氨基酸取代的任一种/或两者。本发明的序列表中包括的特定重组碳酸酐酶多肽的氨基酸序列包括起始甲硫氨酸(M)残基(即，M代表残基位置1)。然而，技术人员理解，该起始甲硫氨酸残基可以通过诸如宿主细胞中或体外翻译系统中的生物加工机制去除，以产生缺乏起始甲硫氨酸残基但在其他方面保留酶的特性的成熟蛋白。因此，如本文使用的术语“相对于SEQ ID NO:2在位置Xn处的氨基酸残基差异”可以指位置“Xn”或已经被加工以便缺少起始甲硫氨酸的参考序列的对应位置(例如，位置(X-1)n)。
如本文使用的，措辞“保守氨基酸取代”是指具有相似侧链的残基的可互换性，并且因此通常包括用相同或相似的氨基酸定义类别内的氨基酸取代多肽中的氨基酸。通过实例且非限制性的方式，在一些实施方案中，具有脂肪族侧链的氨基酸被另一种脂肪族氨基酸(例如，丙氨酸、缬氨酸、亮氨酸和异亮氨酸)取代；具有羟基侧链的氨基酸被另一种具有羟基侧链的氨基酸(例如，丝氨酸和苏氨酸)取代；具有芳香族侧链的氨基酸被另一种具有芳香族侧链的氨基酸(例如，苯丙氨酸、酪氨酸、色氨酸和组氨酸)取代；具有碱性侧链的氨基酸被另一种具有碱性侧链的氨基酸(例如，赖氨酸和精氨酸)取代；具有酸性侧链的氨基酸被另一种具有酸性侧链的氨基酸(例如，天冬氨酸或谷氨酸)取代；和/或疏水性或亲水性氨基酸分别被另一种疏水性或亲水性氨基酸取代。示例性保守取代在表1中提供。
如本文使用的，措辞“非保守取代”是指用具有显著不同的侧链特性的氨基酸取代多肽中的氨基酸。非保守取代可以使用定义的组之间而非定义的组之内的氨基酸，并且影响(a)取代区域中的肽骨架的结构(例如，脯氨酸取代甘氨酸)，(b)电荷或疏水性，或(c)侧链的体积。通过实例且非限制性的方式，示例性非保守取代可以是用碱性氨基酸或脂肪族氨基酸取代的酸性氨基酸；用小氨基酸取代的芳香族氨基酸；以及用疏水性氨基酸取代的亲水性氨基酸。
如本文使用的，“缺失”是指通过从参考多肽去除一个或更多个氨基酸来修饰多肽。缺失可以包括去除1个或更多个氨基酸、2个或更多个氨基酸、5个或更多个氨基酸、10个或更多个氨基酸、15个或更多个氨基酸、或者20个或更多个氨基酸、多达构成多肽的氨基酸总数的10％或多达构成多肽的氨基酸总数的20％，同时保留酶促活性和/或保留工程化酶的改进的特性。缺失可以涉及多肽的内部部分和/或末端部分。在各种实施方案中，缺失可以构成连续的区段，或者可以是不连续的。
如本文使用的，“插入”是指通过向参考多肽添加一个或更多个氨基酸来修饰多肽。在一些实施方案中，改进的工程化酮还原酶包括向天然存在的酮还原酶多肽插入一个或更多个氨基酸，以及向工程化酮还原酶多肽插入一个或更多个氨基酸。插入可以在多肽的内部部分，或在羧基或氨基末端。如本文使用的插入包括如本领域已知的融合蛋白。插入可以是氨基酸的连续区段，或由天然存在的多肽中的一个或更多个氨基酸隔开。
术语“氨基酸取代集”或“取代集”是指与参考序列相比，多肽序列中的一组氨基酸取代。取代集可以具有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个或更多个氨基酸取代。在一些实施方案中，取代集是指存在于实施例中提供的表格中所列的变体KRED的任一个中的氨基酸取代集。
如本文使用的，“片段”是指具有氨基末端和/或羧基末端缺失，但其中剩余的氨基酸序列与序列中的对应位置相同的多肽。片段通常可以具有全长酮还原酶多肽例如多肽SEQ ID NO:4的约80％、约90％、约95％、约98％或约99％。在一些实施方案中，片段是“有生物活性的”(即，它表现出与全长序列相同的酶促活性)。
如本文使用的，“分离的多肽”是指与天然伴随多肽的其他污染物，例如，蛋白、脂质和多核苷酸基本上分离的多肽。该术语包括已经从其天然存在的环境或表达系统(例如，宿主细胞或体外合成)中取出或纯化的多肽。改进的酮还原酶可以存在于细胞内、存在于细胞培养基中，或以各种形式制备，诸如裂解物或分离的制品。如此，在一些实施方案中，本发明的工程化酮还原酶多肽可以是分离的多肽。
如本文使用的，“基本上纯的多肽”是指其中多肽物质是存在的主要物质(即，以摩尔或重量计，其比组合物中的任何其他单独的大分子物质更丰富)的组合物，并且当目标物质以摩尔或％重量计构成存在的大分子物质的至少约50％时，该组合物通常为基本上纯的组合物。通常，以存在于组合物中的所有大分子物质的摩尔或％重量计，基本上纯的工程化酮还原酶多肽组合物将占约60％或更多、约70％或更多、约80％或更多、约90％或更多、约91％或更多、约92％或更多、约93％或更多、约94％或更多、约95％或更多、约96％或更多、约97％或更多、约98％或更多、或约99％。溶剂物质、小分子(<500道尔顿)和元素离子物质不被认为是大分子物质。在一些实施方案中，分离的改进的酮还原酶多肽是基本上纯的多肽组合物。
如本文使用的，当关于核酸或多肽使用时，术语“异源的”是指正常情况下生物体(例如，野生型生物体)不表达及分泌的序列。在一些实施方案中，该术语包括包含两个或更多个子序列的序列，发现所述子序列彼此之间关系与在自然界中正常存在的关系不同，或所述序列被重组工程化，使得其表达水平或与细胞中的其他核酸或其他分子的物理关系或结构不是正常存在于自然界中的。例如，异源核酸通常被重组地产生，具有以自然界中未发现的方式排列的来自不相关的基因的两个或更多个序列(例如，本发明的核酸开放阅读框(ORF)可操作地连接至被插入到表达盒诸如载体中的启动子序列)。在一些实施方案中，“异源多核苷酸”是指通过实验室技术被引入宿主细胞中的任何多核苷酸，并且包括从宿主细胞中取出、经受实验室操作、并且然后重新引入宿主细胞中的多核苷酸。
如本文使用的，“密码子优化”是指编码蛋白的多核苷酸的密码子向特定生物体中优选使用的密码子的改变，使得编码的蛋白在感兴趣的生物体中有效地表达。在一些实施方案中，编码酮还原酶的多核苷酸可以被密码子优化以用于从所选择的用于表达的宿主生物体优化产生。
如本文使用的，“控制序列”在本文中被定义为包括对本发明的多核苷酸和/或多肽的表达是必需或有利的所有组分。每个控制序列对于感兴趣的多核苷酸可以是天然的或外源的。这样的控制序列包括但不限于前导序列、多腺苷酸化序列、前肽序列、启动子、信号肽序列和转录终止子。
如本文使用的，“可操作地连接”在本文中被定义为控制序列被适当地放置(即，以功能性关系)在相对于感兴趣的多核苷酸序列的某一位置处的配置，使得控制序列指导或调控感兴趣的多核苷酸和/或多肽的表达。
如本文使用的，措辞“辅因子再生系统”或“辅因子再循环系统”是指参与还原辅因子的氧化形式的反应(例如，NADP+至NADPH)的一组反应物。由酮还原酶催化的酮底物的还原而氧化的辅因子通过辅因子再生系统以还原的形式再生。辅因子再生系统包括化学计量的还原剂，所述还原剂是还原氢等同物的来源并且能够还原辅因子的氧化形式。辅因子再生系统还可以包含催化剂，例如催化由还原剂还原辅因子的氧化形式的酶催化剂。分别从NAD+或NADP+再生NADH或NADPH的辅因子再生系统是本领域已知的，并且可以用于本文描述的方法。
如本文使用的，“合适的反应条件”是指生物催化反应溶液中的那些条件(例如，酶载量、底物载量、辅因子载量、温度、pH、缓冲剂、共溶剂等的范围)，在这些条件下本发明的酮还原酶多肽能够立体选择性地使底物化合物去外消旋化为产物化合物。在本发明中提供并通过实施例说明了示例性的“合适的反应条件”。
如本文使用的，诸如“化合物载量”、“酶载量”或“辅因子载量”中的“载量”是指在反应开始时反应混合物中组分的浓度或量。
如本文使用的，在生物催化剂介导的方法的上下文中，“底物”是指由生物催化剂作用的化合物或分子。例如，在本文公开的方法中，酮还原酶生物催化剂的示例性底物是化合物(1)。
如本文使用的，在生物催化剂介导的方法的上下文中，“产物”是指由生物催化剂的作用产生的化合物或分子。
如本文使用的，“平衡”是指在化学或酶促反应中产生稳定状态浓度的化学物质的过程(例如，两种物质A和B的相互转化)，包括立体异构体的相互转化，如通过该化学或酶促反应的正向速率常数和逆向速率常数确定的。
如本文使用的，“氧代”是指＝O。
如本文使用的，“氧基”是指二价基团-O-，其可以具有各种取代基以形成不同的氧基基团，包括醚和酯。
如本文使用的，“羧基”是指-COOH。
如本文使用的，“羰基”是指-C(O)-，其可以具有各种取代基以形成不同的羰基基团，包括酸、酰基卤、醛、酰胺、酯和酮。
如本文使用的，“羟基”是指-OH。
如本文使用的，“任选的”和“任选地”意指随后描述的事件或情形可以发生或可以不发生，并且该描述包括其中事件或情形发生的情况和其中事件或情形不发生的情况。本领域普通技术人员将理解，对于被描述为含有一个或更多个任选的取代基的任何分子，仅意图包括空间上可实现的和/或合成上可行的化合物。
如本文使用的，“任选地被取代的”是指术语或一系列化学基团中的所有后续修饰对象(modifier)。例如，在术语“任选地被取代的芳基烷基”中，分子的“烷基”部分和“芳基”部分可以被取代或可以不被取代，并且对于系列“任选地被取代的烷基、环烷基、芳基和杂芳基”，烷基、环烷基、芳基和杂芳基彼此独立地可以被取代或可以不被取代。
工程化酶多肽
酮还原酶(KRED)或羰基还原酶生物催化剂(EC 1.1.1.184)可用于从醛和酮合成醇，以及从对应的前立体异构体酮底物合成光学活性仲醇。KRED也可以催化逆反应(即，醇底物向对应的醛/酮产物的氧化)。KRED对酮和醛的还原以及对醇的氧化使用辅因子，最常见的是还原型烟酰胺腺嘌呤二核苷酸(NADH)或还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)，以及用于氧化反应的烟酰胺腺嘌呤二核苷酸(NAD)或烟酰胺腺嘌呤二核苷酸磷酸(NADP+)。NADH和NADPH充当电子供体，而NAD+和NADP+充当电子受体。
如本领域已知的，KRED酶可见于广泛范围的细菌和酵母中(参见例如，Hummel和Kula Eur.J.Biochem.,184:1-13[1989])。已经报道了许多KRED基因和酶序列，包括木兰假丝酵母(Candida magnoliae)(Genbank登录号JC7338；GI:11360538)；近平滑假丝酵母(Genbank登录号BAA24528.1；GI:2815409)，赭色掷孢酵母(Sporobolomycessalmonicolor)(Genbank登录号AF160799；GI:6539734)，开菲尔乳杆菌(Lactobacilluskefir)(Genbank登录号AAP94029.1；GI:33112056)，短乳杆菌(Lactobacillus brevis)(Genbank登录号1NXQ_A；GI:30749782)和布氏嗜热厌氧菌(Thermoanaerobium brockii)(Genbank登录号P14941；GI:1771790)的那些KRED基因和酶序列。
酮还原酶的立体选择性已经被应用于制备重要的药物构建区块(pharmaceuticalbuilding block)(参见例如，Broussy等人，Org.Lett.,11:305-308[2009])。天然存在或工程化的KRED在产生有用的化学化合物的生物催化过程中的具体应用已针对以下被证明：4-氯乙酰乙酸酯的还原(参见例如，Zhou,J.Am.Chem.Soc.,105:5925-5926[1983]；Santaniello,J.Chem.Res.,(S)132-133[1984]；美国专利第5,559,030号；美国专利第5,700,670号；和美国专利第5,891,685号)、二氧代羧酸的还原(参见例如，美国专利第6,399,339号)、叔丁基(S)-氯-5-羟基-3-氧代己酸酯的还原(参见例如，美国专利第6,645,746号；和WO 01/40450)、基于吡咯并三嗪的化合物的还原(参见例如，美国申请公布第2006/0286646号)；取代的苯乙酮的还原(参见例如，美国专利第6,800,477号和第8,748,143号)、以及酮噻茂烷(ketothiolane)的还原(WO 2005/054491)。
本发明提供了能够在一锅多酶系统中使底物化合物(1)(6,7-二氢-5H-吡咯并[1,2-a]咪唑-7-醇)去外消旋化的工程化酮还原酶，如以下反应和图1中示出的。
本发明还提供了改进的酮还原酶和改进的亚磷酸脱氢酶，以及用于使用工程化酮还原酶和工程化亚磷酸脱氢酶在一锅多酶系统中使手性化合物去外消旋化的方法。
重要的是要注意，只有当氧化反应和还原反应是正交的、相容的且不相互影响时，才能在一锅、一步、多酶系统中获得期望的产物。只有当氧化性酮还原酶及其对应的再循环酶仅仅使用一种辅因子(例如，NAD+)并且还原性酮还原酶及其对应的再循环酶仅仅使用相对的辅因子(即，NADPH)时，这些条件才得到满足。
化合物(1)具有一个手性中心，并且可以以两种不同的非对映异构体形式(1a和1b)存在。通过串联的酮还原酶的去外消旋化反应可以产生两种不同的对映异构体产物(1a-1b)，如图2和下文中示出的。
然而，(1a)是唯一期望的产物。在本发明的开发中使用的演化程序被设计成改进S-选择性酮还原酶的活性，该酶将使外消旋混合物中的S-醇氧化，产生用于R-选择性酮还原酶的酮底物。此外，演化程序被设计成改进R-选择性酮还原酶的选择性、活性和辅因子偏好性。演化还被设计成改进亚磷酸脱氢酶的活性、稳定性和辅因子偏好性，从而能够以一锅、一步、多酶方法使底物(1)去外消旋化为产物(1a)及最小量的酮和(1b)。
选择SEQ ID NO:2的酮还原酶多肽作为用于开发本发明提供的改进的S-选择性酶的初始骨架。选择这种酶作为初始骨架，因为酮(2)仅通过(1b)的氧化产生，留下(1a)。SEQID NO:2的酮还原酶多肽使用NAD+作为辅因子，具有相对于NADP+的大于200:1的效率，并且可以与商业上可得的NADH氧化酶偶联以使辅因子再循环。
选择SEQ ID NO:2的酮还原酶多肽作为开发将酮还原为产物(1a)的R-选择性酶的初始骨架，具有初始选择性为92.7％e.e.。本文中对映选择性值根据以下提供的等式(1)来计算。
(1){[(1a量)-(1b量)]/[(1a量)+(1b量)]}x 100
实际上，本发明的非天然存在的酮还原酶多肽是被工程化为与SEQ IDNO:2的天然存在的酮还原酶相比具有改进的特性的酮还原酶。
选择亚磷酸脱氢酶多肽作为开发改进的PDH酶的初始骨架。这种酶同样有效地使NADH和NADPH两者再循环。
在一些实施方案中，工程化酮还原酶多肽能够在合适的反应条件下以相对于SEQID NO:2的参考多肽的活性增加了至少约1.2倍、1.5倍、2倍、3倍、4倍、5倍、10倍、20倍、30倍、40倍、50倍或100倍的活性将底物化合物转化为产物。在一些实施方案中，工程化酮还原酶多肽能够在合适的反应条件下，以约48h、约36h、约24h的反应时间或甚至更短的时间长度，以至少约40％、至少约50％、至少约60％、至少约70％、至少约80％、或至少约90％、至少约95％、至少约98％、至少约99％的转化率百分比将底物化合物转化为产物。
在一些实施方案中，工程化酮还原酶和亚磷酸脱氢酶能够在一锅、一步、多酶系统中将底物化合物(1)转化为相对于化合物(1b)呈对映异构体过量的产物化合物(1a)。在一些实施方案中，工程化酮还原酶和亚磷酸脱氢酶能够在合适的反应条件下将化合物(1)转化为相对于化合物(1b)呈非对映异构体过量的化合物(1a)。
如本领域技术人员将理解的，除非另外说明，否则以上定义的类别中的一些并不相互排斥。因此，具有显示两种或更多种物理化学特性的侧链的氨基酸可以被包括在多个类别中。对任何氨基酸或残基的适当分类对本领域技术人员来说将是明显的，特别是根据本文提供的详细发明。
在一些实施方案中，改进的工程化酮还原酶和工程化亚磷酸脱氢酶包含天然存在的酮还原酶或亚磷酸脱氢酶多肽中的氨基酸残基缺失、或其他工程化酮还原酶或亚磷酸脱氢酶多肽中的氨基酸残基缺失。因此，在本发明的一些实施方案中，缺失包括酮还原酶多肽的1个或更多个氨基酸、2个或更多个氨基酸、3个或更多个氨基酸、4个或更多个氨基酸、5个或更多的氨基酸、6个或更多个氨基酸、8个或更多个氨基酸、10个或更多个氨基酸、15个或更多个氨基酸、或20个或更多个氨基酸、多达氨基酸总数的10％、多达氨基酸总数的10％、多达氨基酸总数的20％、或多达氨基酸总数的30％的缺失，条件为酮还原酶的功能活性或亚磷酸脱氢酶活性被保持。在一些实施方案中，缺失可以包括1-2个、1-3个、1-4个、1-5个、1-6个、1-7个、1-8个、1-9个、1-10个、1-11个、1-12个、1-14个、1-15个、1-16个、1-18个、1-20个、1-22个、1-24个、1-25个、1-30个、1-35个或约1-40个氨基酸残基的缺失。在一些实施方案中，缺失的数目可以为1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、14个、15个、16个、18个、20个、22个、24个、26个、30个、35个或约40个氨基酸。在一些实施方案中，缺失可以包括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、18个或20个氨基酸残基的缺失。
如本文描述的，本发明的酮还原酶多肽或亚磷酸脱氢酶多肽可以呈融合多肽的形式，其中酮还原酶多肽或亚磷酸脱氢酶多肽与其他多肽诸如抗体标签(例如，myc表位)或纯化序列(例如，His标签)融合。因此，在一些实施方案中，可使用与其他多肽融合或不与其他多肽融合的酮还原酶多肽和/或亚磷酸脱氢酶多肽。
在一些实施方案中，本文描述的多肽不限于遗传编码氨基酸。除了遗传编码氨基酸之外，本文描述的多肽可以完全或部分地包含天然存在的和/或合成的非编码氨基酸。本文描述的多肽可以包含的某些常见的非编码氨基酸包括但不限于：遗传编码氨基酸的D-立体异构体；2,3-二氨基丙酸(Dpr)；α-氨基异丁酸(Aib)；ε-氨基己酸(Aha)；δ-氨基戊酸(Ava)；N-甲基甘氨酸或肌氨酸(MeGly或Sar)；鸟氨酸(Orn)；瓜氨酸(Cit)；叔丁基丙氨酸(Bua)；叔丁基甘氨酸(Bug)；N-甲基异亮氨酸(MeIle)；苯基甘氨酸(Phg)；环己基丙氨酸(Cha)；正亮氨酸(Nle)；萘基丙氨酸(Nal)；2-氯苯丙氨酸(Ocf)；3-氯苯丙氨酸(Mcf)；4-氯苯丙氨酸(Pcf)；2-氟苯丙氨酸(Off)；3-氟苯丙氨酸(Mff)；4-氟苯丙氨酸(Pff)；2-溴苯丙氨酸(Obf)；3-溴苯丙氨酸(Mbf)；4-溴苯丙氨酸(Pbf)；2-甲基苯丙氨酸(Omf)；3-甲基苯丙氨酸(Mmf)；4-甲基苯丙氨酸(Pmf)；2-硝基苯丙氨酸(Onf)；3-硝基苯丙氨酸(Mnf)；4-硝基苯丙氨酸(Pnf)；2-氰基苯丙氨酸(Ocf)；3-氰基苯丙氨酸(Mcf)；4-氰基苯丙氨酸(Pcf)；2-三氟甲基苯丙氨酸(Otf)；3-三氟甲基苯丙氨酸(Mtf)；4-三氟甲基苯丙氨酸(Ptf)；4-氨基苯丙氨酸(Paf)；4-碘苯丙氨酸(Pif)；4-氨基甲基苯丙氨酸(Pamf)；2,4-二氯苯丙氨酸(Opef)；3,4-二氯苯丙氨酸(Mpcf)；2,4-二氟苯丙氨酸(Opff)；3,4-二氟苯丙氨酸(Mpff)；吡啶-2-基丙氨酸(2pAla)；吡啶-3-基丙氨酸(3pAla)；吡啶-4-基丙氨酸(4pAla)；萘-1-基丙氨酸(1nAla)；萘-2-基丙氨酸(2nAla)；噻唑基丙氨酸(taAla)；苯并噻吩基丙氨酸(bAla)；噻吩基丙氨酸(tAla)；呋喃基丙氨酸(fAla)；高苯丙氨酸(hPhe)；高酪氨酸(hTyr)；高色氨酸(hTrp)；五氟苯丙氨酸(5ff)；苯乙烯基丙氨酸(sAla)；蒽基丙氨酸(aAla)；3,3-二苯基丙氨酸(Dfa)；3-氨基-5-苯基戊酸(Afp)；青霉胺(Pen)；1,2,3,4-四氢异喹啉-3-羧酸(Tic)；β-2-噻吩基丙氨酸(Thi)；甲硫氨酸亚砜(Mso)；N(w)-硝基精氨酸(nArg)；高赖氨酸(hLys)；膦酰基甲基苯丙氨酸(pmPhe)；磷酸丝氨酸(pSer)；磷酸苏氨酸(pThr)；高天冬氨酸(hAsp)；高谷氨酸(hGlu)；1-氨基环戊-(2或3)-烯-4-羧酸；哌啶酸(PA)；氮杂环丁烷-3-羧酸(ACA)；1-氨基环戊烷-3-羧酸；烯丙基甘氨酸(aOly)；炔丙基甘氨酸(pgGly)；高丙氨酸(hAla)；正缬氨酸(nVal)；高亮氨酸(hLeu)、高缬氨酸(hVal)；高异亮氨酸(hIle)；高精氨酸(hArg)；N-乙酰赖氨酸(AcLys)；2,4-二氨基丁酸(Dbu)；2,3-二氨基丁酸(Dab)；N-甲基缬氨酸(MeVal)；高半胱氨酸(hCys)；高丝氨酸(hSer)；羟基脯氨酸(Hyp)和高脯氨酸(hPro)。本文描述的多肽可以包含的另外的非编码氨基酸对本领域技术人员是明显的。这些氨基酸可以处于L-构型或D-构型。
本领域技术人员将认识到，带有侧链保护基团的氨基酸或残基也可以构成本文描述的多肽。在这种情况下属于芳香族类别的这些受保护的氨基酸的非限制性实例包括(保护基团在圆括号中列出)但不限于：Arg(tos)、Cys(甲苄基)、Cys(硝基吡啶亚磺酰基)、Glu(δ-苄基酯)、Gln(呫吨基)、Asn(N-δ-呫吨基)、His(bom)、His(苄基)、His(tos)、Lys(fmoc)、Lys(tos)、Ser(O-苄基)、Thr(O-苄基)和Tyr(O-苄基)。
可以构成本文描述的多肽的构象上受限制的非编码氨基酸包括但不限于N-甲基氨基酸(L-构型)；1-氨基环戊-(2或3)-烯-4-羧酸；哌可酸(pipecolic acid)；氮杂环丁烷-3-羧酸；高脯氨酸(hPro)；以及1-氨基环戊烷-3-羧酸。
如以上描述的，被引入天然存在的多肽以产生工程化酮还原酶和工程化亚磷酸脱氢酶的各种修饰可以靶向酶的特定特性。
编码工程化酶的多核苷酸
在另一方面中，本发明提供了编码工程化酮还原酶和工程化亚磷酸脱氢酶的多核苷酸。可以将多核苷酸可操作地连接至控制基因表达的一个或更多个异源调控序列，以创建能够表达多肽的重组多核苷酸。可以将包含编码工程化酮还原酶和/或工程化亚磷酸脱氢酶的异源多核苷酸的表达构建体引入适当的宿主细胞中来表达对应的酮还原酶多肽或工程化亚磷酸脱氢酶多肽。
因为对各种氨基酸所对应的密码子的了解，蛋白序列的可用性提供了对能够编码主题的所有多核苷酸的描述。遗传密码的简并性，其中相同的氨基酸由可替代的密码子或同义密码子编码，允许制备极大数目的核酸，所有这些核酸都编码本文公开的改进的酮还原酶和/或改进的亚磷酸脱氢酶。因此，已经鉴定了特定的氨基酸序列后，本领域技术人员可以通过以不改变蛋白的氨基酸序列的方式简单修改序列的一个或更多个密码子来制备任何数目的不同核酸。在此方面，本发明特别设想了通过选择基于可能的密码子选择的组合可以进行的每个和每一个可能的多核苷酸变化，并且所有这样的变化应被认为针对本文公开的任何多肽(包括实施例中的表格中呈现的氨基酸序列)具体公开。在各种实施方案中，优选地选择密码子来适应在其中产生蛋白的宿主细胞。例如，在细菌中使用的优选的密码子被用于在细菌中表达基因；在酵母中使用的优选的密码子被用于酵母中的表达；并且在哺乳动物中使用的优选的密码子被用于在哺乳动物细胞中表达。
在一些实施方案中，工程化酮还原酶序列或亚磷酸酯脱氢酶序列包括包含被鉴定为有益的位置的序列，如实施例中描述的。
在一些实施方案中，将编码改进的酮还原酶多肽或亚磷酸脱氢酶多肽的分离的多核苷酸以多种方式操作，以提供多肽的改进的表达和/或产生。取决于使用的表达载体，在将分离的多核苷酸插入载体前对分离的多核苷酸的操作可能是期望的或必要的。用于利用重组DNA方法修饰多核苷酸和核酸序列的技术是本领域熟知的。
对于细菌宿主细胞，用于指导本发明的核酸构建体的转录的合适的启动子包括从以下获得的启动子：大肠杆菌(E.coli)lac操纵子、天蓝色链霉菌(Streptomycescoelicolor)琼脂糖酶基因(dagA)、枯草芽孢杆菌(Bacillus subtilis)果聚糖蔗糖酶基因(sacB)、地衣芽孢杆菌(Bacillus licheniformis)α-淀粉酶基因(amyL)、嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)麦芽糖淀粉酶基因(amyM)、解淀粉芽孢杆菌(Bacillusamyloliquefaciens)α-淀粉酶基因(amyQ)、地衣芽孢杆菌青霉素酶基因(penP)、枯草芽孢杆菌xylA和xylB基因、及原核生物β-内酰胺酶基因(参见例如，Villa-Kamaroff等人，Proc.Natl Acad.Sci.USA 75:3727-3731[1978])、以及tac启动子(参见例如，DeBoer等人，Proc.Natl Acad.Sci.USA 80:21-25[1983])。另外的合适的启动子对于本领域技术人员是已知的。
对于丝状真菌宿主细胞，用于指导本发明的核酸构建体的转录的合适的启动子包括从以下的基因获得的启动子：米曲霉(Aspergillus oryzae)TAKA淀粉酶、米黑根毛霉(Rhizomucor miehei)天冬氨酸蛋白酶、黑曲霉(Aspergillus niger)中性α-淀粉酶、黑曲霉酸稳定型α-淀粉酶、黑曲霉或泡盛曲霉(Aspergillus awamori)葡糖淀粉酶(glaA)、米黑根毛霉脂肪酶、米曲霉碱性蛋白酶、米曲霉磷酸丙糖异构酶、构巢曲霉(Aspergillusnidulans)乙酰胺酶和尖孢镰刀菌(Fusarium oxysporum)胰蛋白酶样蛋白酶(WO96/00787)、以及NA2-tpi启动子(来自黑曲霉中性α-淀粉酶基因和米曲霉磷酸丙糖异构酶基因的启动子的杂合体)及其突变启动子、截短启动子和杂合启动子。
在酵母宿主中，可用的启动子包括但不限于来自以下的基因的启动子：酿酒酵母(Saccharomyces cerevisiae)烯醇化酶(ENO-1)、酿酒酵母半乳糖激酶(GAL1)、酿酒酵母醇脱氢酶/甘油醛-3-磷酸脱氢酶(ADH2/GAP)和酿酒酵母3-磷酸甘油酸激酶，以及用于酵母宿主细胞的其它可用的启动子(参见例如，Romanos等人，Yeast 8:423-488[1992])。
控制序列还可以是合适的转录终止子序列，转录终止子序列是由宿主细胞识别以终止转录的序列。终止子序列被可操作地连接至编码多肽的核酸序列的3'末端。在本发明中可以使用在所选择的宿主细胞中起作用的任何终止子。
例如，用于丝状真菌宿主细胞的示例性转录终止子可以从以下的基因获得：米曲霉TAKA淀粉酶、黑曲霉葡糖淀粉酶、构巢曲霉邻氨基苯甲酸合酶、黑曲霉α-葡糖苷酶、和尖孢镰刀菌胰蛋白酶样蛋白酶。
用于酵母宿主细胞的示例性终止子可以从以下的基因获得：酿酒酵母烯醇化酶、酿酒酵母细胞色素C(CYC1)、和酿酒酵母甘油醛-3-磷酸脱氢酶，以及本领域已知的用于酵母宿主细胞的其他可用的终止子(参见例如，Romanos等人，同上)。
控制序列还可以是合适的前导序列，前导序列是对宿主细胞的翻译重要的mRNA的非翻译区。前导序列被可操作地连接至编码多肽的核酸序列的5'末端。可以使用在所选择的宿主细胞中起作用的任何前导序列。用于丝状真菌宿主细胞的示例性前导序列从以下的基因获得：米曲霉TAKA淀粉酶和构巢曲霉磷酸丙糖异构酶。用于酵母宿主细胞的合适的前导序列从以下的基因获得：酿酒酵母烯醇化酶(ENO-1)、酿酒酵母3-磷酸甘油酸激酶、酿酒酵母α-因子、和酿酒酵母醇脱氢酶/甘油醛-3-磷酸脱氢酶(ADH2/GAP)。
控制序列还可以是多腺苷酸化序列，多腺苷酸化序列是被可操作地连接至核酸序列的3'末端的序列，并且其在转录时，被宿主细胞识别为将多腺苷残基添加至转录的mRNA的信号。在本发明中可以使用在所选择的宿主细胞中起作用的任何多腺苷酸化序列。用于丝状真菌宿主细胞的示例性多腺苷酸化序列可以来自以下的基因：米曲霉TAKA淀粉酶、黑曲霉葡糖淀粉酶、构巢曲霉邻氨基苯甲酸合酶、尖孢镰刀菌胰蛋白酶样蛋白酶和黑曲霉α-葡糖苷酶，以及本领域已知的用于酵母宿主细胞的其他可用的多腺苷酸化序列(参见例如，Guo等人，Mol.Cell.Biol.15:5983-5990[1995])。
控制序列还可以是信号肽编码区域，其编码与多肽的氨基末端连接并指导所编码的多肽进入细胞的分泌途径的氨基酸序列。核酸序列的编码序列的5'末端可以固有地包含信号肽编码区，所述信号肽编码区符合翻译阅读框地(in translation reading frame)与编码分泌多肽的编码区的区段天然地连接。可选择地，编码序列的5'末端可以包含对于编码序列是外源的信号肽编码区。当编码序列不天然包含信号肽编码区时可能需要外源信号肽编码区。
可选择地，外源信号肽编码区可以简单替换天然信号肽编码区以增加多肽的分泌。然而，指导所表达的多肽进入所选择的宿主细胞的分泌途径的任何信号肽编码区可以在本发明中使用。
用于细菌宿主细胞的有效的信号肽编码区是从以下的基因获得的信号肽编码区：芽孢杆菌NClB 11837麦芽糖淀粉酶、嗜热脂肪芽孢杆菌α-淀粉酶、地衣芽孢杆菌枯草杆菌蛋白酶、地衣芽孢杆菌β-内酰胺酶、嗜热脂肪芽孢杆菌中性蛋白酶(nprT、nprS、nprM)和枯草芽孢杆菌prsA、以及本领域已知的另外的信号肽(参见例如，Simonen等人，Microbiol.Rev.,57:109-137[1993])。
用于丝状真菌宿主细胞的有效的信号肽编码区包括但不限于从以下的基因获得的信号肽编码区：米曲霉TAKA淀粉酶、黑曲霉中性淀粉酶、黑曲霉葡糖淀粉酶、米黑根毛霉天冬氨酸蛋白酶、特异腐质霉(Humicolainsolens)纤维素酶和Humicola lanuginosa脂肪酶。用于酵母宿主细胞的可用的信号肽可以来自以下的基因：酿酒酵母α因子和酿酒酵母转化酶，以及另外的可用的信号肽编码区(参见例如，Romanos等人，1992，同上)。
控制序列还可以是编码定位于多肽的氨基末端的氨基酸序列的前肽编码区。所得多肽被称为酶原(proenzyme)或多肽原(或在某些情况下称为酶原(zymogen))。多肽原通常是无活性的并且可以通过前肽从多肽原的催化裂解或自动催化裂解转化为成熟的有活性的多肽。前肽编码区可以从以下的基因获得：枯草芽孢杆菌碱性蛋白酶(aprE)、枯草芽孢杆菌中性蛋白酶(nprT)、酿酒酵母α-因子、米黑根毛霉天冬氨酸蛋白酶和嗜热毁丝霉(Myceliophthora thermophila)乳糖酶(WO 95/33836)。
当信号肽区和前肽区两者存在于多肽的氨基末端时，前肽区紧邻多肽的氨基末端定位，并且信号肽区紧邻前肽区的氨基末端定位。
