异戊二烯焦磷酸异构酶的表达及其催化功能验证  

Expression of isopentenyl diphosphate isomerase and confirmation of its catalytic effect

作　　者：刘敏[1,2] 刘建忠[1] 冯红茹[3] 杨建明[2] 苏思正[1,2] 曹玉锦[2] 

机构地区：[1]武汉科技大学化学工程与技术学院,湖北武汉430081 [2]中国科学院青岛生物能源与过程研究所,山东青岛266101 [3]沈阳农业大学生物科学与技术学院,辽宁沈阳110866

出　　处：《武汉科技大学学报》2013年第3期214-218,共5页Journal of Wuhan University of Science and Technology

基　　金：国家"十二五"科技计划农村领域资助项目(2012BAD32B06-2);国家863计划资助项目(SS2013AA050703-2)

摘　　要：以大肠杆菌和枯草芽孢杆菌基因组DNA为模板,采用聚合酶链式反应扩增法分别克隆来自大肠杆菌和枯草芽孢杆菌的异戊二烯焦磷酸异构酶基因,将其在大肠杆菌BL21(DE3)中异源过量表达,验证其催化功能,并考察其对β-胡萝卜素和异戊二烯合成的影响。结果表明,来自枯草芽孢杆菌的异戊二烯焦磷酸异构酶对β-胡萝卜素和异戊二烯的产量有明显的促进作用,该酶的异源表达使β-胡萝卜素的产量由1.08mg/L提高到3.25mg/L,异戊二烯的产量由0.80mg/L提高到2.96mg/L。The isopentenyl diphosphate isomerases from Escherichia coli and Bacillus subtilis were am- plified from their genomic DNA by polymerase chain reaction and then heterologously expressed in E. coli BL21 （DE3） to verify their catalytic activities. The effects of these two enzymes on β-carotene and isoprene biosynthesis were further evaluated. The results show that isopentenyl diphosphate isomer- ase from Bacillus subtilis are helpful for the production of the two compounds. In the engineered strain expressing this enzyme, β-carotene production is raised from 1.08 mg/L to 3.25 mg/L while i- soprene production is raised from 0.80 mg/L to 2.96 mg/L.

关 键 词：异戊二烯焦磷酸异构酶 大肠杆菌 异戊二烯 Β-胡萝卜素 

分 类 号：Q93-33[生物学—微生物学]