Dynamic regulation and enhancement of synthetic network for efficient biosynthesis of monoterpenoid α-pinene in yeast cell factory

Pinene is a plant volatile monoterpenoid which is used in the fragrance, pesticide, and biofuel industries. Although α-pinene has been synthesized in microbial cell factories, the low synthesis efficiency has thus far limited its production. In this study, the cell growth and α-pinene production of the engineered yeast were decoupled by a dynamic regulation strategy, resulting in a 101.1-fold increase in α-pinene production compared to the control. By enhancing the mevalonate pathway and expanding the cytosolic acetyl-CoA pool, α-pinene production was further increased. Overexpression of the transporter Sge1 resulted in a redistribution of global gene transcription, leading to an increased flux of α-pinene synthesis. By optimizing the aeration flow rate in 3-L bioreactors, the α-pinene production reached 1.8 g/L, which is the highest reported α-pinene production in cell factories. Our research provides insights and fundamentals for the efficient synthesis of monoterpenoids in microbial cell factories.