摘要
利用诱变剂甲基磺酸乙酯(EMS)对酿酒酵母(Saccharomyces cerevisiae)W303-1A进行诱变,确定了诱变酿酒酵母最佳的EMS质量浓度为40μg/m L,筛选出1株遗传稳定、可耐3.0%(V/V)异丁醇的优势突变株EMS39。同时在菌株EMS39和W303-1A中过表达L-缬氨酸代谢途径中的关键基因ILV2、ILV5、ILV3和ARO10得到菌株EMS39 23510和W303-1A 23510,将对应的空载体转入菌株W303-1A中得到菌株W303-1A-EP作为对照菌,同时进行发酵性能检测。结果表明,菌株EMS39 23510异丁醇产量达到404.2 mg/L,比对照菌W303-1A-EP提高了529.6%,比菌株W303-1A 23510提高了3.9%,产率比对照菌W303-1A-EP提高了530.0%。其异丁醇及其他发酵产物的产量明显提高,有一定的研究和工业应用价值。
Saccharomyces cerevisiae W303-1A was mutated by chemical mutagen ethyl methane sulfonate(EMS).The optimum EMS mass concentration was determined as 40μg/ml,under the condition,a dominant mutant strain EMS39 with genetic stability and 3.0%(V/V)isobutanol tolerance was obtained.At the same time,strain EMS39 23510 and W303-1A 23510 were obtained by overexpressing key genes including ILV2,ILV5,ILV3 and ARO10 in the L-valine metabolic pathway.The control strain W303-1A-EP was obtained by transfering the corresponding empty vectors into strain W303-1A.Its fermentation performances were simultaneously detected.The results showed that the isobutanol production of strain EMS39 23510 reached 404.2 mg/L,which increased by 529.6%than that of control strain W303-1A-EP and increased by 3.9%than that of strain W303-1A 23510,and the productivity increased by 530.0%than that of strain W303-1A-EP.The production of isobutanol and other metabolic products of strain EMS39 23510 were improved significantly,which had certain research and industrial application value.
作者
温智慧
李敬知
冯瑞琪
苏意德
张爱利
WEN Zhihui;FENG Ruiqi;SU Yide;ZHANG Aili(School of Chemical Engineering and Technology,Hebei University of Technology,Tianjin 300130,China)
出处
《中国酿造》
CAS
北大核心
2018年第10期66-71,共6页
China Brewing
基金
国家自然科学基金资助项目(21206028)
河北省自然科学基金资助项目(B2013202288)
高等学校博士学科点专项科研基金资助项目(20121317120014)
关键词
酿酒酵母
甲基磺酸乙酯
异丁醇
耐受性
Saccharomyces cerevisiae
ethyl methane sulphonate
isobutanol
tolerance
