基于生物量产率的低产乙醇酿酒酵母的筛选  

Screening of Low-ethanol Saccharomyces cerevisiae Based on the Biomass Yield

作　　者：杜青 刘延琳[1,2] 贾鹤 鲁素蕊 刘喜喜 孙晶 管玉清 秦义[1,2] Du Qing;Liu Yanlin;Jia He;Lu Surui;Liu Xixi;Sun Jing;Guan Yuqing;Qin Yi(College of Enology, Northwest A＆F University, Yangling 712100, Shaanxi;Heyang Viticulture Experiment Station of Northwest A＆F University, Weinan 715300, Shaanxi)

机构地区：[1]西北农林科技大学葡萄酒学院,陕西杨凌712100 [2]西北农林科技大学合阳葡萄试验示范站,陕西渭南715300

出　　处：《中国食品学报》2018年第4期30-36,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家重点研发计划项目(2016YFD0400504-01);"十二五"国家科技支撑计划项目(2012BAD31B07);国家自然科学基金项目(31301541);中央高校基本科研业务费项目(含第二批)(QN2013021)

摘　　要：目的:建立一种从酿酒酵母突变库中快速筛选低产乙醇酵母菌株的方法。方法:以转录因子spt15随机突变酿酒酵母文库为研究模型,以高生物量产率作为低产乙醇酿酒酵母的筛选标记,利用24孔板对spt15突变酵母文库进行初筛。对初筛获得的目标菌株,以乙醇产量作为筛选标记进行复筛,最终获得乙醇合成能力较低且生物量较高的酿酒酵母菌株。结果:(1)利用易错PCR技术构建了spt15随机突变文库,并将其转化至酿酒酵母YS59,获得酿酒酵母YS59 spt15突变文库;(2)经初筛,获得14株生物量产率≥70%的酿酒酵母YS59 spt15突变菌株;(3)通过复筛,筛选出编号712的低产乙醇突变菌株,其乙醇合成量降低了28.1%;(4)通过序列比对,发现spt15-712上有9个碱基发生突变,分别是A161G,T426C,T462C,T493A,A506G,T546C,T622C,T658C,A750T9。结论:成功建立了一种高通量筛选低产乙醇酿酒酵母的方法 ,并利用这种方法筛选出乙醇合成量降低28.1%的低产乙醇突变菌株。Objective： Establish a rapid method of screening low-ethanol yeast strain from S. cerevisiae mutation library. Method： The S. cerevisiae random mutation library of transcription factor spt15 was used as a research model, and the high biomass yield was used as screening target of the low-ethanol S.cerevisiae strains for the primary screening. The24-hole plate was used for screening high biomass yield strains from the spt15 mutant yeast library. Then rescreening low-ethanol strains, used low ethanol production as the target, from the strains obtained from primary screening. Result：（1） We successfully constructed a S. cerevisiae YS59 spt15 mutant library. The spt15 random mutation library was established by error-prone PCR technology firstly, and then it was transformed into S. cerevisiae YS59;（2） Compared with the control strain, we obtained 14 S. cerevisiae YS59 spt15 mutant strains with the biomass yields which were more than 70% through primary screening;（3） By rescreening, a low-ethanol mutant strain NO. 712 with the amount of ethanol producyion decreased by 28.1% was got;（4） Comparing with the spt15（Gene Accession No.： NC_001137）,there were 9 bases mutated in spt15-712, which were A161 G, T426 C, T462 C, T493 A, A506 G, T546 C, T622 C,T658 C, A750 T, respectively. Conclusion： We established a high-throughput method of screening low-ethanol S. cerevisiae successfully, and the low-ethanol mutant strain with ethanol production was decreased by 28.1% was selected by this method.

关 键 词：酿酒酵母 生物量产率 低产乙醇 spt15 突变 

分 类 号：TS261.11[轻工技术与工程—发酵工程]