Synergistic acetyl-CoA augmentation strategy (SATS) for improved terpenoid biosynthesis in Saccharomyces cerevisiae

Terpenoids are widely applied in pharmaceuticals, fragrances and biofuels. Acetyl-CoA, as a crucial precursor in the mevalonate pathway, is stringently regulated intracellularly, thereby limiting the biosynthesis of terpenoids. This study develops a synergistic acetyl-CoA augmentation strategy (SATS) to establish a robust platform for terpenoid biosynthesis in Saccharomyces cerevisiae. Optimizing coenzyme A biosynthesis via global regulation of the pantothenic acid module improved intracellular acetyl-CoA levels by 3.26-fold. Enhancing acetyl phosphate supply was achieved by constructing a PPP-PK-PTA pathway, which improved intracellular acetyl-CoA by 1.92-fold. By combining these techniques, acetyl-CoA levels in the SATS-engineered strain increased by 6.03-fold. The titer of amorpha-4,11-diene, a representative terpenoid, rose by 37.77-fold to 188.45 mg/L in shake-flask fermentation and reached 13 g/L in a 5 L bioreactor. Furthermore, the modified strain exhibited enhanced production of other terpenoids. Our research indicates SATS is an effective approach for synthesizing terpenoids and other acetyl-CoA-derived compounds, demonstrating broad applicability.