CN1934261A - 过氧化氢驱动的氧化反应 - Google Patents
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Abstract
本发明提供一种进行由单加氧酶催化及使用过氧化氢作为氧化剂的氧化反应的方法，其中发生单加氧酶的低水平氧化损伤反应，所述方法包括在氧化反应同时产生过氧化氢，其中过氧化氢以低于或等于其在该反应中使用的速率产生，或所述方法包括在H2O2或羟基螯合剂的存在下进行反应，所述螯合剂可控制H2O2或羟基浓度。
Description
技术领域
本发明涉及一种进行氧化反应的方法。
背景技术
单加氧酶催化广泛底物的氧化反应。为催化该反应，单加氧酶通常需要辅助因子及至少一种电子传递配偶蛋白(electron-transfer partnerprotein)(还原酶)。然而，单加氧酶能够使用过氧化氢(H2O2)作为氧化剂，这是因为H2O2可作为分子氧和两个电子的来源。使用H2O2驱动氧化反应被称为“过氧化物旁路(peroxide shunt)”。
发明内容
与过氧化物酶相比，单加氧酶对H2O2通常具有高Km值(例如大约20mM)。因此，当使用过氧化物旁路进行氧化反应时，需要高浓度H2O2以取得单加氧酶活性的可观水平。例如，使用50mM H2O2的单加氧酶活性的起始速率大大低于使用天然辅助因子NAD(P)H及生理电子传递配偶体时单加氧酶活性的起始速率。
本发明提供一种进行氧化反应的更有效的方法，该氧化反应使用过氧化物旁路，通过反应过程中不允许过高水平H2O2存在减少单加氧酶的氧化性损伤而进行。
H2O2以低于或等于其在单加氧酶催化的氧化反应中使用的速率同时产生，结果导致氧化反应效率提高及产物量增加。可使用多种方法以所需速率产生H2O2，例如电化学反应、酶或前体的使用。
因此，本发明提供一种进行氧化反应的方法，该氧化反应由单加氧酶催化并使用过氧化氢作为氧化剂，所述反应中单加氧酶的氧化损伤反应水平低，所述方法包括在氧化反应的同时产生过氧化氢，其中过氧化氢以低于或等于其在该反应中使用的速率产生。
本发明还提供一种单加氧酶催化及使用过氧化氢作为氧化剂进行氧化反应的方法，所述反应中单加氧酶的氧化损伤反应水平低，所述方法包括在H2O2或羟基螯合剂的存在下进行反应，所述螯合剂可控制H2O2或羟基浓度。
序列描述
SEQ ID NO：1显示了恶臭假单胞菌(Pseudomonas putida)的细胞色素P450Cam的核苷酸序列。
SEQ ID NO：2显示了恶臭假单胞菌的细胞色素P450Cam的氨基酸序列。
SEQ ID NO：3显示了巨大芽孢杆菌(Bacillus megaterium)的细胞色素P450BM-3的核苷酸序列。
SEQ ID NO：4显示了巨大芽孢杆菌的细胞色素P450BM-3的氨基酸序列。开始的472个氨基酸残基形成血红素结构域。最后585个氨基酸残基形成还原酶结构域。所有1048个氨基酸残基形成全酶。
在此使用的本领域中指明一个突变体的常规是：参照序列中某一位置的天然氨基酸残基，在其后为突变体中该位置的氨基酸，例如F87是指野生型序列的87位置上的苯丙氨酸，F87A是指在变异体中已改变为丙氨酸的野生型序列87位置上的苯丙氨酸。氨基酸残基编号从起始甲硫氨酸残基之后的氨基酸残基开始。
实例中使用的突变体为F87A(单突变；SEQ ID NO：5和SEQ IDNO：6)和F87V L188Q A74G(三重突变；SEQ ID NO：7和SEQ IDNO：8)。
SEQ ID NO：5显示了巨大芽孢杆菌的细胞色素P450BM-3的F87A突变体的氨基酸序列。
SEQ ID NO：6显示了巨大芽孢杆菌的细胞色素P450BM-3的F87A突变体的核苷酸序列。
SEQ ID NO：7显示了巨大芽孢杆菌的细胞色素P450BM-3的F87V L188Q A74G突变体的氨基酸序列。
SEQ ID NO：8显示了巨大芽孢杆菌的细胞色素P450BM-3的F87V L188Q A74G突变体的核苷酸序列。
SEQ ID NO：9显示了珊瑚诺卡氏菌(Nocardia coralline)的B-276烯烃环氧化酶1亚基的核苷酸序列。
SEQ ID NO：10显示了珊瑚诺卡氏菌的B-276烯烃环氧化酶1亚基的氨基酸序列。
SEQ ID NO：11显示了珊瑚诺卡氏菌的B-276烯烃环氧化酶2亚基的核苷酸序列。
SEQ ID NO：12显示了珊瑚诺卡氏菌的B-276烯烃环氧化酶2亚基的氨基酸序列。
SEQ ID NO：13显示了黄色杆菌属种(Xanthobacta sp.)的Py2烯烃单加氧酶α亚基的核苷酸序列。
SEQ ID NO：14显示了黄色杆菌属种的Py2烯烃单加氧酶α亚基的氨基酸序列。
SEQ ID NO：15显示了黄色杆菌属种的Py2烯烃单加氧酶β亚基的核苷酸序列。
SEQ ID NO：16显示了黄色杆菌属种的Py2烯烃单加氧酶β亚基的氨基酸序列。
SEQ ID NO：17显示了黄色杆菌属种的Py2烯烃单加氧酶γ亚基的核苷酸序列。
SEQ ID NO：18显示了黄色杆菌属种的Py2烯烃单加氧酶γ亚基的氨基酸序列。
SEQ ID NO：19显示了荚膜甲基球菌(Methylococcus capsulatas)可溶性甲烷单加氧酶的α亚基的核苷酸序列。
SEQ ID NO：20显示了荚膜甲基球菌可溶性甲烷单加氧酶α亚基的氨基酸序列。
SEQ ID NO：21显示了荚膜甲基球菌可溶性甲烷单加氧酶β亚基的核苷酸序列。
SEQ ID NO：22显示了荚膜甲基球菌可溶性甲烷单加氧酶β亚基的氨基酸序列。
SEQ ID NO：23显示了荚膜甲基球菌可溶性甲烷单加氧酶γ亚基的核苷酸序列。
SEQ ID NO：24显示了荚膜甲基球菌可溶性甲烷单加氧酶γ亚基的氨基酸序列。
SEQ ID NO：25显示了食油假单胞菌(Pseudomonas oleovorans)Gpo1烷羟化酶(AlkB基因)的核苷酸序列。
SEQ ID NO：26显示了食油假单胞菌Gpo1烷羟化酶的氨基酸序列。
SEQ ID NO：27显示了洋葱伯克霍尔德氏菌(Burkholderiacepacia)甲苯2-单加氧酶α亚基的核苷酸序列。
SEQ ID NO：28显示了洋葱伯克霍尔德氏菌甲苯2-单加氧酶α亚基的氨基酸序列。
SEQ ID NO：29显示了洋葱伯克霍尔德氏菌甲苯2-单加氧酶β亚基的核苷酸序列。
SEQ ID NO：30显示了洋葱伯克霍尔德氏菌甲苯2-单加氧酶β亚基的氨基酸序列。
SEQ ID NO：31显示了洋葱伯克霍尔德氏菌甲苯2-单加氧酶γ亚基的核苷酸序列。
SEQ ID NO：32显示了洋葱伯克霍尔德氏菌甲苯2-单加氧酶γ亚基的氨基酸序列。
SEQ ID NO：33显示了嗜热脂肪芽孢杆菌(Bacillusstearothermophilus)苯酚羟化酶(pheA)基因的核苷酸序列。
SEQ ID NO：34显示了嗜热脂肪芽孢杆菌苯酚羟化酶基因的氨基酸序列。
SEQ ID NO：35显示了向日葵(Helianthus annuus)硬脂酰-ACP去饱和酶的核苷酸序列。
SEQ ID NO：36显示了向日葵硬脂酰-ACP去饱和酶的氨基酸序列。
具体实施方式
应该理解本发明不限于具体实施方案。还应该理解本发明所公开的方法的不同应用可以适合本领域的特定需要。还应该理解在此使用的术语仅为描述本发明具体实施方案使用，而不旨在限制。
另外，除非内容中清楚指出，否则本说明书及附加权利要求书中所使用的单数形式“一”、“一种”、“该”包括复数指示物。因此，例如“一种底物”包括两种或更多种底物，“一种酶”包括引用两种或更多种酶，等等。
本发明引用的所有出版物、专利及专利出版物，无论在上文还是下文，均通过援引的方式完整地纳入本申请。
本发明方法能够使多种底物被氧化。所述底物包括但不限于：烷类、芳香族化合物、萜类化合物、烯烃及脂肪酸。
合适的烷类包括但不限于：甲烷、乙烷、丙烷、丁烷、戊烷、己烷、庚烷、正辛烷、正壬烷、正癸烷、正十二烷及正十六烷。烷类氧化产生醇。甲烷氧化成甲醇在技术上及经济上非常重要。中链醇(如正辛醇)为合成的中间产物，而较长链的醇(如正十二醇)用于脂肪酸衍生物的合成。
合适的芳香族化合物包括但不限于：苯、甲苯、二甲苯、氯苯、苯酚及其取代物。酚及儿茶酚产物用于香味和香料化合物的合成。
合适的萜类化合物包括但不限于：一萜类，如柠檬烯、蒎烯、松油烯及罗勒烯，例如朱栾倍半萜和香橙烯的倍半萜，以及包括甾族化合物类的三萜类。这些产物为用于合成的中间产物、优良香味和香料化学品及药品。
合适的烯烃包括但不限于：简单分子如丙烯、己-1-烯、己-2-烯及苯乙烯，以及复杂分子中的碳-碳双键。烯烃对于单对映异构物的选择性环氧化作用在合成中非常重要。光学纯的氧化丙烯及氧化苯乙烯在合成中是非常有用的中间产物。
羟化脂肪酸为聚合体的前体。
单加氧酶
根据本发明用于进行氧化反应的酶为单加氧酶。本领域普通技术人员使用本领域标准技术能够确定酶是否为单加氧酶。通常情况下，可使用蛋白质晶体学对辅基进行表征，尤其是非血红素铁酶的辅基，这是因为辅基通常不具有发色团。另外，本领域普通技术人员通常会使用序列对比寻找保守基序如活性部位、和铁含量以及亚基组分。
单加氧酶对H2O2的Km值优选为至少15nM、至少20nM、至少25nM、至少30nM、至少35nM、至少40nM、至少45nM或至少50nM。
单加氧酶的实例包括但不限于：细胞色素P450单加氧酶及非血红素二铁单加氧酶。合适的非血红素二铁单加氧酶包括但不限于：甲烷单加氧酶(Colby et al.，Biochem.J.，1977；165：395-402；Dalton，Adv.Appl.Microbiol.，1980；26：71-87；Fox et al.，J.Biol.Chem.，1989；264：10023-10033；Fox et al.，Methods Enzymol.，1990；188：191-202；McDonald et al.，Appl.Environ.Microbiol.，1997；63：1898-1904)，烷羟化酶(van Beilen et al.，Enzyme Microb.Technol.，1994；16：904-911)，甲苯单加氧酶(Luykx et al.，Biochem.Biophys.Res.Commun.，2003；312：373-379；Pikus et al.，Biochemistry，1996；35：9106-9119；Newman & Wackett，Biochemistry，1995；34：14066-14076)，烯烃单加氧酶(Gallagher et al.，Eur.J.Biochem.，1997；247：635-641；Lange & Que，Curr.Opin.Chem.Biol.，1998；2：159-172；Zhou et al.，FEBS Lett.，1998；430：181-185)，酚单加氧酶(Divari et al.，Eur.J.Biochem.，2003；270：2244-2253)以及甾族去饱和酶(Shanklin et al.，Biochemistry，1994；33：12787-12794)。非血红素二铁单加氧酶通常为真核或原核来源，优选地为细菌、真菌、酵母、植物或动物来源。优选的序列示于SEQ ID NO：1至SEQ ID NO：36。
本发明方法使用的酶优选地为细胞色素P450酶，通常为真核或原核来源。细胞色素P450单加氧酶通常以亚铁-一氧化碳络合物的446-450nm亚铁血红素索雷谱带为表征。酶通常为细菌、真菌、酵母、植物或动物来源，因此可来自假单胞菌属(Pseudomonas)细菌。酶可为P450的天然存在形式，例如P450cam，巨大芽孢杆菌的P450BM-3，假单胞菌属种(Pseudomonas sp.)的P450terp，红色糖多孢菌(Saccharopollyspora erythraea)的P450eryF以及灰色链霉菌(Streptomyces griseus)菌株的P450 105 D1(CYP105)。
或者，酶可为P450天然存在形式的突变体。突变体保留天然存在的酶的必需生物活性，即催化使用H2O2的氧化反应的能力。突变体可在酶的活性部位具有一个或多个突变。
“活性部位”的氨基酸是在催化过程中界定或定义结合底物的部位的氨基酸或界定或定义底物在到达催化部位之前所必须经过的部位的氨基酸。因此，这类氨基酸在进入催化部位过程中或催化过程中通常与底物相互作用。所述相互作用通常通过静电相互作用(在带电荷基团或极性基团之间)、疏水性相互作用、氢键或范德华力产生。
可通过本领域普通技术人员使用的常规方法鉴定活性部位的氨基酸。这些方法包括标记研究，其中使酶结合修饰(“标记”)接触底物的氨基酸的底物。另外，可获得酶与结合的底物的晶体结构以推导出活性部位的氨基酸。
单加氧酶可具有1个、2个、3个、4个、5至10个、10至20个或更多其它突变，例如置换、插入或缺失。可在天然存在的酶的氨基酸序列进行氨基酸置换，例如从1、2、3、4或5至10、20或30个置换。例如可根据表1进行保守性置换。第二列中同一格的氨基酸及优选在第三列中同一行的氨基酸可互相置换。
表1保守性氨基酸置换
| 非芳香族 | 非极性 | GAP |
| ILV | ||
| 极性-不带电荷 | CSTM | |
| NQ | ||
| 极性-带电荷 | DE | |
| HKR | ||
| 芳香族 | HFWY |
突变可位于活性部位内或活性部位外。通常突变在影响或接触活性部位一个或多个氨基酸位置或定位的“第二个域”残基中。插入通常在N端和/或C端，因此酶可为嵌合蛋白的一部分。缺失通常包括与催化无关的的氨基酸的缺失，例如那些位于活性部位之外的氨基酸(由此而来的酶为天然产生酶的突变片段)。因此，单加氧酶可仅包括氧化活性所需的那些氨基酸。
活性部位的突变在底物结合于活性部位时通常改变底物的位置和/或构象。突变可使血红素基团更易接近待氧化的底物上的位点。因此，突变可以为对具有较小或较大、或较多或较少极性的侧链的氨基酸进行的置换。
突变通常增加蛋白质的的稳定性，或使蛋白质更易于被纯化。它们通常通过去除蛋白质的半胱氨酸残基(如通过在P450cam的334位置，或在同系物的相同位置半胱氨酸的置换，优选地置换为丙氨酸)以防止蛋白质的二聚体形成。它们通常使得蛋白质可以可溶形式制备，例如通过引入缺失或多组氨酸标签，或通过N端膜锚定序列的突变进行。突变通常抑制蛋白质寡聚化，例如蛋白质表面疏水部位之间接触产生的寡聚化。
突变可影响酶使用H2O2的方式，从而可改进反应的效率。例如，使用正亮氨酸置换P450BM-3的血红素结构域的所有甲硫氨酸残基可使该酶的过氧化物酶活性提高两倍(Cirino et al.，Biotechnol.Bioeng.，2003；83(6)：729-734)。另外，已报道直接进化研究发现酶突变体对过氧化物具有更大抵抗力(Cirino & Arnold，Angew.Chem.Int.Ed.，2003；42：3299-3301)。
因此，在氨基酸同一性基础上突变酶通常与天然存在的酶具有至少70％同源性。
本发明所述的突变蛋白质(即描述为另一蛋白质的突变体)通常在至少20个、优选地至少30个、例如至少40个、60个、或100个或更多连续氨基酸中与相关蛋白质具有至少70％的同源性，或至少80％或90％及更优选地至少95％、97％或99％的同源性。连续氨基酸可包括活性部位。或者可以不通过连续氨基酸测量此同源性，而仅通过活性部位的氨基酸测量。
可使用已知方法测量同源性。例如，UWGCG程序包提供了BESTFIT程序，可用于计算同源性(例如在其缺省设置使用)(Devereux et al.(1984)Nucleic Acids Research 12，p387-395)。PILEUP和BLAST算法可用于计算同源性或排列序列(通常在其缺省设置)，例如Altschul S.F.(1993)J Mol Evol 36：290-300；Altschul，S，F et al.(1990)J Mol Biol 215：403-10中所述。
进行BLAST分析的软件可通过国家生物技术信息中心(http://www.ncbi.nlm.nih.gov/)公开获得。该算法涉及首先通过鉴定查询序列中长度W的短字码鉴定高评分序列对(HSPs)，此短字码当与数据库序列中相同长度字码进行对比时可匹配或满足一些阳性值域值评分T。T被称为邻近字码评分域值(neighbourhood word scorethreshold)(Altschul et al.，supra)。这些起始邻近字码采样数作为起动搜索的种子以发现含有它们的HSPs。字码采样数沿每一序列向两个方向延伸直至累积对比得分可被增加。每一方向字码采样数的延伸在以下情况下被停止：累积对比得分从其最大获得值下降数量X；由于一个或多个负评分残基对比的累积，累积得分到达零或更低；或到达任一序列的末端。BLAST算法参数W、T及X决定对比的灵敏度和速度。BLAST程序使用以下项目作为缺省值：字码长度(W)11、BLOSUM62评分矩阵(见Henikoff and Henikoff(1992)Proc.Natl.Acad.Sci.USA 89：10915-10919)对比(B)50、期望值(E)10、M＝5、N＝4，以及对比两条链。
BLLAST算法实行了两条序列之间相似性的统计学分析，见于例如Karlin and Altschul(1993)Proc.Natl.Acad.Sci.USA 90：5873-5787。BLAST算法提供的一个相似性的量度是最小总概率(P(N))，它表示两条核苷酸或氨基酸序列之间的随机发生的匹配的概率。例如，在第一条序列与第二条序列对比中如果最小总概率小于约1，优选地小于约0.1，更优选地小于约0.01，最优选地小于约0.001，则此序列被认为与另一条序列相似。
突变体包括上述序列的片段。所述片段保留有单加氧酶活性。片段长度可为至少300、至少400或至少450个氨基酸。按照下列更详细描述，所述片段可用于产生嵌合酶。
突变体还包括含有天然存在的酶的片段或部分的嵌合蛋白。一个或多个氨基酸可备选地或额外地添加至上述多肽。可于天然存在的酶或其变异体或片段的N端或C端提供延伸。延伸可以很短，例如1至10氨基酸长度。或者，延伸可较长。可将载体蛋白融合至上述氨基酸序列。因此，合并有上述一种酶的融合蛋白可用于本发明。
天然存在的酶或其变异体还可被化学修饰。许多侧链修饰为本领域已知，并可用于上述酶的侧链。所述修饰包括例如：糖基化、磷酸化、通过与乙醛反应然后使用NaBH4还原的还原烃基化进行的氨基酸修饰、甲基乙酰亚胺酯脒基化(amidination)或使用乙酸酐的酰化作用。所述修饰优选为糖基化。
通常使用本领域已知的方法将本发明讨论的突变引入酶中，例如酶的位点定向诱变，PCR和基因改组方法或使用位点定向诱变周期中多重诱变寡核苷酸。因此，可使用定向或随机方法引入突变。因此，诱变方法可产生编码一个或多个不同突变的一个或多个多核苷酸。通常产生可用于产生突变酶文库的突变寡核苷酸文库。
可使用本领域已知方法通过合成或重组的手段制备酶。单加氧酶的氨基酸序列可被修饰成包括非天然存在的氨基酸或增加酶的稳定性。当通过合成方法产生酶时，可于产生过程中引入所述氨基酸。蛋白质或肽还可以在合成或重组产生之后被修饰。
还可以使用D-氨基酸产生酶。在这种情况下，以由C至N的相反方向序列连接氨基酸。这是本领域产生所述蛋白质或肽的常规方法。
可以通过重组表达载体的多肽的原位表达在细胞中产生酶。表达载体任选地携带诱导型启动子以控制多肽的表达。可以在重组表达之后通过任何蛋白质液相层析系统纯化大规模产生酶。优选的蛋白质液相层析系统包括快速蛋白液相色谱(FPLC)、AKTA系统、Bio-Cad系统，Bio-Cad BioLogic系统及Gilsion高效液相色谱(HPLC)系统。
氧化反应
本发明方法涉及由单加氧酶催化的高效率氧化反应。高效率氧化反应为一种在其发生时没有可观察到的酶转化减少或产物产量减少或单加氧酶失活的反应。优选地，在反应1小时、2小时、6小时、12小时、1天或5天之后单加氧酶显示有反应开始所表现活性的至少70％、至少80％、至少90％、至少95％、或100％。
通常情况下，本发明方法在体外进行，例如在无细胞系统中进行。
通过“过氧化物旁路”驱动反应。在单加氧酶(a)、底物(b)和H2O2(c)存在下进行本发明反应。通常在需氧条件下进行反应，并不需要任何辅助因子。(c)的产生在下文更为详细描述。在此系统中，电子的流动通常为：(c)→(a)→(b)。
在此方法中，(a)和(b)的浓度通常为10-8至10-2M，优选地为10-6至10-4M。(a)∶(b)的浓度比例通常为0.1∶10至1∶10，优选地为1∶0.5至1∶2，或1∶0.8至1∶1.2。优选地，(b)的浓度较(a)的浓度大。(a)的优选浓度为当其与底物反应时产生可通过可用的分析方法例如GC、HPLC检测到足够产物。这通常为uM数量等级。
通常在单加氧酶具有功能的温度和/或pH情况下实施该方法，例如当酶具有峰值活性的至少20％、50％、80％或更多时。通常pH为2-10，例如5-9或6-8，优选地为7至7.8或7.4。可使用合适的缓冲剂如基于磷酸或醋酸的系统维持pH。通常温度为0-80℃，例如25-75℃，30-60℃，或50-80℃。优选地，温度为20-40℃。
通常在所述方法中发生至少20个转化数/分钟，例如至少50、100、200、300、500或更多转化数(按照每纳摩尔酶形成的产物纳摩尔数测量转化数)。
通常，H2O2产生速率为少于或等于每分钟每mg单加氧酶1ug、2ug或3ug。通常，整个反应中H2O2的浓度低于或等于0.1、0.5或1mM。通常，反应持续至少60分钟，至少240分钟，至少6小时或至少10小时。
如果单加氧酶底物在反应条件下为液体，可在单加氧酶底物中实施本发明方法。还可以在溶剂中实施本发明方法。合适的溶剂包括但不限于：水、水缓冲溶液混合水/有机和水缓冲液/有机溶剂系统。优选地，有机溶剂为碳氢化合物例如己烷、苯、乙腈、低脂肪族醇、酮及二噁烷、二甲基酰胺及二甲基亚砜及其混合物。溶剂通常为在其中反应物和产物为高溶解性和维持单加氧酶稳定性和活性的溶剂。
可以在溶液中具有所有组分的均质系统中进行反应。通常，在搅拌釜反应器中将单加氧酶和底物混合在一起，以分批、半分批或连续模式进行反应。
或者，在实施本发明方法之前可于合适的固体支持物例如二氧化硅上固定化单加氧酶。可将固定化单加氧酶充填入固定床反应器并使底物经过酶。在一个实施方案中，可于与单加氧酶相同或不同的材料上固定化产生H2O2的酶(下文较详细讨论)。固定化酶的方法为本领域已知。所述方法实例包括但不限于：共价结合至不溶的有机或无机支持物，于凝胶中捕获及吸附至离子交换树脂或其它吸附性材料。(G.F.Bickerstaff ed.，″Immobilization of Enzymes and Cells，″HumanaPress，Totowa.New Jersey，1997)。
在另一个实施方案中，在“入口”侧的膜允许底物缓慢从“反应物”侧进入，然后在“出口”侧的亲水膜允许亲水化合物流出至流动反应池的“产物”侧。在此情况下，可于膜外产生H2O2，并允许其通过膜流至流动或不流动的酶。
在一个实施方案中，优选地通过下面较详细描述的方法中的一种产生H2O2。在另一个实施方案中，H2O2或羟基螯合剂在氧化反应过程中用于螯合过量的H2O2或羟基。该螯合剂可为一种螯合试剂。在一个实施方案中，螯合剂为EDTA。EDTA抑制羟基的产生，例如痕量铁(或铜)与H2O2反应产生的羟基。
电化学反应产生H2O2
可以使用本发明方法通过一种电化学反应产生H2O2。电化学反应通常为一种将电流引入液体，优选地为溶剂的方法。电化学反应通常为在电流通过的电极处发生的氧化反应或还原反应。电极为能够导电的固体，通常为基于碳的或金属，其使得外部电源或接收器与优选为溶液的液体接触。电极可为带正电荷(负极)或带负电荷(正极)。两个或更多电极可形成电化学池，从电化学池外部电线可从每一电极连接至外部电装置。氧化反应或还原反应可在一个电极发生，而氧化还原反应可于电化学池中发生或直接在液体中发生。
使用电化学反应产生H2O2为具有能效的。通常通过由分子氧至过氧化氢的受控电化学还原产生H2O2。负极的表面积及超电势是由分子氧至过氧化氢的两个电子还原的关键考虑因素。通常使用基于碳的负极，并可使用已知降低该反应超电势的化合物修饰该负极。有效及有效率地实施该方法的电极材料及改性剂为本领域已知。通常可以通过施加于负极的电势控制O2的还原并由此控制过氧化氢的产生。施加于负极的电势根据负极以及对负极进行的任何修饰而改变。
本发明方法中使用的电化学反应可为分子氧的超声电化学还原。该方法为本领域已知(Compton et al.，Electroanalysis，1997；9(7)：509-522)。
酶催化产生H2O2
在本发明方法中可由酶产生H2O2。酶优选地为氧化酶。合适氧化酶的实例包括但不限于：葡萄糖氧化酶(E.C.1.1.3.4)、仲醇氧化酶(E.C.1.1.3.18)、甲醇氧化酶(E.C.1.1.3.31)、草酸氧化酶(E.C.1.2.3.4)、芳香基-乙醛氧化酶(E.C.1.2.3.9)、一氧化碳氧化酶(E.C.1.2.3.10)、胺氧化酶(E.C.1.4.3.4)、乙醇胺氧化酶(E.C.1.4.3.8)、硝基乙烷氧化酶(E.C.1.7.3.1)及亚硫酸盐氧化酶。葡萄糖氧化酶(E.C.1.1.3.4)催化D-葡萄糖转化为D-葡萄糖-1，5-内酯和H2O2。仲醇氧化酶(E.C.1.1.3.18)催化仲醇转化为酮和H2O2。甲醇氧化酶(E.C.1.1.3.31)催化甲醇转化为甲醛和H2O2。草酸氧化酶(E.C.1.2.3.4)催化草酸转化为二氧化碳和H2O2。芳香基-乙醛氧化酶(E.C.1.2.3.9)催化芳族醛转化为芳香酸和H2O2。一氧化碳氧化酶(E.C.1.2.3.10)催化CO和H2O转化为二氧化碳和H2O2。胺氧化酶(E.C.1.4.3.4)催化RCH2NH2和H2O转化为RCHO和NH3和H2O2。乙醇胺氧化酶(E.C.1.4.3.8)催化乙醇胺和H2O转化为羟乙醛和H2O2。硝基乙烷氧化酶(E.C.1.7.3.1)催化硝基乙烷和H2O转化为乙醛和H2O2。亚硫酸盐氧化酶(E.C.1.8.3.1)催化亚硫酸盐和H2O2转化为硫酸盐和H2O2。氧化酶(如葡萄糖氧化酶)可商购。另外，可使用本领域已知方法从已知微生物中提取氧化酶。
氧化酶的底物在本领域内为人熟知。除底物外，产生H2O2的反应还涉及水。通常在反应中还包括H2O2活化金属。合适的金属包括但不限于：铈、铬、钴、铜、铁、锰、钼、银、钛、钨、钒及其混合物。含有上述金属的金属硅酸盐可被制备并用于本发明方法。产生所述金属硅酸盐的方法为本领域已知(Neumann et al.，Journal of Catalysis，1997；166：206-127)。金属硅酸盐优选地为四面体配位钛如硅酸盐-1(TS-1)、硅酸盐-2(TS-2)、沸石-β、ZSM-48的硅类似物和MCM-41(Murugaveland Roesky，Angew.Chem.Int.Ed.Engl.，1997；36(5)：477-479)。
在本发明的一个优选的实施方案中，含金属固体或金属硅酸盐用作固定化产生H2O2的酶的支持物。在另一个优选的实施方案中，还于相同或不同的金属硅酸盐支持物上固定化单加氧酶。
优选地，首先将氧化酶与其它反应成分混合，然后通过添加氧化酶底物起动反应。例如，将单加氧酶、单加氧酶底物和氧化酶全部混合，然后添加氧化酶。在优选的实施方案中，将P450BM3、辛烷和葡萄糖氧化酶混合一起，然后添加葡萄糖。通常可通过控制添加氧化酶底物的速率来实现H2O2产生的控制。
由前体产生H2O2
在本发明的方法中可由前体产生H2O2。通过添加前体至水中产生H2O2在本领域内为人熟知。前体包括但不限于：过硼酸盐、过碳酸盐、过磷酸盐及过亚硝酸盐。优选的前体为钠盐。可通过使用化合物如四乙酰基乙二胺增强前体产生H2O2的性质。添加至含单加氧酶和底物的溶液中的前体量可使与底物的酶促反应最大化，并使H2O2引起的酶失活最小化。优选地，所产生H2O2的浓度不超过酶的Km值，但足够产生酶的反应性种类。
实施例
实施例1
在此实验中，在P450BM3血红素结构域存在下使辛烷与电化学产生的H2O2反应。于100mL玻璃烧杯中室温使用三电极结构进行该实验。网状玻璃碳(RVC)负极、铂网正极及银/氯化银(Ag/AgCl)参照电极装在同一容器中。将RVC负极短暂浸渍于1mM 2-氨基蒽醌乙醇溶液，然后取出并使其在空气中干燥。使用含氧饱和的水Tris缓冲液、0.2MKCl、0.5mM辛烷、及3u M P450BM3 F87V L188Q A74G血红素结构域的反应溶液(50mM，pH 7.4)。将反应溶液搅拌至平衡(5-10分钟)，然后施加相对于Ag/AgCl的-0.55V的电势2小时，整个过程中持续搅拌溶液。气相色谱(GC)分析显示溶剂氯仿、辛烷、2-辛醇、3-辛醇及4-辛醇和内标准1-壬醇的存在。2-辛醇、3-辛醇及4-辛醇的相对比例为1∶1.1∶0.7。所形成的辛醇总浓度为141uM，表示每个酶转化数为47。
使用1.43uM野生型P450BM3血红素结构域进行了相似的实验。所形成的辛醇的总浓度为8.4uM，表示每个酶转化数为6。2-辛醇、3-辛醇及4-辛醇的相对比例为1∶1.7∶2.0。
实施例2
在此实验中，在P450BM3全酶存在下使辛烷与酶作用产生的H2O2发生反应。向玻璃瓶中添加由水Tris缓冲液(50mM，pH 7.4)、0.5mM辛烷、1.6μM P450BM3 F87V L188Q A74G全酶及葡萄糖氧化酶(1.5U)组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加葡萄糖(1×10-6摩尔)起动反应。连续添加葡萄糖(1×10-6摩尔)，每5分钟一次，直至1小时(总共12次添加的当量为1.2×10-5摩尔)。在此时间内持续搅拌反应物，1.5小时之后停止。GC分析显示溶剂氯仿、辛烷、2-辛醇、3-辛醇和4-辛醇及内标准1-壬醇的存在。2-辛醇、3-辛醇和4-辛醇的相对比例为1∶1.1∶0.8。所形成的辛醇的总浓度为17uM，表示每个酶转化数为10。
实施例3
在此实验中，在P450BM3全酶的存在下，使辛烷与过硼酸钠产生的H2O2反应。向玻璃瓶中添加由水Tris缓冲液(40mM，pH 7.4)、0.5mM辛烷、1.3μM P450BM3 F87V L188Q A74G全酶组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加NaBO3.4H2O(1×10-4摩尔)起动反应，连续搅拌1小时。GC分析显示溶剂氯仿、辛烷、2-辛醇、3-辛醇和4-辛醇及内标准1-壬醇的存在。2-辛醇、3-辛醇和4-辛醇的相对比例为1∶1.8∶1.1。所形成的辛醇的总浓度为77uM，表示每个酶转化数为59。
对于实施例1至3，当溶液中无P450酶时观察不到壬醇产物。
实施例4
在此实验中，在P450BM3血红素结构域的存在下，使蒎烯与过硼酸钠产生的H2O2反应。向玻璃瓶中添加由水Tris缓冲液(40mM，pH7.4)、0.63mM蒎烯及3.7uM野生型P450BM3血红素结构域组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加7.8×10-6摩尔NaBO3.4H2O起动反应，持续搅拌1小时。GC分析显示顺式/反式2，3-环氧化物(32％)、(+)-反式马鞭草烯醇(16％)、(+)-顺式马鞭草烯醇(6％)、(+)-马鞭草烯酮/(+)-桃金娘烯醇(13％)、桃金娘烯醛(4％)以及未鉴定的其它氧化产物(29％)的存在。所形成的产物的总浓度为80uM，表示每个酶转化数为22。
实施例5
在此实验中，在过硼酸钠产生的H2O2存在的条件下，使酚单加氧酶与苯酚反应。向玻璃瓶中添加由水Tris缓冲液(40mM，pH 7.4)、0.63mM苯酚及3.7uM野生型酚单加氧酶组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加7.8×10-6摩尔NaBO3.4H2O起动反应，持续搅拌1小时。GC分析显示了氧化产物的存在。
实施例6
在此实验中，在葡萄糖氧化酶产生的H2O2的存在下，使P450BM3与棕榈酸反应。向玻璃瓶中添加由水Tris缓冲液(50mM，pH 7.4)、0.5mM棕榈酸、1.6μM P450BM3全酶及葡萄糖氧化酶(1.5U)组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加葡萄糖(1×10-6摩尔)起动反应。连续添加葡萄糖(1×10-6摩尔)，每5分钟一次，直至1小时(总共12次添加的当量为1.2×10-5摩尔)。在此时间内持续搅拌反应物，1.5小时之后停止。GC分析显示了氧化产物的存在。
实施例7
使植物CYP74C与13S-氢过氧化亚麻酸反应形成化合物3Z-己烯醛(芳香剂)。H2O2由过硼酸钠产生。向玻璃瓶中添加由水Tris缓冲液(40mM，pH 7.4)、0.63mM 13S-氢过氧化亚麻酸及3.7μM野生植物CYP74C组成的溶液(总体积为5mL)。平衡(5分钟)之后，通过添加7.8×10-6摩尔NaBO3.4H2O起动反应，持续搅拌1小时。GC分析显示了氧化产物的存在。
序列表
<110>ISIS创新有限公司
<120>过氧化氢氧化反应
<130>N.91079A SA
<160>36
<170>PatentIn version 3.2
<210>1
<211>1248
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<213>恶臭假单胞菌
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<221>CDS
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atg acg act gaa acc ata caa agc aac gcc aat ctt gcc cct ctg cca 48
Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro
1 5 10 15
ccc cat gtg cca gag cac ctg gta ttc gac ttc gac atg tac aat ccg 96
Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro
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tcg aat ctg tct gcc ggc gtg cag gag gcc tgg gca gtt ctg caa gaa 144
Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu
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tca aac gta ccg gat ctg gtg tgg act cgc tgc aac ggc gga cac tgg 192
Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp
50 55 60
atc gcc act cgc ggc caa ctg atc cgt gag gcc tat gaa gat tac cgc 240
Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg
65 70 75 80
cac ttt tcc agc gag tgc ccg ttc atc cct cgt gaa gcc ggc gaa gcc 288
His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala
85 90 95
tac gac ttc att ccc acc tcg atg gat ccg ccc gag cag cgc cag ttt 336
Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe
100 105 110
cgt gcg ctg gcc aac caa gtg gtt ggc atg ccg gtg gtg gat aag ctg 384
Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu
115 120 125
gag aac cgg atc cag gag ctg gcc tgc tcg ctg atc gag agc ctg cgc 432
Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg
130 135 140
ccg caa gga cag tgc aac ttc acc gag gac tac gcc gaa ccc ttc ccg 480
Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro
145 150 155 160
ata cgc atc ttc atg ctg ctc gca ggt cta ccg gaa gaa gat atc ccg 528
Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro
165 170 175
cac ttg aaa tac cta acg gat cag atg acc cgt ccg gat ggc agc atg 576
His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met
180 185 190
acc ttc gca gag gcc aag gag gcg ctc tac gac tat ctg ata ccg atc 624
Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile
195 200 205
atc gag caa cgc agg cag aag ccg gga acc gac gct atc agc atc gtt 672
Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val
210 215 220
gcc aac ggc cag gtc aat ggg cga ccg atc acc agt gac gaa gcc aag 720
Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys
225 230 235 240
agg atg tgt ggc ctg tta ctg gtc ggc ggc ctg gat acg gtg gtc aat 768
Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn
245 250 255
ttc ctc agc ttc agc atg gag ttc ctg gcc aaa agc ccg gag cat cgc 816
Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg
260 265 270
cag gag ctg atc gag cgt ccc gag cgt att cca gcc gct tgc gag gaa 864
Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu
275 280 285
cta ctc cgg cgc ttc tcg ctg gtt gcc gat ggc cgc atc ctc acc tcc 912
Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser
290 295 300
gat tac gag ttt cat ggc gtg caa ctg aag aaa ggt gac cag atc ctg 960
Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu
305 310 315 320
cta ccg cag atg ctg tct ggc ctg gat gag cgc gaa aac gcc tgc ccg 1008
Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro
325 330 335
atg cac gtc gac ttc agt cgc caa aag gtt tca cac acc acc ttt ggc 1056
Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly
340 345 350
cac ggc agc cat ctg tgc ctt ggc cag cac ctg gcc cgc cgg gaa atc 1104
His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile
355 360 365
atc gtc acc ctc aag gaa tgg ctg acc agg att cct gac ttc tcc att 1152
Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile
370 375 380
gcc ccg ggt gcc cag att cag cac aag agc ggc atc gtc agc ggc gtg 1200
Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val
385 390 395 400
cag gca ctc cct ctg gtc tgg gat ccg gcg act acc aaa gcg gta taa 1248
Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val
405 410 415
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Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro
1 5 10 15
Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro
20 25 30
Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu
35 40 45
Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp
50 55 60
Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg
65 70 75 80
His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala
85 90 95
Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe
100 105 110
Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu
115 120 125
Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg
130 135 140
Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro
145 150 155 160
Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro
165 170 175
His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met
180 185 190
Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile
195 200 205
Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val
210 215 220
Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys
225 230 235 240
Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn
245 250 255
Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg
260 265 270
Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu
275 280 285
Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser
290 295 300
Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu
305 310 315 320
Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro
325 330 335
Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly
340 345 350
His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile
355 360 365
Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile
370 375 380
Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val
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Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val
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agatctttat gaagacatag ctgcagaaga aaaagcaaga gctacatatc aatggttaat 60
tgatatatca gatgatcccg atttaaacga cagcttacga tttttacgag aaagagagat 120
tgttcactca cagcggttcc gcgaggccgt ggagatttta aaagatgaca gagacaggaa 180
gaaaatcttt taactagtaa aaaaacatcc cccttggcga atgcaaacga aaggagggat 240
gttttttgtt gtgactgcgt tgattatgcg ctagaactgc agtgacaaga aacaaccttt 300
aatttccctt caacatcttt ccaaactcgc gtataactgt attcacctcc aatagattca 360
ccggttgcca gtgccccatt taacgctact tttgtaacgg taacggcaag ttcttgaaac 420
agtttaactt cttgttccaa cacttccatg cccgctatat caagactttt tgaacgatga 480
acatttatat cttcttcttt tgacaaccat tgcccaaggt gattcacaaa aataagctca 540
tctgaaagta attcttctaa tagctctatg ttattagaaa gcatggctga gcgaagcatt 600
tcttcgtatt ctataactct tgcttgattc atttttaatc ctcctttacg ccttgtgtaa 660
ctcttttcta tttccacgtt gcttttcctt taaacttctt tcattaataa ttcgtgctaa 720
attatgttaa tagaggggat aagtggacta attttctgta agcactaaat attctgaaat 780
actctgttaa ttacctttaa atggtataaa attagaatga aagaaccttt tctttccact 840
tttctagtta tctttttact attaagatgc agttttttat acttgtaatt gtagcggaat 900
gaacgttcat tccgtttttg aaaagaggtg ataaagtgga atctactcca acaaaacaaa 960
aagcgatttt ttctgcttcg cttctgctgt ttgcagaaag agggtttgat gcaaccacga 1020
tgccaatgat tgcagagaat gccaaagtag gagcaggaac aatttatcgc tactttaaaa 1080
ataaagaaag ccttgtaaat gaattattcc aacagcacgt aaacgagttt ttacagtgca 1140
ttgaaagcgg tctggcaaac gagagagatg gataccgaga tgggtttcat catatctttg 1200
aaggtatggt gacatttact aaaaaccatc ctcgtgctct tggatttatt aaaactcata 1260
gccaaggaac ttttttaaca gaagagagcc gcttagcata tcaaaagctg gtggaatttg 1320
tttgtacgtt cttcagagaa ggacaaaagc aaggtgtgat tagaaatctt cctgaaaatg 1380
cgctaattgc tattttattt ggaagtttca tggaagtata tgaaatgatt gaaaatgact 1440
acttatcttt aactgatgaa cttcttaccg gtgtagaaga gagtctgtgg gcagcactta 1500
gcagacaatc atgaaactta acaagtgaaa gagggataac atg aca att aaa gaa 1555
Met Thr Ile Lys Glu
1 5
atg cct cag cca aaa acg ttt gga gag ctt aaa aat tta ccg tta tta 1603
Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn Leu Pro Leu Leu
10 15 20
aac aca gat aaa ccg gtt caa gct ttg atg aaa att gcg gat gaa tta 1651
Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile Ala Asp Glu Leu
25 30 35
gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta acg cgc tac tta 1699
Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val Thr Arg Tyr Leu
40 45 50
tca agt cag cgt cta att aaa gaa gca tgc gat gaa tca cgc ttt gat 1747
Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu Ser Arg Phe Asp
55 60 65
aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat ttt gca gga gac 1795
Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp Phe Ala Gly Asp
70 75 80 85
ggg tta ttt aca agc tgg acg cat gaa aaa aat tgg aaa aaa gcg cat 1843
Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn Trp Lys Lys Ala His
90 95 100
aat atc tta ctt cca agc ttc agt cag cag gca atg aaa ggc tat cat 189l
Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met Lys Gly Tyr His
105 110 115
gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa aag tgg gag cgt 1939
Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln Lys Trp Glu Arg
120 125 110
cta aat gca gat gag cat att gaa gta ccg gaa gac atg aca cgt tta 1987
Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp Met Thr Arg Leu
135 140 145
acg ctt gat aca att ggt ctt tgc ggc ttt aac tat cgc ttt aac agc 2035
Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr Arg Phe Asn Ser
150 155 160 165
ttt tac cga gat cag cct cat cca ttt att aca agt atg gtc cgt gca 2083
Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser Met Val Arg Ala
170 175 180
ctg gat gaa gca atg aac aag ctg cag cga gca aat cca gac gac cca 2131
Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn Pro Asp Asp Pro
185 190 195
gct tat gat gaa aac aag cgc cag ttt caa gaa gat atc aag gtg atg 2179
Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp Ile Lys Val Met
200 205 210
aac gac cta gta gat aaa att att gca gat cgc aaa gca agc ggt gaa 2227
Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys Ala Ser Gly Glu
215 220 225
caa agc gat gat tta tta acg cat atg cta aac gga aaa gat cca gaa 2275
Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly Lys Asp Pro Glu
230 235 240 245
acg ggt gag ccg ctt gat gac gag aac att cgc tat caa att att aca 2323
Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr Gln Ile Ile Thr
250 255 260
ttc tta att gcg gga cac gaa aca aca agt ggt ctt tta tca ttt gcg 2371
Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu Leu Ser Phe Ala
265 270 275
ctg tat ttc tta gtg aaa aat cca cat gta tta caa aaa gca gca gaa 2419
Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln Lys Ala Ala Glu
280 285 290
gaa gca gca cga gtt cta gta gat cct gtt cca agc tac aaa caa gtc 2467
Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser Tyr Lys Gln Val
295 300 305
aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa gcg ctg cgc tta 2515
Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu Ala Leu Arg Leu
310 315 320 325
tgg cca act gct cct gcg ttt tcc cta tat gca aaa gaa gat acg gtg 2563
Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys Glu Asp Thr Val
330 335 340
ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa cta atg gtt ctg 2611
Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu Leu Met Val Leu
345 350 355
att cct cag ctt cac cgt gat aaa aca att tgg gga gac gat gtg gaa 2659
Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly Asp Asp Val Glu
360 365 370
gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg att ccg cag cat 2707
Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala Ile Pro Gln His
375 380 385
gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt atc ggt cag cag 2755
Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys Ile Gly Gln Gln
390 395 400 405
ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg atg cta aaa cac 2803
Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met Met Leu Lys His
410 415 420
ttt gac ttt gaa gat cat aca aac tac gag ctg gat att aaa gaa act 2851
Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp Ile Lys Glu Thr
425 430 435
tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca aaa tcg aaa aaa 2899
Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala Lys Ser Lys Lys
440 445 450
att ccg ctt ggc ggt att cct tca cct agc act gaa cag tct gct aaa 2947
Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu Gln Ser Ala Lys
455 460 465
aaa gta cgc aaa aag gca gaa aac gct cat aat acg ccg ctg ctt gtg 2995
Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr Pro Leu Leu Val
470 475 480 485
cta tac ggt tca aat atg gga aca gct gaa gga acg gcg cgt gat tta 3043
Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr Ala Arg Asp Leu
490 495 500
gca gat att gca atg agc aaa gga ttt gca ccg cag gtc gca acg ctt 3091
Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln Val Ala Thr Leu
505 510 515
gat tca cac gcc gga aat ctt ccg cgc gaa gga gct gta tta att gta 3139
Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala Val Leu Ile Val
520 525 530
acg gcg tct tat aac ggt cat ccg cct gat aac gca aag caa ttt gtc 3187
Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala Lys Gln Phe Val
535 540 545
gac tgg tta gac caa gcg tct gct gat gaa gta aaa ggc gtt cgc tac 3235
Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys Gly Val Arg Tyr
550 555 560 565
tcc gta ttt gga tgc ggc gat aaa aac tgg gct act acg tat caa aaa 3283
Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr Thr Tyr Gln Lys
570 575 580
gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa ggg gca gaa aac 3331
Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys Gly Ala Glu Asn
585 590 595
atc gct gac cgc ggt gaa gca gat gca agc gac gac ttt gaa ggc aca 3379
Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp Phe Glu Gly Thr
600 605 610
tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta gca gcc tac ttt 3427
Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val Ala Ala Tyr Phe
615 620 625
aac ctc gac att gaa aac agt gaa gat aat aaa tct act ctt tca ctt 3475
Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser Thr Leu Ser Leu
630 635 640 645
caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg aaa atg cac ggt 3523
Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala Lys Met His Gly
650 655 660
gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt caa cag cca ggc 3571
Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu Gln Gln Pro Gly
665 670 675
agt gca cga agc acg cga cat ctt gaa att gaa ctt cca aaa gaa gct 3619
Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu Pro Lys Glu Ala
680 685 690
tct tat caa gaa gga gat cat tta ggt gtt att cct cgc aac tat gaa 3667
Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro Arg Asn Tyr Glu
695 700 705
gga ata gta aac cgt gta aca gca agg ttc ggc cta gat gca tca cag 3715
Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu Asp Ala Ser Gln
710 715 720 725
caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct cat ttg cca ctc 3763
Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala His Leu Pro Leu
730 735 740
gct aaa aca gta tcc gta gaa gag ctt ctg caa tac gtg gag ctt caa 3811
Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr Val Glu Leu Gln
745 750 755
gat cct gtt acg cgc acg cag ctt cgc gca atg gct gct aaa acg gtc 3859
Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala Ala Lys Thr Val
760 765 770
tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt gaa aag caa gcc 3907
Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu Glu Lys Gln Ala
775 780 785
tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg ctt gaa ctg ctt 3955
Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met Leu Glu Leu Leu
790 795 800 805
gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa ttt atc gcc ctt 4003
Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu Phe Ile Ala Leu
810 815 820
ctg cca agc ata cgc ccg cgc tat tac tcg att tct tca tca cct cgt 4051
Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser Ser Ser Pro Arg
825 830 835
gtc gat gaa aaa caa gca agc atc acg gtc agc gtt gtc tca gga gaa 4099
Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val Val Ser Gly Glu
840 845 850
gcg tgg agc gga tat gga gaa tat aaa gga att gcg tcg aac tat ctt 4147
Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala Ser Asn Tyr Leu
855 860 865
gcc gag ctg caa gaa gga gat acg att acg tgc ttt att tcc aca ccg 4195
Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe Ile Ser Thr Pro
870 875 880 885
cag tca gaa ttt acg ctg cca aaa gac cct gaa acg ccg ctt atc atg 4243
Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr Pro Leu Ile Met
890 895 900
gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc ttt gtg cag gcg 4291
Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly Phe Val Gln Ala
905 910 915
cgc aaa cag cta aaa gaa caa gga cag tca ctt gga gaa gca cat tta 4339
Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly Glu Ala His Leu
920 925 930
tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg tat caa gaa gag 4387
Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu Tyr Gln Glu Glu
935 940 945
ctt gaa aac gcc caa agc gaa ggc atc att acg ctt cat acc gct ttt 4435
Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu His Thr Ala Phe
950 955 960 965
tct cgc atg cca aat cag ccg aaa aca tac gtt cag cac gta atg gaa 4483
Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln His Val Met Glu
970 975 980
caa gac ggc aag aaa ttg att gaa ctt ctt gat caa gga gcg cac ttc 4531
Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln Gly Ala His Phe
985 990 995
tat att tgc gga gac gga agc caa atg gca cct gcc gtt gaa gca 4576
Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala Val Glu Ala
1000 1005 1010
