Expression of gene encoding GL-7ACA acylase in Escherichia coli
文献类型：期刊论文
| 作者 | Wang, ED; Zheng, YG; Li, Y; Jiang, WH; Yang, YL |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2002 |
| 卷号 | 34期号:4页码:526-531 |
| 关键词 | GL-7ACA acylase recombinant plasmid expression and purification Pseudomonas Escherichia coli |
| 通讯作者 | , |
| 英文摘要 | Glutaryl 7-amino cephalosporanic acid acylase (GL-7ACA acylase) from Pseudomonas sp. 130 catalyzes hydrolysis of glutaryl 7-amino cephalosporanic acid to produce 7-amino cephalosporanic acid (7-ACA). 7-ACA is the starting material for the industrial production of most cephalosparonic derivatives. Six plasmids for expression of GL-7ACA acylase were constructed and these recombinant plasmids presented different expression characteristics in Escherichia coli. The acylase gene from plasmid pKKCA1 was inserted into plasmid pMFT7-5 and the resulting plasmid pMFT7CA1 has higher expression in E. coli. The specific activity of the crude extract of the transformant JM109(DE3)/MFT7CA1 was near 5 u/g, so the overproduced enzyme was easily purified by a single-step anion exchange column chromatography. The enzyme could be purified by immobilized ion affinity chromatography after fused by 6 X His in the N-terminal of its alpha-subunit. Because plasmid pSMLCA1 brings tc(R) and p15A origin, it is special useful plasmid. in fermentation. Two secretory expression plasmids, pSUCA1S and pETCA1pelB, could secrete the acylase to periplasmic space of bacteria. The whole cells containing the secretory expression plasmid may be used for production of TACA directly. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | TRANSFER-RNA SYNTHETASE ; ACID ACYLASE ; PURIFICATION ; PSEUDOMONAS ; VECTORS |
| 收录类别 | SCI |
| 语种 | 中文 |
| WOS记录号 | WOS:000177017300026 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2488]  |
| 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 | Wang, ED,Zheng, YG,Li, Y,et al. Expression of gene encoding GL-7ACA acylase in Escherichia coli[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2002,34(4):526-531. |
| APA | Wang, ED,Zheng, YG,Li, Y,Jiang, WH,&Yang, YL.(2002).Expression of gene encoding GL-7ACA acylase in Escherichia coli.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,34(4),526-531. |
| MLA | Wang, ED,et al."Expression of gene encoding GL-7ACA acylase in Escherichia coli".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 34.4(2002):526-531. |
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