添加允许相对于宿主细胞的生长来调控多肽的表达的调控序列也可能是期望的。调控系统的实例是引起基因表达响应于化学或物理刺激(包括调控化合物的存在)而开启或关闭的那些。在原核宿主细胞中，合适的调控序列包括lac、tac和trp操纵子系统。在酵母宿主细胞中，合适的调控系统包括例如ADH2系统或GAL1系统。在丝状真菌中，合适的调控序列包括TAKAα-淀粉酶启动子、黑曲霉葡糖淀粉酶启动子和米曲霉葡糖淀粉酶启动子。
调控序列的其他实例是允许基因扩增的那些。在真核系统中，这些包括在氨甲蝶呤存在下扩增的二氢叶酸还原酶基因以及用重金属扩增的金属硫蛋白基因。在这些情况下，编码本发明的KRED多肽或本发明的PDH多肽的核酸序列将与调控序列可操作地连接。
因此，在一些实施方案中，本发明还涉及重组表达载体，所述重组表达载体包含编码工程化酮还原酶多肽或其变体或者工程化亚磷酸脱氢酶多肽或其变体的多核苷酸，以及根据它们待被引入的宿主的类型，一个或更多个表达调控区诸如启动子和终止子、复制起点等。以上描述的各种核酸和控制序列可以被连接在一起以产生重组表达载体，所述重组表达载体可以包括一个或更多个方便的限制性位点，以允许在这些位点插入或取代编码多肽的核酸序列。可选择地，本发明的核酸序列可以通过将包含该序列的核酸序列或核酸构建体插入到用于表达的适当的载体中来表达。在创建表达载体时，编码序列位于载体中以使编码序列与用于表达的适当的控制序列可操作地连接。
重组表达载体可以是任何载体(例如，质粒或病毒)，其可以方便地经受重组DNA程序并且可以引起多核苷酸序列的表达。载体的选择通常将取决于载体与载体待引入的宿主细胞的相容性。载体可以是线性质粒或闭合的环状质粒。
表达载体可以是自主复制载体(即，作为染色体外实体存在的载体)，其复制独立于染色体复制(例如，质粒、染色体外元件、微型染色体或人工染色体)。载体可以包含用于确保自我复制的任何工具(means)。可选择地，载体可以是当被引入宿主细胞中时被整合到基因组中并与其被整合进的染色体一起复制的载体。此外，可以使用单一载体或质粒或者一起包含待引入到宿主细胞基因组中的总DNA的两种或更多种载体或质粒、或转座子。
本发明的表达载体优选地含有一个或更多个可选择的标志物，其允许容易地选择转化的细胞。可选择的标志物可以是基因，其产物提供杀生物剂或病毒抗性、重金属抗性、针对营养缺陷型的原养型等。细菌的可选择的标志物的实例是来自枯草芽孢杆菌或地衣芽孢杆菌的dal基因或赋予抗生素抗性诸如氨苄青霉素、卡那霉素、氯霉素或四环素抗性的标志物。用于酵母宿主细胞的合适的标志物为ADE2、HIS3、LEU2、LYS2、MET3、TRP1和URA3。
用于在丝状真菌宿主细胞中使用的可选择的标志物包括但不限于amdS(乙酰胺酶)、argB(鸟氨酸氨甲酰基转移酶)、bar(膦丝菌素乙酰基转移酶)、hph(潮霉素磷酸转移酶)、niaD(硝酸盐还原酶)、pyrG(乳清酸核苷-5'-磷酸脱羧酶)、sC(硫酸腺苷酰转移酶)和trpC(邻氨基苯甲酸合酶)及其等同物。用于在曲霉属真菌细胞中使用的实施方案包括构巢曲霉或米曲霉的amdS和pyrG基因以及吸水链霉菌(Streptomyces hygroscopicus)的bar基因。
本发明的表达载体可以包含允许载体整合到宿主细胞的基因组中或允许载体在细胞中独立于基因组自主复制的元件。为了整合到宿主细胞基因组中，载体可以依赖于核酸序列，所述核酸序列编码用于通过同源或非同源重组将载体整合到基因组中的多肽或载体的任何其他元件。
可选择地，表达载体可以包含用于指导通过同源重组整合到宿主细胞的基因组中的另外的核酸序列。另外的核酸序列能够使载体在染色体的准确位置整合到宿主细胞基因组中。为了增加在准确位置整合的可能性，整合元件应当优选地包含与对应的靶序列高度同源的足够数目的核酸，诸如100个至10,000个碱基对，优选地400个至10,000个碱基对，并且最优选地800个至10,000个碱基对，以增强同源重组的可能性。整合元件可以是与宿主细胞的基因组中的靶序列同源的任何序列。此外，整合元件可以是非编码的或编码的核酸序列。另一方面，载体可以通过非同源重组整合到宿主细胞的基因组中。
对于自主复制，载体还可以包含能够使载体在所讨论的宿主细胞中自主复制的复制起点。细菌复制起点的实例是允许在大肠杆菌中复制的P15Aori或质粒pBR322、pUC19、pACYCl77(该质粒具有P15A ori)、或pACYC184的复制起点，以及允许在芽孢杆菌属中复制的pUB110、pE194、pTA1060或pAMβ1的复制起点。用于在酵母宿主细胞中使用的复制起点的实例是2微米(2micron)复制起点、ARS1、ARS4、ARS1和CEN3的组合、以及ARS4和CEN6的组合。复制起点可以是具有使其在宿主细胞中的功能对温度敏感的突变的复制起点(参见例如，Ehrlich,Proc.Natl.Acad.Sci.USA 75:1433[1978])。
可以将多于一个拷贝的本发明的核酸序列插入宿主细胞中以增加基因产物的产量。核酸序列的拷贝数的增加可以通过将至少一个另外拷贝的序列整合到宿主细胞基因组中，或通过包括具有核酸序列的可扩增的可选择标志物基因来获得，其中包含可选择标志物基因的扩增拷贝并从而包含核酸序列的另外拷贝的细胞可以通过在适当的选择剂的存在下培养细胞被选择。
用于在本发明中使用的许多表达载体是商业上可得的。合适的商业表达载体包括但不限于p3xFLAGTMTM表达载体(Sigma-Aldrich)，其包括CMV启动子和用于在哺乳动物宿主细胞中表达的hGH多腺苷酸化位点以及用于在大肠杆菌中扩增的pBR322复制起点和氨苄青霉素抗性标志物。其他商业上可得的合适的表达载体包括但不限于pBluescriptII SK(-)和pBK-CMV载体(Stratagene)，以及源自pBR322(Gibco BRL)、pUC(Gibco BRL)、pREP4、pCEP4(Invitrogen)或pPoly的质粒(参见，Lathe等人，Gene57:193-201[1987])。
用于表达工程化多肽的宿主细胞
本发明还提供了包含编码本发明的改进的酮还原酶多肽或改进的亚磷酸脱氢酶多肽的多核苷酸的宿主细胞，所述多核苷酸可操作地连接至用于在宿主细胞中表达酮还原酶或亚磷酸脱氢酶的一个或更多个控制序列。用于在表达由本发明的表达载体编码的KRED多肽或由本发明的表达载体编码的PDH多肽中使用的宿主细胞是本领域熟知的，并且包括但不限于细菌细胞，诸如大肠杆菌、开菲尔乳杆菌、短乳杆菌、微小乳杆菌(Lactobacillusminor)、链霉菌属和鼠伤寒沙门氏菌(Salmonella typhimurium)细胞；真菌细胞，诸如酵母细胞(例如，酿酒酵母或巴斯德毕赤酵母(Pichia pastoris)(ATCC登录号201178))；昆虫细胞，诸如果蝇属(Drosophila)S2和夜蛾属(Spodoptera)Sf9细胞；动物细胞，诸如CHO、COS、BHK、293和Bowes黑素瘤细胞；以及植物细胞。用于以上描述的宿主细胞的适当的培养基和生长条件是本领域熟知的。
用于表达酮还原酶或亚磷酸脱氢酶的多核苷酸可以通过本领域已知的各种方法引入细胞中。技术尤其包括电穿孔、生物弹射颗粒轰击、脂质体介导的转染、氯化钙转染和原生质体融合。用于将多核苷酸引入细胞中的各种方法对技术人员将是明显的。
大肠杆菌W3110是可用于本发明的宿主菌株，尽管并不意图本发明受限于这种特定的宿主菌株。表达载体通过将编码改进的酶的多核苷酸可操作地连接至质粒pCK110900中来创建，所述编码改进的酶的多核苷酸可操作地连接至在lacI阻遏子的控制下的lac启动子。表达载体还包含P15a复制起点和氯霉素抗性基因。大肠杆菌W3110中包含主题多核苷酸的细胞可以通过使细胞经受氯霉素选择来分离。
产生工程化酮还原酶多肽和工程化亚磷酸脱氢酶多肽的方法。
在一些实施方案中，为了制备本发明的改进的KRED多核苷酸和多肽，催化还原反应的天然存在的酮还原酶从近平滑假丝酵母或鲑色锁掷酵母获得(或衍生)。在一些实施方案中，对亲本多核苷酸序列密码子优化以增强酮还原酶在指定宿主细胞中的表达。作为说明，编码鲑色锁掷酵母的野生型KRED多肽的亲本多核苷酸序列由寡核苷酸构建，所述寡核苷酸基于可从Genbank数据库获得的鲑色锁掷酵母KRED序列的已知多肽序列来制备。对亲本多核苷酸序列密码子优化，用于在大肠杆菌中表达，并将密码子优化的多核苷酸克隆到表达载体中，将酮还原酶基因的表达置于lac启动子和lacI阻遏子基因的控制下。鉴定在大肠杆菌中表达活性酮还原酶的克隆，并对基因测序以确认它们的身份。
在一些实施方案中，如以上讨论的，工程化酮还原酶可以通过使编码天然存在的酮还原酶的多核苷酸经受诱变和/或定向演化方法来获得。诱变可以根据本领域已知的任何技术来进行，包括随机诱变和定点诱变。定向演化可以用本领域已知的任何技术包括重排来进行，以筛选改进的启动子变体。诱变和定向演化方法是本领域熟知的(参见例如，美国专利号5,605,793、5,811,238、5,830,721、5,834,252、5,837,458、5,928,905、6,096,548、6,117,679、6,132,970、6,165,793、6,180,406、6,251,674、6,265,201、6,277,638、6,287,861、6,287,862、6,291,242、6,297,053、6,303,344、6,309,883、6,319,713、6,319,714、6,323,030、6,326,204、6,335,160、6,335,198、6,344,356、6,352,859、6,355,484、6,358,740、6,358,742、6,365,377、6,365,408、6,368,861、6,372,497、6,337,186、6,376,246、6,379,964、6,387,702、6,391,552、6,391,640、6,395,547、6,406,855、6,406,910、6,413,745、6,413,774、6,420,175、6,423,542、6,426,224、6,436,675、6,444,468、6,455,253、6,479,652、6,482,647、6,483,011、6,484,105、6,489,146、6,500,617、6,500,639、6,506,602、6,506,603、6,518,065、6,519,065、6,521,453、6,528,311、6,537,746、6,573,098、6,576,467、6,579,678、6,586,182、6,602,986、6,605,430、6,613,514、6,653,072、6,686,515、6,703,240、6,716,631、6,825,001、6,902,922、6,917,882、6,946,296、6,961,664、6,995,017、7,024,312、7,058,515、7,105,297、7,148,054、7,220,566、7,288,375、7,384,387、7,421,347、7,430,477、7,462,469、7,534,564、7,620,500、7,620,502、7,629,170、7,702,464、7,747,391、7,747,393、7,751,986、7,776,598、7,783,428、7,795,030、7,853,410、7,868,138、7,783,428、7,873,477、7,873,499、7,904,249、7,957,912、7,981,614、8,014,961、8,029,988、8,048,674、8,058,001、8,076,138、8,108,150、8,170,806、8,224,580、8,377,681、8,383,346、8,457,903、8,504,498、8,589,085、8,762,066、8,768,871、9,593,326和所有相关的非美国的对应专利；Ling等人，Anal.Biochem.,254(2):157-78[1997]；Dale等人，Meth.Mol.Biol.,57:369-74[1996]；Smith,Ann.Rev.Genet.,19:423-462[1985]；Botstein等人，Science,229:1193-1201[1985]；Carter,Biochem.J.,237:1-7[1986]；Kramer等人，Cell,38:879-887[1984]；Wells等人，Gene,34:315-323[1985]；Minshull等人，Curr.Op.Chem.Biol.,3:284-290[1999]；Christians等人，Nat.Biotechnol.,17:259-264[1999]；Crameri等人，Nature,391:288-291[1998]；Crameri,等人，Nat.Biotechnol.,15:436-438[1997]；Zhang等人，Proc.Nat.Acad.Sci.U.S.A.,94:4504-4509[1997]；Crameri等人，Nat.Biotechnol.,14:315-319[1996]；Stemmer,Nature,370:389-391[1994]；Stemmer,Proc.Nat.Acad.Sci.USA,91:10747-10751[1994]；WO 95/22625；WO 97/0078；WO97/35966；WO 98/27230；WO 00/42651；WO 01/75767和WO 2009/152336；所有这些通过引用并入本文)。
对诱变处理后获得的克隆筛选具有期望的改进的酶特性的工程化酮还原酶。测量来自表达文库的酶活性可以使用标准生物化学技术来进行，所述标准生物化学技术监测NADH或NADPH浓度的降低速率(通过吸光度或荧光的降低)，随着它们被转化为NAD+或NADP+。在这个反应中，当酮还原酶将酮底物还原为对应的羟基基团时，由酮还原酶消耗(氧化)NADH或NADPH。如通过每单位时间吸光度或荧光的降低测量的NADH或NADPH浓度的降低速率指示KRED多肽在固定量的裂解物(或由其制成的冻干粉末)中的相对(酶促)活性。产物的立体化学可以通过各种已知技术来确定，并如实施例中所提供的。当期望的改进的酶特性是热稳定性时，可以使酶制品经受指定的温度后并测量热处理后剩余的酶活性的量来测量酶活性。然后对包含编码酮还原酶的多核苷酸的克隆进行分离、测序以鉴定核苷酸序列的改变(如果有的话)、并且用于在宿主细胞中表达酶。
当已知工程化多肽的序列时，编码该酶的多核苷酸可以根据已知的合成方法通过标准固相方法来制备。在一些实施方案中，多达约100个碱基的片段可以被单独地合成、然后连接(例如，通过酶促连接方法或化学连接方法或聚合酶介导的方法)以形成任何期望的连续序列。例如，本发明的多核苷酸和寡核苷酸可以通过化学合成来制备(例如，使用由Beaucage等人，Tet.Lett.,22:1859-69[1981]描述的经典亚磷酰胺法，或由Matthes等人，EMBO J.,3:801-05[1984]描述的方法，因为它通常以自动化合成方法来实践)。根据亚磷酰胺法，寡核苷酸被合成(例如，在自动DNA合成仪中)、纯化、退火、连接并克隆到适当的载体中。此外，基本上任何核酸可以从多种商业来源中的任一种获得(例如，The MidlandCertified Reagent Company,Midland,TX，The Great American Gene Company,Ramona,CA，ExpressGen Inc.Chicago,IL，Operon Technologies Inc.,Alameda,CA，以及许多其他商业来源)。
在宿主细胞中表达的工程化酮还原酶和工程化亚磷酸脱氢酶可以使用用于蛋白纯化的熟知的技术中的任一种或更多种从细胞和/或培养基来回收，所述技术尤其包括溶菌酶处理、超声、过滤、盐析、超速离心和色谱。用于裂解和从细菌诸如大肠杆菌高效提取蛋白的合适的溶液是以商标名CelLytic BTM(Sigma-Aldrich)商业上可得的。
用于分离酮还原酶多肽和/或亚磷酸脱氢酶多肽的色谱技术尤其包括反相色谱、高效液相色谱、离子交换色谱、凝胶电泳和亲和色谱。用于纯化特定酶的条件将部分地取决于诸如净电荷、疏水性、亲水性、分子量、分子形状等因素，并且对于本领域技术人员将是明显的。
在一些实施方案中，亲和技术被用于分离改进的酮还原酶和/或改进的亚磷酸脱氢酶。对于亲和色谱纯化，可以使用特异性结合酮还原酶多肽或亚磷酸脱氢酶多肽的任何抗体。为了产生抗体，可以通过注射酮还原酶或亚磷酸脱氢酶来免疫各种宿主动物，包括但不限于兔、小鼠、大鼠等。酮还原酶多肽可以通过侧链官能团或附接至侧链官能团的接头的方式被附接至合适的载体诸如BSA。取决于宿主物种，多种佐剂可以被用来增加免疫应答，包括但不限于弗氏佐剂(完全和不完全)、矿物凝胶诸如氢氧化铝、表面活性物质诸如溶血卵磷脂、pluronic多元醇、聚阴离子、肽、油乳剂、钥孔虫戚血兰素、二硝基酚和可能有用的人类佐剂诸如BCG(卡介苗)和短棒杆菌(Corynebacterium parvum)。
酮还原酶和/或亚磷酸脱氢酶可以以表达酶的细胞的形式、作为粗提取物、或作为分离的或纯化的制品来制备及使用。酮还原酶和/或亚磷酸脱氢酶可以被制备为冻干物、呈粉末形式(例如，丙酮粉末)或被制备为酶溶液。在一些实施方案中，酮还原酶或亚磷酸脱氢酶可以呈基本上纯的制品的形式。
在一些实施方案中，酮还原酶多肽和/或亚磷酸脱氢酶多肽可以被附接至固体基底。基底可以是固相、表面和/或膜。固体支持物可以包括有机聚合物，诸如聚苯乙烯、聚乙烯、聚丙烯、聚氟乙烯、聚氧乙烯(polyethyleneoxy)和聚丙烯酰胺以及它们的共聚物和接枝物。固体支持物还可以是无机的，诸如玻璃、二氧化硅(silica)、可控孔隙玻璃(CPG)、反相二氧化硅或金属诸如金或铂。基底的构型可以呈珠、球、微粒(particle)、颗粒(granule)、凝胶、膜或表面的形式。表面可以是平面的、基本上平面的或非平面的。固体支持物可以是多孔的或无孔的，并且可以具有溶胀或非溶胀特征。固体支持物可以被配置为孔、凹陷或其他容器、器皿、特征或位置的形式。多于一种支持物可以被配置在阵列的多个位置上，所述多个位置是试剂的自动递送或通过检测方法和/或仪器可寻址的。
如本领域技术人员已知的，酮还原酶催化的还原反应通常需要辅因子。由本文描述的工程化酮还原酶催化的还原反应通常也需要辅因子，尽管工程化酮还原酶的许多实施方案比用野生型酮还原酶催化的反应需要少得多的辅因子。如本文使用的，术语“辅因子”是指与酮还原酶联合起作用的非蛋白化合物。适合于与本文描述的工程化酮还原酶一起使用的辅因子包括但不限于NADP+(烟酰胺腺嘌呤二核苷酸磷酸)、NADPH(NADP+的还原形式)、NAD+(烟酰胺腺嘌呤二核苷酸)和NADH(NAD+的还原形式)。通常，将还原形式的辅因子添加到反应混合物中。还原的NAD(P)H形式可以使用辅因子再生系统任选地由氧化的NAD(P)+形式再生。术语“辅因子再生系统”是指参与将辅因子的氧化形式还原的反应(例如，NADP+到NADPH)的一组反应物。通过酮还原酶催化的酮底物的还原被氧化的辅因子通过辅因子再生系统以还原形式再生。辅因子再生系统包括化学计量还原剂，所述化学计量还原剂是还原氢等同物的来源，并且能够将辅因子的氧化形式还原。辅因子再生系统还可以包括催化剂，例如，催化由还原剂将辅因子的氧化形式还原的酶催化剂。分别由NAD+或NADP+再生NADH或NADPH的辅因子再生系统是本领域已知的，并且可以用于本文描述的方法中。
实验
本发明的各种特征和实施方案在以下代表性实施例中进行了说明，这些实施例旨在说明而非限制。
在以下实验公开内容中，以下缩写适用：ppm(百万分率)；M(摩尔/升)；mM(毫摩尔/升)，uM和μM(微摩尔/升)；nM(纳摩尔/升)；mol(摩尔)；gm和g(克)；mg(毫克)；ug和μg(微克)；L和l(升)；ml和mL(毫升)；cm(厘米)；mm(毫米)；um和μm(微米)；sec.(秒)；min(分钟)；h和hr(小时)；U(单位)；MW(分子量)；rpm(转每分)；℃(摄氏度)；RT(室温)；CDS(编码序列)；DNA(脱氧核糖核酸)；RNA(核糖核酸)；HPLC(高效液相色谱)；FIOPC(相对于阳性对照的倍数改进)；HTP(高通量)；LB(Luria肉汤)；Sigma-Aldrich(Sigma-Aldrich,St.Louis,MO)；Millipore(Millipore,Corp.,Billerica MA)；Difco(Difco Laboratories,BDDiagnostic Systems,Detroit,MI)；Daicel(Daicel,West Chester,PA)；Genetix(GenetixUSA,Inc.,Beaverton,OR)；Molecular Devices(Molecular Devices,LLC,Sunnyvale,CA)；Applied Biosystems(Applied Biosystems,part of Life Technologies,Corp.,GrandIsland,NY),Agilent(Agilent Technologies,Inc.,Santa Clara,CA)；ThermoScientific(Thermo Fisher Scientific的一部分,Waltham,MA)；Corning(Corning,Inc.,Palo Alto,CA)；和Bio-Rad(Bio-Rad Laboratories,Hercules,CA)。
实施例1
酮还原酶和亚磷酸脱氢酶基因构建及表达载体
从基因组DNA扩增编码野生型近平滑假丝酵母酮还原酶(KRED)的基因，并将其克隆到表达载体pCK110900中(参见，美国专利申请公布第2006/0195947号的图3，该专利申请通过引用并入本文)，处于lac启动子的控制下。表达载体还包含P15a复制起点和氯霉素抗性基因。野生型酮还原酶的活性如WO2008/042876中描述的来确认。将编码本发明的工程化酮还原酶的多核苷酸同样地克隆到载体pCK110900中，用于在大肠杆菌W3110中表达。KRED基因的定向演化通过以下步骤来进行：首先选择亲本基因(即，SEQ ID NO:2、6、104)，随后进行变体基因的文库构建，其中与某些结构特征相关的位置经受诱变。然后，如实施例2、5和12中描述的，将这些文库铺板、使其生长并使用HTP测定来筛选。
基于所报道的酮还原酶的氨基酸序列和如WO2008/042876的实施例1中描述的密码子优化算法，合成了用于在大肠杆菌中表达的编码野生型鲑色锁掷酵母酮还原酶(KRED)的基因，WO2008/042876通过引用并入本文。该基因使用包含42个核苷酸的寡核苷酸来合成，并被克隆到表达载体pCK110900中(参见，美国专利申请公布第2006/0195947号的图3，该专利申请通过引用并入本文)，处于lac启动子的控制下。表达载体还包含P15a复制起点和氯霉素抗性基因。野生型酮还原酶的活性如WO2008/042876中描述的来确认。将编码本发明的工程化酮还原酶的多核苷酸同样地克隆到载体pCK110900中，用于在大肠杆菌W3110中表达。KRED基因的定向演化通过以下步骤来进行：首先选择亲本基因(即，SEQ ID NO:112、124、138)，随后进行变体基因的文库构建，其中与某些结构特征相关的位置经受诱变。然后，如实施例3、6、7、8和12中描述的，将这些文库铺板、使其生长并使用HTP测定来筛选。
将编码野生型斯氏假单胞菌(Pseudomonas stutzeri)亚磷酸脱氢酶(PDH)的基因的变体克隆到表达载体pCK110900中(参见，美国专利申请公布第2006/0195947号的图3，通过引用并入本文)，处于lac启动子的控制下。表达载体还包含P15a复制起点和氯霉素抗性基因。亚磷酸脱氢酶的活性如WO2008/042876中描述的来确认。
将编码本发明的工程化亚磷酸脱氢酶的多核苷酸同样地克隆到载体pCK110900中，用于在大肠杆菌W3110中表达。PDH基因的定向演化通过以下步骤来进行：首先选择亲本基因(即，SEQ ID NO:172、182、200、208、260)，随后进行变体基因的文库构建，其中与某些结构特征相关的位置经受诱变。然后，如实施例4和实施例9至实施例12中描述的，将这些文库铺板、使其生长并使用HTP测定来筛选。
实施例2
用于氧化的工程化KRED多肽的产生及分析
将通过定向演化获得并且含有演化的酮还原酶基因的质粒文库转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，使用自动菌落挑取器(Genetix)将菌落挑取到包含200μL的LB、1％葡萄糖和30μg/ml CAM的96孔浅孔微量滴定板中。使细胞在30℃并以200rpm摇动生长18h-20h。然后将20μL的该培养物转移到360μL的Terrific Broth(TB)、1mMMgCl2、2mM ZnSO4和30μg/ml CAM中。将深孔板在30℃并以250rpm摇动孵育2.5h后(OD6000.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将平板在30℃并以250rpm摇动孵育18h-21h。
将细胞培养物以3500x g沉淀20min，并将其上清液弃去。将细胞沉淀物在300μL的20mM Tris、2mM ZnSO4、1mM MgCl2、pH 7.5、及1g/L溶菌酶和0.5g/L硫酸多粘菌素B中通过在室温摇动2h来裂解。将样品以3500x g离心20min以澄清细胞碎片，并使用上清液来进行实施例5和12中描述的转化。
实施例3
用于还原的工程化KRED多肽的产生及分析
将通过定向演化获得并且含有演化的酮还原酶基因的质粒文库转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，使用自动菌落挑取器(Genetix)将菌落挑取到包含200μL的LB、1％葡萄糖和30μg/ml CAM的96孔浅孔微量滴定板中。使细胞在30℃并以200rpm摇动生长18h-20h。然后将20μL的该培养物转移到360μL的Terrific Broth(TB)、1mM MgSO4和30μg/ml CAM中。将深孔板在30℃并以250rpm摇动孵育2.5h后(OD600 0.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将平板在30℃并以250rpm摇动孵育18h-21h。
将细胞培养物以3500x g沉淀20min，并将其上清液弃去。将细胞沉淀物在300μL的20mM Tris、1mM MgSO4、pH 7.5、及1g/L溶菌酶和0.5g/L硫酸多粘菌素B中通过在室温摇动2h来裂解。将样品以3500x g离心20min以澄清细胞碎片，并使用上清液来进行实施例6至实施例8和实施例12中描述的转化。
实施例4
工程化亚磷酸脱氢酶多肽的产生及分析
将通过定向演化获得并且含有演化的亚磷酸脱氢酶基因的质粒文库转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，使用自动菌落挑取器(Genetix)将菌落挑取到包含200μL的LB、1％葡萄糖和30μg/ml CAM的96孔浅孔微量滴定板中。使细胞在30℃并以200rpm摇动生长18h-20h。然后将20μL的该培养物转移到360μL的Terrific Broth(TB)和30μg/ml CAM中。将深孔板在30℃并以250rpm摇动孵育2.5h后(OD600 0.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将平板在30℃并以250rpm摇动孵育18h-21h。
将细胞培养物以3500x g沉淀20min，并将其上清液弃去。将细胞沉淀物在300μL的20mM Tris、pH 7.5、及1g/L溶菌酶和0.5g/L硫酸多粘菌素B中通过在室温摇动2h来裂解。将样品以3500x g离心20min以澄清细胞碎片，并使用上清液来进行实施例9至实施例12中描述的转化。
实施例5
SEQ ID NO:2的KRED变体
如实施例1中所描述的产生大肠杆菌KRED变体。为了分析变体的活性，将如实施例2中所描述的产生的20μL上清液添加到180μL外消旋醇底物(50g/L)与4g/L NAD+、10g/L商业上可得的NADH氧化酶(NOx-9)和100mM FAD在pH 8.0的100mM亚磷酸钠中的混合物中。使反应物在30℃孵育16h-18h，并通过添加200μL的1M HCl来猝灭。将猝灭的混合物添加到样品中并短暂混合。反应样品通过UPLC来分析，以定量剩余底物和产物，如以上描述的。显著改进的变体在以下表5.1中提供。
实施例6
SEQ ID NO:112的KRED变体
如实施例1中所描述的产生大肠杆菌KRED变体。为了分析变体的活性，将如实施例3中所描述的产生的5μL上清液添加到95μL的含有0.25mM NADPH、19g/L酮底物和5g/L PDH的pH 7.9的0.3M亚磷酸盐缓冲液中。将反应物在室温孵育16-18小时，温和摇动。将反应物通过添加100μL的1M HCl来猝灭。将猝灭的混合物(10μL)稀释到190μL的水中。通过HPLC分析反应样品(10μL)，以定量剩余底物和产物，如以上描述的。显著改进的变体在以下表6.1中提供。
实施例7
SEQ ID NO:124的KRED变体
如实施例1中所描述的产生大肠杆菌KRED变体。为了分析变体的活性，将如实施例3中所描述的产生的7.5μL上清液添加到192.5μL的含有0.25mM NADPH、50g/L酮底物和5g/LPDH的pH 7.9的0.3M亚磷酸盐缓冲液中。将反应物在室温孵育16-18小时，温和摇动。将反应物通过添加100μL的1M HCl来猝灭。将猝灭的混合物(10μL)稀释到190μL的水中。通过HPLC分析反应样品(10μL)，以定量剩余底物和产物，如以上描述的。显著改进的变体在以下表7.1中提供。
实施例8
SEQ ID NO:138的KRED变体
如实施例1中所描述的产生大肠杆菌KRED变体。为了分析变体的辅因子偏好，利用了四种单独测定。第一，将如实施例3中所描述的产生的10μL上清液添加到90μL的含有1g/L酮和1g/L NADPH的pH 7.9的0.2M亚磷酸盐缓冲液中。样品的NADPH消耗的初始速率通过Exλ＝330nm，Emλ＝445nm的荧光来分析，每21秒采集数据，持续180秒。
