公开(公告)号：EP1925670A1

公开(公告)日：2008-05-28

申请号：EP06775269.1

申请日：2006-08-02

申请人： Bioright Worldwide Company Limited

发明人： WANG, Jun ,  TSANG, Waikei ,  YAP, Hongkin ,  LIU, Zhaoming ,  SIU, Yaulung ,  TSANG, Supyin

IPC分类号： C12N15/70 ,  C12N15/72 ,  C12N15/68 ,  C12N9/06 ,  C12N9/14 ,  C12P21/02 ,  C12P35/06

CPC分类号： C12N15/70 ,  C12P21/02

摘要： The present invention discloses Escherichia coli expression vector, as well as a method for steadily, high-efficiently expressing a required target protein in E. coli using the expression vector. The expression vector is constructed from vector pRSET-A, wherein ampicillin resistance gene in said vector pRSET-A is replaced by kanamycin resistance gene. The expression vector can be used to high-efficiently express enzymes required for the production of 7-aminocephalosporanic acid, meanwhile it does not produce β-lactamase by which the productivity of 7-aminocephalosporanic acid is reduced. The present invention also discloses two expression vectors, i.e., one expression vector pHS-GHA whose gene product is D-amino acid oxidase mutant GHA, and another expression vector pT7-kan-ACY whose gene product is glutaryl-7-aminocephalosporanic acid acylase of Pseudomonas sp. SE83.

摘要翻译： 本发明公开了大肠杆菌表达载体，以及使用表达载体在大肠杆菌中稳定地高效表达所需靶蛋白的方法。 表达载体由载体pRSET-A构建，其中所述载体pRSET-A中的氨苄青霉素抗性基因被卡那霉素抗性基因替代。 表达载体可用于高效表达生产7-氨基头孢烷酸所需的酶，同时不产生β-内酰胺酶，其中7-氨基头孢烷酸的产率降低。 本发明还公开了两种表达载体，即其基因产物是D-氨基酸氧化酶突变体GHA的一种表达载体pHS-GHA和其基因产物是戊二酰-7-氨基头孢烷酸酰基转移酶的另一表达载体pT7-kan-ACY 假单胞菌属 SE83。