| 糖多孢红霉菌酮还原酶域在羰基还原中的应用 |
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| 引用本文： | 李凌凌,吕早生,关海燕,贾伟伟.糖多孢红霉菌酮还原酶域在羰基还原中的应用[J].华中科技大学学报(自然科学版),2011(2):72-75. |
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| 作者姓名： | 李凌凌 吕早生 关海燕 贾伟伟 |
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| 作者单位： | 武汉科技大学化学工程与技术学院; |
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| 基金项目： | 湖北省自然科学基金资助项目(2009CAD006); 武汉科技大学校基金资助项目(2006XY14) |
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| 摘 要： | 为研究糖多孢红霉菌聚酮合成酶模块1的酮还原酶域,催化羰基还原的底物特异性和立体选择性,PCR扩增了该酶域的编码基因(eryKR1),并将其克隆到表达载体pET-28a,得到重组质粒pET-eryKR1,转化到Escherichia coli BL21(DE3)后,获得了重组菌株E.coli BL21(pET-eryKR1).将E.coli BL21(pET-eryKR1)和异源表达枯草芽孢杆菌葡萄糖脱氢酶基因的重组大肠杆菌E.coli BL21(pET-gdh1)进行双重组菌耦合,对4-氯乙酰乙酸乙酯、苯乙酮、2-辛酮和环己酮4种底物进行转化还原,利用气相色谱分析转化液,结果显示E.coli BL21(pET-eryKR1)对环己酮的还原效果较好,产物环己醇的得率最高可达93.24%,而对其他3种底物几乎没有还原能力.利用E.coli BL21(pET-eryKR1)2催化2-甲基环己酮不对称还原,产物主要为顺式-2-甲基环己醇,产率可达41.87%,产物的对映体过量值为74.81%.
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| 关 键 词： | 2-甲基环己酮 酮还原酶域 聚酮合成酶 糖多孢红霉菌 生物催化 |
Application of ketoreductase domain in polyketide synthase from Saccharopolyspora erythraea to carbonyl reduction |
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| Li Lingling Lü Zaosheng Guan Haiyan Jia Weiwei.Application of ketoreductase domain in polyketide synthase from Saccharopolyspora erythraea to carbonyl reduction[J].JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY.NATURE SCIENCE,2011(2):72-75. |
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| Authors: | Li Lingling Lü Zaosheng Guan Haiyan Jia Weiwei |
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| Institution: | Li Lingling Lü Zaosheng Guan Haiyan Jia Weiwei(College of Chemical Engineering and Technology,Wuhan University of Science and Technology,Wuhan 430081,China) |
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| Abstract: | In order to define substrate specificity and enantioselectivity of ketoreductase domain in the first module of polyketide synthase from Saccharopolyspora erythraea,the eryKR1 gene coding this domain was amplified by PCR(polymerase chain reaction) and cloned into vector pET-28a to construct recombinant plasmid pET-eryKR1.The plasmid was then transformed into Escherichia coli BL21(DE3) to obtain recombinant E.coli BL21(pET-eryKR1).The recombinant and E.coli BL21(pET-gdh1) harboring Bacillus subtilis glucose d... |
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| Keywords: | 2-methylcyclohexanone ketoreductase domain polyketide synthase Saccharopolyspora erythraea biocatalysis |
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