Growth of E. coli on formate and methanol via the reductive glycine pathway

Engineering a biotechnological microorganism for growth on one-carbon (C₁) intermediates, produced from the abiotic activation of CO₂, is a key synthetic biology step towards the valorization of this greenhouse gas to commodity chemicals. Here, we redesign the central carbon metabolism of the model bacterium Escherichia coli for growth on C₁ compounds using the reductive glycine pathway. Sequential genomic introduction of the four metabolic modules of the synthetic pathway resulted in a strain capable of growth on formate and CO₂ with a doubling time of ~70 hours and growth yield of ~1.5 gCDW / mol-formate. Short-term evolution decreased doubling time to less than 8 hours and improved biomass yield to 2.3 gCDW / mol-formate. Growth on methanol and CO₂ was achieved by the expression of methanol dehydrogenase in the evolved strain. Establishing synthetic formatotrophy and methylotrophy, as demonstrated here, paves the way for sustainable bioproduction rooted in CO₂ and renewable energy.