摘要
以工业酿酒酵母菌株(Saccharomyces cerevisiaeY)为研究对象,针对其复杂的生理生化遗传特性,建立了相对应的转化体系。以pRS41H质粒为基础载体,构建了含有工业酿酒酵母自身的gpd2启动子、终止子和扣囊复膜孢酵母的β-葡萄糖苷酶基因bgl的重组质粒pRS-gb。电击转化进入工业酿酒酵母细胞,潮霉素抗性筛选,获得重组菌。该重组菌可以在以纤维二糖为唯一碳源的培养基中生长,培养36h,β-葡萄糖苷酶酶活达到0.967μ/ml。以纤维二糖为唯一碳源的酒精发酵中,酒精度可以达到0.92g/l。这对工业生产中利用纤维素为原料发酵生产酒精具有重要意义。
The industrial Saccharomyces cerevisiae Y was used as the host strain. According to complex physiological and biochemical genetic characteristics of industrial S. cerevisiae Y, a corresponding transforming system was built. Then a recombinant plasmid （pRS-gb） was constructed and successfully transformed into S. cerevisiae Y. The pRS-gb was based on shuttling expression vector pRS41H and contained the bgll gene from Saccharomycopsis fibuliger , gpd2 promotor and terminator genes from industrial S. cerevisiae Y. The recombined yeast （Saccharomyces cerevisiae BGL-19） could grow on the medium with cellobiose as sole carbon source and the highest enzyme activity （0. 967μ/ml） was achieved at 36h. The ethanol concentration reached 0.92g/l when cellobiose was used as sole carbon source, which was higher than using S. cerevisiae Y. It was important for ethanol production using cellulose in industrial application.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2007年第2期64-69,共6页
China Biotechnology
基金
国家"十五"科技攻关项目(2001BA501A01)
关键词
工业酿酒酵母
潮霉素B
电击转化
Β-葡萄糖苷酶
Industrial Saccharomyces cerevisiae Y Hygromycin B Electroporation β-glucosidase