第二，将如实施例3中所描述的产生的20μL上清液添加到190μL的含有1g/L外消旋醇和2g/L NAD+的pH 7.9的0.2M亚磷酸盐缓冲液中。NAD+消耗的初始速率通过UV 340nm处的动力学读数来分析，每9秒采集数据，持续5分钟。
第三，将如实施例3中所描述的产生的20μL上清液添加到180μL的含有2g/L咪唑酮和16.4mM NADPH的500mM亚磷酸钠中；将样品在室温以300rpm摇动孵育2hr。通过添加200μLMeCN来猝灭反应。摇动5分钟后，将100μL猝灭的反应物转移到Millipore过滤板(45微米孔径)中，用含有100μL水的co-star圆底板收集滤液，并将混合物以4000rpm离心(spin)2分钟。通过HPLC分析反应样品(10μL)，以定量剩余底物和产物，如以上描述的。
第四，将如实施例3中所描述的产生的20μL上清液添加到180μL的含有2g/L咪唑酮和16.4mM NADPH的500mM亚磷酸钠中；将样品在室温以300rpm摇动孵育2hr。通过添加200μLMeCN来猝灭反应。摇动5分钟后，将100μL猝灭的反应物转移到Millipore过滤板(45微米孔径)中，用含有100μL水的co-star圆底板收集滤液，并将混合物以4000rpm离心2分钟。通过HPLC分析反应样品(10μL)，以定量剩余底物和产物，如以上描述的。辅因子特异性被计算为
(用NADPH生成的产物的量)/(用NADH生成的产物的量)
显著改进的变体在以下表8.1中提供。
实施例9
SEQ ID NO:172的PDH变体
如实施例1中所描述的产生大肠杆菌PDH变体。为了分析变体的活性，将如实施例3中所描述的产生的5μL上清液添加到95μL的含有0.25mM NADPH、50g/L酮底物和2g/L SEQID NO:138的KRED的pH 7.9的0.5M亚磷酸钠缓冲液中。将反应物在25℃孵育16-18小时，温和摇动。将反应物通过添加100μL的1M HCl来猝灭。将猝灭的混合物(10μL)稀释到190μL的水中。通过HPLC分析反应样品(10μL)，以定量剩余底物和产物，如以上描述的。显著改进的变体在以下表9.1中提供。
实施例10
SEQ ID NO:208的PDH变体
如实施例1中所描述的产生大肠杆菌PDH变体。为了分析变体的辅因子偏好，将如实施例3中所描述的产生的上清液用pH 7.5的50mMTris-HCl缓冲液稀释4倍。将20μL稀释的裂解物添加到180μL的pH 7.9的0.1M亚磷酸钠缓冲液中，并孵育过夜，以消耗掉裂解物中存在的剩余NAD+和NADP+。然后以分析变体的辅因子特异性的三种单独测定来筛选变体。第一，对于初始速率NADP+测定，添加pH 7.9的0.1M亚磷酸钠缓冲液中的0.2mM NADP+，并通过荧光测定在2分钟内测量初始速率。第二，对于初始速率NAD+测定，添加pH 7.9的0.1M亚磷酸钠缓冲液中的0.2mM NAD+，并通过荧光测定在2分钟内测量初始速率。第三，进行辅因子竞争测定。对于该测定，将含有100uM NADP、1mM NAD和1g/LNADH氧化酶NOx-9的pH 7.9的100mM亚磷酸盐添加到反应中。NOx-9立即消耗掉所有的NADH，只留下通过NADP+和NAD+之间的竞争而减少的NADPH信号。将反应通过添加100μL的1M HCl来猝灭。将猝灭的混合物(10μL)稀释到190μL的水中。通过HPLC分析稀释的反应样品(10μL)，以定量剩余底物和产物，如以上描述的。显著改进的变体在以下表10.1中提供。
实施例11
SEQ ID NO:208的另外的PDH变体
如实施例1中所描述的产生大肠杆菌PDH变体。为了分析变体的辅因子偏好，将如实施例3中所描述的产生的上清液用pH 7.5的50mMTris-HCl缓冲液稀释4倍。将20μL稀释的裂解物添加到180μL的pH 7.9的0.1M亚磷酸钠缓冲液中，并孵育过夜，以消耗掉裂解物中存在的剩余NAD+和NADP+。然后以分析变体的辅因子特异性的三种单独测定来筛选变体。第一，对于初始速率NADP+测定，添加pH 7.9的0.1M亚磷酸钠缓冲液中的0.2mM NADP+，并通过荧光测定在2分钟内测量初始速率。第二，对于初始速率NAD+测定，添加pH 7.9的0.1M亚磷酸钠缓冲液中的0.2mM NAD+，并通过荧光测定在2分钟内测量初始速率。第三，进行辅因子竞争测定。对于该测定，将3μL预孵育的裂解物添加到97μL的含有2mM NAD、0.2mM NADP、2g/LSEQ ID NO:138的KRED、4g/L SEQ IDNO:104的KRED和10g/L酮(2)的pH 7.9的200mM亚磷酸盐中。将反应通过添加100μL的1M HCl来猝灭。将猝灭的混合物(10μL)稀释到190μL的水中。稀释的反应样品(10μL)通过反相HPLC来分析，以定量剩余底物和产物的两种对映异构体，如以上描述的。显著改进的变体在以下表11.1中提供。
实施例12
工程化多肽的产生及性能验证
将包含通过对SEQ ID NO:2的KRED定向演化获得的变体并且含有演化的酮还原酶基因的质粒转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，将单菌落挑取到5mL的LB、1％葡萄糖和30μg/ml CAM中。使细胞在30℃并以250rpm摇动生长18h-20h。然后将该培养物转移到Terrific Broth(TB)、2mM ZnSO4、1mM MgSO4和30μg/ml CAM中，最终OD600为～0.02并且最终体积为250mL。将瓶(flask)在30℃并以250rpm摇动孵育3.5h后(OD600 0.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将瓶在30℃并以250rpm摇动孵育18h-21h。将细胞以3500x g沉淀20min，并将上清液弃去。将细胞沉淀物在50mL冰冷的含2mM ZnSO4和1mM MgSO4的pH 7.5的50mM磷酸钠中洗涤，在30mL相同的缓冲液中重悬浮，并使用细胞破碎仪以18kpsi-20kpsi裂解。将裂解物以10000x g澄清60min，并将澄清的上清液冻干成灰白色粉末。
将包含通过对SEQ ID NO:112和SEQ ID NO:138的KRED定向演化获得的变体并且含有演化的酮还原酶基因的质粒转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，将单菌落挑取到5mL的LB、1％葡萄糖和30μg/ml CAM中。使细胞在30℃并以250rpm摇动生长18h-20h。然后将该培养物转移到Terrific Broth(TB)和30μg/ml CAM中，最终OD600为～0.02并且最终体积为250mL。将瓶在30℃并以250rpm摇动孵育3.5h后(OD600 0.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将瓶在30℃并以250rpm摇动孵育18h-21h。将细胞以3500x g沉淀20min，并将上清液弃去。将细胞沉淀物在50mL冰冷的pH 7.5的50mM磷酸钠中洗涤，在30mL相同的缓冲液中重悬浮，并使用细胞破碎仪以18kpsi-20kpsi裂解。将裂解物以10000x g澄清60min，并将澄清的上清液冻干成灰白色粉末。
将包含通过对SEQ ID NO:172和SEQ ID NO:208的PDH定向演化获得的变体并且含有演化的亚磷酸脱氢酶基因的质粒转化到大肠杆菌W3110中，并放置于含有1％葡萄糖和30μg/ml氯霉素(CAM)的Luria-Bertani(LB)琼脂培养基上。在30℃孵育至少16h后，将单菌落挑取到5mL的LB、1％葡萄糖和30μg/ml CAM中。使细胞在30℃并以250rpm摇动生长18h-20h。然后将该培养物转移到Terrific Broth(TB)和30μg/mlCAM中，最终OD600为～0.02并且最终体积为250mL。将瓶在30℃并以250rpm摇动孵育3.5h后(OD600 0.6-0.8)，通过1mM的终浓度的异丙基硫代糖苷(IPTG)诱导重组基因表达。然后将瓶在30℃并以250rpm摇动孵育18h-21h。将细胞以3500x g沉淀20min，并将上清液弃去。将细胞沉淀物在50mL冰冷的pH 7.5的50mM磷酸钠中洗涤，在30mL相同的缓冲液中重悬浮，并使用细胞破碎仪以18kpsi-20kpsi裂解。将裂解物以10000x g澄清60min，并将澄清的上清液冻干成灰白色粉末。
为了在类似方法的条件下评估最终化合物，将在pH 7.9的500mM亚磷酸钠缓冲液、0.1g/L NAD、0.1g/L NADP、2.5g/L SEQ ID NO:104的KRED、10g/L商业上可得的NADH氧化酶NOx-9、2.5g/L SEQ ID NO:154的KRED、10g/L SEQ ID NO:250的PDH中的50g/L外消旋醇底物，在氧气流和1％v/v防沫剂下，在室温搅拌24小时，产生93％的底物转化率和99.5％的(R)-醇1a对映异构体过量。通过反相HPLC分析反应样品，以定量剩余底物和产物，如以上描述的。
虽然已经说明并描述了多种具体实施方案，但是应当理解，可以进行各种改变，而不偏离本发明的精神和范围。
为了所有目的，本申请中引用的所有出版物、专利、专利申请和其他文件在此通过引用以其整体并入，其程度如同每个单独的出版物、专利、专利申请或其他文件被单独指出为了所有目的通过引用并入一样。
本发明还提供以下项目：
1.一种工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:2、112、124和/或138具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性。
2.一种工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:2具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：37、37/211、37/211/229、37/229、45、52、52/57/110/272/296、52/57/272、52/57/272/274/279/296、52/57/272/279/296、55/57/276、56、57、57/104/114、57/104/114/229、57/286、79/83/275/276、83、83/275/276、83/276、104、110、114、138/146/258/289、211、211/229、228、229、263、268、272、274、275/276、276、279和309，其中所述位置根据SEQ ID NO:2来编号。
3.一种工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:112具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且具有选自以下的至少一个取代或取代集：24/106/136/220/258/260/314/315、24/106/214/250/258/260/314/315、24/220/314/315、122/159/316/318、135、139/207、159/251/272/277/316/318/330和207，其中所述位置根据SEQ ID NO:112来编号。
4.一种工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:124具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下的位置处具有至少一个取代集：2/101/179/182/228/238/282、3/95、3/95/228/314、24/95/228、95、95/135/139/207和159/228/309/330，其中所述位置根据SEQ IDNO:124来编号。
5.一种工程化酮还原酶变体，所述工程化酮还原酶变体与SEQ ID NO:138具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下的位置处具有至少一个取代或取代集：19、24/43/47/49/67/68/70/91/220、24/68/91/218/220、67、72、74/75/78/108、75/78/99/108/215/224、78/107、95、96和114，其中所述位置根据SEQ ID NO:138来编号。
6.如项目1-5中任一项所述的工程化酮还原酶变体，所述工程化酮还原酶变体包含含有与SEQ ID NO:2、112、124和/或138具有至少90％序列同一性的序列的多肽序列。
7.如项目1-6中任一项所述的工程化酮还原酶变体，所述工程化酮还原酶变体包含含有与SEQ ID NO:2、112、124和/或138具有至少95％序列同一性的序列的多肽序列。
8.如项目1-7中任一项所述的工程化酮还原酶变体，所述工程化酮还原酶变体包含SEQ ID NO:2、112、124或138中列出的多肽序列。
9.如项目1-8中任一项所述的工程化酮还原酶变体，其中所述工程化酮还原酶包含编码表5.1、表6.1、表7.1和/或表8.1中提供的变体的多肽序列。
10.如项目1-9中任一项所述的工程化酮还原酶变体，其中所述工程化酮还原酶包含选自SEQ ID NO:4至SEQ ID NO:170中列出的偶数编号的序列的多肽序列。
11.一种工程化多核苷酸序列，所述工程化多核苷酸序列编码如项目1-10中任一项所述的工程化酮还原酶变体。
12.如项目11所述的工程化多核苷酸序列，其中所述序列包含与选自SEQ ID NO:3至SEQ ID NO:169中列出的奇数编号的序列的序列至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多相同的多核苷酸序列。
13.一种载体，所述载体包含如项目11和/或12所述的工程化多核苷酸序列。
14.如项目13所述的载体，所述载体还包含至少一种控制序列。
15.一种宿主细胞，所述宿主细胞包含如项目13和/或14所述的载体。
16.一种用于产生如项目1-10中任一项所述的工程化酮还原酶变体的方法，所述方法包括在项目15的所述宿主细胞藉以产生所述工程化酮还原酶变体的条件下培养所述宿主细胞。
17.如项目16所述的方法，所述方法还包括回收由所述宿主细胞产生的所述工程化酮还原酶变体的步骤。
18.一种组合物，所述组合物包含至少一种如项目1-17的任一项中提供的工程化酮还原酶变体。
19.一种工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQ ID NO:172和/或SEQ ID NO:208具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性。
20.一种工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQ ID NO:172具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：10/73/78/137/323/325、10/73/78/233/323、10/73/137、13/41/63/132/193/195、18/44/119/124/132/137/145/158/175/177/293/317/323、18/44/119/124/132/137/145/158/177/293/323、18/44/119/124/132/137/145/293/323/334/336、32/44/132/137/145/186/233/293/323/336、41/44/88/193/195、44/69/120/132/137/145/175/195/293/323、44/113/132/145、44/119/132/137/145/158/175/177/293/317/323、44/132/135/136/137/145/293、44/132/136/137/145/293、44/132/137/145/233/308/323、44/132/137/145/293/323、44/132/145、44/132/145/195/293/323、137/233/303/323和266，其中所述位置根据SEQ IDNO:172来编号。
21.一种工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体与SEQ ID NO:208具有至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列同一性，并且在选自以下位置中的一个或更多个位置处具有至少一个取代或取代集：32/59/124/177/191/327、78/150/198/327/328、83/266、95/211/213/322、104、178/194/211/213/322、206、211/213/322、215、262、266和323，其中所述位置根据SEQ ID NO:208来编号。
22.如项目19-21中任一项所述的工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体包含含有与SEQ ID NO:172和/或SEQ ID NO:208具有至少90％序列同一性的序列的多肽序列。
23.如项目19-22中任一项所述的工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体包含含有与SEQ ID NO:172和/或SEQ ID NO:208具有至少95％序列同一性的序列的多肽序列。
24.如项目19-23中任一项所述的工程化亚磷酸脱氢酶变体，所述工程化亚磷酸脱氢酶变体包含SEQ ID NO:172或SEQ ID NO:208中列出的多肽序列。
25.如项目19-24中任一项所述的工程化亚磷酸脱氢酶变体，其中所述工程化亚磷酸脱氢酶变体包含编码表9.1、表10.1和/或表11.1中提供的变体的多肽序列。
26.如项目19-25中任一项所述的工程化亚磷酸脱氢酶变体，其中所述工程化亚磷酸脱氢酶变体包含选自SEQ ID NO:172至SEQ ID NO:260中列出的偶数编号的序列的多肽序列。
27.一种组合物，所述组合物包含至少一种如项目18-26中列出的亚磷酸脱氢酶变体。
28.一种工程化多核苷酸序列，所述工程化多核苷酸序列编码如项目19-26中任一项所述的工程化亚磷酸脱氢酶变体。
29.如项目28所述的工程化多核苷酸序列，其中所述序列包含与选自SEQ ID NO:171至SEQ ID NO:259中列出的奇数编号的序列的序列至少85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多相同的多核苷酸序列。
30.一种载体，所述载体包含如项目28和/或29所述的工程化多核苷酸序列。
31.如项目30所述的载体，所述载体还包含至少一种控制序列。
32.一种宿主细胞，所述宿主细胞包含如项目30和/或31所述的载体。
33.一种用于产生如项目19-25中任一项所述的工程化亚磷酸脱氢酶变体的方法，所述方法包括在项目32的所述宿主细胞藉以产生所述工程化亚磷酸脱氢酶变体的条件下培养所述宿主细胞。
34.如项目33所述的方法，所述方法还包括回收由所述宿主细胞产生的所述工程化亚磷酸脱氢酶变体的步骤。
35.一种用于使手性醇去外消旋化的方法，所述方法在使得所述手性醇去外消旋化的条件下，包括至少一种根据项目1-17中任一项的工程化酮还原酶变体，包括至少一种根据项目19-25中任一项的工程化亚磷酸脱氢酶变体、至少一种手性醇和至少一种辅因子。
36.如项目35所述的方法，其中所述方法以一锅反应来进行。
序列表
<110> 科德克希思公司
<120> 酮还原酶多肽及多核苷酸
<130> CX2-166USP1
<140> 尚未分配
<141> 2017-04-27
<160> 260
<210> 1
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 1
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 2
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 2
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 3
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 3
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactt gggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 4
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 4
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 5
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 5
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggacct tggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 6
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 6
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 7
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 7
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctggttgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 8
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 8
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Val
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 9
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 9
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgtatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 10
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 10
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Tyr Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 11
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 11
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggcttttcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 12
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 12
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Phe Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 13
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 13
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggacat tggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 14
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 14
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Ile Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 15
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 15
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttca aagactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 16
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 16
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Lys Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 17
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 17
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgttggacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 18
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 18
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Leu
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 19
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 19
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttca tggactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 20
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 20
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Met Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 21
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 21
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgagtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 22
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 22
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Ser
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 23
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 23
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgttggacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 24
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 24
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Leu
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 25
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 25
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcatgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 26
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 26
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu His
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 27
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 27
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgacagacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 28
