Expression, purification, and characterization of His-tagged penicillin G acylase from Kluyvera citrophila in Escherichia coli
文献类型：期刊论文
| 作者 | Yong, W; Shi, XL; Yuan, ZY; Pei, Z |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2004 |
| 卷号 | 38期号:1页码:24-28 |
| 关键词 | penicillin G acylase Kluyvera citrophila His-tag fusion protein |
| 通讯作者 | Pei, Z (reprint author), Fudan Univ, Sch Pharm, Dept Drug Biosynth, Shanghai 200032, Peoples R China.,pzhou@shmu.edu.cn |
| 英文摘要 | The DNA fragment encoding Kluyvera citrophila penicillin G acylase (KcPGA) was amplified and cloned into the vector pET28b to obtain a C-terminus His-tagged fusion expression plasmid. The fusion protein KcPGA was successfully overexpressed in Escherichia coli BL21(DE3). The optimal induction concentration of isopropylthio-p-D-galactoside (IPTG) was found to be 5 M The fusion protein was purified in a single step by Ni-IDA affinity chromatograph to a specific activity of 35.3 U/mg protein with a final yield of 89% representing a 23-fold purification. The data presented here suggest that the purified fusion protein is stable with respect to pH and temperature. The optimal pH and temperature of recombinant KcPGA are 8.5 and 55 degreesC, respectively. The K-m and V-max are 17.6 muM and 23.8 U/mg, respectively. Therefore, the high yield and high specific activity of recombinant KcPGA produced in E coli, together with other kinetic parameters, represent an excellent basis for further development of recombinant KcPGA as an immobilized biocatalyst for industrial applications. (C) 2004 Elsevier Inc. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | PROTEIN |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000224778800004 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2007]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Yong, W,Shi, XL,Yuan, ZY,et al. Expression, purification, and characterization of His-tagged penicillin G acylase from Kluyvera citrophila in Escherichia coli[J]. PROTEIN EXPRESSION AND PURIFICATION,2004,38(1):24-28. |
| APA | Yong, W,Shi, XL,Yuan, ZY,&Pei, Z.(2004).Expression, purification, and characterization of His-tagged penicillin G acylase from Kluyvera citrophila in Escherichia coli.PROTEIN EXPRESSION AND PURIFICATION,38(1),24-28. |
| MLA | Yong, W,et al."Expression, purification, and characterization of His-tagged penicillin G acylase from Kluyvera citrophila in Escherichia coli".PROTEIN EXPRESSION AND PURIFICATION 38.1(2004):24-28. |
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