Improving the specific synthetic activity of a penicillin G acylase using DNA family shuffling
文献类型：期刊论文
| 作者 | Zhou, Z; Zhang, AH; Wang, JR; Chen, ML; Li, RB; Yang, S; Yuan, ZY |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2003 |
| 卷号 | 35期号:6页码:573-579 |
| 关键词 | penicillin G acylase DNA family shuffling S/H ratio |
| 通讯作者 | , |
| 英文摘要 | Penicillin G Acylas (PGA) of Providencia rettgeri (ATCC 25599) was evolved using a modified DNA family shuffling method. The identity of pga genes from Escherichia coli, Kluyvera citrophila and Providencia rettgeri ranges from 62.5% to 96.9%. The pga genes from above three species were recombined and shuffled to create interspecies pga gene fusion libraries. By substituting assembled chimaeras for corresponding region of pETPPGA, different recombinants were constructed and expressed in E. coli JM109(DE3). Mutants with obvious beta-lactam synthetic activity were selected from the plates and the ratios of synthesis to hydrolysis (S/H) were determined subsequently. It was shown that the primary structures of selected positives exhibited significant diversity among each library. The best mutant possessed 40% higher synthetic activity than the wild type enzyme of PrPGA. It was further proved in this study that the domain of alpha subunit contributed much more to improve the specific activity of synthesis. Results showed a recombinant PGA with higher synthetic activity was acquired by the method of DNA shuffling. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | ESCHERICHIA-COLI ATCC-11105 ; DIRECTED EVOLUTION ; GENES |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000183733400014 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2315]  |
| 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 | Zhou, Z,Zhang, AH,Wang, JR,et al. Improving the specific synthetic activity of a penicillin G acylase using DNA family shuffling[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2003,35(6):573-579. |
| APA | Zhou, Z.,Zhang, AH.,Wang, JR.,Chen, ML.,Li, RB.,...&Yuan, ZY.(2003).Improving the specific synthetic activity of a penicillin G acylase using DNA family shuffling.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,35(6),573-579. |
| MLA | Zhou, Z,et al."Improving the specific synthetic activity of a penicillin G acylase using DNA family shuffling".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 35.6(2003):573-579. |
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