生产β-胡萝卜素大肠杆菌工程菌的构建  

Construction of Engineering Escherichia Coli Accumulating β-Carotene

作　　者：刘敏[1] 李荣贵[1] 杜桂彩[1] 范海[2] 

机构地区：[1]青岛大学天然色素省重点实验室,山东青岛266071 [2]山东师范大学逆境植物研究所,山东济南250014

出　　处：《药物生物技术》2007年第1期1-4,共4页Pharmaceutical Biotechnology

基　　金：山东省中青年科学家奖励基金(2002);青岛市自然科学基金(051JC91)资助项目

摘　　要：将噬夏孢欧文氏菌β-胡萝卜素合成相关基因crtE,crtB,crtI融合后克隆进表达载体pET-15b,构建表达载体pET-15bcrtEIB;将crtY首先克隆进pET-15b,构建pET-15bcrtY,再将crtY连同T7启动子和终止子切下,插入pACYC-184构建pACYC-184crtY。将以上两重组质粒共转化E.coli BL21(DE3)构建工程菌,IPTG诱导工程菌累积橙色色素,经吸收光谱分析,确定工程菌中合成的色素为β-胡萝素。β-胡萝卜素的最高含量可达3.5mg/g.DW,每升发酵液可得6.53g干菌体。To construct engineering E. coli. with accumulating β-carotene, three genes crtE, crtB, crtI related to β-carotene synthesis in Erwinia uredovora were cloned into pET-15b simultaneously to construct pET-15bcrtEIB, and crtY was firstly cloned into pET-15b to construct pET-15bcrtY, then crtY under the control of T7 promoter as well as T7 terminator were cut down from pET-15bcrtY and cloned into pACYC-184 to construct pACYC-184crtY. These two expressing vectors were introduced into E. coli BL21 （DE3） to construct engineering E. coli. The engineering bacteria could accumulate orange-colored pigment when induced by IPTG. This pigment was β-carotene judged by absorbance spectra analysis. The engineering bacteria could grow to 6. 53 g · DW/L in LB medium and accumulate β-carotene to 3. 5 mg/g · DW under culturing conditions.

关 键 词：Β胡萝卜素 工程菌 累积 

分 类 号：Q784[生物学—分子生物学]