| 重组固氮菌腈水合酶催化3-（4-氯苯基）戊二腈的选择性水解 |
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| 引用本文： | 李倩,任杰,冯进辉,吴洽庆,朱敦明.重组固氮菌腈水合酶催化3-（4-氯苯基）戊二腈的选择性水解[J].生物加工过程,2013,11(1):23-28. |
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| 作者姓名： | 李倩 任杰 冯进辉 吴洽庆 朱敦明 |
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| 作者单位： | 中国科学院天津工业生物技术研究所工业酶国家工程实验室 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)资助(2011CB710801) |
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| 摘 要： | 通过基因数据挖掘方法（genome mining）获得了来源于固氮菌Herbaspirillum seropedicae SmR1中的腈水合酶基因hsn1。构建了hsn1／pETDuet-1／BL21的大肠杆菌共表达重组菌，经IPTG诱导获得了具有良好催化能力的Co^2＋依赖型腈水合酶HSN1。利用全细胞反应研究了HSN1的底物谱，发现HSN1对底物3-（4-氯苯基）戊二腈有良好的区域选择性及一定的对映选择性，它可以选择性地水解1个腈基得到3-（4-氯苯基）-4-氰基丁酰胺，该化合物可通过一步化学反应合成巴氯芬。
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| 关 键 词： | 3-（4-氯苯基）戊二腈 水合作用 腈水合酶 |
Recombinant nitrile hydratase from Herbaspirillum seropedicae as biocatalyst for selective hydration of 3-(4-chlorophenyl) glutaronitrile |
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| LI Qian,REN Jie,FENG Jinhui,WU Qiaqing,ZHU Dunming.Recombinant nitrile hydratase from Herbaspirillum seropedicae as biocatalyst for selective hydration of 3-(4-chlorophenyl) glutaronitrile[J].Chinese Journal of Bioprocess Engineering,2013,11(1):23-28. |
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| Authors: | LI Qian REN Jie FENG Jinhui WU Qiaqing ZHU Dunming |
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| Institution: | (National Engineering Laboratory for Industrial Enzymes,Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences,Tianjin 300308,China) |
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| Abstract: | Nitrile hydratase（NHase） is the key enzymes of nitrile metabolism. Genome Mining was utilized to acquire the Nhase gene（hsnl ）from Herbaspirillum seropedicae SmR1. The plasmid hsnl/pETDuet-1 was constructed and expressed in E. coli BL21. The recombinant enzyme （HSN1）was identified as a cobaltdependent type NHase. The substrate profile of HSNI was studied by the whole-cell reactions. HSN1 showed the excellent regioselectivity and the enantioselectivity for the hydration of 3-（ 4-chlorophenyl ） glutaronitrile. The 3-（4-chlorophenyl）4-cyano-butyramide could be given, thus it could be converted to baclofen by one step chemical reaction. |
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| Keywords: | 3 - （ 4-chlorophenyl ） -4-cyano-butyramide hydration nitrile hydratase |
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