High-level production penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli with optimization of carbon sources
- 作者：Shiwei Cheng ; Qingxun Song ; Dongzhi Wei ; Bingxue Gao
- 关键词：Alcaligenes faecalis ; Batch cultivation ; Carbon source ; Overexpression ; Penicillin G acylase
- 刊名：Enzyme and Microbial Technology
- 出版年：2007
- 期刊代码：26_01410229
- 类别：ce
- 出版时间：2 August 2007
- 卷：41
- 期：3
- 页码：326-330
- 文件大小：257 K
摘要
Different carbon sources were investigated for overproduction penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli strains. The results indicated that the enzyme was optimally produced with 45 g/l of dextrin, and about 43,385 and 79,880 U/l for the highest enzyme activities were obtained in batch cultivations of shaken flasks and a 3.7 l bioreactor, respectively. Active site titration and SDS-PAGE electrophoretic analysis demonstrated that the maximum yield of the active enzyme was 2.54 g/l, which was about 40%of total soluble proteins. The highest specific activity of A. faecalis penicillin G acylase obtained was above 10 U/mg protein, and there was almost no plasmid lost in the whole batch cultivations. Furthermore, the cultivation process was relatively simple and suitable for large-scale production.