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 28
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Thr
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 29
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 29
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgacagacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 30
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 30
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Thr
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 31
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 31
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgattgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 32
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 32
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Ile
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 33
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 33
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggacct tggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 34
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 34
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 35
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 35
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgtgggacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 36
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 36
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Trp
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 37
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 37
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaacatcgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 38
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 38
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu His Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 39
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 39
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgcatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 40
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 40
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu His Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 41
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 41
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgrkkgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 42
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<221> unsure
<222> 272
<223> Xaa = Ser, Met, Gly, Arg, Val, Ile
<223> KRED-NADH-102变体
<400> 42
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Xaa
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 43
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 43
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttca tggactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 44
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 44
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Met Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 45
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 45
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtggtttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 46
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 46
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Trp Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 47
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 47
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgcatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 48
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 48
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu His Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 49
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 49
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctggcagacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 50
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 50
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Ala
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 51
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 51
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgcatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 52
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 52
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu His Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 53
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 53
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaaggcgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 54
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 54
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Arg Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 55
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 55
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgacagacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 56
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 56
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Thr
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 57
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 57
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcaagacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 58
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 58
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gln
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 59
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 59
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgcatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 60
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 60
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu His Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 61
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 61
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgcatg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 62
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 62
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu His Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 63
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 63
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctgctttg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 64
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 64
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Leu Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 65
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 65
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tagtggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 66
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 66
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Ser Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 67
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 67
atgtcgatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 68
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 68
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 69
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 69
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttaccaac 240
tacaaatccg gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacaaccg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 70
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 70
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Thr Asn
65 70 75 80
Tyr Lys Ser Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Asn Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 71
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 71
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct agaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 72
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 72
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Arg Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 73
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 73
atgtcgatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgag agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct agaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 74
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 74
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Arg Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Arg Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 75
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 75
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 76
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 76
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 77
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 77
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgacggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgcgttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 78
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 78
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Arg Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 79
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 79
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gccttgactg tggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 80
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 80
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 81
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 81
atgtcgatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgacggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct agaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 82
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 82
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Arg Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 83
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 83
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 84
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 84
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 85
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 85
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 86
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 86
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 87
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 87
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 88
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 88
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 89
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 89
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggcgatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 90
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 90
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 91
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 91
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaatccg gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 92
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 92
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Ser Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 93
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 93
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tgaacatgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 94
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 94
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Asn Met Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 95
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 95
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 96
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 96
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 97
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 97
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctacgaag gcctggactg cggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg gcatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatcttcgga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctgg cgtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat cgtgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 98
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 98
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Cys Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Gly Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ser Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gly Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Val Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 99
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 99
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccattctga tctgcacgtt atctacgaag gcctggacat aggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg gcatcgacaa cgtttgcaaa aacgctttcc atgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggcg ctccgaacct gtcttttaac ctgggtgacc tggctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 100
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 100
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Ile Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Gly Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe His Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 101
