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    Methods for producing biological substances in pigment-deficient mutants of bacillus cells
    1.
    发明申请
    Methods for producing biological substances in pigment-deficient mutants of bacillus cells 有权
    在芽孢杆菌细胞色素缺陷突变体中生产生物物质的方法

    公开(公告)号：US20040096944A1

    公开(公告)日：2004-05-20

    申请号：US10627124

    申请日：2003-07-25

    申请人： Novozymes Biotech, Inc.

    发明人： Maria Tang ,  Alan Sloma ,  David Sternberg ,  Regine Behr

    IPC分类号： C12P021/02 ,  C12N001/21

    CPC分类号： C12N9/0077 ,  C07K14/32 ,  C12P1/04 ,  C12P19/00 ,  C12P21/02

    摘要： The present invention relates to methods of producing a heterologous biological substance, comprising: (a) cultivating a mutant of a parent Bacillus cell under conditions conducive for the production of the heterologous biological substance, wherein (i) the mutant cell comprises a first nucleic acid sequence directing synthesis of the heterologous biological substance and a second nucleic acid sequence comprising a modification of at least one of the genes cypx and yvmC, which are involved in the production of a red pigment, and (ii) the mutant cell is deficient in the production of the red pigment compared to the parent Bacillus cell when cultivated under the same conditions; and (b) recovering the heterologous biological substance from the cultivation medium. The present invention also relates to mutants of Bacillus cells and methods for producing the mutants.

    摘要翻译： 本发明涉及产生异源生物物质的方法，其包括：（a）在有利于产生异源生物物质的条件下培养亲本芽孢杆菌细胞的突变体，其中（i）突变细胞包含第一核酸 序列指导异源生物物质的合成和包含参与红色素生产的至少一种基因cypx和yvmC的修饰的第二核酸序列，和（ii）突变体细胞在 在相同条件下培养时，与母体芽孢杆菌细胞相比，产生红色素; 和（b）从培养基中回收异源生物物质。 本发明还涉及芽孢杆菌细胞的突变体和产生突变体的方法。

    Methods for producing a polypeptide in a bacillus cell
    2.
    发明申请
    Methods for producing a polypeptide in a bacillus cell 有权
    在芽孢杆菌细胞中产生多肽的方法

    公开(公告)号：US20030170876A1

    公开(公告)日：2003-09-11

    申请号：US09834271

    申请日：2001-04-12

    申请人： Novozymes Biotech, Inc.

    发明人： William Widner ,  Alan Sloma ,  Michael D. Thomas

    IPC分类号： C12N001/20

    CPC分类号： C12N15/75

    摘要： The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and alternatively also (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium. The present invention also relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a nullconsensusnull promoter having the sequence TTGACA for the nullnull35null region and TATAAT for the nullnull10null region operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and (ii) an mRNA processing/stabilizing sequence located downstream of the nullconsensusnull promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium.

    摘要翻译： 本发明涉及产生多肽的方法，其包括：（a）在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞，其中所述芽孢杆菌细胞包含核酸构建体，其包含（i）串联启动子，其中 串联启动子的每个启动子序列可操作地连接到编码多肽的核酸序列的单拷贝，或者还有（ii）位于串联启动子下游的mRNA加工/稳定序列和编码多肽的核酸序列的上游 ; 和（b）从培养基中分离多肽。 本发明还涉及产生多肽的方法，其包括：（a）在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞，其中所述芽孢杆菌细胞包含核酸构建体，其包含（i）“共有” 具有“-35”区域的TTGACA序列的启动子和“-10”区域的TATAAT可操作地连接到编码多肽的核酸序列的单拷贝，和（ii）位于“ 共有“启动子和编码多肽的核酸序列的上游; 和（b）从培养基中分离多肽。

    Methods for producing hyaluronan in a recombinant host cell
    4.
    发明申请
    Methods for producing hyaluronan in a recombinant host cell 有权
    在重组宿主细胞中产生透明质酸的方法

    公开(公告)号：US20030175902A1

    公开(公告)日：2003-09-18

    申请号：US10326185

    申请日：2002-12-20

    申请人： Novozymes Biotech, Inc.

    发明人： Alan Sloma ,  Regine Behr ,  William Widner ,  Maria Tang ,  David Sternberg ,  Stephen Brown

    IPC分类号： C12P019/26 ,  C12N009/24 ,  C12N001/21 ,  C12N015/74

    CPC分类号： C12P19/26 ,  C12N9/1051 ,  C12N15/52

    摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase

    摘要翻译： 本发明涉及透明质酸的制备方法，其包括：（a）在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞，其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体 涉及透明质酸合酶编码序列外源的启动子序列; 和（b）从培养基中回收透明质酸。 本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列，以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因， UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。 本发明还涉及编码UDP-葡萄糖-6-脱氢酶，UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列

    Methods for producing secreted polypeptides having L-asparaginase activity
    5.
    发明申请
    Methods for producing secreted polypeptides having L-asparaginase activity 审中-公开
    用于产生具有L-天冬酰胺酶活性的分泌多肽的方法

    公开(公告)号：US20030186380A1

    公开(公告)日：2003-10-02

    申请号：US10406025

    申请日：2003-04-01

    申请人： Novozymes Biotech, Inc.

    发明人： Michael D. Thomas ,  Alan Sloma

    IPC分类号： C12P021/02 ,  C12N001/21 ,  C07H021/04 ,  C12N009/64 ,  C12N015/74

    CPC分类号： C12N9/82 ,  C07K2319/02 ,  C12Y305/01001

    摘要： The present invention relates to recombinant methods for producing a secreted polypeptide having L-asparaginase activity, comprising (a) cultivating under conditions conducive for production of the polypeptide a host cell comprising a nucleic acid construct comprising a first nucleic acid sequence encoding a secretory signal peptide operably linked to second nucleic acid sequence encoding the polypeptide having L-asparaginase activity, wherein the signal peptide directs the polypeptide into the cell's secretory pathway; and (b) recovering the secreted polypeptide. The present invention also relates to isolated polypeptides having L-asparaginase activity and nucleic acids thereof.

    摘要翻译： 本发明涉及用于产生具有L-天冬酰胺酶活性的分泌多肽的重组方法，其包括（a）在有助于产生多肽的条件下培养宿主细胞，宿主细胞包含核酸构建体，其包含编码分泌信号肽的第一核酸序列 与编码具有L-天冬酰胺酶活性的多肽的第二核酸序列可操作地连接，其中所述信号肽将多肽引导到细胞的分泌途径中; 和（b）回收分泌的多肽。 本发明还涉及具有L-天冬酰胺酶活性的分离的多肽及其核酸。