acg ctt atg aaa agc tat gct gac gtt cac caa gtg agt gaa gca 4621
Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val Ser Glu Ala
1015 1020 1025
gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa ggc cga tac 4666
Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys Gly Arg Tyr
1030 1035 1040
gca aaa gac gtg tgg gct ggg taa attaaaaaga ggctaggata aaagtagttt 4720
Ala Lys Asp Val Trp Ala Gly
1045
agttggttga aggaagatcc gaacgatgaa tcgttcggat ctttttattg gtagagtaaa 4780
cgtagatttc atctatttag tgacttgtag cggttgattg gagggcaagg tgaagactcc 4840
aatcaaccgc ggtgtcacat gcaagccata cgaaattcat ttctcccatt tattcgtctt 4900
ttgtccccac ttaattttta tagcgcctta acgtttcttc tgcgtgacag cagatct 4957
<210>4
<211>1049
<212>PRT
<213>巨大芽孢杆菌
<400>4
Met Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys
1 5 10 15
Asn Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys
20 25 30
Ile Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg
35 40 45
Val Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp
50 55 60
Glu Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg
65 70 75 80
Asp Phe Ala Gly Asp Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn
85 90 95
Trp Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala
100 105 110
Met Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val
115 120 125
Gln Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu
130 135 140
Asp Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn
145 150 155 160
Tyr Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr
165 170 175
Ser Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala
180 185 190
Asn Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu
195 200 205
Asp Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg
210 215 220
Lys Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn
225 230 235 240
Gly Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg
245 250 255
Tyr Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly
260 265 270
Leu Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu
275 280 285
Gln Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro
290 295 300
Ser Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn
305 310 315 320
Glu Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala
325 330 335
Lys Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp
340 345 350
Glu Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp
355 360 365
Gly Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser
370 375 380
Ala Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala
385 390 395 400
Cys Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly
405 410 415
Met Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu
420 425 430
Asp Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys
435 440 445
Ala Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr
450 455 460
Glu Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn
465 470 475 480
Thr Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly
485 490 495
Thr Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro
500 505 510
Gln Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly
515 520 525
Ala Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn
530 535 540
Ala Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val
545 550 555 560
Lys Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala
565 570 575
Thr Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala
580 585 590
Lys Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp
595 600 605
Asp Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp
610 615 620
Val Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys
625 630 635 640
Ser Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu
645 650 655
Ala Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu
660 665 670
Leu Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu
675 680 685
Leu Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile
690 695 700
Pro Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly
705 710 715 720
Leu Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu
725 730 735
Ala His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln
740 745 750
Tyr Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met
755 760 765
Ala Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu
770 775 780
Leu Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr
785 790 795 800
Met Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser
805 810 815
Glu Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile
820 825 830
Ser Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser
835 840 845
Val Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile
850 855 860
Ala Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys
865 870 875 880
Phe Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu
885 890 895
Thr Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg
900 905 910
Gly Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu
915 920 925
Gly Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr
930 935 940
Leu Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr
945 950 955 960
Leu His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val
965 970 975
Gln His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp
980 985 990
Gln Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro
995 1000 1005
Ala Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln
1010 1015 1020
Val Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu
1025 1030 1035
Lys Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly
1040 1045
<210>5
<211>3147
<212>DNA
<213>人工序列
<220>
<223>细胞色素P450BM-3突变体
<220>
<221>CDS
<222>(1)..(3147)
<400>5
aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat 48
Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn
1 5 10 15
tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att 96
Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile
20 25 30
gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta 144
Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val
35 40 45
acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa 192
Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu
50 55 60
tca cgc ttt gat aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat 240
Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp
65 70 75 80
ttt gca gga gac ggg tta gct aca agc tgg acg cat gaa aaa aat tgg 288
Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp
85 90 95
aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg 336
Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met
100 105 110
aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa 384
Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln
115 120 125
aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac 432
Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp
130 135 140
atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat 480
Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr
145 150 155 160
cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt 528
Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser
165 170 175
atg gtc cgt gca ctg gat gaa gca atg aac aag ctg cag cga gca aat 576
Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn
180 185 190
cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat 624
Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp
195 200 205
atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa 672
Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys
210 215 220
gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga 720
Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly
225 230 235 240
aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat 768
Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr
245 250 255
caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt 816
Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu
260 265 270
tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa 864
Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln
275 280 285
aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc 912
Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser
290 295 300
tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa 960
Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu
305 310 315 320
gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa 1008
Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys
325 330 335
gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa 1056
Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu
340 345 350
cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga 1104
Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly
355 360 365
gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg 1152
Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala
370 375 380
att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt 1200
Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys
385 390 395 400
atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg 1248
Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met
405 410 415
atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat 1296
Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp
420 425 430
att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca 1344
Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala
435 440 445
aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa 1392
Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu
450 455 460
cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg 1440
Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr
465 470 475 480
ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg 1488
Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr
485 490 495
gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag 1536
Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln
500 505 510
gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct 1584
Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala
515 520 525
gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca 1632
Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala
530 535 540
aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa 1680
Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys
545 550 555 560
ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act 1728
Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr
565 570 575
acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa 1776
Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys
580 585 590
ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac 1824
Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp
595 600 605
ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta 1872
Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val
610 615 620
gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct 1920
Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser
625 630 635 640
act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg 1968
Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala
645 650 655
aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt 2016
Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu
660 665 670
caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt 2064
Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu
675 680 685
cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct 2112
Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro
690 695 700
cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta 2160
Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu
705 710 715 720
gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct 2208
Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala
725 730 735
cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac 2256
His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr
740 745 750
gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct 2304
Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala
755 760 765
gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt 2352
Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu
770 775 780
gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg 2400
Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met
785 790 795 800
ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa 2448
Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu
805 810 815
ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct 2496
Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser
820 825 830
tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt 2544
Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val
835 840 845
gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg 2592
Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala
850 855 860
tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt 2640
Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe
865 870 875 880
att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg 2688
Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr
885 890 895
ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc 2736
Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly
900 905 910
ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga 2784
Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly
915 920 925
gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg 2832
Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu
930 935 940
tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt 2880
Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu
945 950 955 960
cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag 2928
His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln
965 970 975
cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa 2976
His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln
980 985 990
gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc 3024
Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala
995 1000 1005
gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg 3069
Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val
1010 1015 1020
agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa 3114
Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys
1025 1030 1035
ggc cga tac gca aaa gac gtg tgg gct ggg taa 3147
Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly
1040 1045
<210>6
<211>1048
<212>PRT
<213>人工序列
<220>
<223>细胞色素P450BM-3突变体
<400>6
Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn
1 5 10 15
Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile
20 25 30
Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val
35 40 45
Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu
50 55 60
Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp
65 70 75 80
Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp
85 90 95
Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met
100 105 110
Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln
115 120 125
Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp
130 135 140
Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr
145 150 155 160
Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser
165 170 175
Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn
180 185 190
Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp
195 200 205
Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys
2l0 215 220
Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly
225 230 235 240
Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr
245 250 255
Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu
260 265 270
Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln
275 280 285
Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser
290 295 300
Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu
305 310 315 320
Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys
325 330 335
Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu
340 345 350
Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly
355 360 365
Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala
370 375 380
Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys
