Generation of an Escherichia coli strain growing on methanol via the ribulose monophosphate cycle

Philipp Keller, Michael A. Reiter, Patrick Kiefer, Thomas Gassler, Lucas Hemmerle, Philipp Christen, Elad Noor and Julia A. Vorholt ()
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Philipp Keller: ETH Zurich
Michael A. Reiter: ETH Zurich
Patrick Kiefer: ETH Zurich
Thomas Gassler: ETH Zurich
Lucas Hemmerle: ETH Zurich
Philipp Christen: ETH Zurich
Elad Noor: Weizmann Institute of Science
Julia A. Vorholt: ETH Zurich

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: Abstract Methanol is a liquid with high energy storage capacity that holds promise as an alternative substrate to replace sugars in the biotechnology industry. It can be produced from CO2 or methane and its use does not compete with food and animal feed production. However, there are currently only limited biotechnological options for the valorization of methanol, which hinders its widespread adoption. Here, we report the conversion of the industrial platform organism Escherichia coli into a synthetic methylotroph that assimilates methanol via the energy efficient ribulose monophosphate cycle. Methylotrophy is achieved after evolution of a methanol-dependent E. coli strain over 250 generations in continuous chemostat culture. We demonstrate growth on methanol and biomass formation exclusively from the one-carbon source by 13C isotopic tracer analysis. In line with computational modeling, the methylotrophic E. coli strain optimizes methanol oxidation by upregulation of an improved methanol dehydrogenase, increasing ribulose monophosphate cycle activity, channeling carbon flux through the Entner-Doudoroff pathway and downregulating tricarboxylic acid cycle enzymes. En route towards sustainable bioproduction processes, our work lays the foundation for the efficient utilization of methanol as the dominant carbon and energy resource.

Date: 2022
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DOI: 10.1038/s41467-022-32744-9

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