Cytidine 5'-monophosphate (5'-CMP) is a key intermediate in various nucleotide derivatives and is widely used in the food and pharmaceutical industries. In this study, the titer of 5'-CMP in engineered Escherichia coli CM006, which blocks the degradation pathway of 5'-CMP, increased 134.7-fold compared to E. coli MG1655. Integrated expression of the 5'-CMP diphosphate hydrolase gene nudG significantly enhanced the 5'-CMP titer, reduced orotic acid accumulation, and alleviated feedback inhibition in the 5'-CMP synthesis pathway. By blocking cytidine degradation and increasing carbon flux from cytidine to 5'-CMP, the 5'-CMP titer in E. coli CM011 was increased to 338.2 mg/L. Furthermore, by enhancing the supply of the precursor 5-phospho-α-D-ribose 1-diphosphate (PRPP), the 5'-CMP titer in E. coli CM012 reached 417.9 mg/L, the highest value reported to date. The results and methods presented in this study are of great significance for the sustainable industrial production of 5'-CMP.
Keywords: 5′-CMP; Escherichia coli; green production; systematic metabolic engineering.