Cellular and Molecular Life Sciences

, Volume 76, Issue 21, pp 4319–4340 | Cite as

Surface glycan-binding proteins are essential for cereal beta-glucan utilization by the human gut symbiont Bacteroides ovatus

  • Kazune Tamura
  • Matthew H. Foley
  • Bernd R. Gardill
  • Guillaume Dejean
  • Matthew Schnizlein
  • Constance M. E. Bahr
  • A. Louise Creagh
  • Filip van Petegem
  • Nicole M. KoropatkinEmail author
  • Harry BrumerEmail author
Original Article


The human gut microbiota, which underpins nutrition and systemic health, is compositionally sensitive to the availability of complex carbohydrates in the diet. The Bacteroidetes comprise a dominant phylum in the human gut microbiota whose members thrive on dietary and endogenous glycans by employing a diversity of highly specific, multi-gene polysaccharide utilization loci (PUL), which encode a variety of carbohydrases, transporters, and sensor/regulators. PULs invariably also encode surface glycan-binding proteins (SGBPs) that play a central role in saccharide capture at the outer membrane. Here, we present combined biophysical, structural, and in vivo characterization of the two SGBPs encoded by the Bacteroides ovatus mixed-linkage β-glucan utilization locus (MLGUL), thereby elucidating their key roles in the metabolism of this ubiquitous dietary cereal polysaccharide. In particular, molecular insight gained through several crystallographic complexes of SGBP-A and SGBP-B with oligosaccharides reveals that unique shape complementarity of binding platforms underpins specificity for the kinked MLG backbone vis-à-vis linear β-glucans. Reverse-genetic analysis revealed that both the presence and binding ability of the SusD homolog BoSGBPMLG-A are essential for growth on MLG, whereas the divergent, multi-domain BoSGBPMLG-B is dispensable but may assist in oligosaccharide scavenging from the environment. The synthesis of these data illuminates the critical role SGBPs play in concert with other MLGUL components, reveals new structure–function relationships among SGBPs, and provides fundamental knowledge to inform future (meta)genomic, biochemical, and microbiological analyses of the human gut microbiota.


Microbiota Microbiome Dietary fiber Bacteroidetes Beta-glucan Cereal 



We thank Associate Professor Russ Algar (Dept. Chemistry, University of British Columbia) for use of a fluorescence microplate reader. We thank Associate Professor Eric Martens and his laboratory for the use of a microplate reader in the anaerobic chamber and qPCR thermal cycler. We thank Prof. Charles Haynes (Michael Smith Laboratories, UBC) for access to ITC equipment and invaluable technical advice. We thank the Canadian Macromolecular Crystallography Facility for access to beamline 08B1-1 at the Canadian Light Source, which is supported by the Canada Foundation for Innovation, Natural Sciences and Engineering Research Council of Canada, the University of Saskatchewan, the Government of Saskatchewan, Western Economic Diversification Canada, the National Research Council Canada, and the Canadian Institutes of Health Research. We thank the Life Sciences Collaborative Access Team for access to beamline 21-ID-F and 21-ID-G at the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory (Argonne, IL, USA) under Contract No. DE-AC02-06CH11357. We thank the Stanford Synchrotron Radiation Lightsource at the SLAC National Accelerator Laboratory (Menlo Park, CA, USA) for access to beamline 9-2, the use of which is supported by the U.S. DOE, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences (including P41GM103393). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH.

Author contributions

KT cloned, expressed and purified recombinant SGBPs, GFP-fusions, and site-directed mutants, conducted AGE and pull-down depletion isotherm analyses, produced and purified MLG partial digest oligosaccharides, solved crystal structures of BoSGBPMLG-A, BoSGBPMLG-A_MLG7 and BoSGBPMLG-B_MLG7 with BRG, and co-wrote the article. MHF conducted reverse genetics and growth phenotype analyses, and co-wrote the article. BRG solved crystal structures of BoSGBPMLG-A, BoSGBPMLG-A_MLG7 and BoSGBPMLG-B_MLG7 with KT. GD conducted ITC experiments. MS expressed, purified and crystallized SeMet BoSGBPMLG-B_cellohexaose. CMEB crystallized BoSGBPMLG-A_cellohexaose and native BoSGBPMLG-B_cellohexaose. ALC assisted with ITC data collection and analysis. FVP, NMK, and HB designed and directed research, and co-wrote the article with input from all authors.


Work in the Brumer group was supported by operating grants from the Canadian Institutes of Health Research (MOP-137134 and MOP-142472) and infrastructure support from the Canadian Foundation for Innovation (Project #30663) and the British Columbia Knowledge Development Fund. Work in the Koropatkin group was funded by the National Institutes of Health (NIH R01 GM118475). Work in the van Petegem group was supported by the Canadian Institutes of Health Research (MOP-119404). K.T. was partially supported by a four-year doctoral fellowship from the University of British Columbia. M.H.F. was partially supported by a predoctoral fellowship from the Cellular Biotechnology Training Program (T32GM008353).

Supplementary material

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Supplementary material 1 (PDF 2054 kb)


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Michael Smith LaboratoriesUniversity of British ColumbiaVancouverCanada
  2. 2.Department of Biochemistry and Molecular BiologyUniversity of British ColumbiaVancouverCanada
  3. 3.Department of Microbiology and ImmunologyUniversity of Michigan Medical SchoolAnn ArborUSA
  4. 4.Department of Chemical and Biological EngineeringUniversity of British ColumbiaVancouverCanada
  5. 5.Department of ChemistryUniversity of British ColumbiaVancouverCanada
  6. 6.Department of BotanyUniversity of British ColumbiaVancouverCanada

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