385 390 395 400
Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met
405 410 415
Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp
420 425 430
Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala
435 440 445
Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu
450 455 460
Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr
465 470 475 480
Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr
485 490 495
Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln
500 505 510
Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala
515 520 525
Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala
530 535 540
Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys
545 550 555 560
Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr
565 570 575
Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys
580 585 590
Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp
595 600 605
Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val
610 615 620
Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser
625 630 635 640
Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala
645 650 655
Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu
660 665 670
Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu
675 680 685
Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro
690 695 700
Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu
705 710 715 720
Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala
725 730 735
His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr
740 745 750
Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala
755 760 765
Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu
770 775 780
Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met
785 790 795 800
Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu
805 810 815
Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser
820 825 830
Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val
835 840 845
Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala
850 855 860
Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe
865 870 875 880
Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr
885 890 895
Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly
900 905 910
Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly
915 920 925
Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu
930 935 940
Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu
945 950 955 960
His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln
965 970 975
His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln
980 985 990
Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala
995 1000 1005
Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val
1010 1015 1020
Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys
1025 1030 1035
Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly
1040 1045
<210>7
<211>3147
<212>DNA
<213>人工序列
<220>
<223>细胞色素P450BM-3突变体
<220>
<221>CDS
<222>(1)..(3147)
<400>7
aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat 48
Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn
1 5 10 15
tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att 96
Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile
20 25 30
gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta 144
Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val
35 40 45
acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa 192
Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu
50 55 60
tca cgc ttt gat aaa aac tta agt caa ggg ctt aaa ttt gta cgt gat 240
Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp
65 70 75 80
ttt gca gga gac ggg tta gtt aca agc tgg acg cat gaa aaa aat tgg 288
Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp
85 90 95
aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg 336
Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met
100 105 110
aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa 384
Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln
115 120 125
aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac 432
Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp
130 135 140
atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat 480
Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr
145 150 155 160
cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt 528
Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser
165 170 175
atg gtc cgt gca ctg gat gaa gca atg aac aag cag cag cga gca aat 576
Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn
180 185 190
cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat 624
Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp
195 200 205
atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa 672
Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys
210 215 220
gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga 720
Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly
225 230 235 240
aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat 768
Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr
245 250 255
caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt 816
Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu
260 265 270
tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa 864
Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln
275 280 285
aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc 912
Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser
290 295 300
tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa 960
Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu
305 310 315 320
gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa 1008
Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys
325 330 335
gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa 1056
Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu
340 345 350
cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga 1104
Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly
355 360 365
gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg 1152
Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala
370 375 380
att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt 1200
Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys
385 390 395 400
atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg 1248
Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met
405 410 415
atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat 1296
Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp
420 425 430
att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca 1344
Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala
435 440 445
aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa 1392
Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu
450 455 460
cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg 1440
Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr
465 470 475 480
ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg 1488
Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr
485 490 495
gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag 1536
Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln
500 505 510
gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct 1584
Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala
515 520 525
gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca 1632
Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala
530 535 540
aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa 1680
Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys
545 550 555 560
ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act 1728
Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr
565 570 575
acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa 1776
Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys
580 585 590
ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac 1824
Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp
595 600 605
ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta 1872
Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val
610 615 620
gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct 1920
Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser
625 630 635 640
act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg 1968
Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala
645 650 655
aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt 2016
Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu
660 665 670
caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt 2064
Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu
675 680 685
cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct 2112
Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro
690 695 700
cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta 2160
Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu
705 710 715 720
gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct 2208
Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala
725 730 735
cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac 2256
His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr
740 745 750
gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct 2304
Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala
755 760 765
gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt 2352
Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu
770 775 780
gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg 2400
Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met
785 790 795 800
ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa 2448
Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu
805 810 815
ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct 2496
Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser
820 825 830
tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt 2544
Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val
835 840 845
gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg 2592
Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala
850 855 860
tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt 2640
Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe
865 870 875 880
att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg 2688
Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr
885 890 895
ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc 2736
Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly
900 905 910
ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga 2784
Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly
915 920 925
gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg 2832
Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu
930 935 940
tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt 2880
Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu
945 950 955 960
cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag 2928
His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln
965 970 975
cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa 2976
His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln
980 985 990
gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc 3024
Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala
995 1000 1005
gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg 3069
Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val
1010 1015 1020
agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa 3114
Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys
1025 1030 1035
ggc cga tac gca aaa gac gtg tgg gct ggg taa 3147
Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly
1040 1045
<210>8
<211>1048
<212>PRT
<213>人工序列
<220>
<223>细胞色素P450BM-3突变体
<400>8
Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn
1 5 10 15
Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile
20 25 30
Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val
35 40 45
Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu
50 55 60
Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp
65 70 75 80
Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp
85 90 95
Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met
100 105 110
Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln
115 120 125
Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp
130 135 140
Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr
145 150 155 160
Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser
165 170 175
Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn
180 185 190
Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp
195 200 205
Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys
210 215 220
Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly
225 230 235 240
Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr
245 250 255
Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu
260 265 270
Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln
275 280 285
Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser
290 295 300
Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu
305 310 315 320
Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys
325 330 335
Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu
340 345 350
Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly
355 360 365
Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala
370 375 380
Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys
385 390 395 400
Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met
405 410 415
Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp
420 425 430
Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala
435 440 445
Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu
450 455 460
Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr
465 470 475 480
Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr
485 490 495
Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln
500 505 510
Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala
515 520 525
Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala
530 535 540
Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys
545 550 555 560
Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr
565 570 575
Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys
580 585 590
Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp
595 600 605
Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val
610 615 620
Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser
625 630 635 640
Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala
645 650 655
Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu
660 665 670
Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu
675 680 685
Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro
690 695 700
Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu
705 710 715 720
Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala
725 730 735
His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr
740 745 750
Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala
755 760 765
Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu
770 775 780
Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met
785 790 795 800
Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu
805 810 815
Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser
820 825 830
Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val
835 840 845
Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala
850 855 860
Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe
865 870 875 880
Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr
885 890 895
Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly
900 905 910
Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly
915 920 925
Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu
930 935 940
Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu
945 950 955 960
His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln
965 970 975
His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln
980 985 990
Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala
995 1000 1005
Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val
1010 1015 1020
Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys
1025 1030 1035
Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly
1040 1045
<210>9
<211>1032
<212>DNA
<213>珊瑚诺卡氏菌
<220>
<221>CDS
<222>(1)..(1032)
<400>9
atg acg aca gag gcg acg gtg gcc cga ccg gtg gag ctc gaa ggt cac 48
Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His
1 5 10 15
cgg aca ttc acc tgg ttc acg ccc gcc agg cga aag ccg acg gag tac 96
Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr
20 25 30
gag ctc tac acc gtg ggt caa cag tcc act ccg gac gag tgg ctg cat 144
Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His
35 40 45
gtg gac tgg ccg ctg cgc ttc gac gac ggc cgc gcc ccg tgg gag gag 192
Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu
50 55 60
gag tcg agt gcg gta cgg acc tcg gag tgg tcg gct tac cgc gac cca 240
Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro
65 70 75 80
cac caa ctg tgg cag cgt ccc tac gtc agc acg tgc aac cag gac cag 288
His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln
85 90 95
cag gcc ctc gcg cgg ctg gtc ccc gtc ctg acc atg ggg tcg gcg gcg 336
Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala
100 105 110
atc acg ccc atc tgg tcg cag aag atc ctc gcc agg tcc tac gcc gcc 384
Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala
115 120 125
tgg cca ttc gtc gag tac ggg ctc ttc ctg agc ctg gcc tac gcc gtg 432
Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val
130 135 140
cgc cag gcc atg tcc gac acg gtc cag ttc agc gtg gtg ttc cag gcc 480
Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala
145 150 155 160
gtg gac cgc atg cgg ctg ctc cag gac atc gtc cac cac ctg gac cac 528
Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His
165 170 175
ctg cag gag tcg ccg gaa ttc agc gac gcc ggg gcc cgc gag gcc tgg 576
Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp
180 185 190
atg tcc gac tcc acc ctg gtc ccg atc cgg gaa gtg atc gag cgc atc 624
Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile
195 200 205
gcc gcc agc cag gac tgg gtg gag atc ctg gtc gcc ggc acg ctc gtc 672
Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val
210 215 220
ttc gag cct ctg gtc ggc cac ctg gcg aag gcc gag ttg ttc agc cgc 720
Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg
225 230 235 240
cgt gcg cca atg ttc ggg gac ggg acc acg ccg gcg gtg ctg gcg tcg 768
Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser
245 250 255
gcc ctg ctg gac agc ggc agg cac ctc gaa tcg gtc cag gcg ctc gtc 816
Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val
260 265 270
cgc ctc gtc tgc caa gac ccc gtc cat ggc gac cag aac cag gcg act 864
Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr
275 280 285
gtg cgg cgg tgg atc gag gaa tgg cag ccg cgg tgc aag gcg gcg gcc 912
Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala
290 295 300
cag tcc ttc ctg ccg acg ttc tcc gac tgc ggc atc gac gcc aag gaa 960
Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu
305 310 315 320
agc gcc aac gcg ctg tcc cgg gcg ctg gcg aac cag cgg gcc gcc gtc 1008
Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val
325 330 335
gag ggc gcc ggc atc acg gca tga 1032
Glu Gly Ala Gly Ile Thr Ala
340
<210>10
<211>343
<212>PRT
<213>珊瑚诺卡氏菌
<400>10
Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His
1 5 10 15
Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr
20 25 30
Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His
35 40 45
Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu
50 55 60
Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro
65 70 75 80
His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln
85 90 95
Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala
100 105 110
Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala
115 120 125
Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val
130 135 140
Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala
145 150 155 160
Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His
165 170 175
Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp
180 185 190
Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile
195 200 205
Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val
210 215 220
Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg
225 230 235 240
Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser
245 250 255
Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val
260 265 270
Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr
275 280 285
Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala
290 295 300
Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu
305 310 315 320
Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val
325 330 335
Glu Gly Ala Gly Ile Thr Ala
340
<210>11
<211>1506
<212>DNA
<213>珊瑚诺卡氏菌
<220>
<221>CDS
<222>(1)..(1506)
<400>11
atg gca tcg aac ccc acc cag ctc cac gag aag tcg aag tcc tac gac 48
Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp
1 5 10 15
tgg gac ttc acc tcc gtc gag cgg cgc ccc aag ttc gag acg aag tac 96
Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr
20 25 30
aag atg ccc aag aag ggc aag gac ccg ttc cgc gtc ctg atc cgt gac 144
Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp
35 40 45
tac atg aag atg gaa gcg gag aag gac gac cgg acc cat ggc ttc ctc 192
Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu
50 55 60
gac ggc gcc gtg cgg acg cgt gag gcc acc agg att gag ccg cgg ttc 240
Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe
65 70 75 80
gct gag gcc atg aag atc atg gtg ccg cag ctg acc aac gcc gag tac 288
Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr
85 90 95
cag gcg gtg gcg ggc tgc gga atg atc atc tcg gcc gtc gag aac cag 336
Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln
100 105 110
gag ctc cgt cag ggc tac gcc gct cag atg ctc gat gag gtg cgg cac 384
Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His
115 120 125
gcg cag ctc gag atg acg cta cgc aac tac tac gcg aag cac tgg tgc 432
Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys
130 135 140
gat ccc tcc ggc ttc gac atc ggt cag cgc ggc ctg tac cag cac ccc 480
Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro
145 150 155 160
gcg ggg ctg gtg tcc atc ggc gag ttc cag cac ttc aat act ggt gac 528
Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp
165 170 175
ccg ctt gac gtc atc atc gat ctc aac atc gtg gcc gag acg gcg ttc 576
Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe
180 185 190
acg aac atc ctg ctg gtg gcc act cca cag gtc gcc gtg gcc aac ggg 624
Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly
195 200 205
gac aac gcg atg gcc agc gtg ttc ctc tcg atc cag tcg gac gag gcc 672
Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala
210 215 220
agg cac atg gcc aac ggg tac ggc tcg gtc atg gcg ctg ctg gag aac 720
Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn
225 230 235 240
gag gac aac ctc ccg ctg ctc aac cag tct ctc gat cgg cac ttc tgg 768
Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp
245 250 255
cgt gcc cac aag gcc ttg gac aac gcg gtc gga tgg tgt tcg gag tat 816
Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr
260 265 270
ggc gcc cgc aag cgg cca tgg agc tac aag gcc cag tgg gag gaa tgg 864
Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp
275 280 285
gtc gtc gac gac ttc gtg ggc ggc tac atc gac cga ctc agc gag ttc 912
Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe
290 295 300
ggc gtt cag gct ccg gcc tgc ctt ggc gcg gcc gcc gac gag gtc aag 960
Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys
305 310 315 320
tgg tcg cac cac acg ctc ggt cag gtg ctg tcg gcg gtg tgg ccg ctg 1008
Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu
325 330 335
aac ttc tgg cgc tcg gac gcc atg gga ccg gcg gac ttc gag tgg ttc 1056
Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe
340 345 350
gag aac cac tac ccg ggc tgg agc gcg gcc tac cag ggt tac tgg gag 1104
Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu
355 360 365
ggc tac aag gcg ctc gcc gac cca gca ggc gga cgc atc atg ctc cag 1152
Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln
370 375 380
gag ctg ccg ggt ctg ccg ccg atg tgt cag gtg tgc cag gtg ccg tgc 1200
Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys
385 390 395 400
gtg atg ccg cgg ctg gat atg aac gcc gcg cgg atc atc gag ttc gag 1248
Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu
405 410 415
ggg cag aaa atc gcg ctg tgc agc gaa ccc tgc cag cgg atc ttc acc 1296
Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr
420 425 430
aac tgg ccg gag gcg tac cgc cac cgc aag caa tac tgg gcc cgc tac 1344
Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr
435 440 445
cac gga tgg gac ctg gcg gac gtc atc gtt gat ctc ggc tac atc cgc 1392
His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg
450 455 460
ccg gac ggc aag acc ctc atc ggc cag ccg ctg ctc gag atg gag cgg 1440
Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg
465 470 475 480
ctg tgg acc atc gac gac atc cgg gcc ctt cag tac gaa gtc aag gac 1488
Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp
485 490 495
ccg ttg cag gag gcg tga 1506
Pro Leu Gln Glu Ala
500
<210>12
<211>501
<212>PRT
<213>珊瑚诺卡氏菌
<400>12
Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp
1 5 10 15
Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr
20 25 30
Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp
35 40 45
Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu
50 55 60
Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe
65 70 75 80
Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr
85 90 95
Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln
100 105 110
Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His
115 120 125
Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys
130 135 140
Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro
145 150 155 160
Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp
165 170 175
Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe
180 185 190
Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly
195 200 205
Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala
210 215 220
Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn
225 230 235 240
Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp
245 250 255
Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr
260 265 270
Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp
275 280 285
Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe
290 295 300
Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys
305 310 315 320
Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu
325 330 335
Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe
340 345 350
Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu
355 360 365
Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln
370 375 380
Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys
385 390 395 400
Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu
405 410 415
Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr
420 425 430
Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr
435 440 445
His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg
450 455 460
Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg
465 470 475 480
Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp
485 490 495
Pro Leu Gln Glu Ala
500
<210>13
<211>1494
<212>DNA
<213>黄色杆菌属种
<220>
<221>CDS
<222>(1)..(1494)
<400>13
atg gcg ctc ttg aat cgg gac gat tgg tac gac atc gcg cgc gat gtc 48
Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val
1 5 10 15
gac tgg acg ctc agc tat gtc gac cgc gcg gtc gcc ttt ccc gag gag 96
Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu
20 25 30
tgg aaa ggc gaa aag gac att tgc ggc acg gcc tgg gac gat tgg gac 144
Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp
35 40 45
gag ccc ttc cgg gtc tcc ttc cgc gaa tat gtg atg gtc cag cgc gac 192
Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp
50 55 60
aag gaa gcg agc gtc ggc gcc atc cgc gag gcc atg gtc cgc gcc aag 240
Lys Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys
65 70 75 80
gcc tat gag aag ctc gac gac ggc cac aag gcc acc tcg cac ctg cac 288
Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His
85 90 95
atg ggc acc atc acc atg gtg gag cac atg gcg gtc acc atg cag agc 336
Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser
100 105 110
cgg ttc gtg cgc ttc gcg ccg tcc gcc cgc tgg cgc agc ctc ggg gcg 384
Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala
115 120 125
ttc ggc atg ctg gac gag acc cgc cac acc cag ctc gac ctg cgc ttc 432
Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe
130 135 140
agc cac gat ctg ctc aac gat tcc ccg agc ttc gac tgg agc cag cgg 480
Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg
145 150 155 160
gcg ttc cac acc gac gaa tgg gcg gtt ctc gcc acc cgc aac ctg ttc 528
Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe
165 170 175
gac gac atc atg ctc aac gcc gac tgc gtg gag gcg gcg ctc gcc acc 576
Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr
180 185 190
agc ctg acg ctg gag cac ggc ttc acc aac atc cag ttc gtg gcg ctc 624
Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu
195 200 205
gcc tcc gac gcc atg gaa gcc ggc gac gtg aac ttc tcc aac ctc ttg 672
Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu
210 215 220
tcg agc atc cag acc gac gag gcg cgg cac gcc cag ttg ggc ttt ccc 720
Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro
225 230 235 240
acc ctc gac gtg atg atg aag cac gac ccc aag cgc gcc cag cag atc 768
Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile
245 250 255
ctg gac gtc gcc ttc tgg cgc tcc tac cgc atc ttc cag gcg gtg acc 816
Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr
260 265 270
ggc gtc tcc atg gac tac tac acg ccg gtc gcc aag cgg cag atg tcg 864
Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser
275 280 285
ttc aag gag ttc atg ctg gag tgg atc gtc aag cat cat gag cgc atc 912
Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile
290 295 300
ctg cgc gac tac ggc ctc cag aag ccc tgg tac tgg gac acg ttc gag 960
Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu
305 310 315 320
aag acc ctc gat cac ggc cac cac gcg ctg cac atc ggc acc tgg ttc 1008
Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe
325 330 335
tgg cgc ccg acc ctg ttc tgg gat ccc aat ggc ggc gtc tcg cgc gag 1056
Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu
340 345 350
gag cgg cgc tgg ctg aac cag aag tat ccg aac tgg gaa gag agc tgg 1104
Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp
355 360 365
ggc gtc ctg tgg gac gag atc atc tcc aac atc aat gcg ggc aac att 1152
Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile
370 375 380
gaa aag acc ttg ccc gag acg ctg ccg atg ctg tgc aac gtc acc aac 1200
Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn
385 390 395 400
ctg ccc atc ggc tcg cac tgg gac cgc ttc cac ctg aag ccc gag cag 1248
Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln
405 410 415
ctc gtc tac aag ggg cgg ctc tac acc ttc gac agc gac gtc tcc aag 1296
Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys
420 425 430
tgg atc ttc gag ctc gat ccg gag cgc tat gcc ggc cac acc aac gtg 1344
Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val
435 440 445
gtc gac cgc ttc atc ggc ggg cag atc cag ccc atg acc atc gag ggc 1392
Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly
450 455 460
gtg ctc aac tgg atg ggc ctg acg ccc gaa gtc atg ggc aag gac gtg 1440
Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val
465 470 475 480
ttc aac tac cgt tgg gcc ggc gat tac gcc gag aac cgg atc gcc gcc 1488
Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala
485 490 495
gag taa 1494
Glu
<210>14
<211>497
<212>PRT
<213>黄色杆菌属种
<400>14
Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val
1 5 10 15
Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu
20 25 30
Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp
