Methanol-dependent Escherichia coli strains with a complete ribulose monophosphate cycle

Philipp Keller, Elad Noor, Fabian Meyer, Michael A. Reiter, Stanislav Anastassov, Patrick Kiefer and Julia A. Vorholt ()
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Philipp Keller: Institute of Microbiology, Department of Biology, ETH Zurich
Elad Noor: Institute of Molecular Systems Biology, Department of Biology, ETH Zurich
Fabian Meyer: Institute of Microbiology, Department of Biology, ETH Zurich
Michael A. Reiter: Institute of Microbiology, Department of Biology, ETH Zurich
Stanislav Anastassov: Institute of Microbiology, Department of Biology, ETH Zurich
Patrick Kiefer: Institute of Microbiology, Department of Biology, ETH Zurich
Julia A. Vorholt: Institute of Microbiology, Department of Biology, ETH Zurich

Nature Communications, 2020, vol. 11, issue 1, 1-10

Abstract: Abstract Methanol is a biotechnologically promising substitute for food and feed substrates since it can be produced renewably from electricity, water and CO2. Although progress has been made towards establishing Escherichia coli as a platform organism for methanol conversion via the energy efficient ribulose monophosphate (RuMP) cycle, engineering strains that rely solely on methanol as a carbon source remains challenging. Here, we apply flux balance analysis to comprehensively identify methanol-dependent strains with high potential for adaptive laboratory evolution. We further investigate two out of 1200 candidate strains, one with a deletion of fructose-1,6-bisphosphatase (fbp) and another with triosephosphate isomerase (tpiA) deleted. In contrast to previous reported methanol-dependent strains, both feature a complete RuMP cycle and incorporate methanol to a high degree, with up to 31 and 99% fractional incorporation into RuMP cycle metabolites. These strains represent ideal starting points for evolution towards a fully methylotrophic lifestyle.

Date: 2020
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DOI: 10.1038/s41467-020-19235-5

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