Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli
文献类型:期刊论文
作者 | Wang, Ziqiang1,2; Wang, Yunshan1; Su, Zhiguo1 |
刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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出版日期 | 2013-03-01 |
卷号 | 97期号:6页码:2433-2441 |
关键词 | Cis-epoxysuccinic acid hydrolase Nocardia tartaricans Prokaryotic expression Enantioselectivity |
英文摘要 | A highly enantioselective cis-epoxysuccinic acid hydrolase from Nocardia tartaricans was purified to electrophoretic homogeneity. The enzyme was purified 184-fold with a yield of 18.8 %. The purified cis-epoxysuccinic acid hydrolase had a monomeric molecular weight of 28 kDa, and its optimum conditions were 37 A degrees C and pH 7-9. With sodium cis-epoxysuccinate as the substrate, Michaelis-Menten enzyme kinetics analysis gave a Km value of 35.71 mM and a Vmax of 2.65 mM min(-1). The enzyme was activated by Ni2+ and Al3+, while strongly inhibited by Fe3+, Fe2+, Cu2+, and Ag+. The cis-epoxysuccinic acid hydrolase gene was cloned, and its open reading frame sequence predicted a protein composed of 253 amino acids. A pET11a expression plasmid carrying the gene under the control of the T7 promoter was introduced into Escherichia coli, and the cis-epoxysuccinic acid hydrolase gene was successfully expressed in the recombinant strains. |
WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
类目[WOS] | Biotechnology & Applied Microbiology |
研究领域[WOS] | Biotechnology & Applied Microbiology |
关键词[WOS] | AGROBACTERIUM-RADIOBACTER AD1 ; MICROBIAL EPOXIDE HYDROLASES ; ASPERGILLUS-NIGER M200 ; DIRECTED EVOLUTION ; ENANTIOSELECTIVITY ; HYDROLYSIS ; MECHANISM ; CLONING ; ENZYME ; CELLS |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000315650100013 |
公开日期 | 2015-05-27 |
源URL | [http://ir.ipe.ac.cn/handle/122111/13741] ![]() |
专题 | 过程工程研究所_研究所(批量导入) |
作者单位 | 1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
推荐引用方式 GB/T 7714 | Wang, Ziqiang,Wang, Yunshan,Su, Zhiguo. Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2013,97(6):2433-2441. |
APA | Wang, Ziqiang,Wang, Yunshan,&Su, Zhiguo.(2013).Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,97(6),2433-2441. |
MLA | Wang, Ziqiang,et al."Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 97.6(2013):2433-2441. |
入库方式: OAI收割
来源:过程工程研究所
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