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Using one carbon compounds as feedstock is a promising approach in abating climate change. Here, the authors report the conversion of E. coli into a synthetic methylotroph that assimilates methanol via the ribulose monophosphate cycle and a set of distinctive mutations.

The engineering of methanol-dependent growth in Escherichia coli is challenging. Here, the authors predict and experimentally validate methanol-dependent strains with a complete RuMP cycle and high potential for the development of a methylotrophic platform organism.

Engineering synthetic methylotrophy remains challenging. Here, the authors engineer a methanol-essential E. coli by an in silico-guided multiple knockout approach and show a laboratory evolved strain can incorporate up to 24% methanol into core metabolites during growth.