首页> 外国专利> PROTEINS WITH ENZYME ACTIVITY FROM S. EPIDERMIS, THEIR PREPARATION AND THEIR USE IN BIOSYNTHESIS OF COMPOUNDS CONTAINING DEHYDROAMINO AND RESIDUES AND/OR THIOETHER BRIDGES

PROTEINS WITH ENZYME ACTIVITY FROM S. EPIDERMIS, THEIR PREPARATION AND THEIR USE IN BIOSYNTHESIS OF COMPOUNDS CONTAINING DEHYDROAMINO AND RESIDUES AND/OR THIOETHER BRIDGES

机译:来自表皮葡萄球菌的具有酶活性的蛋白质,其制备及其在生物合成中的作用,该化合物包含脱水亚氨基和残基和/或硫醚桥

摘要

Novel recombinant DNA molecule (I) has a sequence corresp. to one or more DNA sequences contd. in a Bgl II fragment of the 54 Rpb episomal element pTii32 from staphylococcus epidermis and codes for at least one protein having EpiB, EpiC, EpiD, EpiP or EpiQ enzymatic activity; or (I) is capable of hybridising to the BglII fragment and codes for at least one protein with the above enzymatic activities. Also claimed are: the protein EpiB, EpiC, EpiD, EpiP or EpiQ in the absence of impurities with which it is normally associated; a fusion protein compising one of EpiB, C, D, P or Q and an auxiliary protein, a fusion protein in which one of the Epi B, C, D, P or Q is fused to an auxiliary protein such that the junction between the Epi protein and the auxiliary protein can be cleaved by an enzyme; and a method of producing a protein with at least one of the structural features of a lantibiotic. In the novel method comprising inserting a DNA sequence coding for a predetermined aminoacid sequence in a vector plasmid such that it is under the control of a regulator promoter, transforming the resulting plasmid vector into a host and cultivating the host so as to express the DNA and isolating the protein, the improvement comprises also transforming the host by a DNA sequence or sequences comprising functionally operable genes coding for EpiB, C, D, P or Q (or an active variant), the genes being expressed during cultivation of the host so that the resulting protein has at least one of the structural features of a lantibiotic.
机译:新的重组DNA分子(I)具有相应的序列。一个或多个DNA序列。在来自葡萄球菌表皮的54个Rpb附加型元件pTii32的Bgl II片段中,编码至少一种具有EpiB,EpiC,EpiD,EpiP或EpiQ酶活性的蛋白质;或(I)能够与BglII片段杂交并编码具有上述酶活性的至少一种蛋白质。还要求保护的是:蛋白质EpiB,EpiC,EpiD,EpiP或EpiQ在没有杂质的情况下通常与蛋白质结合;包含EpiB,C,D,P或Q之一和辅助蛋白的融合蛋白,其中Epi B,C,D,P或Q之一与辅助蛋白融合在一起的融合蛋白, Epi蛋白和辅助蛋白可以被酶裂解。以及生产具有羊毛硫抗生素的至少一种结构特征的蛋白质的方法。在该新方法中,包括在载体质粒中插入编码预定氨基酸序列的DNA序列,使其处于调节启动子的控制下,将所得的质粒载体转化入宿主中,并培养该宿主以表达该DNA。分离蛋白质,改进还包括通过DNA序列或包含编码EpiB,C,D,P或Q的功能可操作基因的序列(或活性变体)转化宿主,所述基因在宿主培养期间表达,从而所得蛋白质具有羊毛硫抗生素的至少一种结构特征。

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