From toxin to biofuel: engineering microbes for methanol biomanufacturing.

Methanol, a promising one-carbon (C1) feedstock for biofuels, faces challenges in bioconversion due to its cellular toxicity. This review summarizes recent advances in methanol-based biosynthesis of biofuels, such as short-chain alcohols, fatty acid derivatives, and terpenoids, in both native and synthetic methylotrophs. We also discuss the mechanisms of methanol cytotoxicity and systematically examine engineering strategies to enhance methanol utilization and tolerance, including metabolic pathway rewiring, compartmentalization, and adaptive evolution. Finally, we highlight that integrating systems biology and synthetic biology can pave the way toward sustainable methanol-based biomanufacturing.