Elucidating dynamic anaerobic metabolism in obligate anaerobes with living cells
Leo Cheng1, Aidan Pavao2, Brintha Girinathan2, Johann Peltier3, Pamela Altamirano Silva4, Bruno Dupuy5, Isabella Muti1, Craig Malloy6, and Lynn Bry2
1Radiology and Pathology, Massachusetts General HospitalMassachusetts General Hospital, Harvard Medical School, Charlestown, MA, United States, 2Massachusetts Host-Microbiome Center, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, United States, 3Institut Pasteur, Université de Paris, Paris, France, 4Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, San Jose, Costa Rica, 5Laboratory of the Pathogenesis of Bacterial Anaerobes, Institut Pasteur, Université de Paris, Paris, France, 6Radiology, The University of Texas Southwestern Medical Center, Dallas, TX, United States
Anaerobic microbial metabolism drives critical functions within global ecosystems, host-microbiota interactions, and industrial applications, yet remains ill-defined. We resolved dynamic metabolism in living cells of the anaerobic pathogen Clostridioides difficile using High-Resolution Magic Angle Spinning (HRMAS) Nuclear Magnetic Resonance (NMR) spectroscopy to inform genome-scale predictions of cellular metabolism. Analyses leveraged the sensitivity of 13C NMR spectroscopy to simultaneously track cellular carbon and nitrogen flow from fermentable 13C and 15N-labeled substrates. We illustrate a versatile approach to elaborate complex anaerobic metabolism for clinical, scientific, and industrial applications.
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