Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown

Lucy I Crouch; Marcelo V Liberato; Paulina A Urbanowicz; Arnaud Baslé; Christopher A Lamb; Christopher J Stewart; Katie Cooke; Mary Doona; Stephanie Needham; Richard R Brady; Janet E Berrington; Katarina Madunic; Manfred Wuhrer; Peter Chater; Jeffery P Pearson; Robert Glowacki; Eric C Martens; Fuming Zhang; Robert J Linhardt; Daniel I R Spencer; David N Bolam

doi:10.1038/s41467-020-17847-5

Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown

Lucy I Crouch, Marcelo V Liberato, Paulina A Urbanowicz, Arnaud Baslé, Christopher A Lamb, Christopher J Stewart, Katie Cooke, Mary Doona, Stephanie Needham, Richard R Brady, Janet E Berrington, Katarina Madunic, Manfred Wuhrer, Peter Chater, Jeffery P Pearson, Robert Glowacki, Eric C Martens, Fuming Zhang, Robert J Linhardt, Daniel I R SpencerDavid N Bolam

Biosciences

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Abstract

The thick mucus layer of the gut provides a barrier to infiltration of the underlying epithelia by both the normal microbiota and enteric pathogens. Some members of the microbiota utilise mucin glycoproteins as a nutrient source, but a detailed understanding of the mechanisms used to breakdown these complex macromolecules is lacking. Here we describe the discovery and characterisation of endo-acting enzymes from prominent mucin-degrading bacteria that target the polyLacNAc structures within oligosaccharide side chains of both animal and human mucins. These O-glycanases are part of the large and diverse glycoside hydrolase 16 (GH16) family and are often lipoproteins, indicating that they are surface located and thus likely involved in the initial step in mucin breakdown. These data provide a significant advance in our knowledge of the mechanism of mucin breakdown by the normal microbiota. Furthermore, we also demonstrate the potential use of these enzymes as tools to explore changes in O-glycan structure in a number of intestinal disease states.

Original language	English
Article number	4017
Journal	Nature Communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-17847-5
Publication status	Published - 11 Aug 2020

Bibliographical note

Funding Information: We thank Carl Morland (Newcastle University, UK) for his expert technical assistance. We thank Dr. Mirjam Czjzek for her expert advice on the structural data and Prof Robert Hirt for his insightful conversations about phylogenetics. We would like to thank Diamond Light Source (Oxfordshire, UK) for beamtime (proposal mx18598) and staff of beamline I03, I04-1 and I24. We are grateful to Newcastle Biobank and NIHR Newcastle Biomedical Research Centre. Dr. Jose Muñoz-Muñoz kingly gifted the arabinogalactan substrates used. The NEC samples were collected as part of the ethically approved SERVIS study (REC 10/H0908/39). The colorectal cancer cell lines were from the Department of Surgery, Leiden University Medical Centre (LUMC), Leiden. The work was funded by a BBSRC/Innovate UK IB catalyst award ‘Glycoenzymes for Bioindustries’ (BB/M029018/1).

Publisher Copyright:
© 2020, The Author(s).

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

Animals
Bacteria/classification
Crystallography, X-Ray
Gastrointestinal Microbiome
Glycoside Hydrolases/genetics
Hexosaminidases/chemistry
Humans
Membrane Glycoproteins/chemistry
Molecular Structure
Mucins/chemistry
Phylogeny
Polysaccharides/chemistry
Structure-Activity Relationship
Substrate Specificity

ASJC Scopus subject areas

Physics and Astronomy(all)
Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/s41467-020-17847-5Licence: Creative Commons: Attribution (CC BY)

CrouchLI2020Prominent
Crouch, L.I., Liberato, M.V., Urbanowicz, P.A. et al. Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown. Nat Commun 11, 4017 (2020). https://doi.org/10.1038/s41467-020-17847-5
Final published version, 1.64 MBLicence: Creative Commons: Attribution (CC BY)

