产甘油重组大肠杆菌的构建及其耐高渗透压性能的测定

Engineering Escherichia coli for production of glycerol and assay for its tolerance to high osmosis

克隆了酿酒酵母(Saccharomyces cerevisiae)中的3-磷酸甘油脱氢酶和3-磷酸甘油酯酶基因,通过融合PCR构建了双基因共表达载体,将酵母细胞内应答渗透压变化的甘油合成途径引入大肠杆菌(Escherichia coli)。以葡萄糖为底物对重组大肠杆菌进行摇瓶发酵培养,该重组菌的甘油产量为1g/L。渗透压胁迫测试证明该重组菌的耐渗透压性能较出发菌株有明显提高。

A pathway for glycerol synthesis has been introduced into Escherichia coli BL-21 by constructing a co-expression vector harbouring genes encoding glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase that govern the glycerol production in Saccharomyces cerevisiae.As a result,a glycerol concentration of 1g/L was achieved in flask culture and the recombinant species could regulate intracellular osmotic pressure and resist environmental stress in the synthesis of glycerol in vivo.