RNA支架系统介导大肠杆菌生产甲羟戊酸

Increasing mevalonate production mediated by RNA scaffolds system in Escherichia coli

利用0D RNA支架系统可共聚集酶的优势提高大肠杆菌甲羟戊酸的产量。通过RNA-EMSA证明RNA适配体与RBDs(RNA binding domains)存在相互作用;将RBDs与荧光蛋白融合表达以评估RNA支架系统效率。相比对照菌株BLCCG,表达0D RNA支架的菌株BLPCG及表达RBDs的菌株BLCPG荧光强度分别提高126%和129%。表达0D PP7 RNA支架系统的菌株BLPPG提高375%;应用0D RNA支架系统生产甲羟戊酸。相比对照菌株BLCCES,表达0D PP7 RNA支架系统菌株BLPPES的甲羟戊酸产量提高84%,达3.13 g/L。因此,RNA支架系统是提高多酶代谢途径效率的有效工具。

In this study,the advantage of 0D RNA scaffold system was used to co-localize the three-step reaction of mevalonate to improve the yield of mevalonate in E.coli.Firstly,RNA-EMSA was used to demonstrate the interaction between RNA aptamers and RBDs(RNA binding domains).Subsequently,RBDs were fused with fluorescent protein to evaluate the efficiency of RNA scaffold system.Compared with that of the control strain BLCGG,the relative fluorescence intensities of BLPCG and BLCPG were increased by 126%and 129%,respectively.The fluorescence intensity of BLPPG was increased by 375%in the strain expressed 0D PP7 RNA scaffold system.Finally,mevalonate was produced by 0D RNA scaffold system.Compared with that of the control strain BLCCES,the mevalonate production of BLPPES expressing 0D PP7 RNA scaffold system was increased by 84%,achieved 3.13 g/L.Therefore,RNA scaffold system is an effective tool to improve the efficiency of multi-enzyme metabolic pathway.