title: Characterization of an α-l-fucosidase from the periodontal pathogen Tannerella forsythia.
abstract: The periodontal pathogen Tannerella forsythia expresses several glycosidases which are linked to specific growth requirements and are involved in the invasion of host tissues. α-l-Fucosyl residues are exposed on various host glycoconjugates and, thus, the α-l-fucosidases predicted in the T. forsythia ATCC 43037 genome could potentially serve roles in host-pathogen interactions. We describe the molecular cloning and characterization of the putative fucosidase TfFuc1 (encoded by the bfo_2737 = Tffuc1 gene), previously reported to be present in an outer membrane preparation. In terms of sequence, this 51-kDa protein is a member of the glycosyl hydrolase family GH29. Using an artificial substrate, p-nitrophenyl-α-fucose (KM 670 μM), the enzyme was determined to have a pH optimum of 9.0 and to be competitively inhibited by fucose and deoxyfuconojirimycin. TfFuc1 was shown here to be a unique α(1,2)-fucosidase that also possesses α(1,6) specificity on small unbranched substrates. It is active on mucin after sialidase-catalyzed removal of terminal sialic acid residues and also removes fucose from blood group H. Following knock-out of the Tffuc1 gene and analyzing biofilm formation and cell invasion/adhesion of the mutant in comparison to the wild-type, it is most likely that the enzyme does not act extracellularly. Biochemically interesting as the first fucosidase in T. forsythia to be characterized, the biological role of TfFuc1 may well be in the metabolism of short oligosaccharides in the periplasm, thereby indirectly contributing to the virulence of this organism. TfFuc1 is the first glycosyl hydrolase in the GH29 family reported to be a specific α(1,2)-fucosidase.
authors.affiliation: a Department of NanoBiotechnology; NanoGlycobiology unit; Universität für Bodenkultur Wien ; Vienna , Austria.
authors.forename: Z A
authors.initials: ZA
authors.lastname: Megson
authors.forename: A
authors.initials: A
authors.lastname: Koerdt
authors.forename: H
authors.initials: H
authors.lastname: Schuster
authors.forename: R
authors.initials: R
authors.lastname: Ludwig
authors.forename: B
authors.initials: B
authors.lastname: Janesch
authors.forename: A
authors.initials: A
authors.lastname: Frey
authors.forename: K
authors.initials: K
authors.lastname: Naylor
authors.forename: I B H
authors.initials: IB
authors.lastname: Wilson
authors.forename: G P
authors.initials: GP
authors.lastname: Stafford
authors.forename: P
authors.initials: P
authors.lastname: Messner
authors.forename: C
authors.initials: C
authors.lastname: Schäffer
mesh_terms: D000818:Animals; D041963:Bacteroidetes / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D003001:Cloning, Molecular; D005643:Fucose / Q000378:metabolism; D054884:Host-Pathogen Interactions; D007700:Kinetics; D009077:Mucins / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010518:Periodontitis / Q000382:microbiology*; D013379:Substrate Specificity; D005644:alpha-L-Fucosidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D009077:Mucins; D009844:Oligosaccharides; D005643:Fucose; D005644:alpha-L-Fucosidase
keywords: 2) fucosidase; 4-nitrophenyl-α-l-fucopyranoside; Amp, ampicillin; BHI, brain heart infusion medium; CBB, Coomassie brilliant blue G 250; DFJ, deoxyfuconojirimycin; Erm, erythromycin; FDH, fucose dehydrogenase; HPAEC, high-performance anion-exchange chromatography with pulsed amperometric detection; LC-ESI-MS, liquid chromatography-electrospray ionisation-mass spectrometry; NAM, N-acetylmuramic acid; PBS, phosphate-buffered saline; SDS-PAGE, sodium dodecylsulphate polyacrylamide gel electrophoresis; T. forsythia, Tannerella forsythia ATCC 43037; TfFuc1, T. forsythia ATCC 43037 fucosidase-1 encoded by the bfo_2737 gene, equally Tffuc1; WT, wild-type bacterium.; enzyme activity; enzyme specificity; oral pathogen; pNP-fucose; periodontitis; rTfFuc-1, recombinant TfFuc1 enzyme; tannerella forsythia; α(1
