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    Production of active Pseudomonas glumae lipase in homologous or heterologous hosts
    4.
    发明授权
    Production of active Pseudomonas glumae lipase in homologous or heterologous hosts 失效
    在同源或异源宿主中产生活性的假单胞菌脂肪酶

    公开(公告)号：US5804409A

    公开(公告)日：1998-09-08

    申请号：US449015

    申请日：1995-05-24

    申请人： Jannetje Wilhelmina Bos ,  Leon Gerardus Frenken ,  Cornelis Theodorus Verrips ,  Christiaan Visser

    发明人： Jannetje Wilhelmina Bos ,  Leon Gerardus Frenken ,  Cornelis Theodorus Verrips ,  Christiaan Visser

    IPC分类号： C12N1/21 ,  C12N9/20 ,  C12N15/55 ,  C12N15/75 ,  C07K14/195 ,  C12N15/31 ,  C12P21/02

    CPC分类号： C12N15/75 ,  C12N9/20

    摘要： The production of an active Pseudomonas glumae lipase isolated from P. glumae PG1 (CBS 322.89) and other lipases suitable for application in detergent systems is described in homologous, but particularly in heterologous hosts, e.g. Bacillus subtilis. For the latter a "helper function" or "lipase-specific stabilization/translocation protein" is needed, for which a gene is provided which, when expressed in concert with the lipase gene, can improve the stabilization of the intermediates involved in the production and translocation/secretion of the lipase. The hosts are transformed by recombinant DNA methods and modified lipases can be made by site-directed mutation or classical mutation techniques. The lipase gene and the gene encoding the helper function can be part of one operon that can be transcribed to form a polycistronic messenger or be present as separate genes yielding two mRNA's on transcription. The production level can be further improved by optimising the regulation sequences. It can be advantageous to use at least part of the signal sequence of the lipase gene in addition to a signal sequence homologous to the host in which the lipase is produced.

    摘要翻译： 分离自P.glumae PG1（CBS322.89）和适用于洗涤剂体系的其它脂肪酶的活性假单胞菌脂肪酶的生产描述为同源的，但特别是异源宿主，例如， 枯草芽孢杆菌 对于后者，需要“辅助功能”或“脂肪酶特异性稳定/易位蛋白”，其中提供了一种基因，当与脂肪酶基因一致表达时，可以改善参与生产的中间体的稳定性， 易位/分泌脂肪酶。 通过重组DNA方法转化宿主，修饰的脂肪酶可以通过定点突变或经典突变技术进行。 脂肪酶基因和编码辅助功能的基因可以是一个可以被转录以形成多顺反子信使的一个操纵子的一部分，或作为在转录上产生两个mRNA的单独基因存在。 通过优化调节序列可以进一步提高生产水平。 除了与其中产生脂肪酶的宿主同源的信号序列之外，使用脂肪酶基因的至少部分信号序列是有利的。

    Production of active Pseudomonas glumae lipase in homologous or heterologous hosts
    5.
    发明授权
    Production of active Pseudomonas glumae lipase in homologous or heterologous hosts 失效
    在同源或异源宿主中产生活性的假单胞菌脂肪酶

    公开(公告)号：US5641671A

    公开(公告)日：1997-06-24

    申请号：US34650

    申请日：1993-03-22

    申请人： Jannetje Wilhelmina Bos ,  Leon Gerardus Frenken ,  Cornelis Theodorus Verrips ,  Christiaan Visser

    发明人： Jannetje Wilhelmina Bos ,  Leon Gerardus Frenken ,  Cornelis Theodorus Verrips ,  Christiaan Visser

    IPC分类号： C12N1/21 ,  C12N9/20 ,  C12N15/55 ,  C12N15/75 ,  C12N1/20 ,  C12N1/15 ,  C12N1/19

    CPC分类号： C12N15/75 ,  C12N9/20

    摘要： The production of an active Pseudomonas glumae lipase isolated from P. glumae PG1 (CBS 322.89) and other lipases suitable for application in detergent systems is described in homologous, but particularly in heterologous hosts, e.g. Bacillus subtilis. For the latter a "helper function" or "lipase-specific stabilization/translocation protein" is needed, for which a gene is provided which, when expressed in concert with the lipase gene, can improve the stabilization of the intermediates involved in the production and translocation/secretion of the lipase. The hosts are transformed by recombinant DNA methods and modified lipases can be made by site-directed mutation or classical mutation techniques. The lipase gene and the gene encoding the helper function can be part of one operon that can be transcribed to form a polycistronic messenger or be present as separate genes yielding two mRNA's on transcription. The production level can be further improved by optimizing the regulation sequences. It can be advantageous to use at least part of the signal sequence of the lipase gene in addition to a signal sequence homologous to the host in which the lipase is produced.

    摘要翻译： 分离自P.glumae PG1（CBS322.89）和适用于洗涤剂体系的其它脂肪酶的活性假单胞菌脂肪酶的生产描述为同源的，但特别是异源宿主，例如， 枯草芽孢杆菌 对于后者，需要“辅助功能”或“脂肪酶特异性稳定/易位蛋白”，其中提供了一种基因，当与脂肪酶基因一致表达时，可以改善参与生产的中间体的稳定性， 易位/分泌脂肪酶。 通过重组DNA方法转化宿主，修饰的脂肪酶可以通过定点突变或经典突变技术进行。 脂肪酶基因和编码辅助功能的基因可以是一个可以被转录以形成多顺反子信使的一个操纵子的一部分，或作为在转录上产生两个mRNA的单独基因存在。 通过优化调节序列可以进一步提高生产水平。 除了与其中产生脂肪酶的宿主同源的信号序列之外，使用脂肪酶基因的至少部分信号序列是有利的。