Elucidating dynamic anaerobe metabolism with HRMAS 13 C NMR and genome-scale modeling.

Autor: Pavao A; Massachusetts Host-Microbiome Center, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA., Girinathan B; Massachusetts Host-Microbiome Center, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.; Ginkgo Bioworks, The Innovation and Design Building, Boston, MA, USA., Peltier J; Laboratoire Pathogenèse des Bactéries Anaérobies, F-75015, Institut Pasteur, Université Paris-Cité, UMR-CNRS 6047, Paris, France.; Institute for Integrative Biology of the Cell (I2BC), 91198, University of Paris-Saclay, CEA, CNRS, Gif-sur-Yvette, France., Altamirano Silva P; Centre for Investigations in Tropical Diseases, Faculty of Microbiology, University of Costa Rica, San José, Costa Rica., Dupuy B; Laboratoire Pathogenèse des Bactéries Anaérobies, F-75015, Institut Pasteur, Université Paris-Cité, UMR-CNRS 6047, Paris, France., Muti IH; Departments of Radiology and Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA., Malloy C; Department of Radiology, The University of Texas Southwestern Medical Center, Dallas, TX, USA., Cheng LL; Departments of Radiology and Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. lcheng@mgh.harvard.edu., Bry L; Massachusetts Host-Microbiome Center, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. lbry@bwh.harvard.edu.; Clinical Microbiology Laboratory, Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. lbry@bwh.harvard.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2023 May; Vol. 19 (5), pp. 556-564. Date of Electronic Publication: 2023 Mar 09.
DOI: 10.1038/s41589-023-01275-9
Abstrakt: Anaerobic microbial metabolism drives critical functions within global ecosystems, host-microbiota interactions, and industrial applications, yet remains ill-defined. Here we advance a versatile approach to elaborate cellular metabolism in obligate anaerobes using the pathogen Clostridioides difficile, an amino acid and carbohydrate-fermenting Clostridia. High-resolution magic angle spinning nuclear magnetic resonance (NMR) spectroscopy of C. difficile, grown with fermentable 13 C substrates, informed dynamic flux balance analysis (dFBA) of the pathogen's genome-scale metabolism. Analyses identified dynamic recruitment of oxidative and supporting reductive pathways, with integration of high-flux amino acid and glycolytic metabolism at alanine's biosynthesis to support efficient energy generation, nitrogen handling and biomass generation. Model predictions informed an approach leveraging the sensitivity of 13 C NMR spectroscopy to simultaneously track cellular carbon and nitrogen flow from [U- 13 C]glucose and [ 15 N]leucine, confirming the formation of [ 13 C, 15 N]alanine. Findings identify metabolic strategies used by C. difficile to support its rapid colonization and expansion in gut ecosystems.
(© 2023. The Author(s).)
Databáze: MEDLINE
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