扣囊复膜孢酵母BGL1基因在工业酒精酵母中的整合表达-微生物学报.PDF

微生物学通报 MAR 20, 2008, 35(3): 321~326 Microbiology © 2008 by Institute of Microbiology, CAS tongbao@ 研究报告 扣囊复膜孢酵母BGL 1 基因在工业 酒精酵母中的整合表达 张 梁 周 衍 石贵阳* (1. 214122) (2. 214122) 摘 要: 构建了含有工业酿酒酵母自身 GPD2 启动子和终止子、扣囊复膜孢酵母β-葡萄糖苷酶基 因(BGL 1)和潮霉素选择性标记 hyg 的重组质粒 pPIC-gpd-bgl-hyg, 通过酵母染色体同源重组, 将 BGL 1 基因整合进入工业酒精酵母的染色体上。重组酵母可以在以纤维二糖为唯一碳源的培养基 上生长, 48 h 时β-葡萄糖苷酶酶活达到0.764 U/mL 。在玉米浓醪酒精发酵实验中, 与宿主菌株相比, 重组酵母醪液中纤维二糖含量减少约80 ％, 达到了消耗醪液中纤维二糖含量的目的。 关键词: 工业酒精酵母, β- 葡萄糖苷酶, 同源重组, 潮霉素B Integration and Expression of BGL 1 Gene from Saccharomy- copsis fibuligera in Industrial Saccharomyces cerevisiae * ZHANG Liang ZHOU Yan SHI Gui-Yang (1. Institute of Biomass Refinery Processing, School of Biotechnology, Jiangnan University , Wuxi 214122) (2. The Key Laboratory of Industry Biotechnology, Ministry of Education, Jiangnan University , Wuxi 214122) Abstract: The recombinant plasmid pPIC-gpd-bgl-hyg was constructed, which contained GPD2 promotor and terminator from industrial yeasts Saccharomyces cerevisiae, β-glucosidase gene (BGL 1) from Sac- charomycopsis fibuligera and hyg from hygromycin as the selected marker. With the yeast’s high efficiency of homologous integrated, the BGL 1 gene was successfully integrated into industrial yeasts S. cerevisiae. The recombined yeast could grow on the cultures with the cel