米曲霉(Aspergillus oryzae)黄酒小曲4转化体系的构建及脂肪酶的异源表达  

Construction of transformation system of Aspergillus oryzae H4 and heterologous expression of lipase

作　　者：李童 李燕萍[1,2,3] LI Tong;LI Yanping(State Key Laboratory of Food Science and Technology,Nanchang University Nanchang 330006,China;Sino-German Joint Research Institute,Nanchang University Nanchang 330006,China;School of Food Science and Technology,Nanchang University Nanchang 330006,China)

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330006 [2]南昌大学中德联合研究院,江西南昌330006 [3]南昌大学食品学院,江西南昌330006

出　　处：《南昌大学学报（理科版）》2019年第3期246-250,256,共6页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(31260389)

摘　　要：以米曲霉(Aspergillus oryzae)H4为研究对象,初步探究了其对潮霉素的敏感度及原生质体的制备时间。在0.1mmol·L^-1氯丙嗪和150μg·mL^-1潮霉素B共同存在的条件下,H4的生长被完全抑制;在酶解时间为3.5h时,H4的原生质体有最佳的再生率33%。成功构建了重组表达载体pUC18-hygB-LIP并转化至米曲霉H4中。在液态发酵条件下,脂肪酶活力最高的转化子的酶活力比出发菌株H4高10.16U·mL^-1。The sensitivity of Aspergillus oryzae H4 to hygromycin and the preparation conditions of protoplasts were preliminarily explored.Under the Czapek-Dox plate contain with 0.1mmol·L^-1 chlorpromazine and 150μg·mL^-1 hygromycin B,H4 was completely inhibited from growing;at the enzymatic hydrolysis time of 3.5 h,the protoplast of H4 had the best regeneration rate of 33%.The recombinant expression vector pUC18-hygB-LIP was successfully constructed and transformed into Aspergillus oryzae H4.Under liquid fermentation conditions,the enzyme activity of the transformant with the highest lipase activity was 10.16 U· mL^-1 higher than that of H4.

关 键 词：米曲霉 异源表达 液态发酵 脂肪酶 

分 类 号：Q815[生物学—生物工程]