标题:
代谢工程大肠杆菌过量生产番茄红素Overproduction of Lycopene by Metabolic Engineering Escherichia coli
作者:
翁志明, 王玥, 刘建忠
关键字:
番茄红素, 大肠杆菌, 代谢工程, 启动子置换, 组成型质粒Lycopene; Escherichia coli; Metabolic Engineering; Promoter Replacement; Constitutive Plasmid
期刊名称:
《Bioprocess》, Vol.2 No.2, 2012-06-25
摘要:
番茄红素是一种高效抗氧化剂和潜在抗癌药物。gdhA，aceE和fdhF基因敲除促进了重组大肠杆菌番茄红素的合成，其中gdhA和aceE双基因敲除和gdhA，aceE和fdhF三基因敲除具有相似的作用。在gdhA和aceE双基因敲除的基础上，dxs基因天然启动子被T5启动子置换后，重组大肠杆菌番茄红素的产量提高了103%。为了避免采用诱导剂进行基因表达，构建了一系列组成型质粒，最终构建的代谢工程大肠杆菌BW25113(rgdhAraceE, PT5-dxs，pAC316-WZM4R)在不需诱导条件下摇床发酵可产番茄红素15.6 mg/g DCW。
Lycopene is an effective antioxidant and a potential pharmaceutical drug with anticancer. The knockout of gdhA, aceE or fdhF was beneficial to lycopene production in engineered E. coli. The double (gdhA and aceE) gene knockouts showed a similar effect to the triple (gdhA, aceE and fdhF) on lycopene production. Replacement of native promoter of dxs gene in the double gene knockout strain resulted in 103% increase in lycopene content. In order to avoid application of expensive inducer, we also constructed some constitutive plasmids containing heterologous carotenoid genes. The final engineered E. coli BW25113 (rgdhAraceE, PT5-dxs, pAC316-WZM4R) produced lycopene of 15.6 mg/g DCW without inducer in a batch shake flask.