α-淀粉酶和糖化酶的表达及酿酒酵母工程菌的构建  

Expression of α-amylase and Glucoamylase Genes in S cerevisiae and Construction of Starch Degrading Yeast Strain

作　　者：吴晓萍[1] 李文清[1] 罗进贤[1] 后茂立 林资诚 

机构地区：[1]中山大学生命科学学院 [2]广州市微生物研究所

出　　处：《中山大学学报（自然科学版）》1999年第2期80-84,共5页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：广东省自然科学基金资助项目(940628);广州市重点项目基金资助项目

摘　　要：将切除了５′端非编码区５０碱基对片段的黑曲霉糖化酶ＧＡＩｃＤＮＡ与大麦α淀粉酶基因重组进大肠杆菌酵母穿梭载体，构建重组表达质粒ｐＭＡＧ１１，转化酿酒酵母ＧＲＦ１８，获得含α淀粉酶和糖化酶双基因的酵母工程菌ＧＲＦ１８（ｐＭＡＧ１１）．在酵母ＰＧＫ基因启动子和终止信号的调控下，α淀粉酶和糖化酶基因获得高效表达，９９％的表达产物分泌至胞外．在含ｗ＝１５％的可溶性淀粉的ＹＰＳ培养基中培养４４ｈ，淀粉水解率达到９９％。Noncoding region deleted glucoamylase GAI cDNA of A niger were cloned into an E coli yeast shuttle vector resulting in recombinant plasmid pMAG11 which was then transformed into Saccharomyes cerevisiae GRF18 The α amylase and glucoamylase genes under the control of PGK promoter and terminator were efficiently expressed in engineered strain GRF18(pMAG11) and 99% of the expressed product secreted into the medium The engineered strain hydrolysed 99% of the starch in YPS medium containing 15% soluble starch within 44 hours The starch hydrolysate was used for fermentation and ethanol production

关 键 词：Α-淀粉酶 糖化酶 基因表达 酿酒酵母工程菌 分解淀粉 

分 类 号：Q556[生物学—生物化学] TS261.11[轻工技术与工程—发酵工程]