谷胱甘肽生产菌重组巴斯德毕赤氏酵母的构建

The construction of recombinant pichia pastoris for production of glutathione

目的:构建一株新型的产谷胱甘肽的重组巴斯德毕赤氏酵母,并研究该重组菌合成谷胱甘肽的能力;方法:利用PCR技术克隆来源于酿酒酵母的γ-L-谷氨酰-L-半胱氨酸合成酶编码基因GSH1和谷胱甘肽合成酶的基因GSH2,串联插入表达载体pGAPZA,转化Pichia pastoris GS115,在Zeo-cin平板上挑选阳性转化子,并对阳性转化重组菌进行摇瓶培养筛选。结果:得到一株GSH的产量高达162.2mg/L重组菌,说明串联表达酿酒酵母GSH1和GSH2基因,重组甲醇酵母生产谷胱甘肽可以达到较高的水平。

Objective：Construction of one novel recombinant Pichia pastoris for production of glutathione, and to study its glutathione synthesis capacity. Method： The gene encoding γ-glutamyl-cysteine synthetase （GSH1） and synthetase （GSH2） were cloned to vector pGAPZA and then transformed into Pichia pastoris GS115 strain. Recombinants resistant to zeocin were selected and identified. The GSH production was tested through fermentation in 250 mL flasks. Conclusion：The GSHI and GSH2 gene can be co-expressed. The GSH yield of the recombinant Pichia pastoris was 162.2 mg/L.