Metabolic Engineering of <i>Yarrowia lipolytica</i> for Enhanced De Novo Biosynthesis of Icaritin

Icaritin (ICT) is a naturally occurring flavonoid compound with notable anticancer properties, recently recognized for its efficacy in treating advanced hepatic carcinoma. Traditional methods of ICT production, including plant extraction and chemical synthesis, face challenges such as low yield and environmental concerns. This study leverages synthetic biology to construct a microbial cell factory using <i>Yarrowia lipolytica</i> for de novo ICT synthesis. We engineered the yeast by integrating the ICT synthesis pathway involving EsPT from <i>Epimedium sagittatum</i> and OsOMTm from <i>Oryza sativa</i>. By optimizing the metabolic pathways, including enhancing the supply of DMAPP via mevalonate pathway modifications, and fine-tuning the expression and catalytic efficiency of EsPT through truncation strategies, we significantly improved ICT yield to 247.02 mg/L─the highest microbial ICT titer reported to date. These findings lay a solid foundation for the large-scale industrial production of ICT and offer valuable insights into the biosynthesis of other flavonoid plant natural products.