title: Role of glycoside phosphorylases in mannose foraging by human gut bacteria.
abstract: To metabolize both dietary fiber constituent carbohydrates and host glycans lining the intestinal epithelium, gut bacteria produce a wide range of carbohydrate-active enzymes, of which glycoside hydrolases are the main components. In this study, we describe the ability of phosphorylases to participate in the breakdown of human N-glycans, from an analysis of the substrate specificity of UhgbMP, a mannoside phosphorylase of the GH130 protein family discovered by functional metagenomics. UhgbMP is found to phosphorolyze β-D-Manp-1,4-β-D-GlcpNAc-1,4-D-GlcpNAc and is also a highly efficient enzyme to catalyze the synthesis of this precious N-glycan core oligosaccharide by reverse phosphorolysis. Analysis of sequence conservation within family GH130, mapped on a three-dimensional model of UhgbMP and supported by site-directed mutagenesis results, revealed two GH130 subfamilies and allowed the identification of key residues responsible for catalysis and substrate specificity. The analysis of the genomic context of 65 known GH130 sequences belonging to human gut bacteria indicates that the enzymes of the GH130_1 subfamily would be involved in mannan catabolism, whereas the enzymes belonging to the GH130_2 subfamily would rather work in synergy with glycoside hydrolases of the GH92 and GH18 families in the breakdown of N-glycans. The use of GH130 inhibitors as therapeutic agents or functional foods could thus be considered as an innovative strategy to inhibit N-glycan degradation, with the ultimate goal of protecting, or restoring, the epithelial barrier.
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Simon
authors.initials: S
authors.lastname: Ladevèze
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Laurence
authors.initials: L
authors.lastname: Tarquis
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Davide A
authors.initials: DA
authors.lastname: Cecchini
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Juliette
authors.initials: J
authors.lastname: Bercovici
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Isabelle
authors.initials: I
authors.lastname: André
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Topham
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Morel
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Laville
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse.
authors.forename: Pierre
authors.initials: P
authors.lastname: Monsan
authors.affiliation: the Architecture et Fonction des Macromolécules Biologiques, Aix-Marseille Université, CNRS UMR 7257, 163 Avenue de Luminy, F-13288 Marseille, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Lombard
authors.affiliation: the Architecture et Fonction des Macromolécules Biologiques, Aix-Marseille Université, CNRS UMR 7257, 163 Avenue de Luminy, F-13288 Marseille, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Henrissat
authors.affiliation: From the Institut National des Sciences Appliquées, Université Paul Sabatier, Institut National Polytechnique, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse,; the CNRS, UMR5504, F-31400 Toulouse,; the Institut National de Recherche Agronomique, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse. Electronic address: veronese@insa-toulouse.fr.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Potocki-Véronèse
mesh_terms: D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D008358:Mannose / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054892:Metagenome / Q000502:physiology; D016297:Mutagenesis, Site-Directed; D006005:Phosphorylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D006005:Phosphorylases; D008358:Mannose
keywords: Carbohydrate Metabolism; Enzyme Mechanisms; GH130; Glycoprotein; Human Gut; Intestinal Epithelium; N-Glycan; Phosphorylase
references.citation: Genome Res. 2010 Nov;20(11):1605-12
references.pmid: 20841432
references.citation: ISME J. 2008 Jul;2(7):716-27
references.pmid: 18401439
references.citation: Nat Rev Microbiol. 2012 Apr 11;10(5):323-35
references.pmid: 22491358
references.citation: Nat Protoc. 2010 Apr;5(4):725-38
references.pmid: 20360767
references.citation: Nature. 2010 Mar 4;464(7285):59-65
references.pmid: 20203603
references.citation: Nat Chem Biol. 2007 Jun;3(6):313-20