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 101
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccattctga tctgcacgtt atctacgaag gcctggactt aggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg gcatcgacaa cgtttgcaaa aacgctttcc atgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgcgttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggcg ctccgaacct gtcttttaac ctgggtgacc tggcgttgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttctgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 102
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 102
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Tyr Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Gly Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe His Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Arg Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu Gly
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 103
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 103
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctcagaag gcctggattt gggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcatgacc ttgctctgcg tgaacaccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgtttttaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgatct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 104
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 104
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Ser Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu His
260 265 270
Asp Leu Ala Leu Arg Glu His Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Phe Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 105
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 105
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atcgatgaag gcctggattt gggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcatgacc ttgctctgcg tgaaatacgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttttgaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 106
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 106
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Asp Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu His
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Leu Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 107
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 107
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctcagaag gcctggattt gggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaaa aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcatgacg ttgctctgcg tgaacaccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttttcaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgatct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 108
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 108
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Ser Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Lys Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu His
260 265 270
Asp Val Ala Leu Arg Glu His Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Phe Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 109
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 109
atgtctatcc cgtcttctca gtatggtttc gtatttaaca aacagtctgg tctgaaactg 60
cgcaacgatc tgccagtaca caagccgaaa gctggtcaac tgctgctgaa agtagatgca 120
gtaggtctgt gccactctga tctgcacgtt atctcagaag gcctggattt gggtgacaac 180
tacgttatgg gtcacgaaat cgctggtacc gttgctgctg ttggtgatga cgttatcaac 240
tacaaagtag gtgaccgtgt agcttgtgtt ggtccgaacg gttgcggtgg ttgcaaatac 300
tgccgtggtg ctatcgacaa cgtttgcaca aacgctttcg gtgactggtt cggtctgggt 360
tatgatggtg gctatcaaca atacctgctg gtaactcgcc cgcgtaacct gtctcgtatc 420
ccggataacg tatctgctga tgttgctgct gcttctaccg acgctgttct gactccgtac 480
cacgctatca aaatggctca ggtttctccg acctctaaca tcctgctgat cggtgctggt 540
ggtctgggtg gtaacgctat ccaggttgct aaagcattcg gtgctaaagt taccgttctg 600
gacaaaaaaa aagaagctcg tgaccaggct aaaaaactgg gtgctgacgc tgtttacgaa 660
accctgccgg aatctatctc tccgggttct ttctctgctt gcttcgactt cgtttctgtt 720
caggctacct tcgacgtttg ccagaaatac gttgaaccga aaggtgttat catgccggtt 780
ggtctgggtg ctccgaacct gtcttttaac ctgcatgacc ttgctctgcg tgaaatccgt 840
atcctgggtt ctttctgggg taccaccaac gacctggacg acgttttcaa actggtttct 900
gaaggtaaag ttaaaccggt tgtacgttct gctaaactga aagaactgcc ggaatacatc 960
gaaaaactgc gtaacaacgc ttacgaaggt cgtgttgttt tcaacccgta a 1011
<210> 110
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-NADH-102变体
<400> 110
Met Ser Ile Pro Ser Ser Gln Tyr Gly Phe Val Phe Asn Lys Gln Ser
1 5 10 15
Gly Leu Lys Leu Arg Asn Asp Leu Pro Val His Lys Pro Lys Ala Gly
20 25 30
Gln Leu Leu Leu Lys Val Asp Ala Val Gly Leu Cys His Ser Asp Leu
35 40 45
His Val Ile Ser Glu Gly Leu Asp Leu Gly Asp Asn Tyr Val Met Gly
50 55 60
His Glu Ile Ala Gly Thr Val Ala Ala Val Gly Asp Asp Val Ile Asn
65 70 75 80
Tyr Lys Val Gly Asp Arg Val Ala Cys Val Gly Pro Asn Gly Cys Gly
85 90 95
Gly Cys Lys Tyr Cys Arg Gly Ala Ile Asp Asn Val Cys Thr Asn Ala
100 105 110
Phe Gly Asp Trp Phe Gly Leu Gly Tyr Asp Gly Gly Tyr Gln Gln Tyr
115 120 125
Leu Leu Val Thr Arg Pro Arg Asn Leu Ser Arg Ile Pro Asp Asn Val
130 135 140
Ser Ala Asp Val Ala Ala Ala Ser Thr Asp Ala Val Leu Thr Pro Tyr
145 150 155 160
His Ala Ile Lys Met Ala Gln Val Ser Pro Thr Ser Asn Ile Leu Leu
165 170 175
Ile Gly Ala Gly Gly Leu Gly Gly Asn Ala Ile Gln Val Ala Lys Ala
180 185 190
Phe Gly Ala Lys Val Thr Val Leu Asp Lys Lys Lys Glu Ala Arg Asp
195 200 205
Gln Ala Lys Lys Leu Gly Ala Asp Ala Val Tyr Glu Thr Leu Pro Glu
210 215 220
Ser Ile Ser Pro Gly Ser Phe Ser Ala Cys Phe Asp Phe Val Ser Val
225 230 235 240
Gln Ala Thr Phe Asp Val Cys Gln Lys Tyr Val Glu Pro Lys Gly Val
245 250 255
Ile Met Pro Val Gly Leu Gly Ala Pro Asn Leu Ser Phe Asn Leu His
260 265 270
Asp Leu Ala Leu Arg Glu Ile Arg Ile Leu Gly Ser Phe Trp Gly Thr
275 280 285
Thr Asn Asp Leu Asp Asp Val Phe Lys Leu Val Ser Glu Gly Lys Val
290 295 300
Lys Pro Val Val Arg Ser Ala Lys Leu Lys Glu Leu Pro Glu Tyr Ile
305 310 315 320
Glu Lys Leu Arg Asn Asn Ala Tyr Glu Gly Arg Val Val Phe Asn Pro
325 330 335
<210> 111
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 111
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 112
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 112
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 113
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 113
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttcggc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagggc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc gggcgctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 114
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 114
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Gly Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Arg Ala Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 115
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 115
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttatgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttccgcc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgttgcggc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagggc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cgtgggtgca 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc gggcgctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 116
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 116
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Pro Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ala Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Gly Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Val Gly Ala Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Arg Ala Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 117
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 117
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttatgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttccgcc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tagatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgtg gttgatattg gcctgctgca cgtgggtgca 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc gggcgctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 118
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 118
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Pro Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Leu Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Val Val Asp Ile Gly Leu Leu
245 250 255
His Val Gly Ala Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Arg Ala Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 119
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 119
atggctaaaa tcgataacgc agttcttccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aatctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctgagccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccgttgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gtagatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccgaaga acttctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 120
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 120
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Glu Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Val Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Glu Glu Leu Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 121
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 121
atggctaaaa tcgataacgc agttcttccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccgttgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacaccatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct caggatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttttggta cggctggtcc gttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccgaaga acttctgaaa 960
tctctgggtc gcccaggttg gcgttccctt gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 122
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 122
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Val Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Gln Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Phe
260 265 270
Gly Thr Ala Gly Pro Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Glu Glu Leu Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Leu Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 123
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 123
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 124
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 124
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 125
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 125
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 126
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 126
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 127
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 127
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattttgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 128
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 128
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 129
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 129
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 130
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 130
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 131
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 131
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccatc gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattttgaaa 960
tctttgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 132
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 132
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Ser Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 133
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 133
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggccctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 134
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 134
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 135
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 135
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg atacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gacgtccctg tttaacggcg aggtttcccc ggccctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 136
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 136
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Thr Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 137
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 137
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 138
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 138
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 139
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 139
atggcataca tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggccctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 140
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 140
Met Ala Tyr Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 141
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 141
atggcataca tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg atacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gacgtccctg tttaacggcg aggtttcccc ggccctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc ggtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 142
