Structural analysis of the O‐antigenic polysaccharide from Vibrio mimicus N‐1990

Abstract

The O-antigenic polysaccharide part of the lipopolysaccharide from Vibrio mimicus N-1990 has been investigated. Sugar and methylation analysis of native and dephosphorylated polysaccharide together with NMR spectroscopy show that the polysaccharide is composed of tetrasaccharide repeating units. The structure of the repeating unit of the polysaccharide from V. mimicus N-1990 could be determined as :The V. mimicus N-1990 strain cross-reacts with antibodies elicited against the Vibrio cholerae O139. The nature of this cross-reactivity resides in the partial structure comprising the galactosyl residue substituted with a cyclic phosphate. This element is present in the cell-wall-associated polysaccharides of both strains.Keywords : Vibrio mimicus ; polysaccharide ; structure ; cyclic phosphate ; NMR.Vibrio cholerae O139 Bengal emerged as a second aetiologic MATERIALS AND METHODS agent of epidemic cholera in the Indian subcontinent in 1992General methods. Anion-exchange chromatography was [1]. It has a lipopolysaccharide (LPS) with a short side chain [2] performed on a DEAE-Sepharose CL column (3 cmϫ20 cm, and produces a polysaccharide capsule [3]. Serological and ge-Pharmacia) eluted with a gradient from 0.1 M to 1 M NaCl. Gelnetic studies suggested that the capsular polysaccharide has the permeation chromatography (GPC) was performed on a Bio-Gel same repeating unit as the O-antigen chain [4, 5] and, thus, car-P-2 column irrigated with 0.4% acetic acid and 1 % pyridine ries the determinants of O-specificity. The capsular polysaccha-in water. Column eluents were monitored using a differential ride (CPS) has two unique constituents, a 3,6-dideoxy-L-xylo-refractometer (Waters) or by detection of total carbohydrates hexose (colitose) and a cyclic phosphate diester [6].using the phenol/sulfuric acid assay [10]. Alditol acetates and V. cholerae O139 cross-reacts with a number of bacteria in-partially methylated alditol acetates were separated on an HP-5 cluding V. cholerae serogroups O22 and O155 [7] and Aeromo-fused-silica column (0.25 mmϫ30 m) with a temperature pronas trota [8]. Until now, only the A. trota O-antigen structure gram of 180°C for 1 min followed by 3°C change/min to 210°C. has been determined [9]. The epitope that is responsible for the Hydrogen was used as carrier gas. The column was fitted to a above mentioned cross-reactivity involves the colitose residues Hewlett-Packard model 5890 gas chromatograph (Hewlett-Packpresent in the antigens of both strains. While searching for bac-ard) equipped with a flame ionisation detector. Gas-chromatogteria that cross-react with V. cholerae O139, we detected two raphy/mass-spectrometry (GC/MS) was performed using a Hewstrains of Vibrio mimicus that agglutinated with specific antise-lett-Packard model 5970 mass spectrometer. The GC/MS separarum to V. cholerae O139 in slide and tube agglutination tests tions were performed on an HP-5-MS fused-silica column (Ansaruzzaman, M. and Albert, M. J., unpublished data). We (0.25 mmϫ30 m) with a temperature program of 170°C f...

The Vibrio cholerae O139 O-antigen polysaccharide is essential for Ca 2+ -dependent biofilm development in sea water

“…The mechanism of Ca 2ϩ -dependent biofilm development and dissolution in these various Vibrio strains has not yet been examined. Cyclic phosphate groups, which we have suggested may play a role in Ca 2ϩ -dependent biofilm formation in V. cholerae O139, are a component of the O antigens of a few Vibrio strains (18,39). However, such a group is not known to be present in the O antigen of V. cholerae O1.…”

Metabolite Cross-Feeding between Rhodococcus ruber YYL and Bacillus cereus MLY1 in the Biodegradation of Tetrahydrofuran under pH Stress

“…3). Among these nutrients, 2-phenyl butyramide acts as an inhibitor of cell receptors, farnesylcysteine, 5-methylcytidine, and (R)-5-phosphomevalonate are required for cell survival, sedoheptulose is associated with the cell structure, farnesylcysteine and its analogs have high affinity for methyltransferase and function as specific inhibitor of the COOH-terminal S-farnesyl-cysteine methyltransferase, stearamide and cresol are antibacterial compounds, and (R)-5-phosphomevalonate is the synthetic precursor of some biomolecules in the phosphomevalonate pathway (27,28). All of these metabolites play important roles in the basal metabolism of microorganisms.…”

Carbohydrate binding, quaternary structure and a novel hydrophobic binding site in two legume lectin oligomers from Dolichos biflorus 1 1Edited by R. Huber

“…The Forssman disaccharide is present in tumor-associated Forssman glycolipid (Hakomori, 1984) and in lipopolysaccharides from certain pathogenic bacteria, e.g. Vibrio mimicus (Landersjo È et al, 1998).…”