Methane (CH 4 ) is recognized not only as a greenhouse gas but also as a promising feedstock for carbon-based chemicals. Biological methane conversion has gained attention for operating under mild conditions. Efforts have focused on enhancing methane-oxidizing enzymes, including methane monooxygenases and cytochrome P450, for both in vivo and in vitro applications. Methanol dehydrogenase is traditionally used for methanol conversion, whereas recombinant alcohol oxidase has emerged as an attractive alternative due to cofactor-independent oxidation with molecular oxygen, reduced cofactor demand, and favorable thermodynamics. Formaldehyde, produced by methanol oxidation, serves as a versatile C1 intermediate, enabling condensation reactions to form higher-value compounds. This review summarizes recent enzymatic advances, multi-step methanol upgrading pathways, and design principles, and discusses current challenges and future directions for sustainable methane valorization. • Enzyme systems enable selective methane-to-chemical conversion under mild conditions. • Engineering MMOs and P450s expands biological methane activation capacity. • Alcohol oxidases bypass cofactor limits and improve methanol oxidation cascades. • Formaldehyde is repurposed from toxic intermediate to a versatile C1 building block. • In vitro enzyme cascades provide modular routes for efficient C1 upgrading.
Jeong et al. (Thu,) studied this question.