Development of an efficient yeast platform for cannabigerolic acid biosynthesis.

Cannabinoids are important therapeutical molecules for human ailments, cancer treatment, and SARS-CoV-2. The central cannabinoid, cannabigerolic acid (CBGA), is generated from geranyl pyrophosphate and olivetolic acid by Cannabis sativa prenyltransferase (CsPT4). Despite efforts to engineer microorganisms such as Saccharomyces cerevisiae (S. cerevisiae) for CBGA production, their titers remain suboptimal because of the low conversion of hexanoate into olivetolic acid and the limited activity and stability of the CsPT4. To address the low hexanoate conversion, we eliminated hexanoate consumption by the beta-oxidation pathway and reduced its incorporation into fatty acids. To address CsPT4 limitations, we expanded the endoplasmic reticulum and fused an auxiliary protein to CsPT4. Consequently, the engineered S. cerevisiae chassis showed a marked improvement of 78.64-fold in CBGA production, reaching a titer of 510.32 ± 10.70 mg l−1 from glucose and hexanoate. • Commercial production of cannabigerolic acid (CBGA) in S. cerevisiae presents challenges. • Blocking hexanoate degradation and optimizing pathway enzyme expression resulted in improved CBGA titers. • Fatty acid synthase confirmed cryptical utilization of hexanoyl-CoA. • Compartmentalization expansion and auxiliary partner fusion elevated activity and stability of prenyltransferase CsPT4. • Engineered CBGA-producing yeast yielded in 510 mg l−1 CBGA from cost-effective carbon source glucose and hexanoate. [ABSTRACT FROM AUTHOR]