CAJ | 학술논문

以工业酒精酵母菌株(Saccharomyces cerevisiae Y)为对象,通过生孢子法获得其a型和α型单倍体.根据同源重组原理构建整合载体pUC19-GPD1::Q Km 线性化后电击转化进入a型单倍体.G418抗性筛选缺失GPD1基因的转化子S.cerevisiae Y(a,△gpd1).重组菌S.cerevisiae Y(a.△gpd1)与α型单倍体杂交获得二倍体,采用PCR扩增法鉴定得到GPD1全突变菌株S.cerevisiae Y(△gpd1,△gpd1).分别以20 g/L和150 g/L的初糖在摇瓶中进行发酵实验考察.结果表明,S.cerevisiae Y(a,△gpd1)的葡萄糖转甘油率分别从12.43%和6.04%降低到9.35%和5.54%,葡萄糖转酒精率分别从36.4.9%和39.96%提高到36.91%和40.29%;而S.cerevisiae Y(△god1,△gpd1)的葡萄糖转甘油率分别从14.16%和7.49%降低到10.46%和5.79%,葡萄糖转酒精率分别从36.23%和39.89%提高到38.52%和41.50%.
이공업주정효모균주(Saccharomyces cerevisiae Y)위대상,통과생포자법획득기a형화α형단배체.근거동원중조원리구건정합재체pUC19-GPD1::Q Km 선성화후전격전화진입a형단배체.G418항성사선결실GPD1기인적전화자S.cerevisiae Y(a,△gpd1).중조균S.cerevisiae Y(a.△gpd1)여α형단배체잡교획득이배체,채용PCR확증법감정득도GPD1전돌변균주S.cerevisiae Y(△gpd1,△gpd1).분별이20 g/L화150 g/L적초당재요병중진행발효실험고찰.결과표명,S.cerevisiae Y(a,△gpd1)적포도당전감유솔분별종12.43%화6.04%강저도9.35%화5.54%,포도당전주정솔분별종36.4.9%화39.96%제고도36.91%화40.29%;이S.cerevisiae Y(△god1,△gpd1)적포도당전감유솔분별종14.16%화7.49%강저도10.46%화5.79%,포도당전주정솔분별종36.23%화39.89%제고도38.52%화41.50%.