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    • 4. 发明授权
    • Recombinant yeast PDI and process for production thereof
    • 重组酵母PDI及其制备方法
    • US06387683B1
    • 2002-05-14
    • US09368588
    • 1999-08-05
    • Nobuyoshi IshiiYasuo SuzukiKohji UchidaYushi MatuoHideo Tanaka
    • Nobuyoshi IshiiYasuo SuzukiKohji UchidaYushi MatuoHideo Tanaka
    • C12N990
    • C12N9/90Y10S435/942
    • The present invention provides a process for producing biologically active recombinant yeast protein disulfide isomerase, which includes the steps of: a) deleting, substituting, or adding one or more bases in a region encoding an endoplasmic reticulum retention signal in a gene encoding protein disulfide isomerase of yeast to modify the gene so as not to encode part or all of the endoplasmic reticulum retention signal; b) incorporating the modified gene into an expression vector; c) transforming host cells with the expression vector; and d) culturing the host cells transformed with the expression vector in a culture medium kept at a nearly neutral pH, thereby causing protein disulfide isomerase to be secreted in an active state outside the host cells. According to the process of the invention, recombinant yeast protein disulfide isomerase can be secreted in a large amount in a biologically active state into a culture medium, and can be collected by a simple purification method.
    • 本发明提供生产生物活性重组酵母蛋白二硫键异构酶的方法,其包括以下步骤:a)在编码蛋白质二硫键异构酶的基因的编码内质网保持信号的区域中缺失,取代或添加一个或多个碱基 的酵母修饰基因,以免编码部分或全部内质网保持信号; b)将修饰的基因并入表达载体中; c)用表达载体转化宿主细胞; 和d)在保持接近中性pH的培养基中培养用表达载体转化的宿主细胞,从而使蛋白质二硫键异构酶以宿主细胞外的活性状态分泌。 根据本发明的方法,可以将重组酵母蛋白二硫键异构酶以大量的生物活性状态分泌到培养基中,并且可以通过简单的纯化方法收集。