采用pHsh载体克隆与表达N-乙酰鸟氨酸脱乙酰基酶基因  

Cloning and high-level active expression of N-acetyl-L-ornithine deacetylase gene

作　　者：张琛[1] 李环[1] 吴圆丽[1] 韦萍[1] 

机构地区：[1]南京工业大学生物与制药工程学院,南京211800

出　　处：《生物加工过程》2012年第5期67-71,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展计划(973计划)资助项目(2003CB716004)

摘　　要：利用质粒pHsh为表达载体,构建高效表达N-乙酰鸟氨酸脱乙酰基酶的基因工程菌E.coli DH10B/argE-pHsh。为提高酶活并降低生产成本,优化了诱导条件。结果表明:NAOase可在pHsh系统中高活性表达,诱导起始OD600为0.6,在空气摇床中42℃热激诱导5 h重组菌比酶活达到152 U/mL。Using pHsh as the expressing vector, the genetic engineering strain argE-pHsh producing acetylomithine deacetylase was successfully constructed. In order to improve the NAOase activity and to reduce the cost, the inducement conditions and culture medium compoments were optimized. The result showed that NAOase was over-expressed in E. coli using pHsh expression vector in fermentors with the ＂flow-in-heat＂ method. The NAOase activity reached 152 U/mL after the cells harboring pHsh-argE were heat-shocked and continued to grow at 42 ℃ for 5 h.

关 键 词：N-乙酰鸟氨酸脱乙酰基酶 大肠杆菌 克隆 

分 类 号：Q786[生物学—分子生物学]