Single cell fluorescence imaging of glycan uptake by intestinal bacteria

Jan Hendrik Hehemann (Lead / Corresponding author), Greta Reintjes, Leeann Klassen, Adam D. Smith, Didier Ndeh, Carol Arnosti, Rudolf Amann, D. Wade Abbott (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)
8 Downloads (Pure)

Abstract

Microbes in the intestines of mammals degrade dietary glycans for energy and growth. The pathways required for polysaccharide utilization are functionally diverse; moreover, they are unequally dispersed between bacterial genomes. Hence, assigning metabolic phenotypes to genotypes remains a challenge in microbiome research. Here we demonstrate that glycan uptake in gut bacteria can be visualized with fluorescent glycan conjugates (FGCs) using epifluorescence microscopy. Yeast α-mannan and rhamnogalacturonan-II, two structurally distinct glycans from the cell walls of yeast and plants, respectively, were fluorescently labeled and fed to Bacteroides thetaiotaomicron VPI-5482. Wild-type cells rapidly consumed the FGCs and became fluorescent; whereas, strains that had deleted pathways for glycan degradation and transport were non-fluorescent. Uptake of FGCs, therefore, is direct evidence of genetic function and provides a direct method to assess specific glycan metabolism in intestinal bacteria at the single cell level.

Original languageEnglish
Pages (from-to)1883-1889
Number of pages7
JournalISME Journal
Volume13
Issue number7
Early online date1 Apr 2019
DOIs
Publication statusPublished - Jul 2019

Keywords

  • Biological techniques
  • Metabolomics

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