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重组酿酒酵母合成谷胱甘肽的研究

, PP. 160-165

Keywords: γ,-谷氨酰半胱氨酸合成酶,谷胱甘肽合成酶,谷胱甘肽,酿酒酵母

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Abstract:

利用PCR方法以酿酒酵母SaccharomycescerevisiaeS288c基因组DNA为模板,PCR扩增γ-谷氨酰半胱氨酸合成酶(GSH1)及谷胱甘肽合成酶(GSH2)基因,以乙醇脱氢酶(ADH1)启动子进行表达,并以Kanr基因和Hygr基因作为筛选标记,构建了两个重组表达质粒YEplace181AK-GSH1和YEplace181AH-GSH2。采用电击法将这两个质粒分别和同时转入工业酿酒酵母菌株SaccharomycescerevisiaeTS013,在含有G418或(和)Hygromycin的平板上筛选阳性克隆S.TS013/GSH1、S.TS013/GSH2和S.TS013/GSH1+GSH2。重组菌进行摇瓶发酵,采用四氧嘧啶法测定培养细胞合成谷胱甘肽含量。结果显示,不同转化子重组菌的胞内谷胱甘肽产量比宿主菌分别提高了44.11%、29.79%和56.47%。

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