Engineering Saccharomyces cerevisiae for Efficient LiquiritigeninProduction

Production of liquiritigenin, a plant-derived significant flavonoid traditionally extracted from licorice plants, is constrained by ecological and operational inefficiencies. Despite efforts to achieve heterologous reconstruction of liquiritigenin synthesis pathway in microorganisms, the liquiritigenin titers remain low and the process is still at the proof-of-concept stage, insufficient to replace plant extraction. Herein, to achieve the efficient production of liquiritigenin, the galactose induction system in Saccharomyces cerevisiae was reengineered for better decoupling of production and growth stages, making it more suitable for heterologous pathways, and then applied to a naringenin-producing strain and modified to redirect the pathway for liquiritigenin production. To improve liquiritigenin ratio, a dual NADPH supply system was developed to enhance production capabilities. Subsequently, the concept of using endogenous metabolites to regulate the simplification and optimization of heterologous natural product biosynthetic pathways was proposed, and a general metabolic strategy model for flavonoid compounds, the aromatic ester model, was introduced. The final engineered strain achieved 867.67 mg/L liquiritigenin in the 5 L fermenter. These results demonstrated the innovative use of genetic and metabolic modifications to overcome conventional extraction limitations, providing valuable insights for synthesizing flavonoids and other natural products.