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 142
Met Ala Tyr Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Thr Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Arg Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 143
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 143
atgactaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
cctgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctctgtcc 540
aagatggaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gaggtccctg tttaacggcg aggtttcccc gttgctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacgaagttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 144
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 144
Met Thr Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Pro Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Leu Ser Lys Met Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Arg Ser Leu Phe Asn Gly Glu Val Ser Pro Leu Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Glu Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 145
<211> 1032
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 145
atggctaaaa tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttgtttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccgtttccgc tctgattccg 420
aagccgaacg ttgaaggtat ctacctggac gaaaagtcct ggaacctgga atccgttgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcatcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccagg gtacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gctgtccctg tttaacggcg aggtttcgcc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctcaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccctt gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgctt aa 1032
<210> 146
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 146
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Val Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Val Ser Ala Leu Ile Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Val Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Gly Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Leu Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Gln Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Leu Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 147
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 147
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgtgc gttccgctga aaaaaatgct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgt ggaagttcca gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac gttgcttccg taacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aacccagggc 660
gggtccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 148
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 148
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Val Arg Ser Ala Glu Lys
35 40 45
Asn Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Val Glu Val Pro Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Val Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Gly Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 149
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 149
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttca ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgcgc gttccgcttc aaaacttgct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc ggaagttgag gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac gttgcttccg taacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aaaccagggc 660
gggtccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 150
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 150
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Ile Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Glu Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Val Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Asn Gln Gly Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 151
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 151
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctta agcaaggtgc ttttgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ttccaacaaa 300
tacgacgagg ttgttaccgg tgcgatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactatct ttgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 152
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 152
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Phe Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Gly Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 153
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 153
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgcttg cagaaggtgc ttttgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgagg ttgttacccc cgtgatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct cgggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcaatacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 154
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 154
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Ala Glu Gly Ala Phe Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Val Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 155
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 155
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctta aggaaggtgc ttttgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctcccccaaa 300
tacgacgagg ttgttacccc cgtgatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactatct tttctccgga aactcagtcc 660
ggttccaccg cgggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 156
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 156
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Glu Gly Ala Phe Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Pro Lys Tyr Asp Glu Val Val Thr Pro Val Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Ser Pro Glu Thr Gln Ser Gly Ser Thr Ala
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 157
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 157
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt tacctcggct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 158
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 158
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Ser Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 159
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 159
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg tttgttcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 160
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 160
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Cys Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 161
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 161
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacagggtt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 162
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 162
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Gly
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 163
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 163
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt tacctcggct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 164
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 164
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Ser Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 165
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 165
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gaccagctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 166
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 166
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Gln Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 167
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 167
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaacctg ggttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttga acgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 168
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 168
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Trp Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Asn Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 169
<211> 1029
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 169
atggcaaaga tcgataacgc agttctgccg gaaggttccc tggttctggt taccggtgct 60
aacggtttcg ttggttccca cgttgttgaa cagctgctgg aacacggtta caaagttcgt 120
ggtaccgctc gttccgcttc caaactggct aacctgcaga aacgttggga cgctaaatac 180
ccgggtcgtt tcgaaaccgc tgttgttgaa gacatgctga aacagggtgc ttacgacgaa 240
gttatcaaag gtgctgctgg tgttgctcac atcgcttccg ttacatcctt ctccaacaaa 300
tacgacgaag ttgttacccc ggctatcggt ggtaccttgg ttgctctgcg tgctgctgct 360
gctaccccgt ccgttaaacg tttcgttctg acctcctcca ccttttccgc tctggttccg 420
aaaccgaacg ttgaaggtat ctacctggac gaaaaatcct ggaacctgga atccatcgac 480
aaagctaaaa ccctgccgga atccgacccg cagaaatccc tgtgggtata cgctgcgtcc 540
aagaccgaag ctgaactggc tgcatggaaa tttatggatg agaacaagcc acacttcact 600
ctgaacgctg tactgccaaa ctacactatt ggcactattt tcgatccgga aactcagtcc 660
ggttccacct ccggttggat gatgtccctg tttaacggcg aggtttcccc ggctctggct 720
ctgatgccac cgcagtacta cgtttccgct gttgatattg gcctgctgca cctgggttgc 780
ctggttctgc cacaaatcga acgtcgtcgt gtttacggta cggctggtac tttcgattgg 840
aacaccgttc tggctacctt ccgtaaactg tacccgtcca aaaccttccc ggctgacttc 900
ccagatcaag gtcaggacct gtctaaattc gacaccgctc cgtccctgga aattctgaaa 960
tctctgggtc gcccaggttg gcgttccatc gaagaatcca tcaaagacct ggttggttcc 1020
gaaaccgct 1029
<210> 170
<211> 343
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> KRED-101变体
<400> 170
Met Ala Lys Ile Asp Asn Ala Val Leu Pro Glu Gly Ser Leu Val Leu
1 5 10 15
Val Thr Gly Ala Asn Gly Phe Val Gly Ser His Val Val Glu Gln Leu
20 25 30
Leu Glu His Gly Tyr Lys Val Arg Gly Thr Ala Arg Ser Ala Ser Lys
35 40 45
Leu Ala Asn Leu Gln Lys Arg Trp Asp Ala Lys Tyr Pro Gly Arg Phe
50 55 60
Glu Thr Ala Val Val Glu Asp Met Leu Lys Gln Gly Ala Tyr Asp Glu
65 70 75 80
Val Ile Lys Gly Ala Ala Gly Val Ala His Ile Ala Ser Val Thr Ser
85 90 95
Phe Ser Asn Lys Tyr Asp Glu Val Val Thr Pro Ala Ile Gly Gly Thr
100 105 110
Leu Val Ala Leu Arg Ala Ala Ala Ala Thr Pro Ser Val Lys Arg Phe
115 120 125
Val Leu Thr Ser Ser Thr Phe Ser Ala Leu Val Pro Lys Pro Asn Val
130 135 140
Glu Gly Ile Tyr Leu Asp Glu Lys Ser Trp Asn Leu Glu Ser Ile Asp
145 150 155 160
Lys Ala Lys Thr Leu Pro Glu Ser Asp Pro Gln Lys Ser Leu Trp Val
165 170 175
Tyr Ala Ala Ser Lys Thr Glu Ala Glu Leu Ala Ala Trp Lys Phe Met
180 185 190
Asp Glu Asn Lys Pro His Phe Thr Leu Asn Ala Val Leu Pro Asn Tyr
195 200 205
Thr Ile Gly Thr Ile Phe Asp Pro Glu Thr Gln Ser Gly Ser Thr Ser
210 215 220
Gly Trp Met Met Ser Leu Phe Asn Gly Glu Val Ser Pro Ala Leu Ala
225 230 235 240
Leu Met Pro Pro Gln Tyr Tyr Val Ser Ala Val Asp Ile Gly Leu Leu
245 250 255
His Leu Gly Cys Leu Val Leu Pro Gln Ile Glu Arg Arg Arg Val Tyr
260 265 270
Gly Thr Ala Gly Thr Phe Asp Trp Asn Thr Val Leu Ala Thr Phe Arg
275 280 285
Lys Leu Tyr Pro Ser Lys Thr Phe Pro Ala Asp Phe Pro Asp Gln Gly
290 295 300
Gln Asp Leu Ser Lys Phe Asp Thr Ala Pro Ser Leu Glu Ile Leu Lys
305 310 315 320
Ser Leu Gly Arg Pro Gly Trp Arg Ser Ile Glu Glu Ser Ile Lys Asp
325 330 335
Leu Val Gly Ser Glu Thr Ala
340
<210> 171
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 171
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 172
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 172
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 173
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 173
atgctgccga aactcgttat aactcacaaa gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggcgctg cgctcaaggg ctttgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccaca gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaaa ttgaacgttg cgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 174
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 174
Met Leu Pro Lys Leu Val Ile Thr His Lys Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Ala Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Gln Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 175
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 175
atgctgccga aactcgttat aactcacaaa gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggcgctg cgctcaaggg ctatgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggccatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttatta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg cgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 176
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 176
Met Leu Pro Lys Leu Val Ile Thr His Lys Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Ala Ala Leu Lys Gly Tyr Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Ile Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 177
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 177
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg ctttgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggccatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccaca gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttatta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggcaa ttgaacgttg cgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 178
