Profiling human gut bacterial metabolism and its kinetic using [U-(13)C]glucose and NMR

A.A. de Graaf, A. Maathuis, P. de Waard, N.E.P. Deutz, C. Dijkema, W.M. de Vos, K. Venema

Research output: Contribution to journalArticleAcademicpeer-review

55 Citations (Scopus)

Abstract

This study introduces a stable-isotope metabolic approach employing [U-(13)C]glucose that, as a novelty, allows selective profiling of the human intestinal microbial metabolic products of carbohydrate food components, as well as the measurement of the kinetics of their formation pathways, in a single experiment. A well-established, validated in vitro model of human intestinal fermentation was inoculated with standardized gastrointestinal microbiota from volunteers. After culture stabilization, [U-(13)C]glucose was added as an isotopically labeled metabolic precursor. System lumen and dialysate samples were taken at regular intervals. Metabolite concentrations and isotopic labeling were determined by NMR, GC, and enzymatic methods. The main microbial metabolites were lactate, acetate, butyrate, formate, ethanol, and glycerol. They together accounted for a (13)C recovery rate as high as 91.2%. Using an NMR chemical shift prediction approach, several minor products that showed (13)C incorporation were identified as organic acids, amino acids, and various alcohols. Using computer modeling of the (12)C contents and (13)C labeling kinetics, the metabolic fluxes in the gut microbial pathways for synthesis of lactate, formate, acetate, and butyrate were determined separately for glucose and unlabeled background substrates. This novel approach enables the study of the modulation of human intestinal function by single nutrients, providing a new rational basis for achieving control of the short-chain fatty acids profile by manipulating substrate and microbiota composition in a purposeful manner.
Original languageEnglish
Pages (from-to)2-12
JournalNMR in Biomedicine
Volume23
Issue number1
DOIs
Publication statusPublished - 2010

Keywords

  • chain fatty-acids
  • amino-acids
  • fermentation products
  • large-intestine
  • colon-cancer
  • human feces
  • butyrate
  • ethanol
  • spectrometry
  • acetaldehyde

Fingerprint Dive into the research topics of 'Profiling human gut bacterial metabolism and its kinetic using [U-(13)C]glucose and NMR'. Together they form a unique fingerprint.

Cite this