title: Synthesis and utility of sulfated chromogenic carbohydrate model substrates for measuring activities of mucin-desulfating enzymes.
abstract: A chromogenic substrate, 4-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside 6-sodium sulfate was synthesized and used in combination with beta-N-acetylhexosaminidase for detection of the sulfatase, MdsA, by release of 4-nitrophenol. MdsA was originally isolated from the bacterium Prevotella strain RS2 and is believed to be involved in desulfation of sulfomucins, major components of the mucus barrier protecting the human colon surface. The exo nature of the MdsA sulfatase was indicated by its inability to de-esterify the disaccharide 4-nitrophenyl beta-D-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-D-glucopyranoside 6-sodium sulfate. This latter compound was prepared from monosaccharide precursors by two different methods, the shorter requiring just six steps from 4-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside and giving an overall yield of 26.4%. The syntheses of 4-nitrophenyl beta-D-galactopyranoside 3-triethylammonium sulfate and 6-triethylammonium sulfate and their use in combination with beta-galactosidase as chromogenic substrates for detecting Bacteroides fragilis sulfatases with different specificities was also demonstrated.
authors.affiliation: Industrial Research Ltd., Gracefield Road, PO Box 31-310, Lower Hutt, New Zealand. k.clinch@irl.cri.nz
authors.forename: Keith
authors.initials: K
authors.lastname: Clinch
authors.forename: Gary B
authors.initials: GB
authors.lastname: Evans
authors.forename: Richard H
authors.initials: RH
authors.lastname: Furneaux
authors.forename: Phillip M
authors.initials: PM
authors.lastname: Rendle
authors.forename: Phillippa L
authors.initials: PL
authors.lastname: Rhodes
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Roberton
authors.forename: Douglas I
authors.initials: DI
authors.lastname: Rosendale
authors.forename: Peter C
authors.initials: PC
authors.lastname: Tyler
authors.forename: Damian P
authors.initials: DP
authors.lastname: Wright
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology; D001441:Bacteroides fragilis / Q000201:enzymology; D002863:Chromogenic Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D008956:Models, Chemical; D015394:Molecular Structure; D009077:Mucins / Q000737:chemistry / Q000378:metabolism*; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry; D013429:Sulfatases / Q000378:metabolism*; D013431:Sulfates / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: D002863:Chromogenic Compounds; D009077:Mucins; D009844:Oligosaccharides; D013431:Sulfates; C012564:sulfomucin; D013429:Sulfatases