Cite this

Crouch, L. I., Liberato, M. V., Urbanowicz, P. A., Baslé, A., Lamb, C. A., Stewart, C. J., Cooke, K., Doona, M., Needham, S., Brady, R. R., Berrington, J. E., Madunic, K., Wuhrer, M., Chater, P., Pearson, J. P., Glowacki, R., Martens, E. C., Zhang, F., Linhardt, R. J., ... Bolam, D. N. (2020). Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown. Nature Communications, 11(1), Article 4017. https://doi.org/10.1038/s41467-020-17847-5

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abstract = "The thick mucus layer of the gut provides a barrier to infiltration of the underlying epithelia by both the normal microbiota and enteric pathogens. Some members of the microbiota utilise mucin glycoproteins as a nutrient source, but a detailed understanding of the mechanisms used to breakdown these complex macromolecules is lacking. Here we describe the discovery and characterisation of endo-acting enzymes from prominent mucin-degrading bacteria that target the polyLacNAc structures within oligosaccharide side chains of both animal and human mucins. These O-glycanases are part of the large and diverse glycoside hydrolase 16 (GH16) family and are often lipoproteins, indicating that they are surface located and thus likely involved in the initial step in mucin breakdown. These data provide a significant advance in our knowledge of the mechanism of mucin breakdown by the normal microbiota. Furthermore, we also demonstrate the potential use of these enzymes as tools to explore changes in O-glycan structure in a number of intestinal disease states.",

keywords = "Animals, Bacteria/classification, Crystallography, X-Ray, Gastrointestinal Microbiome, Glycoside Hydrolases/genetics, Hexosaminidases/chemistry, Humans, Membrane Glycoproteins/chemistry, Molecular Structure, Mucins/chemistry, Phylogeny, Polysaccharides/chemistry, Structure-Activity Relationship, Substrate Specificity",

author = "Crouch, {Lucy I} and Liberato, {Marcelo V} and Urbanowicz, {Paulina A} and Arnaud Basl{\'e} and Lamb, {Christopher A} and Stewart, {Christopher J} and Katie Cooke and Mary Doona and Stephanie Needham and Brady, {Richard R} and Berrington, {Janet E} and Katarina Madunic and Manfred Wuhrer and Peter Chater and Pearson, {Jeffery P} and Robert Glowacki and Martens, {Eric C} and Fuming Zhang and Linhardt, {Robert J} and Spencer, {Daniel I R} and Bolam, {David N}",

note = "Funding Information: We thank Carl Morland (Newcastle University, UK) for his expert technical assistance. We thank Dr. Mirjam Czjzek for her expert advice on the structural data and Prof Robert Hirt for his insightful conversations about phylogenetics. We would like to thank Diamond Light Source (Oxfordshire, UK) for beamtime (proposal mx18598) and staff of beamline I03, I04-1 and I24. We are grateful to Newcastle Biobank and NIHR Newcastle Biomedical Research Centre. Dr. Jose Mu{\~n}oz-Mu{\~n}oz kingly gifted the arabinogalactan substrates used. The NEC samples were collected as part of the ethically approved SERVIS study (REC 10/H0908/39). The colorectal cancer cell lines were from the Department of Surgery, Leiden University Medical Centre (LUMC), Leiden. The work was funded by a BBSRC/Innovate UK IB catalyst award {\textquoteleft}Glycoenzymes for Bioindustries{\textquoteright} (BB/M029018/1). Publisher Copyright: {\textcopyright} 2020, The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2020",

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doi = "10.1038/s41467-020-17847-5",

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Crouch, LI, Liberato, MV, Urbanowicz, PA, Baslé, A, Lamb, CA, Stewart, CJ, Cooke, K, Doona, M, Needham, S, Brady, RR, Berrington, JE, Madunic, K, Wuhrer, M, Chater, P, Pearson, JP, Glowacki, R, Martens, EC, Zhang, F, Linhardt, RJ, Spencer, DIR & Bolam, DN 2020, 'Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown', Nature Communications, vol. 11, no. 1, 4017. https://doi.org/10.1038/s41467-020-17847-5

