Engineering new-to-nature biochemical conversions by combining fermentative metabolism with respiratory modules

Helena Schulz-Mirbach, Jan Lukas Krüsemann, Theofania Andreadaki, Jana Natalie Nerlich, Eleni Mavrothalassiti, Simon Boecker, Philipp Schneider, Moritz Weresow, Omar Abdelwahab, Nicole Paczia, Beau Dronsella, Tobias J. Erb, Arren Bar-Even, Steffen Klamt and Steffen N. Lindner ()
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Helena Schulz-Mirbach: Max Planck Institute for Terrestrial Microbiology
Jan Lukas Krüsemann: Max Planck Institute for Terrestrial Microbiology
Theofania Andreadaki: Max Planck Institute of Molecular Plant Physiology
Jana Natalie Nerlich: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität
Eleni Mavrothalassiti: Max Planck Institute of Molecular Plant Physiology
Simon Boecker: Max Planck Institute for Dynamics of Complex Technical Systems
Philipp Schneider: Max Planck Institute for Dynamics of Complex Technical Systems
Moritz Weresow: Max Planck Institute of Molecular Plant Physiology
Omar Abdelwahab: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität
Nicole Paczia: Max Planck Institute for Terrestrial Microbiology
Beau Dronsella: Max Planck Institute for Terrestrial Microbiology
Tobias J. Erb: Max Planck Institute for Terrestrial Microbiology
Arren Bar-Even: Max Planck Institute of Molecular Plant Physiology
Steffen Klamt: Max Planck Institute for Dynamics of Complex Technical Systems
Steffen N. Lindner: Max Planck Institute of Molecular Plant Physiology

Nature Communications, 2024, vol. 15, issue 1, 1-15

Abstract: Abstract Anaerobic microbial fermentations provide high product yields and are a cornerstone of industrial bio-based processes. However, the need for redox balancing limits the array of fermentable substrate-product combinations. To overcome this limitation, here we design an aerobic fermentative metabolism that allows the introduction of selected respiratory modules. These can use oxygen to re-balance otherwise unbalanced fermentations, hence achieving controlled respiro-fermentative growth. Following this design, we engineer and characterize an obligate fermentative Escherichia coli strain that aerobically ferments glucose to stoichiometric amounts of lactate. We then re-integrate the quinone-dependent glycerol 3-phosphate dehydrogenase and demonstrate glycerol fermentation to lactate while selectively transferring the surplus of electrons to the respiratory chain. To showcase the potential of this fermentation mode, we direct fermentative flux from glycerol towards isobutanol production. In summary, our design permits using oxygen to selectively re-balance fermentations. This concept is an advance freeing highly efficient microbial fermentation from the limitations imposed by traditional redox balancing.

Date: 2024
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DOI: 10.1038/s41467-024-51029-x

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