摘要
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Saccharomyces 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 (hor2 ) ,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 p GEM- Cgpd1- Chor2 ,后者转入E. coli JM10 9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。
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.
出处
《化学通报》
CAS
CSCD
北大核心
2004年第12期925-929,共5页
Chemistry
基金
国家"8 6 3"计划资助项目 (2 0 0 3AA0 0 110 39)
关键词
甘油
杂合
葡萄糖
构建
研究结果
表达
重组质粒
重组菌株
基因工程菌
磷酸
Glycerol, Glycerol 3-phosphate dehydrogenase, Glycerol 3-phosphatase, Two expression cassettes, Recombinant organism
