Abstract: 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.

Key words: mevalonate, MVA pathway, RNA scaffold system, E. coli, GFP