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

Keller, Philipp; Noor, Elad; Meyer, Fabian; Reiter, Michael A; Anastassov, Stanislav; Kiefer, Patrick; Vorholt, Julia A · Nat Commun · 2020

basic_science · Level V

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Abstract

Methanol is a biotechnologically promising substitute for food and feed substrates since it can be produced renewably from electricity, water and CO<sub>2</sub>. 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.

Medical subject headings