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| Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli | |
| Wang, Ziqiang1,2; Wang, Yunshan1; Su, Zhiguo1 | |
| 2013-03-01 | |
| Source Publication | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| Volume | 97Issue:6Pages:2433-2441 |
| Abstract | 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. |
| Keyword | Cis-epoxysuccinic Acid Hydrolase Nocardia Tartaricans Prokaryotic Expression Enantioselectivity |
| Subtype | Article |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Indexed By | SCI |
| Language | 英语 |
| WOS Keyword | AGROBACTERIUM-RADIOBACTER AD1 ; MICROBIAL EPOXIDE HYDROLASES ; ASPERGILLUS-NIGER M200 ; DIRECTED EVOLUTION ; ENANTIOSELECTIVITY ; HYDROLYSIS ; MECHANISM ; CLONING ; ENZYME ; CELLS |
| WOS Research Area | Biotechnology & Applied Microbiology |
| WOS Subject | Biotechnology & Applied Microbiology |
| WOS ID | WOS:000315650100013 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ipe.ac.cn/handle/122111/13741 |
| Collection | 研究所（批量导入） |
| Affiliation | 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 |
| Recommended Citation 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. |
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| File Name/Size | DocType | Version | Access | License | ||
| Purification and cha（307KB） | 期刊论文 | 出版稿 | 限制开放 | CC BY-NC-SA | Application Full Text | |
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