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Methanol-essential growth of Escherichia coli

Author

Listed:
  • Fabian Meyer

    (ETH Zurich)

  • Philipp Keller

    (ETH Zurich)

  • Johannes Hartl

    (ETH Zurich)

  • Olivier G. Gröninger

    (ETH Zurich)

  • Patrick Kiefer

    (ETH Zurich)

  • Julia A. Vorholt

    (ETH Zurich)

Abstract

Methanol represents an attractive substrate for biotechnological applications. Utilization of reduced one-carbon compounds for growth is currently limited to methylotrophic organisms, and engineering synthetic methylotrophy remains a major challenge. Here we apply an in silico-guided multiple knockout approach to engineer a methanol-essential Escherichia coli strain, which contains the ribulose monophosphate cycle for methanol assimilation. Methanol conversion to biomass was stoichiometrically coupled to the metabolization of gluconate and the designed strain was subjected to laboratory evolution experiments. Evolved strains incorporate up to 24% methanol into core metabolites under a co-consumption regime and utilize methanol at rates comparable to natural methylotrophs. Genome sequencing reveals mutations in genes coding for glutathione-dependent formaldehyde oxidation (frmA), NAD(H) homeostasis/biosynthesis (nadR), phosphopentomutase (deoB), and gluconate metabolism (gntR). This study demonstrates a successful metabolic re-routing linked to a heterologous pathway to achieve methanol-dependent growth and represents a crucial step in generating a fully synthetic methylotrophic organism.

Suggested Citation

  • Fabian Meyer & Philipp Keller & Johannes Hartl & Olivier G. Gröninger & Patrick Kiefer & Julia A. Vorholt, 2018. "Methanol-essential growth of Escherichia coli," Nature Communications, Nature, vol. 9(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-03937-y
    DOI: 10.1038/s41467-018-03937-y
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    Cited by:

    1. Liang-Yu Nieh & Frederic Y.-H. Chen & Hsin-Wei Jung & Kuan-Yu Su & Chao-Yin Tsuei & Chun-Ting Lin & Yue-Qi Lee & James C. Liao, 2024. "Evolutionary engineering of methylotrophic E. coli enables fast growth on methanol," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    2. Cláudio J. R. Frazão & Nils Wagner & Kenny Rabe & Thomas Walther, 2023. "Construction of a synthetic metabolic pathway for biosynthesis of 2,4-dihydroxybutyric acid from ethylene glycol," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    3. Philipp Keller & Michael A. Reiter & Patrick Kiefer & Thomas Gassler & Lucas Hemmerle & Philipp Christen & Elad Noor & Julia A. Vorholt, 2022. "Generation of an Escherichia coli strain growing on methanol via the ribulose monophosphate cycle," Nature Communications, Nature, vol. 13(1), pages 1-13, December.

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