植物甜蛋白monellin基因工程菌表达条件的研究  

Research of Expression Condition of Monellin in the Recombinant E.coli Strain

作　　者：蔡恒[1] 陈忠军[2] 万红贵[1] 

机构地区：[1]南京工业大学制药与生命科学学院,南京210009 [2]内蒙古农业大学食品科学与工程学院,呼和浩特010018

出　　处：《食品与发酵工业》2006年第6期23-26,共4页Food and Fermentation Industries

摘　　要：将带有monellin基因的重组分泌型表达载体pETMO转入大肠杆菌BL21(DE3),得到重组工程菌株BMC33。经IPTG诱导,其所含有的甜蛋白基因可高效表达。研究不同的表达条件对甜蛋白表达水平的影响,得到优化发酵条件:装液量为50 mL/250 mL三角瓶,当培养液OD值达0.8时,添加诱导剂IPTG至终浓为0.9mmol/L,32℃诱导5 h。此时,甜蛋白monellin表达量可高达细菌可溶性蛋白的44.8%。A expression vector pETMO harboring the single-chain monellin gene was transferred into E. coli BL21（DE3）. When inducing by IPTG, the sweet protein monellin could be produced at a high level in the recombinant E.coli. In this study, the optimal condition of expression was obtained. The recombinant E.coli was grown at 37℃ with shaking at 200 r/min. When the culture OD600 nm reached 0.8, IPTG was added to give a final concentration of 0.9 mmol/L. The culture was then incubated at 32 ℃ for 5 h and the monellin content reach 44.8 % of total soluble proteins. The E. coli -expressed monellin was 3 000 times sweeter than sucrose.

关 键 词：大肠杆菌 甜蛋白MONELLIN 重组菌株 表达条件 

分 类 号：Q943.2[生物学—植物学]