乙醇脱氢酶I基因敲除的酿酒酵母重组菌构建的初步研究  

Preliminary Study on Deletion of Saccharomyces cerevisiae Alcohol Dehydrogeniase I Gene

作　　者：葛菁萍[1] 宋刚[1] 孙宗祥[1] 凌宏志[1] 蔡柏岩[1] 刘松梅[1] 平文祥[1] 

机构地区：[1]黑龙江大学微生物黑龙江省高校重点实验室,黑龙江哈尔滨150080

出　　处：《食品科学》2008年第2期210-212,共3页Food Science

基　　金：黑龙江省科学技术厅青年基金项目(QCO4C33);黑龙江省教育厅一般项目(10551233);黑龙江大学青年基金项目(QL200435)

摘　　要：本实验根据酿酒酵母乙醇代谢途径,构建一株低乙醇产量的酿酒酵母基因工程菌株,以满足人们对低醇啤酒的需要。利用抗性基因筛选基因敲除突变体的方法,通过引物L1和L2扩增潮霉素B基因(两翼与酿酒酵母同源),按常规醋酸锂法转化酵母细胞后,筛选标记与酵母adhI基因发生同源重组,得到一株ADHI酶活性降低的工程菌株。发酵实验结果表明,转化菌株乙醇含量平均值为1.8%(V/V),较原始菌株低了65%。说明转化菌株体内乙醇生成途径受到干扰。The main purpose of this research is to construct a low alcohol producing strain according to the alcohol metabolic pathway of Saccharomyces cerevisiae, so as to satisfy the people who prefer to drink low-alcohol beer. Hygromycin B resistant gene was used to screen mutants with adh Ⅰ gene knocked out. After Hygromycin B resistant gene was amplified with primers L1 and L2 （the flanking fragments were complement with Saccharomyces cerevisiae gene）, it was transformed into yeast HDY- 01 by LiAc method and the alcohol dehydrogenase Ⅰ （ADH Ⅰ） in Saccharomyces cerevisiae was deleted through homologous recombination. A transformant was obtained with low ADH Ⅰ activity. The fermentation tests showed that the average alcohol content of the transformant is 1.8%（V/V）, 65% lower than the origin one. The alcohol metabolic pathway in this transformant is interfered.

关 键 词：酿酒酵母 基因敲除 乙醇脱氢酶Ⅰ 

分 类 号：TS262.5[轻工技术与工程—发酵工程]