35 40 45
Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp
50 55 60
Lya Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys
65 70 75 80
Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His
85 90 95
Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser
100 105 110
Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala
115 120 125
Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe
130 135 140
Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg
145 150 155 160
Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe
165 170 175
Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr
180 185 190
Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu
195 200 205
Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu 210
215 220
Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro
225 230 235 240
Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile
245 250 255
Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr
260 265 270
Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser
275 280 285
Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile
290 295 300
Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu
305 310 315 320
Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe
325 330 335
Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu
340 345 350
Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp
355 360 365
Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile
370 375 380
Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn
385 390 395 400
Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln
405 410 415
Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys
420 425 430
Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val
435 440 445
Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly
450 455 460
Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val
465 470 475 480
Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala
485 490 495
Glu
<210>15
<211>1026
<212>DNA
<213>黄色杆菌属种
<220>
<221>CDS
<222>(1)..(1026)
<400>15
atg aca cag cag cgc ccc acc cgc acg cgc gag cgc aag aag acc tgg 48
Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp
1 5 10 15
acg gct ttc ggc aat ctc gga cgc aag ccg acc gac tac gag gtc gtc 96
Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val
20 25 30
acc cac aac atg aac cac acc atg cgc ggc acg ccc ctg gag ctg tcg 144
Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser
35 40 45
ccg acg gtg cac gcc aat gtg tgg ctc aag aag aac cgc gac gag atc 192
Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile
50 55 60
gcg ctc aag gtc gac agc tgg gat ctg ttc cgc gat ccc gac cgc acc 240
Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr
65 70 75 80
acc tac gac acc tac gtc aag atg cag gac gac cag gag acc tat gtc 288
Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val
85 90 95
gac aac ctg ctc ctg tcc tac acc ggc gag ggc cgc tac gac gag gag 336
Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu
100 105 110
ctt tcc tcg cgc agc ctc gac ctc ctg tcc gcg ggg ctg acg ccg acc 384
Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr
115 120 125
cgc tat ctg ggc cat ggg ctg cag atg ctc gcg gcc tat atc cag cag 432
Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln
130 135 140
ctc gcc ccg tcg gcc tat gtg ggc aat tgc gcg gtg ttc cag acc tcc 480
Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser
145 150 155 160
gac gcg ctg cgc cgc gtg cag cgc gtc gcc tac cgc acc cgc cag ctc 528
Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu
165 170 175
gcc gac gcc cat ccg gcc cgc ggc ttc ggc tcc ggc gac cgg gcg gtg 576
Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val
180 185 190
tgg gag aag tcc ccg gac tgg cag ccc atc cgc aag gcc atc gag gag 624
Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu
195 200 205
ctg ctc gtc acc ttc gaa tgg gac aag gcg ctc gcc ggc acc aat ttc 672
Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe
210 215 220
gtg gtg aag ccg atc ctc gac gag ctg ttc ctc aac cac ctg gcg cgc 720
Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg
225 230 235 240
ctg ctc cac gtg gag ggc gac gag ctc gac agc ctc gtg ctg cgg aac 768
Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn
245 250 255
ctt cac ggc gac gcc cag cgc cac gcc cgc tgg acg gcc gcg ctc ggc 816
Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly
260 265 270
cgc ttc gcc gtc gag cag aac gtg aac aac cgc acg gtc ctg cgc gac 864
Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp
275 280 285
gcc atc gcc ggc tgg cac gag acc ggc gag gcg gtc ctc gcc gcg ggc 912
Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly
290 295 300
gcc ggg atg ctt gcg agc cgc gcc ccc agc gcg gat gcg gcc aag atc 960
Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile
305 310 315 320
gcc gac gag gtc cgc gcc acg ctc gcg cag ctg cac gcc aat gcg ggc 1008
Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly
325 330 335
ctc ggg cac gat gcc tga 1026
Leu Gly His Asp Ala
340
<210>16
<211>341
<212>PRT
<213>黄色杆菌属种
<400>16
Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp
1 5 10 15
Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val
20 25 30
Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser
35 40 45
Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile
50 55 60
Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr
65 70 75 80
Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val
85 90 95
Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu
100 105 110
Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr
115 120 125
Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln
130 135 140
Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser
145 150 155 160
Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu
165 170 175
Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val
180 185 190
Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu
195 200 205
Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe
210 215 220
Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg
225 230 235 240
Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn
245 250 255
Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly
260 265 270
Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp
275 280 285
Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly
290 295 300
Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile
305 310 315 320
Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly
325 330 335
Leu Gly His Asp Ala
340
<210>17
<211>267
<212>DNA
<213>黄色杆菌属种
<220>
<221>CDS
<222>(1)..(267)
<400>17
atg tct ttg ttc ccc atc gtg ggc cgc ttc gtg ggg gat ttc gtc ccc 48
Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro
1 5 10 15
cac ctg gtg gcg gtg gac acc tct gac acc atc gat cag atc gcc gag 96
His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu
20 25 30
aag gtg gcg gtc cac acg gtc ggg cgg cgc ttg ccg ccc gat ccc acc 144
Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr
35 40 45
gcc acc ggc tat gag gtg ctc ctc gac ggc gag acc ctg gac ggg ggc 192
Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly
50 55 60
gcc acc ctg gag gcc atc atg acc aag cgc gag atg ctg ccc ctg cag 240
Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln
65 70 75 80
tgg ttc gac gtg agg ttc aag aag tga 267
Trp Phe Asp Val Arg Phe Lys Lys
85
<210>18
<211>88
<212>PRT
<213>黄色杆菌属种
<400>18
Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro
1 5 10 15
His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu
20 25 30
Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr
35 40 45
Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly
50 55 60
Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln
65 70 75 80
Trp Phe Asp Val Arg Phe Lys Lys
85
<210>19
<211>1584
<212>DNA
<213>荚膜甲基球菌
<220>
<221>CDS
<222>(1)..(1584)
<400>19
atg gca ctt agc acc gca acc aag gcc gcg acg gac gcg ctg gct gcc 48
Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala
1 5 10 15
aat cgg gca ccc acc agc gtg aat gca cag gaa gtg cac cgt tgg ctc 96
Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu
20 25 30
cag agc ttc aac tgg gat ttc aag aac aac cgg acc aag tac gcc acc 144
Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr
35 40 45
aag tac aag atg gcg aac gag acc aag gaa cag ttc aag ctg atc gcc 192
Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala
50 55 60
aag gaa tat gcg cgc atg gag gca gtc aag gac gaa agg cag ttc ggt 240
Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly
65 70 75 80
agc ctg cag gat gcg ctg acc cgc ctc aac gcc ggt gtt cgc gtt cat 288
Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His
85 90 95
ccg aag tgg aac gag acc atg aaa gtg gtt tcg aac ttc ctg gaa gtg 336
Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val
100 105 110
ggc gaa tac aac gcc atc gcc gct acc ggg atg ctg tgg gat tcc gcc 384
Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala
115 120 125
cag gcg gcg gaa cag aag aac ggc tat ctg gcc cag gtg ttg gat gaa 432
Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu
130 135 140
atc cgc cac acc cac cag tgt gcc tac gtc aac tac tac ttc gcg aag 480
Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys
145 150 155 160
aac ggc cag gac ccg gcc ggt cac aac gat gct cgc cgc acc cgt acc 528
Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr
165 170 175
atc ggt ccg ctg tgg aag ggc atg aag cgc gtg ttt tcc gac ggc ttc 576
Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe
180 185 190
att tcc ggc gac gcc gtg gaa tgc tcc ctc aac ctg cag ctg gtg ggt 624
Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly
195 200 205
gag gcc tgc ttc acc aat ccg ctg atc gtc gca gtg acc gaa tgg gct 672
Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala
210 215 220
gcc gcc aac ggc gat gaa atc acc ccg acg gtg ttc ctg tcg atc gag 720
Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu
225 230 235 240
acc gac gaa ctg cgc cac atg gcc aac ggt tac cag acc gtc gtt tcc 768
Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser
245 250 255
atc gcc aac gat ccg gct tcc gcc aag tat ctc aac acg gac ctg aac 816
Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn
260 265 270
aac gcc ttc tgg acc cag cag aag tac ttc acg ccg gtg ttg ggc atg 864
Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met
275 280 285
ctg ttc gag tat ggc tcc aag ttc aag gtc gag ccg tgg gtc aag acg 912
Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr
290 295 300
tgg gac cgc tgg gtg tac gag gac tgg ggc ggc atc tgg atc ggc cgt 960
Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg
305 310 315 320
ctg ggc aag tac ggg gtg gag tcg ccg cgc agc ctc aag gac gcc aag 1008
Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys
325 330 335
cag gac gct tac tgg gct cac cac gac ctg tat ctg ctg gct tat gcg 1056
Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala
340 345 350
ctg tgg ccg acc ggc ttc ttc cgt ctg gcg ctg ccg gat cag gaa gaa 1104
Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu
355 360 365
atg gag tgg ttc gag gcc aac tac ccc ggc tgg tac gac cac tac ggc 1152
Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly
370 375 380
aag atc tac gag gaa tgg cgc gcc cgc ggt tgc gag gat ccg tcc tcg 1200
Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser
385 390 395 400
ggc ttc atc ccg ctg atg tgg ttc atc gaa aac aac cat ccc atc tac 1248
Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr
405 410 415
atc gat cgc gtg tcg caa gtg ccg ttc tgc ccg agc ttg gcc aag ggc 1296
Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly
420 425 430
gcc agc acc ctg cgc gtg cac gag tac aac ggc gag atg cac acc ttc 1344
Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe
435 440 445
agc gac cag tgg ggc gag cgc atg tgg ctg gcc gag ccg gag cgc tac 1392
Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr
450 455 460
gag tgc cag aac atc ttc gaa cag tac gaa gga cgc gaa ctg tcg gaa 1440
Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu
465 470 475 480
gtg atc gcc gaa ctg cac ggg ctg cgc agt gat ggc aag acc ctg atc 1488
Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile
485 490 495
gcc cag ccg cat gtc cgt ggc gac aag ctg tgg acg ttg gac gat atc 1536
Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile
500 505 510
aaa cgc ctg aac tgc gtc ttc aag aac ccg gtg aag gca ttc aat tga 1584
Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn
515 520 525
<210>20
<21l>527
<212>PRT
<213>荚膜甲基球菌
<400>20
Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala
1 5 10 15
Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu
20 25 30
Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr
35 40 45
Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala
50 55 60
Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly
65 70 75 80
Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His
85 90 95
Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val
100 105 110
Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala
115 120 125
Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu
130 135 140
Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys
145 150 155 160
Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr
165 170 175
Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe
180 185 190
Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly
195 200 205
Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala
210 215 220
Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu
225 230 235 240
Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser
245 250 255
Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn
260 265 270
Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met
275 280 285
Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr
290 295 300
Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg
305 310 315 320
Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys
325 330 335
Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala
340 345 350
Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu
355 360 365
Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly
370 375 380
Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser
385 390 395 400
Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr
405 410 415
Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly
420 425 430
Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe
435 440 445
Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr
450 455 460
Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu
465 470 475 480
Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile
485 490 495
Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile
500 505 510
Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn
515 520 525
<210>21
<211>1170
<212>DNA
<213>荚膜甲基球菌
<220>
<221>CDS
<222>(1)..(1170)
<400>21
atg agc atg tta gga gaa aga cgc cgc ggt ctg acc gat ccg gaa atg 48
Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met
1 5 10 15
gcg gcc gtc att ttg aag gcg ctt cct gaa gct ccg ctg gac ggc aac 96
Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn
20 25 30
aac aag atg ggt tat ttc gtc acc ccc cgc tgg aaa cgc ttg acg gaa 144
Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu
35 40 45
tat gaa gcc ctg acc gtt tat gcg cag ccc aac gcc gac tgg atc gcc 192
Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala
50 55 60
ggc ggc ctg gac tgg ggc gac tgg acc cag aaa ttc cac ggc ggc cgc 240
Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg
65 70 75 80
cct tcc tgg ggc aac gag acc acg gag ctg cgc acc gtc gac tgg ttc 288
Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe
85 90 95
aag cac cgt gac ccg ctc cgc cgt tgg cat gcg ccg tac gtc aag gac 336
Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp
100 105 110
aag gcc gag gaa tgg cgc tac acc gac cgc ttc ctg cag ggt tac tcc 384
Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser
115 120 125
gcc gac ggt cag atc cgg gcg atg aac ccg acc tgg cgg gac gag ttc 432
Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe
130 135 140
atc aac cgg tat tgg ggc gcc ttc ctg ttc aac gaa tac gga ttg ttc 480
Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe
145 150 155 160
aac gct cat tcg cag ggc gcc cgg gag gcg ctg tcg gac gta acc cgc 528
Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg
165 170 175
gtc agc ctg gct ttc tgg ggc ttc gac aag atc gac atc gcc cag atg 576
Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met
180 185 190
atc caa ctc gaa cgg ggt ttc ctc gcc aag atc gta ccc ggt ttc gac 624
Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp
195 200 205
gag tcc aca gcg gtg ccg aag gcc gaa tgg acg aac ggg gag gtc tac 672
Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr
210 215 220
aag agc gcc cgt ctg gcc gtg gaa ggg ctg tgg cag gag gtg ttc gac 720
Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp
225 230 235 240
tgg aac gag agc gct ttc tcg gtg cac gcc gtc tat gac gcg ctg ttc 768
Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe
245 250 255
ggt cag ttc gtc cgc cgc gag ttc ttt cag cgg ctg gct ccc cgc ttc 816
Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe
260 265 270
ggc gac aat ctg acg cca ttc ttc atc aac cag gcc cag aca tac ttc 864
Gly Asp Asn Leu Thr Pro Phe Phe Ile Asn Gln Ala Gln Thr Tyr Phe
275 280 285
cag atc gcc aag cag ggc gta cag gat ctg tat tac aac tgt ctg ggt 912
Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly
290 295 300
gac gat ccg gag ttc agc gat tac aac cgt acc gtg atg cgc aac tgg 960
Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp
305 310 315 320
acc ggc aag tgg ctg gag ccc acg atc gcc gct ctg cgc gac ttc atg 1008
Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met
325 330 335
ggg ctg ttt gcg aag ctg ccg gcg ggc acc act gac aag gaa gaa atc 1056
Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile
340 345 350
acc gcg tcc ctg tac cgg gtg gtc gac gac tgg atc gag gac tac gcc 1104
Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala
355 360 365
agc gcg atc gac ttc aag gcg gac cgc gat cag atc gtt aaa gcg gtt 1152
Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val
370 375 380
ctg gca gga ttg aaa taa 1170
Leu Ala Gly Leu Lys
385
<210>22
<211>389
<212>PRT
<213>荚膜甲基球菌
<400>22
Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met
1 5 10 15
Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn
20 25 30
Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu
35 40 45
Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala
50 55 60
Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg
65 70 75 80
Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe
85 90 95
Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp
100 105 110
Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser
115 120 125
Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe
130 135 140
Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe
145 150 155 160
Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg
165 170 175
Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met
180 185 190
Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp
195 200 205
Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr
210 215 220
Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp
225 230 235 240
Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe
245 250 255
Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe
260 265 270
Gly Asp Asn Leu Thr Pro Phe Phe Ile Asn Gln Ala Gln Thr Tyr Phe
275 280 285
Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly
290 295 300
Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp
305 310 315 320
Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met
325 330 335
Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile
340 345 350
Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala
355 360 365
Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val
370 375 380
Leu Ala Gly Leu Lys
385
<210>23
<211>513
<212>DNA
<213>荚膜甲基球菌
<220>
<221>CDS
<222>(1)..(513)
<400>23
atg gcg aaa ctg ggt ata cac agc aac gac acc cgc gac gcc tgg gtg 48
Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val
1 5 10 15
aac aag atc gcg cag ctc aac acc ctg gaa aaa gcg gcc gag atg ctg 96
Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu
20 25 30
aag cag ttc cgg atg gac cac acc acg ccg ttc cgc aac agc tac gaa 144
Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu
35 40 45
ctg gac aac gac tac ctc tgg atc gag gcc aag ctc gaa gag aag gtc 192
Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val
50 55 60
gcc gtc ctc aag gca cgc gcc ttc aac gag gtg gac ttc cgt cat aag 240
Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys
65 70 75 80
acc gct ttc ggc gag gat gcc aag tcc gtt ctg gac ggc acc gtc gcg 288
Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala
85 90 95
aag atg aac gcg gcc aag gac aag tgg gag gcg gag aag atc cat atc 336
Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile
100 105 110
ggt ttc cgc cag gcc tac aag ccg ccg atc atg ccg gtg aac tat ttc 384
Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe
115 120 125
ctg gac ggc gag cgt cag ttg ggg acc cgg ctg atg gaa ctg cgc aac 432
Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn
130 135 140
ctc aac tac tac gac acg ccg ctg gaa gaa ctg cgc aaa cag cgc ggt 480
Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly
145 150 155 160
gtg cgg gtg gtg cat ctg cag tcg ccg cac tga 513
Val Arg Val Val His Leu Gln Ser Pro His
165 170
<210>24
<211>170
<212>PRT
<213>荚膜甲基球菌
<400>24
Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val
1 5 10 15
Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu
20 25 30
Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu
35 40 45
Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val
50 55 60
Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys
65 70 75 80
Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala
85 90 95
Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile
100 105 110
Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe
115 120 125
Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn
130 135 140
Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly
145 150 155 160
Val Arg Val Val His Leu Gln Ser Pro His
165 170
<210>25
<211>1206
<212>DNA
<213>食油假单胞菌
<220>
<221>CDS
<222>(1)..(1206)
<400>25
atg ctt gag aaa cac aga gtt ctg gat tcc gct cca gag tac gta gat 48
Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp
1 5 10 15
aaa aag aaa tat ctc tgg ata cta tca act ttg tgg ccg gct act ccg 96
Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro
20 25 30
atg atc gga atc tgg ctt gca aat gaa act ggt tgg ggg att ttt tat 144
Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr
35 40 45
ggg ctg gta ttg ctc gta tgg tac ggc gca ctt cca ttg ctt gat gcg 192
Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala
50 55 60
atg ttt ggt gag gac ttt aat aat ccg cct gaa gaa gtg gtg ccg aaa 240
Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys
65 70 75 80
cta gag aag gag cgg tac tat cga gtt ttg aca tat cta aca gtt cct 288
Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro
85 90 95
atg cat tac gct gca tta att gtg tca gca tgg tgg gtc gga act cag 336
Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln
100 105 110
cca atg tct tgg ctt gaa att ggt gcg ctt gcc ttg tca ctg ggt atc 384
Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile
115 120 125
gtg aac gga cta gcg ctc aat aca gga cac gaa ctc ggt cac aag aag 432
Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys
130 135 140
gag act ttt gat cgt tgg atg gcc aaa att gtg ttg gct gtc gta ggg 480
Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly
145 150 155 160
tac ggt cac ttc ttt att gag cat aat aag ggt cat cac cgt gat gtc 528
Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val
165 170 175
gct aca ccg atg gat cct gca aca tcc cgg atg gga gaa agc att tat 576
Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr
180 185 190
aag ttt tca atc cgt gag atc cca gga gca ttt att cgt gct tgg ggg 624
Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly
195 200 205
ctt gag gaa caa cgc ctt tcg cgc cgt ggc caa agc gtt tgg agt ttc 672
Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe
210 215 220
gat aat gaa atc ctc caa cca atg atc atc aca gtt att ctt tac gcc 720
Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala
225 230 235 240
gtt ctc ctt gcc ttg ttt gga cct aag atg ctg gtg ttc ctg ccg att 768
Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile
245 250 255
caa atg gct ttc ggt tgg tgg cag ctg acc agt gcg aac tat att gaa 816
Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu
260 265 270
cat tac ggc ttg ctc cgt caa aaa atg gag gac ggt cga tat gag cat 864
His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His
275 280 285
caa aag ccg cac cat tct tgg aat agt aat cac atc gtc tct aat cta 912
Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu
290 295 300
gtg ctg ttc cac ctt cag cgg cac tcg gat cac cac gcg cat cca aca 960
Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr
305 310 315 320
cgt tct tat cag rca crt cgg gat ttt ccc ggc ctg ccg gct ctt ccg 1008
Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro
325 330 335
acg ggt tac cct ggt gca ttt ttg atg gcg atg att cct cag tgg ttt 1056
Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe
340 345 350
aga tca gtt atg gat ccc aag gta gta gat tgg gct ggt ggt gac ctt 1104
Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu
355 360 365
aat aag atc caa att gat gat tcg atg cga gaa acc tat ttg aaa aaa 1152
Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys
370 375 380
ttt ggc act agt agt gct ggt cat agt tcg agt acc tct gcg gta gca 1200
Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala
385 390 395 400
tcg tag 1206
Ser
<210>26
<211>401
<212>PRT
<213>食油假单胞菌
<400>26
Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp
1 5 10 15
Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro
20 25 30
Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr
35 40 45
Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala
50 55 60
Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys
65 70 75 80
Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro
85 90 95
Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln
100 105 110
Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile
115 120 125
Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys
130 135 140
Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly
145 150 155 160
Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val
165 170 175
Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr
180 185 190
Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly
195 200 205
Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe
210 215 220
Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala
225 230 235 240
Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile
245 250 255
Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu
260 265 270
His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His
275 280 285
Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu
290 295 300
Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr
305 310 315 320
Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro
325 330 335
Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe
340 345 350
Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu
355 360 365
Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys
370 375 380
Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala
385 390 395 400
Ser
<210>27
<211>1560
<212>DNA
<213>洋葱伯克霍尔德氏菌
<220>
<221>CDS
<222>(1)..(1560)
<400>27
atg gac act tct gtg cag aag aag aaa ctc ggt tta aag aat cgc tac 48
Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr
1 5 10 15
gca gcg atg acc cgc ggt ctt ggc tgg cag acc agc tac cag ccg atg 96
Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met
20 25 30
gag aaa gtg ttt ccg tac gac aag tac gaa ggc atc aag atc cac gat 144
Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp
35 40 45
tgg gat aaa tgg gaa gac ccc ttc cgc ctg acc atg gac gcc tac tgg 192
Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp
50 55 60
aaa tat cag ggc gag aag gaa aaa aag ctt tac gcc gtc atc gac gct 240
Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala
65 70 75 80
ttc gcg cag aac aac ggg cag ttg agc att tcc gac gcg cga tat gtc 288
Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val
85 90 95
aac gca ctc aag gtg ttt atc cag ggt gtg aca ccg ttg gag tat atg 336
Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met
100 105 110
gca cac cga ggt ttt gcc cac att ggt cgg cat ttt acg ggt gaa ggg 384
Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly
115 120 125
gca cgt gtt gct tgc cag atg cag tcc atc gac gag ctg cgt cac ttc 432
Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe
130 135 140
cag acc gaa atg cat gct ctc tcg cac tac aac aag tat ttt aac ggt 480
Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly
145 150 155 160
ctg cac aac tcc atc cat tgg tac gac cgg gtt tgg tat ttg tcg gtg 528
Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val
165 170 175
ccc aag tca ttt ttt gaa gac gcg gcc acc ggt gga ccg ttc gag ttt 576
Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe
180 185 190
ctt acc gcg gtg agc ttt tcg ttc gaa tat gtg ttg acc aac ctg ctg 624
Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu
195 200 205
ttt gtc ccc ttc atg tcg ggt gct gct tac aac ggg gac atg tct acg 672
Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr
210 215 220
gtc act ttc ggt ttt tcg gcg caa agt gac gaa tcg cgc cac atg aca 720
Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr
225 230 235 240
ctc ggc atc gaa tgc atc aag ttc atg cta gaa cag gat ccg gac aac 768
Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn
245 250 255
gtg ccc atc gtg cag cgc tgg atc gac aag tgg ttc tgg cgc ggc tat 816
Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr
260 265 270
cgg ctg ttg agc atc gtg gcc atg atg cag gac tac atg ctg ccc aac 864
Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn
275 280 285
cgg gtg atg agc tgg cgc gag agc tgg gag atg tac gtc gag cag aac 912
Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn
290 295 300
ggc ggc gcg ctg ttc aag gat ctt gcg cgt tat ggc atc cgc aag ccc 960
Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro
305 310 315 320
aag ggc tgg gac cag gct tgc gaa ggc aag gac cac atc agc cat cag 1008
Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln
325 330 335
acc ttc gcc gta ttc tat aac tat aac gcc gcg gcc ccc atc cac acc 1056
Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr
340 345 350
tgg gtt ccc aca aaa gaa gaa atg gga tgg ctg tcg gag aag tac ccc 1104
Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro
355 360 365
gag acg ttc gac aag tat tac cgt ccg cgt tgg gac tac tgg cgc gag 1152
Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu
370 375 380
cag gcc gcc aag ggc aac cgt ttc tac aac aag acg ctg ccg atg ctc 1200
Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu
385 390 395 400
tgc act acc tgc cag att ccg atg ata ttc acc gag cct ggc gac gca 1248
Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala
405 410 415
acc aag atc tgc tat cgc gag tcg gcc tac ctc ggc gac aag tat cac 1296
Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His
420 425 430
ttc tgc agc gac cac tgc aag gag att ttt gac aac gaa ccc gaa aag 1344
Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys
435 440 445
ttc gtg cag tca tgg ctt ccg ccg cag caa gtg tat caa gga aac tgt 1392
Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys
450 455 460
ttc aag ccg gat gcc gat ccg acc aag gag ggt ttt gat ccc ttg atg 1440
Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met
465 470 475 480
gcc ttg ctc gac tac tac aac ctg aat gta ggc cgg gac aac ttc gat 1488
Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp
485 490 495
ttc gag gga tcg gaa gac caa aag aac ttt gct gcc tgg cgt gga gag 1536
Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu
500 505 510
gtc ttg caa gga gaa gcc aaa tga 1560
Val Leu Gln Gly Glu Ala Lys
515
<210>28
<211>519
<212>PRT
<213>洋葱伯克霍尔德氏菌
<400>28
Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr
1 5 10 15
Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met
20 25 30
Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp
35 40 45
Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp
50 55 60
Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala
65 70 75 80
Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val
85 90 95
Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met
100 105 110
Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly
115 120 125
Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe
130 135 140
Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly
145 150 155 160
Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val
165 170 175
Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe
180 185 190
Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu
195 200 205
Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr
210 215 220
Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr
225 230 235 240
Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn
245 250 255
Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr
260 265 270
Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn
275 280 285
Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn
290 295 300
Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro
305 310 315 320
Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln
325 330 335
Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr
340 345 350
Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro
355 360 365
Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu
370 375 380
Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu
385 390 395 400
Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala
405 410 415
Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His
420 425 430
Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys
435 440 445
Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys
450 455 460
Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met
465 470 475 480
Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp
485 490 495
Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu
500 505 510
Val Leu Gln Gly Glu Ala Lys
515
<210>29
<211>996
<212>DNA
<213>洋葱伯克霍尔德氏菌
<220>
<221>CDS
<222>(1)..(996)
<400>29
atg acc atc gat ttg aag acg cgg gaa atc aaa cca ctg cgt cac acc 48
Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr
1 5 10 15
tac acg cac gtg gct caa tac atc ggg gcc gat aaa gcc gct tcg cgc 96
Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg
20 25 30
tat cag gaa ggc act gta ggt gct caa ccc gca gcg aat ttt cat tac 144
Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr
35 40 45
cgg ccc acg tgg gat ccc gag cat gaa ctg ttc gac acg tcg cgt acc 192
Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr
50 55 60
gcg att caa atg aag gac tgg tat gcg ctg aaa gac ccg cgt cag ttc 240
Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe
65 70 75 80
tac tac gcg tcg tgg acg atg acc cga gcg cgg cag caa gac gcg atg 288
Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met
85 90 95
gaa tcc aac ttc gag ttt gtc gag tcg cgc ggc atg atc gat ctc gtt 336
Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val
100 105 110
tcc gat gag gtt cga caa cgg gcg ctt tcc gtt ctc gtg cct ttg cgt 384
Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg
115 120 125
cac gcg gcc tgg ggc gcg aac atg aac aac tcc cag atc tgt gcc cta 432
His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu
130 135 140
ggt tat ggc acg acc ttc act gcg ccg gct atg ttc cac gca atg gac 480
Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp
145 150 155 160
aat ctg ggt gta gcg cag tat ctc aca cga ctg gcg ctg gta atg tct 528
Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser
165 170 175
gga ccc gat ctt ctt gac gaa gcc aag caa gcc tgg atg acg agt ccc 576
Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro
180 185 190
gat tgg caa ccg ttg cgt cgt tat gtg gaa aac act ctg gtg ctg caa 624
Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln
195 200 205
gat ccg gtg gaa ctg ttc atc gcc caa aat ctg gcg ctc gac ggt ctt 672
Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu
210 215 220
ctt tat ccc atg atc tac ggc gct ttc gtc gac gat tac atc gca ctc 720
Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu
225 230 235 240
aac ggt ggt agc gca gtg gca atg cta acc act ttc atg ccc gag tgg 768
Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp
245 250 255
cat gac gaa tcc agt cgc tgg gtc gat gcg gta gta aag acc atg gcg 816
His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala
260 265 270
acg gaa tcg gag gat aac aaa gcg ctg ctc att cac tgg ttg cgt acc 864
Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr
275 280 285
tgg gaa gat cag gcg gcg tca gcg ttg ttg cct gtc gct gaa atg gct 912
Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala
290 295 300
ttg gcg gaa aac ggc cac gac gcc ttg gaa gaa gta agg cag caa ctt 960
Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu
305 310 315 320
cgt gcc cgc gtt gcg aag gcc ggg att gtt ctg taa 996
Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu
325 330
<210>30
<211>331
<212>PRT
<213>洋葱伯克霍尔德氏菌
<400>30
Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr
1 5 10 15
Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg
20 25 30
Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr
35 40 45
Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr
50 55 60
Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe
65 70 75 80
Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met
85 90 95
Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val
100 105 110
Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg
115 120 125
His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu
130 135 140
Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp
145 150 155 160
Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser
165 170 175
Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro
180 185 190
Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln
195 200 205
Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu
210 215 220
Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu
225 230 235 240
Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp
245 250 255
His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala
260 265 270
Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr
275 280 285
Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala
290 295 300
Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu
305 310 315 320
Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu
325 330
<210>31
<211>357
<212>DNA
<213>洋葱伯克霍尔德氏菌
<220>
<221>CDS
<222>(1)..(357)
<400>31
atg agc gtt gtt gcc ctc aaa ccc tac aag ttc ccg gca cga gac gcg 48
Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala
1 5 10 15
cgc gaa aac ttt ccg gcg ccg ttg ctg ttt atc ggc tgg gaa gac cat 96
Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His
20 25 30
ctg ttg ttt gcg gca cct gtt gcc ttg ccc ctg ccg tcg gac acg ttg 144
Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu
35 40 45
ttc ggt gcg ctg tgc acc cag gtg ttg ccc ggc act tat ggc tat cac 192
Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His
50 55 60
ccc gat ttc tca aag atc gac tgg agc cag gtg cag tgg ttt aag tcc 240
Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser
65 70 75 80
ggc cag ccg tgg cat ccc gac ccg gcg aag tcg ctg gct gaa aac ggt 288
Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly
85 90 95
ctg acg cac aaa gac gtg atc cgc ttt cgc acg cct ggc ttg aac ggt 336
Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly
100 105 110
ctg agc ggt tcc tgc aat tga 357
Leu Ser Gly Ser Cys Asn
115
<210>32
<211>118
<212>PRT
<213>洋葱伯克霍尔德氏菌
<400>32
Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala
1 5 10 15
Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His
20 25 30
Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu
35 40 45
Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His
50 55 60
Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser
65 70 75 80
Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly
85 90 95
Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly
100 105 110
Leu Ser Gly Ser Cys Asn
115
<210>33
<211>1143
<212>DNA
<213>嗜热脂肪芽孢杆菌
<220>
<221>CDS
<222>(1)..(1143)
<400>33
atg gaa aaa aat aaa atg tta ata gaa gaa aag ttg gac act gct gct 48
Met Glu Lys Asn Lys Met Leu Ile Glu Glu Lys Leu Asp Thr Ala Ala
1 5 10 15
ctt ctt gct aag gcg gag gaa ata ggc cgg att gct gag gaa gag gcg 96
Leu Leu Ala Lys Ala Glu Glu Ile Gly Arg Ile Ala Glu Glu Glu Ala
20 25 30
ggt gaa gcg gac cgc aat gcc tgt ttc tcc gac cgg gtg gct agg gcc 144
Gly Glu Ala Asp Arg Asn Ala Cys Phe Ser Asp Arg Val Ala Arg Ala
35 40 45
att aaa gaa gct gga ttc cac aag ctc atg cgt ccc aag cag tac gga 192
Ile Lys Glu Ala Gly Phe His Lys Leu Met Arg Pro Lys Gln Tyr Gly
50 55 60
gga ctg caa gta gac ttg cga act tac ggg gag att gtc cgc aca gtg 240
Gly Leu Gln Val Asp Leu Arg Thr Tyr Gly Glu Ile Val Arg Thr Val
65 70 75 80
gcc cgg tac agt gtt gcc gca gga tgg ctg acc tat ttt tat tcc atg 288
Ala Arg Tyr Ser Val Ala Ala Gly Trp Leu Thr Tyr Phe Tyr Ser Met
85 90 95
cat gag gtt tgg gct gca tat ctg cct cca aaa ggc aga gaa gaa att 336
His Glu Val Trp Ala Ala Tyr Leu Pro Pro Lys Gly Arg Glu Glu Ile
100 105 110
ttt gga caa gga ggg ctg ttg gca gac gtc gtt gcc cct gtt ggc cgg 384
Phe Gly Gln Gly Gly Leu Leu Ala Asp Val Val Ala Pro Val Gly Arg
115 120 125
gtg gag aag gac ggg gac ggc tac cgt ctc tat ggg cag tgg aac ttc 432
Val Glu Lys Asp Gly Asp Gly Tyr Arg Leu Tyr Gly Gln Trp Asn Phe
130 135 140
tgt agc ggt gtc ctc cat agt gac tgg atc gga ctt ggc gcc atg atg 480
Cys Ser Gly Val Leu His Ser Asp Trp Ile Gly Leu Gly Ala Met Met
145 150 155 160
gag ctg cct gac ggc aat agt cct gag tac tgt ttg tta gtg ctg cct 528
Glu Leu Pro Asp Gly Asn Ser Pro Glu Tyr Cys Leu Leu Val Leu Pro
165 170 175
aag tcg gat gtc cag atc gta gaa aat tgg gat acc atg ggc ctc cgc 576
Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg
180 185 190
gct tcg gga agc aac ggg gta tta gtt gaa ggt gct tat gtt cca tta 624
Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu
195 200 205
cac cgg atc ttt ccg gct ggc cgg gtg atg gct cat ccg ctt ttc ttg 672
His Arg Ile Phe Pro Ala Gly Arg Val Met Ala His Pro Leu Phe Leu
210 215 220
ctt ggg ttc cct tta gta tct tta ggc ggc gac gaa cga ttg gtg tca 720
Leu Gly Phe Pro Leu Val Ser Leu Gly Gly Asp Glu Arg Leu Val Ser
225 230 235 240
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Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg
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Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu
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Gln Leu Glu Ala Cys Gln Lys Glu Gly Lys Thr Val Met Asn Asp Met
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Glu Arg Glu Gln Leu Phe Ala Trp Arg Gly Tyr Val Ala Lys Ala Ser
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Ala Asn Ile Ala Val Arg Thr Leu Leu Thr Leu Gly Gly Asn Ser Ile
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Phe Lys Gly Asp Pro Val Glu Leu Phe Thr Arg Asp Leu Leu Ala Val
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Ala Ala His Pro Asn Ser Leu Trp Glu Asp Ala Met Ala Ala Tyr Gly
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Arg Thr Ile Phe Gly Leu Pro Gly Asp Pro Val Trp
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Ser Phe Ala Phe Pro Gln Pro Lys Pro Leu Arg Ser Pro Lys Phe Ala
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Claims (17)
1.一种进行氧化反应的方法，该氧化反应由单加氧酶催化并使用过氧化氢作为氧化剂，其中单加氧酶的氧化损伤反应水平低，所述方法包括在氧化反应同时产生过氧化氢，其中所述过氧化氢以低于或等于其在该反应中使用的速率产生。
2.根据权利要求1的方法，其中所述的单加氧酶对于H2O2具有至少15nM的Km值。
3.根据权利要求1或2的方法，其中所述的单加氧酶为P450酶。
4.根据前述权利要求任一项的方法，其中所述的H2O2产生速率为小于或等于每mg酶3ug。
5.根据前述权利要求任一项的方法，其中所述整个反应过程中H2O2的浓度小于或等于1mM。
6.根据前述权利要求任一项的方法，其中所述反应持续至少240分钟。
7.根据前述权利要求任一项的方法，其中所述的H2O2由电化学反应产生。
8.根据权利要求1至6任一项的方法，其中所述的H2O2由酶反应产生。
9.根据权利要求8的方法，其中所述的酶为葡萄糖氧化酶。
10.根据权利要求1至6任一项的方法，其中所述的H2O2由H2O2前体产生。
11.根据权利要求10的方法，其中所述的H2O2前体为过硼酸盐、过碳酸盐或过磷酸盐。
12.根据前述权利要求任一项的方法，其中所述的由单加氧酶氧化的底物为烷、芳香族化合物、萜类化合物、烯烃或脂肪酸。
13.电极用于产生H2O2以驱动权利要求7所限定的氧化反应的用途。
14.酶用于产生H2O2以驱动权利要求8或9所限定的氧化反应的用途。
15.过硼酸盐、过碳酸盐或过磷酸盐用于产生H2O2以驱动权利要求10所限定的氧化反应的用途。
16.一种进行氧化反应的方法，该氧化反应由单加氧酶催化并使用过氧化氢作为氧化剂，其中单加氧酶氧化损伤反应水平低，所述方法包括在H2O2或羟基螯合剂的存在下进行反应，所述螯合剂可控制H2O2或羟基浓度。
17.根据权利要求16的方法，其中所述螯合剂为EDTA。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB0403992.1 | 2004-02-23 | ||
| GBGB0403992.1A GB0403992D0 (en) | 2004-02-23 | 2004-02-23 | Oxidation by hydrogen peroxide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1934261A true CN1934261A (zh) | 2007-03-21 |
Family
ID=32050716
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2005800057244A Pending CN1934261A (zh) | 2004-02-23 | 2005-02-23 | 过氧化氢驱动的氧化反应 |
Country Status (6)
| Country | Link |
|---|---|
| US (1) | US20080044882A1 (zh) |
| EP (1) | EP1725670A1 (zh) |
| JP (1) | JP2007522815A (zh) |
| CN (1) | CN1934261A (zh) |
| GB (1) | GB0403992D0 (zh) |
| WO (1) | WO2005083100A1 (zh) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106497956A (zh) * | 2016-10-25 | 2017-03-15 | 上海交通大学 | Cyp101酶重组载体及构建方法、cyp101酶高效表达纯化方法 |
| CN108271374A (zh) * | 2015-07-07 | 2018-07-10 | 科德克希思公司 | 具有改进的活性的新颖p450-bm3变体 |
| CN113106139A (zh) * | 2020-01-20 | 2021-07-13 | 中国科学院青岛生物能源与过程研究所 | 一种催化苯乙烯及其衍生物制备手性有机化合物的方法 |
| CN116790529A (zh) * | 2023-05-24 | 2023-09-22 | 天津大学 | P450 bm3蛋白突变体及其应用 |
| CN121065119A (zh) * | 2025-11-06 | 2025-12-05 | 南京大学 | 一种生成(4r)-4-羟基异佛尔酮的单加氧酶突变体及其制备方法 |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CL2008002960A1 (es) | 2007-10-04 | 2009-05-08 | Bio Arch Lab Inc | Método para producir 2-ceto-3-desoxi d-gluconato (kdg), basado en proporciopnar acido-desoxi l-eritro hexocelulosa uronico (dehu), y contactar dicha molecula con un microorganismo recombinante que codofica una dehu-hidrogenasa. |
| GB0719620D0 (en) | 2007-10-08 | 2007-11-14 | Isis Innovation | Mutant Enzymes |
| CN102747052B (zh) * | 2012-07-24 | 2013-10-30 | 浙江大学宁波理工学院 | 细胞色素p450 bm-3(l148s/q229r)变体酶及其编码基因和用途 |
| EP3450549B1 (en) | 2012-10-29 | 2020-09-30 | University Of Rochester | Artemisinin derivatives, methods for their preparation and their use as antimalarial agents |
| CN110128364B (zh) * | 2018-02-08 | 2022-01-04 | 中国科学院青岛生物能源与过程研究所 | 一种激活酶催化反应的双功能小分子化合物及其应用 |
| KR20230044012A (ko) * | 2021-07-05 | 2023-03-31 | 와커 헤미 아게 | 설핀산의 설폰산으로의 효소적 산화 방법 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1375671A1 (en) * | 2002-06-28 | 2004-01-02 | Eidgenössische Technische Hochschule Zürich | Selective functionalization of hydrocarbons with isolated oxygenases and mediator-based regeneration |
-
2004
- 2004-02-23 GB GBGB0403992.1A patent/GB0403992D0/en not_active Ceased
-
2005
- 2005-02-23 WO PCT/GB2005/000674 patent/WO2005083100A1/en not_active Ceased
- 2005-02-23 US US10/590,426 patent/US20080044882A1/en not_active Abandoned
- 2005-02-23 JP JP2006553685A patent/JP2007522815A/ja not_active Withdrawn
- 2005-02-23 CN CNA2005800057244A patent/CN1934261A/zh active Pending
- 2005-02-23 EP EP05717769A patent/EP1725670A1/en not_active Withdrawn
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108271374A (zh) * | 2015-07-07 | 2018-07-10 | 科德克希思公司 | 具有改进的活性的新颖p450-bm3变体 |
| CN106497956A (zh) * | 2016-10-25 | 2017-03-15 | 上海交通大学 | Cyp101酶重组载体及构建方法、cyp101酶高效表达纯化方法 |
| CN113106139A (zh) * | 2020-01-20 | 2021-07-13 | 中国科学院青岛生物能源与过程研究所 | 一种催化苯乙烯及其衍生物制备手性有机化合物的方法 |
| CN113106139B (zh) * | 2020-01-20 | 2022-07-26 | 中国科学院青岛生物能源与过程研究所 | 一种催化苯乙烯及其衍生物制备手性有机化合物的方法 |
| CN116790529A (zh) * | 2023-05-24 | 2023-09-22 | 天津大学 | P450 bm3蛋白突变体及其应用 |
| CN121065119A (zh) * | 2025-11-06 | 2025-12-05 | 南京大学 | 一种生成(4r)-4-羟基异佛尔酮的单加氧酶突变体及其制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2007522815A (ja) | 2007-08-16 |
| GB0403992D0 (en) | 2004-03-31 |
| WO2005083100A1 (en) | 2005-09-09 |
| US20080044882A1 (en) | 2008-02-21 |
| EP1725670A1 (en) | 2006-11-29 |
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