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    MODIFIED MESSENGER RNA STABILIZING SEQUENCES FOR EXPRESSING GENES IN BACTERIAL CELLS
    3.
    发明公开
    MODIFIED MESSENGER RNA STABILIZING SEQUENCES FOR EXPRESSING GENES IN BACTERIAL CELLS 有权
    改进的mRNA稳定的基因表达序列菌体

    公开(公告)号：EP2104739A2

    公开(公告)日：2009-09-30

    申请号：EP07874376.2

    申请日：2007-12-19

    申请人： Novozymes Inc.

    发明人： THOMAS, Michael ,  ERICHSEN, Gloria ,  WIDNER, William

    IPC分类号： C12N15/75

    CPC分类号： C07K14/32 ,  C12N15/75

    摘要： The present invention relates to methods of producing a polypeptide having biological activity in a bacterial cell, comprising: (a) cultivating a bacterial host cell in a medium conducive for production of the polypeptide, wherein the bacterial host cell comprises a nucleic acid construct comprising a promoter region operably linked to a polynucleotide sequence encoding the polypeptide and a modified mRNA processing/stabilizing sequence located downstream of the promoter region and upstream of the ribosome binding site of the polynucleotide sequence encoding the polypeptide, wherein the modified mRNA processing/stabilizing sequence promotes higher expression of the polynucleotide sequence compared to an unmodified mRNA processing/stabilizing sequence; and (b) isolating the polypeptide having biological activity from the cultivation medium. The present invention also relates to such modified mRNA processing/stabilizing sequences, nucleic acid constructs, and bacterial host cells and to methods of obtaining such bacterial host cells.

    METHODS FOR PRODUCING HETEROLOGOUS POLYPEPTIDES IN THIOL-DISULFIDE OXIDOREDUCTASE-DEFICIENT BACTERIAL MUTANT CELLS
    5.
    发明公开
    METHODS FOR PRODUCING HETEROLOGOUS POLYPEPTIDES IN THIOL-DISULFIDE OXIDOREDUCTASE-DEFICIENT BACTERIAL MUTANT CELLS 审中-公开
    工艺用于生产突变细菌中异源多肽与巯基二硫化物OXIDOREDUKTASEMANGEL

    公开(公告)号：EP2516636A1

    公开(公告)日：2012-10-31

    申请号：EP10801045.5

    申请日：2010-12-17

    申请人： Novozymes Inc. ,  Novozymes A/S

    发明人： WIDNER, William ,  CHRISTENSEN, Bjarke

    IPC分类号： C12N9/02 ,  C12N15/75 ,  C12N15/53 ,  C12P21/02 ,  C07K14/32

    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.