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    ESCHERICHIA COLI EXPRESSION VECTOR AND THE USES THEREOF
    1.
    发明公开
    ESCHERICHIA COLI EXPRESSION VECTOR AND THE USES THEREOF 审中-公开
    ESCHERICHIA COLI-EXPRESSIONSVEKTOR UND VERWENDUNGEN DAVON

    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。

    TWO-STEPS ENZYME METHOD FOR PREPARING 7-AMINOCEPHALOSPORANIC ACID
    3.
    发明公开
    TWO-STEPS ENZYME METHOD FOR PREPARING 7-AMINOCEPHALOSPORANIC ACID 审中-公开
    ZWEISCHRITT-ENZYMVERFAHREN ZUR HERSTELLUNG VON 7-AMINOCEPHALOSPORANSÄURE

    公开(公告)号：EP1925663A1

    公开(公告)日：2008-05-28

    申请号：EP06775270.9

    申请日：2006-08-02

    申请人： Bioright Worldwide Company Limited

    发明人： WANG, Jun ,  TSANG, Waikei ,  YAP, Hongkin ,  CHEN, Junmin ,  SIU, Yaulung ,  TSANG, Supyin

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

    CPC分类号： C12P35/02 ,  C12N9/0024 ,  C12P35/06

    摘要： The present invention discloses a two-step enzyme method for preparing 7-aminocephalosporanic acid from cephalosporin C, wherein D-amino acid oxidase used is purified D-amino acid oxidase mutant of yeast Trigonopsis variabilis, having a specific activity of 105% higher than that of parent D-amino acid oxidase. The method has no need of addition of hydrogen peroxide, β-lactamase inhibitor, catalase inhibitor, catalase and the like commonly used in the prior art. The productivity of the method can reach more than 93%. Thus, the method is simple, low in cost and high in productivity.

    摘要翻译： 本发明公开了一种用于从头孢菌素C制备7-氨基头孢烷酸的两步酶方法，其中使用D-氨基酸氧化酶是酵母Trigonopsis variabilis的纯化D-氨基酸氧化酶突变体，比活性比其活性高105％ 的亲代D-氨基酸氧化酶。 该方法不需要添加现有技术中通常使用的过氧化氢，β-内酰胺酶抑制剂，过氧化氢酶抑制剂，过氧化氢酶等。 该方法的生产率可达93％以上。 因此，该方法简单，成本低，生产率高。