Single cell fluorescence imaging of glycan uptake by intestinal bacteria

TY - JOUR

T1 - Single cell fluorescence imaging of glycan uptake by intestinal bacteria

AU - Hehemann, Jan Hendrik

AU - Reintjes, Greta

AU - Klassen, Leeann

AU - Smith, Adam D.

AU - Ndeh, Didier

AU - Arnosti, Carol

AU - Amann, Rudolf

AU - Abbott, D. Wade

N1 - Funding Information: Acknowledgements This work was supported by funding from the Beef and Cattle Research Council awarded to DWA (Grant: FDE.13.15). J-HH, GR, and RA were supported by the Max Planck Society. J-HH acknowledges support by the Deutsche For-schungsgemeinschaft (DFG) grant HE 7217/1-1. GR acknowledges support by a Marie Skłodowska-Curie IF-Grant (RUMIC No. 840804). CA was supported by the U.S. National Science Foundation (OCE-1332881 and OCE-1736772). We wish to thank Dr. Malcolm O’Neill (Complex Carbohydrate Research Centre, University of Georgia) for providing purified wine RGII and Dr. Elizabeth Lowe (Newcastle University) for the BtΔMAN1/2/3 mutant. Publisher Copyright: © 2019, The Author(s). Copyright: Copyright 2019 Elsevier B.V., All rights reserved.

PY - 2019/7

Y1 - 2019/7

N2 - 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.

AB - 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.

KW - Biological techniques

KW - Metabolomics

UR - https://www.scopus.com/pages/publications/85063755010

U2 - 10.1038/s41396-019-0406-z

DO - 10.1038/s41396-019-0406-z

M3 - Article

C2 - 30936421

AN - SCOPUS:85063755010

SN - 1751-7362

VL - 13

SP - 1883

EP - 1889

JO - ISME Journal

JF - ISME Journal

IS - 7

ER -