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    Methods for Producing Heterologous Polypeptides in Thiol-Disulfide Oxidoreductase-Deficient Bacterial Mutant Cells
    1.
    发明申请
    Methods for Producing Heterologous Polypeptides in Thiol-Disulfide Oxidoreductase-Deficient Bacterial Mutant Cells 审中-公开
    在硫代二硫化物氧化还原酶缺陷型细菌突变体细胞中产生异源多肽的方法

    公开(公告)号：US20120329090A1

    公开(公告)日：2012-12-27

    申请号：US13516475

    申请日：2010-12-17

    申请人： William Widner ,  Bjarke Christensen

    发明人： William Widner ,  Bjarke Christensen

    IPC分类号： C12N1/21 ,  C12N15/87 ,  C12P21/00

    CPC分类号： C12N15/75 ,  C12N9/0036 ,  C12N9/0051 ,  C12Y108/04002

    摘要： The present invention relates to methods of producing a heterologous polypeptide, comprising: (a) cultivating a mutant of a parent bacterial cell in a medium for the production of the heterologous polypeptide, wherein (i) the mutant cell comprises a first polynucleotide encoding the heterologous polypeptide which comprises two or more (several) cysteines, and a second polynucleotide comprising a modification of a gene encoding a thiol-disulfide oxidoreductase that incorrectly catalyzes the formation of one or more (several) disulfide bonds between the two or more (several) cysteines of the heterologous polypeptide, and (ii) the mutant cell is deficient in production of the thiol-disulfide oxidoreductase compared to the parent bacterial cell when cultivated under the same conditions; and (b) recovering the heterologous polypeptide from the cultivation medium. The present invention also relates to such bacterial mutants and methods for producing such bacterial mutants.

    摘要翻译： 本发明涉及产生异源多肽的方法，其包括：（a）在用于产生异源多肽的培养基中培养母细菌细胞的突变体，其中（i）突变细胞包含编码异源多肽的第一个多核苷酸 包括两个或多个（几个）半胱氨酸的多肽，以及第二多核苷酸，其包含编码硫醇 - 二硫键氧化还原酶的基因的修饰，其不正确地催化两个或多个（几个）半胱氨酸之间的一个或多个（几个）二硫键的形成 的异源多肽，和（ii）当在相同条件下培养时，与母体细菌细胞相比，突变细胞缺乏硫醇 - 二硫键氧化还原酶的产生; 和（b）从培养基中回收异源多肽。 本发明还涉及这种细菌突变体及其生产方法。