title: α-N-acetylgalactosaminidase from infant-associated bifidobacteria belonging to novel glycoside hydrolase family 129 is implicated in alternative mucin degradation pathway.
abstract: Bifidobacteria inhabit the lower intestine of mammals including humans where the mucin gel layer forms a space for commensal bacteria. We previously identified that infant-associated bifidobacteria possess an extracellular membrane-bound endo-α-N-acetylgalactosaminidase (EngBF) that may be involved in degradation and assimilation of mucin-type oligosaccharides. However, EngBF is highly specific for core-1-type O-glycan (Galβ1-3GalNAcα1-Ser/Thr), also called T antigen, which is mainly attached onto gastroduodenal mucins. By contrast, core-3-type O-glycans (GlcNAcβ1-3GalNAcα1-Ser/Thr) are predominantly found on the mucins in the intestines. Here, we identified a novel α-N-acetylgalactosaminidase (NagBb) from Bifidobacterium bifidum JCM 1254 that hydrolyzes the Tn antigen (GalNAcα1-Ser/Thr). Sialyl and galactosyl core-3 (Galβ1-3/4GlcNAcβ1-3(Neu5Acα2-6)GalNAcα1-Ser/Thr), a major tetrasaccharide structure on MUC2 mucin primarily secreted from goblet cells in human sigmoid colon, can be serially hydrolyzed into Tn antigen by previously identified bifidobacterial extracellular glycosidases such as α-sialidase (SiaBb2), lacto-N-biosidase (LnbB), β-galactosidase (BbgIII), and β-N-acetylhexosaminidases (BbhI and BbhII). Because NagBb is an intracellular enzyme without an N-terminal secretion signal sequence, it is likely involved in intracellular degradation and assimilation of Tn antigen-containing polypeptides, which might be incorporated through unknown transporters. Thus, bifidobacteria possess two distinct pathways for assimilation of O-glycans on gastroduodenal and intestinal mucins. NagBb homologs are conserved in infant-associated bifidobacteria, suggesting a significant role for their adaptation within the infant gut, and they were found to form a new glycoside hydrolase family 129.
authors.affiliation: Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan; Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa 921-8836, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Kiyohara
authors.affiliation: Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakatomi
authors.affiliation: Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Kurihara
authors.affiliation: Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Fushinobu
authors.affiliation: Graduate School of Science and Technology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: Graduate School of Science and Technology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Tomonari
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Graduate School of Engineering, Tohoku University, Aoba-ku, Sendai 980-8579, Japan.
authors.forename: Shin-Ichiro
authors.initials: SI
authors.lastname: Shoda
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Motomitsu
authors.initials: M
authors.lastname: Kitaoka
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa 921-8836, Japan.
authors.forename: Takane
authors.initials: T
authors.lastname: Katayama
authors.affiliation: Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan; Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa 921-8836, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: ashida@lif.kyoto-u.ac.jp.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Ashida
mesh_terms: D001644:Bifidobacterium / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D055162:Biocatalysis; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D006801:Humans; D007223:Infant; D042541:Intracellular Space / Q000201:enzymology; D008969:Molecular Sequence Data; D009077:Mucins / Q000378:metabolism*; D010802:Phylogeny; D059748:Proteolysis; D048809:alpha-N-Acetylgalactosaminidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D009077:Mucins; D048809:alpha-N-Acetylgalactosaminidase
references.citation: Carbohydr Res. 2001 Feb 28;330(4):487-93
references.pmid: 11269400
references.citation: Arch Biochem Biophys. 2000 Dec 15;384(2):305-10
references.pmid: 11368317
references.citation: J Biol Chem. 2008 May 9;283(19):13165-73
references.pmid: 18332142
references.citation: Anal Biochem. 2000 Dec 15;287(2):337-9
references.pmid: 11112283
references.citation: Appl Environ Microbiol. 2005 Jun;71(6):3158-62
references.pmid: 15933016
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 15;108 Suppl 1:4659-65
references.pmid: 20615996
references.citation: J Biol Chem. 1994 Jan 28;269(4):2440-6
references.pmid: 8300571
references.citation: FEMS Microbiol Lett. 2008 Aug;285(2):278-83
references.pmid: 18564335
references.citation: J Biochem. 2009 Sep;146(3):389-98
references.pmid: 19502354
references.citation: J Biol Chem. 1977 Dec 10;252(23):8609-14
references.pmid: 21877
references.citation: Biochemistry. 2007 Mar 20;46(11):3319-30
references.pmid: 17323919
references.citation: Biosci Biotechnol Biochem. 2007 Aug;71(8):2101-4
references.pmid: 17690443
references.citation: Chem Commun (Camb). 2008 May 7;(17):2016-8
references.pmid: 18536806
references.citation: J Mol Biol. 2002 Apr 26;318(2):443-53
references.pmid: 12051850
references.citation: Glycobiology. 2010 Nov;20(11):1402-9
references.pmid: 20581010
references.citation: Arch Biochem Biophys. 2000 Jan 15;373(2):394-400
references.pmid: 10620364
references.citation: Carbohydr Res. 2009 Dec 14;344(18):2573-6
references.pmid: 19850285
references.citation: Nat Protoc. 2009;4(3):363-71
references.pmid: 19247286
references.citation: Biosci Biotechnol Biochem. 1998;62(8):1643-6
references.pmid: 27388851
references.citation: Glycobiology. 2009 Jul;19(7):756-66
references.pmid: 19321523
references.citation: Biochem Biophys Res Commun. 2008 Oct 31;375(4):441-6
references.pmid: 18725192
references.citation: Biosci Biotechnol Biochem. 2009 May;73(5):1175-9
references.pmid: 19420691
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):15064-9
references.pmid: 18806221
references.citation: Biosci Biotechnol Biochem. 2010;74(12):2374-84
references.pmid: 21150123
references.citation: Appl Environ Microbiol. 2008 Jul;74(13):3996-4004
references.pmid: 18469123
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 2;105(48):18964-9
references.pmid: 19033196
references.citation: J Biol Chem. 2004 Dec 31;279(53):55097-103
references.pmid: 15491996
references.citation: Glycobiology. 2008 Sep;18(9):727-34
references.pmid: 18559962
references.citation: Proc Natl Acad Sci U S A. 2010 Nov 9;107(45):19514-9
references.pmid: 20974960
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D233-8
references.pmid: 18838391
references.citation: Glycobiology. 2008 Oct;18(10):799-805
references.pmid: 18635885
references.citation: J Biol Chem. 2007 Jun 22;282(25):18497-18509
references.pmid: 17459873
references.citation: FEMS Microbiol Lett. 2002 Aug 27;214(1):77-80
references.pmid: 12204375
references.citation: J Bacteriol. 2004 Aug;186(15):4885-93
references.pmid: 15262925
references.citation: Aliment Pharmacol Ther. 2005 Sep 15;22(6):495-512
references.pmid: 16167966
references.citation: Glycobiology. 2011 Apr;21(4):437-47
references.pmid: 21036948
references.citation: Agric Biol Chem. 1990 Jan;54(1):233-4
references.pmid: 1369985
references.citation: J Biol Chem. 2005 Feb 11;280(6):4602-8
references.pmid: 15590624
references.citation: Glycobiology. 2009 Sep;19(9):1010-7
references.pmid: 19520709
references.citation: Glycoconj J. 2010 Jan;27(1):125-32
references.pmid: 19562481
references.citation: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Sep 1;63(Pt 9):751-3
references.pmid: 17768345
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14422-7
references.pmid: 12381787
references.citation: Org Lett. 2005 Sep 29;7(20):4415-8
references.pmid: 16178547
references.citation: Nat Biotechnol. 2007 Apr;25(4):454-64
references.pmid: 17401360
references.citation: J Biochem. 1976 Jul;80(1):1-8
references.pmid: 9374
references.citation: Appl Microbiol Biotechnol. 2009 Mar;82(3):471-7
references.pmid: 19005653
references.citation: J Org Chem. 2009 Mar 6;74(5):2210-2
references.pmid: 19203234
references.citation: J Biol Chem. 2005 Nov 11;280(45):37415-22
references.pmid: 16141207