CYS3基因的敲除及其对Saccharomy cescerevisiae3-甲硫基丙醇合成代谢的影响  

Effects of Knockout of Gene CYS3 on Methionol Anabolism in Saccharomyces cerevisiae

作　　者：刘丽[1] 张婵[1,2] 梁婧如 王成涛[1,2] 赵吉兴 

机构地区：[1]北京工商大学北京市食品添加剂工程技术研究中心,北京100048 [2]北京市食品风味化学重点实验室,北京100048 [3]山东中惠食品有限公司,山东惠民251706

出　　处：《食品科学》2014年第5期139-143,共5页Food Science

基　　金：国家自然科学基金面上项目(31071593);教育部科学技术研究重点项目(211101);北京市教委科技面上项目(KM201110011001)

摘　　要：研究胱硫醚γ-裂解酶基因CYS3敲除对S.cerevisiae 3-甲硫基丙醇合成代谢的影响。将编码胱硫醚-γ-裂解酶的CYS3基因和抗性标记基因Zeocin克隆,构建敲除组件CYS3Δ:Zeocin,醋酸锂法将其转化导入S.cerevisiae S288C表达,构建CYS3基因敲除的工程菌。结果表明:摇瓶发酵120 h时,工程菌S.cerevisiae C3和S.cerevisiae S288C的3-甲硫基丙醇生成量分别为0.60 g/L和0.94 g/L,S.cerevisiae C3较野生型S288C的3-甲硫基丙醇生成量降低36.2%。说明CYS3基因敲除对S.cerevisiae的3-甲硫基丙醇有较大影响,并呈现负调节作用。This work studied effects of knockout of gene CYS3 on methionol anabolism in Saccharomyces cerevisiae （S. cerevisiae）. A fragment of gene CYS3 and the resistance marker gene Zeocin were cloned and a construct of knockout of gene CYS3 CYS3 A ：Zeocin was built. Using chemical lithium acetate method, the component （CYS3△：Zeocin） was introduced into S. cerevisiae S288C and an engineering bacterium with the knockout of gene CYS3 was obtained. The results showed that methionol yields ofS. cerevisiae C3 and S. cerevisiae S288C were 0.60 g/L and 0.94 g/L in shake flask fermentation at 120 h, respectively. Compared to that ofS. cerevisiae S288C, the methionol yield ofS. cerevisiae C3 was decreased by 36.2%. These results suggested that knockout of gene CYS3 had a great influence on methionol anabolism, which appeared to play a negatively regulatory role.

关 键 词：酿酒酵母 胱硫醚-Γ-裂解酶 基因敲除 3-甲硫基丙醇合成 负调节 

分 类 号：Q815[生物学—生物工程]