Monoterpenoid production in yeast coupled with cell-free monooxygenase catalysation

Monoterpenoids are versatile biomolecules with applications in health care, agriculture, and consumer products. However, applications of monoterpenoids remain rare, as traditional production methods are too expansive at an industrial scale. Microbial production of monoterpenoids is a promising alternative, but poor precursor availability and complicated expression of cytochrome P450 monooxygenases (CYPs) and reductases (CPRs) limit their potential. We are overcoming current limitations by increasing precursor availability in S. cerevisiae and Y. lipolytica by extensive metabolic engineering, compartmentalization, and creating a cell-free, electricity-driven CYP system. As a proof-of-concept, we will produce the monoterpenoid verbenone, which has the proven capability to save forests from bark beetle invasion. Additionally, our project will establish an adaptable platform for monoterpenoid production, allowing further deployment of monoterpenoids in unexplored areas.