(735l) Advancing Plant Cytochrome P450 Functionality in Yeast

Plant natural products (PNPs) are important for drug discovery, yet their complex structures and low abundance pose challenges for sourcing. Cytochrome P450s (CYPs), which catalyze various oxidation in plants, are key players in PNP biosynthesis. While yeast fermentation has emerged as a promising alternative for PNP production, achieving efficient functional reconstitution of CYPs in yeast remains a major challenge. Traditional strategies for enhancing CYPs, such as protein engineering and electron transfer optimization, do not consistently yield effective results and are context dependent. Recently, the role of the microenvironment, especially membrane composition, was recognized in supporting CYP function. In our study, we identified plant membrane steroid binding proteins as a general enhancer for CYP activities in baker’s yeast. By investigating the function of these steroid binding proteins in yeast, we illuminates the scaffold protein-mediated regulation of CYP activities. Through targeted engineering of relevant microenvironment, we successfully enhanced CYP activities in yeast. This work provides new insights for improving CYP activities in yeast.