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 178
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Gln Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Ile Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Ala Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 179
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 179
atgctgccga aactcgttat aactcacaaa gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggcgctg cgctcaaggg ctatgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccaca gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcgacg ctgcgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 180
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 180
Met Leu Pro Lys Leu Val Ile Thr His Lys Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Ala Ala Leu Lys Gly Tyr Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Gln Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Ala Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 181
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 181
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcaaccacg gttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 182
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 182
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 183
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 183
atgctgccga aactcgttat aactcaccga gtacacgatg agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
gcacgctgtc gggatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagat 180
tttcttgctg cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtgtcgt gcgaagaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 184
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 184
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Asp Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Ala Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Ala Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ser Cys Glu Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 185
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 185
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
gcacgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagat 180
tttcttcagg cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tcggcgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtgtcgt gcgaagaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 186
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 186
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Ala Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Arg Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ser Cys Glu Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 187
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 187
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggttga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 188
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 188
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Val Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 189
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 189
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatgtggga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 190
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 190
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Trp Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 191
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 191
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cgccgctgtc gcgatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcccgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccggggct ggcaaccacg gttctacggc 420
acggggctgg ataacgctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatgtctga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaatcaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 192
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 192
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Arg Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Arg Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Asn Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Ser Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 193
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 193
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcaggacca gttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcggcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 194
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 194
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Asp Gln Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 195
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 195
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttatta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgtggcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 196
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 196
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Ile Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Val Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 197
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 197
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct gggctgacat tttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 198
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 198
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Ala Asp Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 199
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 199
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 200
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 200
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 201
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 201
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcaaccacg gttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcgaggaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtta acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 202
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 202
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Glu Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 203
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 203
atgctgccga aactcgttat aactcaccga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgcg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcgtcgg ggctggggcg gcatctgcgg 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcaaccacg gttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctcggta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcacatag ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattaacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 204
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 204
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Ser Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Arg Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Ile Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 205
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 205
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca aatgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg aattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg actcgccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagccca aggcccgctg a 1011
<210> 206
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 206
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Met Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Glu Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Lys Ala Arg
325 330 335
<210> 207
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 207
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 208
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 208
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 209
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 209
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca aatgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg aattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 210
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 210
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Met Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Glu Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 211
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 211
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca aatgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg aattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagtgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 212
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 212
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Met Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Glu Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 213
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 213
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gactcgacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgaaagta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctggtt 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acacccggaa ggctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcgaggaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtta acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 214
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 214
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Lys Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Val Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Thr Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Glu Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 215
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 215
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacgtcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgagagta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatctgcgc 360
gcagcagatg cgttcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagctac ggtcggcttc cttggcatgg gcgccatcgg actggccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgcccggaa ggctctggat 540
acacaaactg agcaaaccct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg tccgggcgct ctgcttatta acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattggc aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcaagctg a 1011
<210> 216
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 216
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Val
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Leu Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Leu Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ala Arg
165 170 175
Lys Ala Leu Asp Thr Gln Thr Glu Gln Thr Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Ile Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Ala Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Ser
325 330 335
<210> 217
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 217
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg ctatgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcatt ctgggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttagccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaacag cccgcccaag gccgagcctg ccgcatgttg a 1011
<210> 218
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 218
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Tyr Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Ile Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Leu Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Ser Pro Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 219
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 219
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga cctttgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg gcggcccaga ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccacaagacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 220
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 220
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Ala Ala Gln Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Gln Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 221
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 221
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga cctttgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac ccctctggat 540
acacaaacag agcaacggct cggcctgcgc caggtggcgc ttagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg gcggcccaga ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccacaagacg ctgtgaaccg tctgcccaag gccgagcccg ccgcatgttg a 1011
<210> 222
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 222
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Pro Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Leu Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Ala Ala Gln Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Gln Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 223
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 223
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccattgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg gcggcccaga ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaatggacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 224
