Developing the E. coli platform for efficient production of UMP-derived chemicals

5-Methyluridine (5 -MU) is a prominent intermediate for industrial synthesis of several antiviral -drugs, however, its availability over the past decades has overwhelmingly relied on chemical and enzymatic strategies. Here, we have realized efficient production of 5 -MU in E. coli, for the first time, via a designer artificial pathway consisting of a two -enzyme cascade (UMP 5-methylase and phosphatase). More importantly, we have engineered the E. coli cell factory to boost 5 -MU production by systematic evaluation of multiple strategies, and as a proof of concept, we have further developed an antibiotic -free fermentation strategy to realize 5 -MU production (10.71 g/L) in E. coli MB229 (a Delta thyA strain). Remarkably, we have also established a versatile and robust platform with exploitation of the engineered E. coli for efficient production of diversified UMP-derived chemicals. This study paves the way for future engineering of E. coli as a synthetic biology platform for acceleratively accessing UMPderived chemical diversities.