MetabolicEngineering and Strain Mating of Yarrowia lipolytica for Sustainable Production ofPrenylated Aromatic Compounds

Prenylated aromatic compounds possess great pharmaceutical and nutraceutical significance. However, their low abundance in nature hampers their exploitation. Here, we harnessed Yarrowia lipolytica for the production of drupanin, artepillin C, and 8-prenylnaringenin. Specially, we first modularly engineered supply of prenyl donor dimethylallyl diphosphate and aromatic acceptors in parallel in separate haploid strains through engineering prenyltransferases, strengthening the donor or acceptor biosynthesis pathway, tuning the competing pathway, and removing feedback regulation. Then, the mating strategy was developed to mate two haploid strains into diploid strains for <i>de novo</i> biosynthesis of drupanin and artepillin C. The production of drupanin and artepillin C was 52.26 and 7.45 mg/L, respectively, the total production reaching the highest reported titer for <i>de novo</i> synthesis in microbial cell factories. Using a similar approach, we also achieved the biosynthesis of 8-prenylnaringenin in Y. lipolytica for the first time. Our work provides a valuable precedent for the sustainable production of prenylated aromatic natural products, and the mating strategy can extend to synthesis of other similar products.