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 224
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Ile Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Ala Ala Gln Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Met Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 225
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 225
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacgtcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatatggac 180
tttcttcaag cctgccctga gctgagagta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg acatttccgg 360
gcagcagatg aattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggag cgctctggat 540
acacaaaccg agcaacggct cggcctgcgc catgtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaacga tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 226
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 226
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Val
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Met Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Glu Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Ser Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg His Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Asp Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 227
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 227
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccccgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 228
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 228
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Pro His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 229
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 229
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgaatgc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 230
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 230
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Asn Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 231
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 231
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccgt ttgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 232
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 232
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Phe Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 233
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 233
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccgc ttgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 234
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 234
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Leu Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 235
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 235
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatgtctga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 236
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 236
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Ser Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 237
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 237
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatcctttcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 238
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 238
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Pro Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 239
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 239
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgatttcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 240
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 240
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Asp Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 241
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 241
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgcgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggcgga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 242
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 242
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Ala Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Ala Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 243
<211> 1011
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 243
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattaatg cggtgaaccg tctgcccaag gccgagcctg ccgcatgttg a 1011
<210> 244
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 244
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asn Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 245
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 245
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgcgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccgt tggccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgaatgc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtgttcg aaatggagga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 246
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 246
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Ala Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Leu Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Asn Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 247
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 247
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgta ccctcaaggg cttcgacaat 240
ttcgatgcgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgaatgc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtgttcg aaatggagga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 248
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 248
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Thr Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Ala Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Asn Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 249
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 249
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccgt tggccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatctgttcg aaatggagga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 250
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 250
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Leu Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Leu Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 251
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 251
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg ccctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccgt tggccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgaatgc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtgttcg aaatggagga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 252
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 252
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Leu Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Asn Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 253
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 253
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggcgggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 254
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 254
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Arg Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 255
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 255
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgggcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 256
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 256
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Gly
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 257
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 257
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga tggccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc actcgcggac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 258
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 258
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Met Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Ser Arg
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335
<210> 259
<211> 1008
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 259
atgctgccga aactcgttat aactcacaga gtacacgaag agatcctgca actgctggcg 60
ccacattgcg agctgataac caaccagacc gacagcacgc tgacgcgcga ggaaattctg 120
cggcgctgtg cagatgctca ggcgatgatg gcgttcatgc ccgatcgggt cgatgcagac 180
tttcttcaag cctgccctga gctgcgtgta atcggctgtg cgctcaaggg cttcgacaat 240
ttcgatgtgg acgcctgtac tgcacgcggg gtctggctga ccttcgtgcc tgatctgttg 300
acggtcccga ctgccgagct ggcgatcgga ctggcggtgg ggctggggcg gcatttccgg 360
gcagcagatg cattcgtccg ctctggcaag ttccagggct ggcagccaat tttctacggc 420
acggggctgg atggagcaac ggtcggcttc cttggcatgg gcgccatcgg aaaagccatg 480
gctgatcgct tgcagggatg gggcgcgacc ctgcagtacc acgaagcgac agctctggat 540
acacaaaccg agcaacggct cggcctgcgc caggtggcgt gcagcgaact cttcgccagc 600
tcggacttca tcctgctggc gcttcccttg aatgccgata ccctgcatct ggtcaacgcc 660
gagctgcttg ccctcgtacg gccgggcgct ctgcttgtaa acccctgtcg tggctctgta 720
gtggatgaag ccgccgtgct cgcggcgctt gagcgaggcc agctcggcgg gtatgcggcg 780
gatgtattcg aaatggaaga ctgggctcgc gcggaccggc cgcagcagat cgatcctgcg 840
ctgctcgcgc atccgaatac gctgttcact ccgcaccttg ggtcggcagt gcgcgcggtg 900
cgcctggaga ttgaacgttg tgcagcgcag aacatcctcc aggcattgcg aggtgagcgc 960
ccaattgacg ctgtgaaccg tctgcccaag gccgagcctg ccgcatgt 1008
<210> 260
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> PDH-101变体
<400> 260
Met Leu Pro Lys Leu Val Ile Thr His Arg Val His Glu Glu Ile Leu
1 5 10 15
Gln Leu Leu Ala Pro His Cys Glu Leu Ile Thr Asn Gln Thr Asp Ser
20 25 30
Thr Leu Thr Arg Glu Glu Ile Leu Arg Arg Cys Ala Asp Ala Gln Ala
35 40 45
Met Met Ala Phe Met Pro Asp Arg Val Asp Ala Asp Phe Leu Gln Ala
50 55 60
Cys Pro Glu Leu Arg Val Ile Gly Cys Ala Leu Lys Gly Phe Asp Asn
65 70 75 80
Phe Asp Val Asp Ala Cys Thr Ala Arg Gly Val Trp Leu Thr Phe Val
85 90 95
Pro Asp Leu Leu Thr Val Pro Thr Ala Glu Leu Ala Ile Gly Leu Ala
100 105 110
Val Gly Leu Gly Arg His Phe Arg Ala Ala Asp Ala Phe Val Arg Ser
115 120 125
Gly Lys Phe Gln Gly Trp Gln Pro Ile Phe Tyr Gly Thr Gly Leu Asp
130 135 140
Gly Ala Thr Val Gly Phe Leu Gly Met Gly Ala Ile Gly Lys Ala Met
145 150 155 160
Ala Asp Arg Leu Gln Gly Trp Gly Ala Thr Leu Gln Tyr His Glu Ala
165 170 175
Thr Ala Leu Asp Thr Gln Thr Glu Gln Arg Leu Gly Leu Arg Gln Val
180 185 190
Ala Cys Ser Glu Leu Phe Ala Ser Ser Asp Phe Ile Leu Leu Ala Leu
195 200 205
Pro Leu Asn Ala Asp Thr Leu His Leu Val Asn Ala Glu Leu Leu Ala
210 215 220
Leu Val Arg Pro Gly Ala Leu Leu Val Asn Pro Cys Arg Gly Ser Val
225 230 235 240
Val Asp Glu Ala Ala Val Leu Ala Ala Leu Glu Arg Gly Gln Leu Gly
245 250 255
Gly Tyr Ala Ala Asp Val Phe Glu Met Glu Asp Trp Ala Arg Ala Asp
260 265 270
Arg Pro Gln Gln Ile Asp Pro Ala Leu Leu Ala His Pro Asn Thr Leu
275 280 285
Phe Thr Pro His Leu Gly Ser Ala Val Arg Ala Val Arg Leu Glu Ile
290 295 300
Glu Arg Cys Ala Ala Gln Asn Ile Leu Gln Ala Leu Arg Gly Glu Arg
305 310 315 320
Pro Ile Asp Ala Val Asn Arg Leu Pro Lys Ala Glu Pro Ala Ala Cys
325 330 335

Claims (15)

1.一种工程化酮还原酶变体，其中所述工程化酮还原酶变体的多肽序列与SEQ ID NO:124之间的氨基酸残基差异为V95T或者所述工程化酮还原酶变体的多肽序列与SEQ ID NO:124之间的氨基酸残基差异为V95T和选自以下的1-12个氨基酸残基差异：K3Y、G19S、V24I、A43V、S47E、L49N、A67V/W、V68E、E70P、M72Q、K74A、Q75E、Y78F、I91V、S96G、N99P、P107G、A108V、N114V、V135F、I139V、G207N、D215S、T218N、S220G、S224A、M228T和P314R；其中位置根据SEQ ID NO:124来编号；并且其中所述工程化酮还原酶变体具有酮还原酶活性。
2.如权利要求1所述的工程化酮还原酶变体，其中所述工程化酮还原酶变体的多肽序列与SEQ ID NO:124之间的氨基酸残基差异为K3Y/V95T；K3Y/V95T/M228T/P314R；V24I/V95T/M228T；或者V95T/V135F/I139V/G207N，其中位置根据SEQ ID NO:124来编号。
3.如权利要求1-2中任一项所述的工程化酮还原酶变体，所述工程化酮还原酶变体由SEQ ID NO:138中列出的多肽序列组成。
4.如权利要求1-2中任一项所述的工程化酮还原酶变体，其中所述工程化酮还原酶变体由SEQ ID NO:134、136、138、140、142、148、150、152、154、156、158、162、164、166、168或170中列出的多肽序列组成。
5.一种工程化多核苷酸，所述工程化多核苷酸编码如权利要求1-4中任一项所述的工程化酮还原酶变体。
6.如权利要求5所述的工程化多核苷酸，其中所述工程化多核苷酸包含SEQ ID NO:133、135、137、139、141、147、149、151、153、155、157、161、163、165、167或169中列出的多核苷酸序列。
7.一种载体，所述载体包含如权利要求5和/或6所述的工程化多核苷酸的序列。
8.如权利要求7所述的载体，所述载体还包含至少一种控制序列。
9.一种宿主细胞，所述宿主细胞包含如权利要求7和/或8所述的载体。
10.一种用于产生如权利要求1-4中任一项所述的工程化酮还原酶变体的方法，所述方法包括在权利要求9的所述宿主细胞藉以产生所述工程化酮还原酶变体的条件下培养所述宿主细胞。
11.如权利要求10所述的方法，所述方法还包括回收由所述宿主细胞产生的所述工程化酮还原酶变体的步骤。
12.一种组合物，所述组合物包含至少一种如权利要求1-11的任一项中提供的工程化酮还原酶变体。
13.一种用于使手性醇去外消旋化的方法，所述方法包括在使得所述手性醇去外消旋化的条件下，提供至少一种根据权利要求1-11的任一项中提供的工程化酮还原酶变体，提供至少一种工程化亚磷酸脱氢酶变体、至少一种手性醇和至少一种辅因子；
其中所述工程化亚磷酸脱氢酶变体与SEQ ID NO:172具有至少94％的序列同一性并且所述工程化亚磷酸脱氢酶变体的多肽序列与SEQ ID NO:172之间的氨基酸残基差异为选自以下的一个或更多个氨基酸残基差异：R10K、E13D、L18M、S32V、R41A、R44A、A59M、Q63A、R69K、C73A、A74T、F78Y、V83A、A88R、F95I、V96G、T104F/L/M、V113S、L119F、R120V、A124E、R132Q、Q135A、P136D、R137Q/I、N145G、F150I、L158K、A175S/T、K177T/S、A178P、R186T、Q191H、A193S、C194L、S195E、F198L、L206N、N211A、D213Q、L215P、V233I、V262P/D/L、E266V/W/S/A、I293L、S295R、E303A、A308V、A317R、I322Q/M、N323D/N、V325A、R327S/D、L328P、A334K、C336R/S；或者其中所述工程化亚磷酸脱氢酶变体由SEQ ID NO:172或260中列出的多肽序列组成；并且
其中所述工程化亚磷酸脱氢酶变体具有亚磷酸脱氢酶活性。
14.如权利要求13所述的方法，其中所述工程化亚磷酸脱氢酶变体由选自SEQ ID NO:174至258中列出的偶数编号的序列的多肽序列组成。
15.如权利要求13或14所述的方法，其中所述方法以一锅反应来进行。
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