MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration

Mycofactocin is a redox cofactor essential for the alcohol metabolism of mycobacteria. While the biosynthesis of mycofactocin is well established, the gene mftG, which encodes an oxidoreductase of the glucose-methanol-choline superfamily, remained functionally uncharacterized. Here, we show that Mft...

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Main Authors: Ana Patrícia Graça, Vadim Nikitushkin, Mark Ellerhorst, Cláudia Vilhena, Tilman E Klassert, Andreas Starick, Malte Siemers, Walid K Al-Jammal, Ivan Vilotijevic, Hortense Slevogt, Kai Papenfort, Gerald Lackner
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Language:English
Published: eLife Sciences Publications Ltd 2025-01-01
Series:eLife
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Online Access:https://elifesciences.org/articles/97559
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author Ana Patrícia Graça
Vadim Nikitushkin
Mark Ellerhorst
Cláudia Vilhena
Tilman E Klassert
Andreas Starick
Malte Siemers
Walid K Al-Jammal
Ivan Vilotijevic
Hortense Slevogt
Kai Papenfort
Gerald Lackner
author_facet Ana Patrícia Graça
Vadim Nikitushkin
Mark Ellerhorst
Cláudia Vilhena
Tilman E Klassert
Andreas Starick
Malte Siemers
Walid K Al-Jammal
Ivan Vilotijevic
Hortense Slevogt
Kai Papenfort
Gerald Lackner
author_sort Ana Patrícia Graça
collection DOAJ
description Mycofactocin is a redox cofactor essential for the alcohol metabolism of mycobacteria. While the biosynthesis of mycofactocin is well established, the gene mftG, which encodes an oxidoreductase of the glucose-methanol-choline superfamily, remained functionally uncharacterized. Here, we show that MftG enzymes are almost exclusively found in genomes containing mycofactocin biosynthetic genes and are present in 75% of organisms harboring these genes. Gene deletion experiments in Mycolicibacterium smegmatis demonstrated a growth defect of the ∆mftG mutant on ethanol as a carbon source, accompanied by an arrest of cell division reminiscent of mild starvation. Investigation of carbon and cofactor metabolism implied a defect in mycofactocin reoxidation. Cell-free enzyme assays and respirometry using isolated cell membranes indicated that MftG acts as a mycofactocin dehydrogenase shuttling electrons toward the respiratory chain. Transcriptomics studies also indicated remodeling of redox metabolism to compensate for a shortage of redox equivalents. In conclusion, this work closes an important knowledge gap concerning the mycofactocin system and adds a new pathway to the intricate web of redox reactions governing the metabolism of mycobacteria.
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spelling doaj-art-b80f6a38d33a49a2924d4528ebf0cea62025-01-29T12:28:20ZengeLife Sciences Publications LtdeLife2050-084X2025-01-011310.7554/eLife.97559MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respirationAna Patrícia Graça0Vadim Nikitushkin1Mark Ellerhorst2Cláudia Vilhena3Tilman E Klassert4Andreas Starick5Malte Siemers6Walid K Al-Jammal7Ivan Vilotijevic8Hortense Slevogt9Kai Papenfort10https://orcid.org/0000-0002-5560-9804Gerald Lackner11https://orcid.org/0000-0002-0307-8319Leibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Junior Research Group Synthetic Microbiology, Jena, GermanyLeibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Junior Research Group Synthetic Microbiology, Jena, Germany; University of Bayreuth, Chair of Biochemistry of Microorganisms, Kulmbach, GermanyLeibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Junior Research Group Synthetic Microbiology, Jena, Germany; University of Bayreuth, Chair of Biochemistry of Microorganisms, Kulmbach, GermanyLeibniz Institute for Natural Product Research and Infection Biology– Hans Knöll Institute, Department of Infection Biology, Jena, GermanyRespiratory Infection Dynamics, Helmholtz Centre for Infection Research - HZI Braunschweig, Braunschweig, Germany; Department of Respiratory Medicine and Infectious Diseases, Hannover Medical School, German Center for Lung Research (DZL), BREATH, Hannover, GermanyFriedrich Schiller University Jena, Institute of Microbiology, Jena, Germany; Microverse Cluster, Friedrich Schiller University Jena, Jena, GermanyFriedrich Schiller University Jena, Institute of Microbiology, Jena, Germany; Microverse Cluster, Friedrich Schiller University Jena, Jena, GermanyFriedrich Schiller University Jena, Institute of Organic Chemistry and Macromolecular Chemistry, Jena, GermanyFriedrich Schiller University Jena, Institute of Organic Chemistry and Macromolecular Chemistry, Jena, GermanyRespiratory Infection Dynamics, Helmholtz Centre for Infection Research - HZI Braunschweig, Braunschweig, Germany; Department of Respiratory Medicine and Infectious Diseases, Hannover Medical School, German Center for Lung Research (DZL), BREATH, Hannover, GermanyFriedrich Schiller University Jena, Institute of Microbiology, Jena, Germany; Microverse Cluster, Friedrich Schiller University Jena, Jena, GermanyLeibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Junior Research Group Synthetic Microbiology, Jena, Germany; University of Bayreuth, Chair of Biochemistry of Microorganisms, Kulmbach, Germany; Microverse Cluster, Friedrich Schiller University Jena, Jena, GermanyMycofactocin is a redox cofactor essential for the alcohol metabolism of mycobacteria. While the biosynthesis of mycofactocin is well established, the gene mftG, which encodes an oxidoreductase of the glucose-methanol-choline superfamily, remained functionally uncharacterized. Here, we show that MftG enzymes are almost exclusively found in genomes containing mycofactocin biosynthetic genes and are present in 75% of organisms harboring these genes. Gene deletion experiments in Mycolicibacterium smegmatis demonstrated a growth defect of the ∆mftG mutant on ethanol as a carbon source, accompanied by an arrest of cell division reminiscent of mild starvation. Investigation of carbon and cofactor metabolism implied a defect in mycofactocin reoxidation. Cell-free enzyme assays and respirometry using isolated cell membranes indicated that MftG acts as a mycofactocin dehydrogenase shuttling electrons toward the respiratory chain. Transcriptomics studies also indicated remodeling of redox metabolism to compensate for a shortage of redox equivalents. In conclusion, this work closes an important knowledge gap concerning the mycofactocin system and adds a new pathway to the intricate web of redox reactions governing the metabolism of mycobacteria.https://elifesciences.org/articles/97559Mycolicibacterium smegmatisMycofactocinmycobacteriarespirationredox-cofactoroxidoreductase
spellingShingle Ana Patrícia Graça
Vadim Nikitushkin
Mark Ellerhorst
Cláudia Vilhena
Tilman E Klassert
Andreas Starick
Malte Siemers
Walid K Al-Jammal
Ivan Vilotijevic
Hortense Slevogt
Kai Papenfort
Gerald Lackner
MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
eLife
Mycolicibacterium smegmatis
Mycofactocin
mycobacteria
respiration
redox-cofactor
oxidoreductase
title MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
title_full MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
title_fullStr MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
title_full_unstemmed MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
title_short MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
title_sort mftg is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration
topic Mycolicibacterium smegmatis
Mycofactocin
mycobacteria
respiration
redox-cofactor
oxidoreductase
url https://elifesciences.org/articles/97559
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