Metabolic Engineering of Synechocystis sp. PCC 6803 for Olivetolic acid Production

As a pivotal aromatic precursor for phytocannabinoids, olivetolic acid (OLA) is traditionally sourced from resource-intensive plant extraction or complex chemical synthesis. In this study, we established a sustainable photosynthetic production platform by engineering the model cyanobacterium Synechocystis sp. PCC 6803. We successfully reconstituted the OLA biosynthetic pathway by heterologously expressing Cannabis sativa-derived tetraketide synthase (CsTKS), olivetolic acid cyclase (CsOAC), and acyl-activating enzyme (CsAAE1). To overcome metabolic bottlenecks, the pathway was optimized using the strong promoter P _cpc560 and carbon flux redirection. Disruption of the glycogen storage pathway and reinforcement of the malonyl-CoA pool via maeB overexpression significantly enhanced precursor availability. Furthermore, increasing CO ₂ supplementation achieved a maximum OLA titer of 9.41 mg/L. This work demonstrates the feasibility of utilizing photosynthetic chassis for the production of olivetolic acid, providing a robust foundation for the sustainable manufacturing of cannabinoid-related metabolites.