A formate-based yeast chassis for efficient formate and CO2 assimilation

Here, we reprogrammed Yarrowia lipolytica into a synthetic formatotrophic yeast capable of stable growth on formate alone. Adaptive laboratory evolution yielded a strain with a biomass yield of 5.22 +/- 0.12 g CDW mol-1 formate, exceeding all previously reported formate-assimilating systems. Multi-omics and allelic reconstruction revealed a coupled adaptive program that redistributes ATP and reducing power, reinforces protein homeostasis via enhanced 26S proteasome assembly, and mitigates oxidative and carbonyl stress through coordinated membrane lipid remodeling and activation of the cellular integrity system.