Summary of Study ST002433
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001566. The data can be accessed directly via it's Project DOI: 10.21228/M88M5G This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
Study ID | ST002433 |
Study Title | Elucidating dynamic anaerobe metabolism with HRMAS 13C NMR and genome-scale modeling |
Study Summary | 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 (HRMAS) Nuclear Magnetic Resonance (NMR) spectroscopy of C. difficile, grown with fermentable 13C 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 13C NMR spectroscopy to simultaneously track cellular carbon and nitrogen flow from [U-13C]glucose and [15N]leucine, confirming the formation of [13C,15N]alanine. Findings identify metabolic strategies used by C. difficile to support its rapid colonization and expansion in gut ecosystems. |
Institute | Brigham and Women's Hospital |
Department | Pathology |
Laboratory | Bry Lab, Massachusetts Host-Microbiome Center; Cheng Lab, Massachusetts General Hospital |
Last Name | Pavao |
First Name | Aidan |
Address | 221 Longwood Ave, EBRC-411, Boston, MA, 02115, USA |
apavao2@bwh.harvard.edu | |
Phone | 617-525-7184 |
Submit Date | 2023-01-05 |
Num Groups | 8 |
Total Subjects | 18 |
Raw Data Available | Yes |
Raw Data File Type(s) | fid |
Analysis Type Detail | NMR |
Release Date | 2023-01-20 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Collection:
Collection ID: | CO002515 |
Collection Summary: | For live cell time series, labeled MMM with 10% D2O was inoculated with 100,000 vegetative Clostridioides difficile ATCC43255 delPaLoc cells in a Kel-F HRMAS rotor insert under anaerobic atmosphere. The sample was spun at 3600 Hz at 37°C in a 4mm zirconia rotor and successive spectra were acquired over 36+ hours. For static culture supernatants, labeled MMM was inoculated with 100,000 vegetative Clostridioides difficile ATCC43255 delPaLoc cells in an anaerobic chamber and incubated at 37°C for 48 hours. Cultures were centrifuged and supernatants collected and lyophilized, then resuspended in D2O prior to NMR spectra acquisition. |
Sample Type: | Bacterial cells |
Volumeoramount Collected: | 30 µL |
Collection Vials: | Kel-F inserts for 4mm MAS rotor |
Collection Tube Temp: | 37°C |