gTME改造野生酿酒酵母利用木糖产乙醇的初步研究  

Ethanol fermentation using xylose by gTME reconstructing wild Saccharomyces scerevisiae

作　　者：裴建新[1] 韦宇拓[1] 杜丽琴[1] 杨登峰[2] 曹博[1] 黄日波[1,2] 

机构地区：[1]广西大学生命科学与技术学院,南宁530004 [2]广西科学院,南宁530007

出　　处：《工业微生物》2010年第4期18-22,共5页Industrial Microbiology

基　　金：国家自然基金(No.20666002);广西科学院(No.08YJ16SW03);桂科自(NO.0991077)

摘　　要：利用全转录工程(Global transcription machinery engineering,gTME)方法对野生型酿酒酵母(Saccharomyces scerevisiae)GX7菌株进行了改造。通过PCR方法克隆获得酿酒酵母转录因子spt15基因,与表达载体pYX418连接,构建了重组子pYX418-spt15,并对SPT15蛋白的177、195和217位点进行了定点突变,然后醋酸锂转化入酿酒酵母GX7中,利用不同浓度的G418抗性筛选得到重组突变酿酒酵母菌株GX7-1,经初步研究表明:重组菌株细胞表型发生了变化,生长速度加快,并可以发酵木糖生成木糖醇和乙醇。为构建新型工业酿酒酵母奠定了理论基础。The reform of the wild Saccharomyces scerevisiae GX7 by global transcription machinery engineering （gTME） was carried out. The first step was to get transcription factor Saccharornyces scerevisiae sptl5 gene by PCR, and insert the gene with expression vector- pYX418;then to construct recombinant pYX418-spt 15, site-directed mutations of three sites, and convert into Saccharomyces scerevisiae GX7 by LiAc, select recombinant Saccharornyces scerevisiae strain GX7-1 according to different generation of G418 resistance. The results showed that the cell phenotypic of recombinant Saccharomyces scerevisiae strain changed, the speed of which accelerated, and it could use xylose to produce xylitol and ethanol. This study will provide theory base for reform Saccharomyces scerevisiae to be a new industrial strain.

关 键 词：全局转录工程 重组酿酒酵母 生物乙醇 定点突变 

分 类 号：TQ223.122[化学工程—有机化工] TQ926[轻工技术与工程—发酵工程]