产甘油基因工程菌的构建

利用途径工程的基本原理，在大肠杆菌中构建一条产甘油的新代谢途径。从酿酒酵母Sacchdromyces cerevisiae INVSc1菌株总DNA克隆3-磷酸甘油脱氢酶基因(gpdl)和3-磷酸甘油酯酶基因(hot2)，构建由两个杂合启动子trc启动基因的双表达盒的重组质粒pGEM-Cgpd1-Chor2，后者转入E．coli JM109菌株，构建的重组菌株就具有一条直接将葡萄糖转化为甘油的新代谢途径，将该重组菌株以葡萄糖为底物进行摇瓶发酵，甘油产率为1．18g／L。该研究结果为进一步构建生产1，3-丙二醇工程菌打下了基础。

Construction of Recombinant Organism for Producing Glycerol

Based on the principle of the metabolic engineering, a novel pathway of producing glycerol was built in E.coli. The gene gpd1 encoding glycerol 3-phosphate dehydrogenase and the gene hor2 encoding glycerol 3-phosphatase were cloned from the total DNA of Saccharomyces cerevisiae INVSc1 strain, respectively. A recombinant plasmid pGEM-Cgpd1-Chor2 with two expression cassettes was constructed. After its transformation into E.coli JM109, a new metabolic pathway was constructed in the strain, who could convert glucose to glycerol directly. And the recombinant organism was fermented to product glycerol from D-glucose in shake-flask for 24h. Glycerol production could reach 1.18g/L. The result is useful for further working involving construction of recombinant organism for producing 1,3-propanediol.