产白藜芦醇大肠杆菌基因工程菌的构建  

Construction of genetically engineering Escherichia coli for resveratrol production

作　　者：梁景龙[1] 郭丽琼[1] 孙萍[1] 林俊芳[1] 何杨[1] 

机构地区：[1]华南农业大学食品学院生物工程系,广东广州510640

出　　处：《食品工业科技》2013年第23期138-142,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(31272217;31071837)

摘　　要：利用降落重叠延伸PCR和连接酶连接的方法,构建了组成型表达载体pMD18-VvRs-At4cl,该表达载体带有大肠杆菌甘油醛三磷酸脱氢酶启动子(pGAP),拟南芥(Arabidopsis thalianna)4-香豆酸辅酶A连接酶(At4CL)基因和葡萄(Vitis vinifera)白藜芦醇合酶(VvRS)基因。表达载体转化大肠杆菌BW27784,并通过PCR鉴定和酶切鉴定后,获得重组菌株。对重组菌株进行发酵培养,对发酵液进行HPLC检测,发酵液中白藜芦醇含量为4.51mg/L。这表明,拟南芥的4cl基因与金手指葡萄的rs基因在重组大肠杆菌中成功得到了表达,并且表达产物利用对香豆酸为前体物质合成了白藜芦醇。To construction of expression plasmid pMD18-Vvrs-Atcl by touchdown overlap extension PCR and ligase connection.The constitutive vector containing a gap promoter（ pGAP） of Escherichia coil and 4-coumaroyl： CoA ligase gene of Arabidopsis thalianna（At4CL） and resveratrol synthase gene of Vitis vinifera, （ VvRS）, which transformed into host strain E coil BW2T784.Next,with fermentation and HPLC detection,created an recombination Ecofi strain caple of bioproducing resveratrol by additionally supplemented with p-coumaric acid and resulted in a final titer of 4.51mg/L resveratrol.

关 键 词：白藜芦醇 大肠杆菌 原核表达 pGAP 组成型表达载体 

分 类 号：TS201.3[轻工技术与工程—食品科学]