Prenylflavonoids are interesting bioactive compounds. Microbial cell factories are an efficient and cheap method for their production. In this study, a pathway to produce prenylnaringenins (PNs) in Escherichia coli was constructed. To produce naringenin (689.5 mg/L), tyrosine ammonia-lyase, 4-coumarate-CoA ligase, chalcone synthase, and chalcone isomerase were expressed. Several prenyltransferase (PT) were then evaluated to produce PNs. Since dimethylallyl pyrophosphate (DMAPP) (PT extended substrate) ) is limiting, synthetic biology approaches were used to build boosted DMAPP-E. coli strains. PNs were successfully produced in these strains, marking a significant achievement in E. coli.