references.citation: Lancet. 2012 Dec 15;380(9859):2163-96
references.pmid: 23245607
references.citation: Glycobiology. 2002 Apr;12(4):273-82
references.pmid: 12042250
references.citation: J Biol Chem. 2003 Apr 18;278(16):14112-20
references.pmid: 12562767
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Gene. 2006 Apr 12;371(1):102-11
references.pmid: 16488557
references.citation: Periodontol 2000. 2011 Feb;55(1):16-35
references.pmid: 21134226
references.citation: J Bacteriol. 2009 Jun;191(11):3623-8
references.pmid: 19304852
references.citation: Microbiology. 2009 Jun;155(Pt 6):1912-22
references.pmid: 19389765
references.citation: Appl Environ Microbiol. 2011 Jan;77(2):703-5
references.pmid: 21097595
references.citation: Biomolecules. 2012 Oct 12;2(4):467-82
references.pmid: 24970146
references.citation: Microbiology. 2003 Dec;149(Pt 12):3617-27
references.pmid: 14663093
references.citation: Microbiology. 2007 Mar;153(Pt 3):866-76
references.pmid: 17322207
references.citation: J Bacteriol. 2004 Aug;186(15):4885-93
references.pmid: 15262925
references.citation: Science. 2005 Mar 18;307(5716):1778-81
references.pmid: 15774760
references.citation: Mol Oral Microbiol. 2012 Feb;27(1):11-22
references.pmid: 22230462
references.citation: Arch Microbiol. 2012 Jun;194(6):525-39
references.pmid: 22273979
references.citation: Adv Microb Physiol. 2014;65:257-335
references.pmid: 25476768
references.citation: Semin Immunopathol. 2012 May;34(3):383-99
references.pmid: 22461019
references.citation: Biochem J. 1990 Jan 1;265(1):277-82
references.pmid: 2137330
references.citation: J Biol Chem. 2013 Aug 23;288(34):24410-28
references.pmid: 23824194
references.citation: Microb Pathog. 2007 Apr;42(4):156-66
references.pmid: 17363213
references.citation: Microbiology. 2011 Nov;157(Pt 11):3195-202
references.pmid: 21885482
references.citation: J Proteome Res. 2009 Sep;8(9):4279-92
references.pmid: 19663511
references.citation: Microbiology. 2003 Feb;149(Pt 2):279-94
references.pmid: 12624191
references.citation: Biotechnol Lett. 2006 Jan;28(1):55-9
references.pmid: 16369876
references.citation: Science. 1993 Dec 17;262(5141):1892-5
references.pmid: 8018146
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):7194-9
references.pmid: 21482772
references.citation: Oral Microbiol Immunol. 2008 Jun;23(3):196-205
references.pmid: 18402605
references.citation: J Biol Chem. 2006 Feb 24;281(8):4867-75
references.pmid: 16339150
references.citation: Mol Oral Microbiol. 2013 Oct;28(5):366-78
references.pmid: 23714361
references.citation: Infect Immun. 2006 Sep;74(9):5023-8
references.pmid: 16926393
references.citation: J Biol Chem. 2009 Sep 11;284(37):24673-7
references.pmid: 19553672
references.citation: Biochemistry. 2014 Mar 11;53(9):1447-55
references.pmid: 24527659
references.citation: J Clin Periodontol. 1998 Feb;25(2):134-44
references.pmid: 9495612
references.citation: Mol Oral Microbiol. 2012 Dec;27(6):409-19
references.pmid: 23134607
references.citation: ISME J. 2014 Aug;8(8):1659-72
references.pmid: 24599074
references.citation: J Biol Chem. 2011 Nov 4;286(44):38714-24
references.pmid: 21911490
references.citation: Nature. 2014 Feb 27;506(7489):498-502
references.pmid: 24463512
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Mol Oral Microbiol. 2014 Dec;29(6):307-20
references.pmid: 24943676
references.citation: Adv Nutr. 2012 May;3(3):383S-91S
references.pmid: 22585916
references.citation: Infect Immun. 2011 Jan;79(1):393-401
references.pmid: 21078857
references.citation: Aust Dent J. 2009 Sep;54 Suppl 1:S62-9
references.pmid: 19737269