references.pmid: 17510649
references.citation: Biotechnol J. 2010 Dec;5(12):1324-38
references.pmid: 21154671
references.citation: Protein Expr Purif. 2003 Aug;30(2):194-202
references.pmid: 12880768
references.citation: J Nutr. 2010 Nov;140(11):1943-8
references.pmid: 20861211
references.citation: PLoS One. 2012;7(11):e49138
references.pmid: 23209564
references.citation: Front Cell Infect Microbiol. 2012 Aug 09;2:104
references.pmid: 22919693
references.citation: Nucleic Acids Res. 2004 Mar 19;32(5):1792-7
references.pmid: 15034147
references.citation: Curr Opin Chem Biol. 2013 Apr;17(2):301-9
references.pmid: 23403067
references.citation: Gut. 2011 Oct;60(10):1412-25
references.pmid: 21228430
references.citation: Gut Microbes. 2013 Jan-Feb;4(1):28-40
references.pmid: 23022739
references.citation: Nat Chem Biol. 2010 Feb;6(2):125-32
references.pmid: 20081828
references.citation: Science. 2003 Mar 28;299(5615):2074-6
references.pmid: 12663928
references.citation: Biochimie. 2009 May;91(5):632-8
references.pmid: 19327384
references.citation: Appl Environ Microbiol. 2008 Oct;74(20):6333-7
references.pmid: 18723650
references.citation: Oral Microbiol Immunol. 2003 Oct;18(5):309-12
references.pmid: 12930523
references.citation: Structure. 2004 Jun;12(6):937-47
references.pmid: 15274915
references.citation: Carbohydr Res. 2011 Sep 27;346(13):1860-7
references.pmid: 21798524
references.citation: J Biochem. 2005 Jan;137(1):87-93
references.pmid: 15713887
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D233-8
references.pmid: 18838391
references.citation: Biochim Biophys Acta. 1999 Jun 28;1428(1):88-98
references.pmid: 10366763
references.citation: J Biol Chem. 2004 Sep 10;279(37):38555-62
references.pmid: 15247239
references.citation: BMC Evol Biol. 2012 Sep 20;12:186
references.pmid: 22992189
references.citation: Biochimie. 2010 Dec;92(12):1818-26
references.pmid: 20678539
references.citation: Biochim Biophys Acta. 2011 Dec;1814(12):1720-9
references.pmid: 21867780
references.citation: Syst Biol. 2012 Dec 1;61(6):1061-7
references.pmid: 22780991
references.citation: Appl Microbiol Biotechnol. 2008 May;79(2):165-78
references.pmid: 18385995
references.citation: Glycoconj J. 1994 Apr;11(2):105-10
references.pmid: 7804000
references.citation: Science. 2011 Oct 7;334(6052):105-8
references.pmid: 21885731
references.citation: Crit Rev Biotechnol. 2009;29(3):214-24
references.pmid: 19708823
references.citation: J Biol Chem. 2012 Dec 7;287(50):42389-99
references.pmid: 23093406
references.citation: Appl Environ Microbiol. 2011 Oct;77(19):6939-44
references.pmid: 21803886
references.citation: Gut. 2006 Feb;55(2):205-11
references.pmid: 16188921
references.citation: Nat Struct Biol. 2003 Jan;10(1):59-69
references.pmid: 12483203
references.citation: Chem Biol. 2008 Oct 20;15(10):1058-67
references.pmid: 18848471
references.citation: J Gastroenterol Hepatol. 2012 Jan;27(1):28-38
references.pmid: 21913981
references.citation: Cell Host Microbe. 2008 Nov 13;4(5):447-57
references.pmid: 18996345
references.citation: Carbohydr Res. 1999 Jun 30;319(1-4):184-91
references.pmid: 10520265
references.citation: Protein Eng Des Sel. 2006 Dec;19(12):555-62
references.pmid: 17085431
references.citation: Nucleic Acids Res. 2007 Jan;35(Database issue):D301-3
references.pmid: 17142228
references.citation: Proc Natl Acad Sci U S A. 2012 Jan 17;109(3):781-6
references.pmid: 22219371
references.citation: J Biol Chem. 2013 Sep 20;288(38):27366-27374
references.pmid: 23943617
references.citation: J Biol Chem. 2007 Apr 13;282(15):11291-9
references.pmid: 17287210
references.citation: Br J Nutr. 2005 Jul;94(1):1-11
references.pmid: 16115326
references.citation: FEMS Microbiol Lett. 2011 Jul;320(1):25-32
references.pmid: 21488939
references.citation: Nature. 2011 May 12;473(7346):174-80
references.pmid: 21508958
references.citation: Genome Res. 2009 Dec;19(12):2317-23
references.pmid: 19819907
references.citation: Biochem Biophys Res Commun. 2011 May 20;408(4):701-6
references.pmid: 21539815
references.citation: Science. 2006 Jun 2;312(5778):1355-9
references.pmid: 16741115
references.citation: Protein Expr Purif. 2005 May;41(1):207-34
references.pmid: 15915565
references.citation: Biosci Biotechnol Biochem. 1997 Jul;61(7):1059-66
references.pmid: 9255967