|
|||||
|
|
| 雷氏普罗威登斯菌青霉素G酰化酶基因在大肠杆菌中的克隆与表达 | |
| 引用本文： | 周政,张爱晖,周丽萍,杨晟,姜涌明,袁中一.雷氏普罗威登斯菌青霉素G酰化酶基因在大肠杆菌中的克隆与表达[J].工业微生物,2002,32(3):1-5. |
| 作者姓名： | 周政 张爱晖 周丽萍 杨晟 姜涌明 袁中一 |
| 作者单位： | 1. 中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031 2. 扬州大学生物科学与技术学院 3. 江苏大学 |
| 基金项目： | 国家自然科学基金(No.30100029)，“863”国家高科技资助项目(No.2001AA235081) |
| 摘 要： | 利用PCR和分子克隆技术从雷氏普罗威登斯菌（Prouidencia rettgeri)(ATCC29944)的基因组DNA中获得一个青霉素G酰化酶（penicillinGacylase,PGA)基因并将其装入表达质粒pET24a。携带有重组质粒pETPGA的Escherichia coli基因工程菌BL21（DE3）/pETPGA实现了PGA的高效表达，对发酵条件的研究表明基因工程菌在24℃，添加5g/L甘油条件下以1.0mmol/LIPTG诱导1.5h酶活力即达到993.4U/L，比野生菌酶活力（15U/L）提高了66倍。 |
| 关 键 词： | 雷氏普罗威登斯菌 青霉素G 酰化酶 基因 大肠杆菌 克隆 表达 |
Cloning and expression of penicillin G acylase gene from Providencia rettgeri in Escherichia coli |
|
| ZHOU Zheng,ZHANG Ai-hui,ZHOU Li-ping,YANG Sheng,JIANG Yong-ming,YUAN Zhong-yi.Cloning and expression of penicillin G acylase gene from Providencia rettgeri in Escherichia coli[J].Industrial Microbiology,2002,32(3):1-5. | |
| Authors: | ZHOU Zheng ZHANG Ai-hui ZHOU Li-ping YANG Sheng JIANG Yong-ming YUAN Zhong-yi |
| Institution: | ZHOU Zheng,ZHANG Ai-hui,ZHOU Li-ping,YANG Sheng,JIANG Yong-ming2,YUAN Zhong-yi |
| Abstract: | Based on PCR techniques, a gene encoding penicillin G acylase(PGA) was obtained from genomic DNA of Providencia rettgeri(A.TCC29944). The pET24a vector combined with the PGA gene was transformed into BL21(DE3) and the resulting recombinant Escherichia coli gave a high level expression of PGA. Several factors, including IPTG concentration, temperature and carbon source, were optimized for enzyme overproduction. The optimization significantly increased PGA expression by 66-fold compared to the native expression (15U/L) in P. rettgeri. After inducing with 1. 0mmol/L IPTG at 24! and fermentation in medium with additive of 5g/L glycerol for 1.5 hour, The enzyme expression of recombinant E. coli reached 993.4U/L. |
| Keywords: | penicillin G acylase Providencia rettgeri cloning and expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |