大肠杆菌丁醇生产菌的构建及检测  

Construction of Butanol Generating E.coli and Its Identification

作　　者：李晓丹[1] 姬艳丽[1] 吴日[1] 屠培培[1] 段会坤 尼再中 王海松[1] 李淼[1] 姜苹哲 李明刚[1] 

机构地区：[1]南开大学教育部活性材料重点实验室,生命科学学院,天津300071

出　　处：《吉林大学学报（理学版）》2016年第2期391-395,共5页Journal of Jilin University:Science Edition

基　　金：天津市科技支撑项目(批准号:14ZCZDSY00013)

摘　　要：用合成生物学和代谢工程技术,筛选并克隆丁醇合成途径中酶活性较高的atoB,hbd,crt,ter,adhE2等5个关键酶基因,先通过重组PCR构建pUC18-ato B-hbd-crt-ter质粒,再将串联基因atoB-hbd-crt-ter和基因adhE2克隆至质粒pET-21b中,构建含丁醇合成途径的表达载体p ET-21b-ato B-hbd-crt-ter-adh E2,将其转入大肠杆菌E.coli BL21(DE3)中制备工程菌株.对样品进行IPTG诱导表达检测与色谱分析的结果表明,已合成出关键酶蛋白并产生一定量的丁醇.Using synthetic biology and metabolic engineering techniques,we screened and cloned 5 key enzyme gene ato B,hbd,crt,adhE2 and ter with relatively high activity in butanol synthesis pathway. Firstly,we constructed the pUC18-ato B-hbd-crt-ter plasmid by recombinant PCR,and then coloned tandem gene ato Bhbd-crt-ter and gene adhE2 into the plasmid pET-21 b and eventually constructed the expression vector of pET-21b-ato B-hbd-crt-ter-adhE2 containing butanol synthesis pathway,and transferred it into E.coli BL21（ DE3）for the preparation of engineering strains. We performed IPTG induced expression detection and gas chromatograph（ GC） analysis of samples. The results show that key enzyme proteins have been synthesized and produced a certain amount of butanol.

关 键 词：大肠杆菌 生物燃料 丁醇 发酵 

分 类 号：Q819[生物学—生物工程]