TY - JOUR

T1 - Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown

AU - Crouch, Lucy I

AU - Liberato, Marcelo V

AU - Urbanowicz, Paulina A

AU - Baslé, Arnaud

AU - Lamb, Christopher A

AU - Stewart, Christopher J

AU - Cooke, Katie

AU - Doona, Mary

AU - Needham, Stephanie

AU - Brady, Richard R

AU - Berrington, Janet E

AU - Madunic, Katarina

AU - Wuhrer, Manfred

AU - Chater, Peter

AU - Pearson, Jeffery P

AU - Glowacki, Robert

AU - Martens, Eric C

AU - Zhang, Fuming

AU - Linhardt, Robert J

AU - Spencer, Daniel I R

AU - Bolam, David N

N1 - Funding Information: We thank Carl Morland (Newcastle University, UK) for his expert technical assistance. We thank Dr. Mirjam Czjzek for her expert advice on the structural data and Prof Robert Hirt for his insightful conversations about phylogenetics. We would like to thank Diamond Light Source (Oxfordshire, UK) for beamtime (proposal mx18598) and staff of beamline I03, I04-1 and I24. We are grateful to Newcastle Biobank and NIHR Newcastle Biomedical Research Centre. Dr. Jose Muñoz-Muñoz kingly gifted the arabinogalactan substrates used. The NEC samples were collected as part of the ethically approved SERVIS study (REC 10/H0908/39). The colorectal cancer cell lines were from the Department of Surgery, Leiden University Medical Centre (LUMC), Leiden. The work was funded by a BBSRC/Innovate UK IB catalyst award ‘Glycoenzymes for Bioindustries’ (BB/M029018/1). Publisher Copyright: © 2020, The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/8/11

Y1 - 2020/8/11

N2 - The thick mucus layer of the gut provides a barrier to infiltration of the underlying epithelia by both the normal microbiota and enteric pathogens. Some members of the microbiota utilise mucin glycoproteins as a nutrient source, but a detailed understanding of the mechanisms used to breakdown these complex macromolecules is lacking. Here we describe the discovery and characterisation of endo-acting enzymes from prominent mucin-degrading bacteria that target the polyLacNAc structures within oligosaccharide side chains of both animal and human mucins. These O-glycanases are part of the large and diverse glycoside hydrolase 16 (GH16) family and are often lipoproteins, indicating that they are surface located and thus likely involved in the initial step in mucin breakdown. These data provide a significant advance in our knowledge of the mechanism of mucin breakdown by the normal microbiota. Furthermore, we also demonstrate the potential use of these enzymes as tools to explore changes in O-glycan structure in a number of intestinal disease states.

AB - The thick mucus layer of the gut provides a barrier to infiltration of the underlying epithelia by both the normal microbiota and enteric pathogens. Some members of the microbiota utilise mucin glycoproteins as a nutrient source, but a detailed understanding of the mechanisms used to breakdown these complex macromolecules is lacking. Here we describe the discovery and characterisation of endo-acting enzymes from prominent mucin-degrading bacteria that target the polyLacNAc structures within oligosaccharide side chains of both animal and human mucins. These O-glycanases are part of the large and diverse glycoside hydrolase 16 (GH16) family and are often lipoproteins, indicating that they are surface located and thus likely involved in the initial step in mucin breakdown. These data provide a significant advance in our knowledge of the mechanism of mucin breakdown by the normal microbiota. Furthermore, we also demonstrate the potential use of these enzymes as tools to explore changes in O-glycan structure in a number of intestinal disease states.

KW - Animals

KW - Bacteria/classification

KW - Crystallography, X-Ray

KW - Gastrointestinal Microbiome

KW - Glycoside Hydrolases/genetics

KW - Hexosaminidases/chemistry

KW - Humans

KW - Membrane Glycoproteins/chemistry

KW - Molecular Structure

KW - Mucins/chemistry

KW - Phylogeny

KW - Polysaccharides/chemistry

KW - Structure-Activity Relationship

KW - Substrate Specificity

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U2 - 10.1038/s41467-020-17847-5

DO - 10.1038/s41467-020-17847-5

M3 - Article

C2 - 32782292

SN - 2041-1723

VL - 11

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 4017

ER -

Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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