Stress-driven regulation of cellular robustness for improving terpenoid production in Ogataea polymorpha

Yeast cell factories are widely employed for terpenoid biosynthesis. The non-conventional Crabtree-negative yeast Ogataea polymorpha is emerging as a promising platform due to its thermotolerance, broad substrate utilization, and suitability for high-density fermentation. However, the limited terpenoid biosynthesis regulators in O. polymorpha presents a critical challenge, as blindly copying Saccharomyces cerevisiae strategies often proves ineffective. Here, we identified two key genes (RSC9 and NUC-2) regulating terpenoid biosynthesis and cell growth in O. polymorpha using weighted gene co-expression network analysis (WGCNA), which could up-regulate genes in the mevalonate (MVA) pathway, glycolysis, and acetyl-CoA synthesis at the transcriptional levels. Furthermore, RSC9 improved thermotolerance while NUC-2 enhanced oxidative stress resistance, collectively boosting cellular robustness for significantly increased biomass and terpenoid titers in engineered strains. Consequently, this study establishes an effective platform for industrial-scale terpenoid production.