METABOLIC ENGINEERING OF YARROWIA LIPOLYTICA AS A MICROBIAL CELL FACTORY FOR GLYCYRRHETINIC ACID BIOSYNTHESIS

The rising demand for plant-derived bioactive compounds has prompted the exploration of sustainable production alternatives. Glycyrrhetinic acid (GA), a pharmacologically valuable triterpenoid derived from licorice roots, faces production challenges due to low natural yields and complex extraction processes. This study focuses on engineering Yarrowia lipolytica, an oleaginous yeast with high acetyl-CoA flux, as a microbial cell factory for GA biosynthesis. A heterologous pathway incorporating four key genes—GgbAS1, CYP88D6, GuCPR1, and CYP72A63—was designed, codon-optimized, and assembled under strong constitutive promoters for expression in Y. lipolytica. The final construct (pCV316) was successfully integrated into the yeast genome using auxotrophic selection and validated by PCR and sequencing. GC-MS analysis confirmed the production of glycyrrhetinic acid in engineered strains. This work demonstrates the feasibility of microbial GA production using Y. lipolytica and lays the groundwork for future pathway optimization and scale-up, providing a promising alternative to traditional plant extraction methods.