Structure of the O-specific, sialic acid containing polysaccharide chain and its linkage to the core region in lipopolysaccharide from Hafnia alvei strain 2 as elucidated by chemical methods, gas-liquid chromatography/mass spectrometry, and 1H NMR spectroscopy

Mild acid hydrolysis of Hafnia alvei strain 2 lipopolysaccharide released no O-specific polysaccharide but instead gave a monomeric octasaccharide repeating unit with N-acetylneuraminic acid as the reducing terminus. In addition, a dimer of the octasaccharide repeating unit, and also a decasaccharide composed of a fragment of the O-specific polysaccharide chain and the core region, were obtained in minute amounts. On the basis of the sugar and methylation analyses, periodate oxidation, and 1H NMR spectroscopy of the lipopolysaccharide hydrolytic products, the biological repeating unit of the O-specific polysaccharide was shown to be a branched octasaccharide: (Formula; see text) The linkage between the O-specific polysaccharide chain and core region has also been determined and has yield strong evidence that N-acetylneuraminic acid is an inherent lipopolysaccharide component. The lipopolysaccharide of H. alvei strain 2 is the first lipopolysaccharide reported to contain 4-substituted neuraminic acid in its O-specific polysaccharide region.     

The structure of O-specific polysaccharide from Yersinia enterocolitica serotype O:6.31 lipopolysaccharide

The serologically active O-specific polysaccharide has been isolated from the lipopolysaccharide of Yersinia enterocolitica, serovar O: 6.31. Using methylation, partial acid hydrolysis and 13C NMR spectroscopy, the main structural moiety of the O-specific polysaccharide is shown to be the following disaccharide repeating unit: (Formula: see text).     

Structure of the O-specific polysaccharide from the lipopolysaccharide of Citrobacter gillenii O11, strain PCM 1540

The O-specific polysaccharide of the lipopolysaccharide of Citrobacter gillenii PCM 1540 (serogroup O11) consists of D-Glc, D-Man, D-GalNAc, D-GlcNAc, 2-acetamido-2,6-dideoxy-D-galactose (D-FucNAc) and O-acetyl groups in the ratios 2:1:1:1:1:1. On the basis of sugar and methylation analyses and Smith-degradation along with 1D and 2D 1H and 13C NMR spectroscopy, the following structure of the branched hexasaccharide repeating unit was established: [structure: see text]. Citrobacter werkmanii PCM 1541 belonging to the same serogroup O11 was found to have an R-form lipopolysaccharide devoid of the O-specific polysaccharide.     

Structure of O-specific polysaccharide isolated from the Yersinia pseudotuberculosis serotype 1A lipopolysaccharide

An O-specific polysaccharide from the lipopolysaccharide Yersinia pseudotuberculosis 1A serovar has been isolated and characterized. This compound was shown to contain residues of paratose, 6-deoxy-D-manno-heptose, D-galactose and 2-amino-2-deoxy-D-glucose in equimolar ratios. Using methylation studies, partial acid hydrolysis and 13C NMR spectroscopy, the following structure was proposed for the repeating unit of the O-specific polysaccharide: (Formula: see text).     

O-specific polysaccharide of the marine bacterium "Alteromonas marinoglutinosa" NCIMB 1770

The O-specific polysaccharide of the marine bacterium "Alteromonas marinoglutinosa" NCIMB 1770 was obtained by mild acid degradation of the corresponding lipopolysaccharide and found to contain D-galactose, N-acetyl-D-glucosamine, and N-acetyl-D-mannosamine residues in equimolar ratio. Based on methylation analysis, periodate oxidation, and 13C-NMR spectroscopy data of native and modified polysaccharides, the following structure of the trisaccharide repeating unit of the O-specific polysaccharide was established: [structure: see text]     

Somatic antigens of the Brucella genus. The structure of the O-specific polysaccharide chain of Brucella melitensis lipopolysaccharide

The phenol-phase soluble antigenic lipopolysaccharide was isolated from Brucella melitensis, strain 565, by the routine phenol/water procedure followed by chromatography on Sepharose 4B. After mild acid hydrolysis and chromatography on Sephadex G-50, the lipopolysaccharide yielded a linear O-specific polysaccharide built up from 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl units. The structure of the polysaccharide was deduced mainly from the nuclear magnetic resonance and methylation analyses. The phenol-soluble lipopolysaccharide, isolated from commercial vaccine strain B. abortus 19-BA, on mild hydrolysis afforded material, 13C and 1H-NMR spectra of which were identical to those of the O-specific polysaccharide from B. melitensis 565.     

Structure of the O-specific polysaccharide chain of the Yersinia pseudotuberculosis lipopolysaccharide (serovar II C)]

An O-specific polysaccharide has been isolated on mild acid hydrolysis of lipopolysaccharide from Yersinia pseudotuberculosis serovar IIc and shown to consist of abequose, D-mannose and 2-acetamido-2-deoxy-D-galactose residues in the ratio 0.8:3:1. From the results of acid hydrolysis, 13C NMR, methylation and periodate oxidation studies the structure of the repeating unit of the O-specific polysaccharide is deduced as follows: (formula; see text)     

Somatic antigens of Shigella. Structural investigation on the O-specific polysaccharide chain of Shigella dysenteriae type-2 lipopolysaccharide.

The O-specific polysaccharide obtained from Shigella dysenteriae type-2 lipopolysaccharide by mild acid hydrolysis consisted of N-acetylgalactosamine, N-acetylglucosamine, D-galactose, D-glucose, and O-acetyl group in the ratio of 2:1:1:1:1. A number of oligosaccharides were obtained by deamination of the N-deacetylated polysaccharide and by Smith degradation of the both native and O-deacetylated polysaccharides. The identification of oligosaccharides along with methylation analysis and chromic anhydride oxidation showed that the polysaccharide was built up of the repeating pentasaccharide units whose proposed structure is given below: (see article) Serological properties of Sh. dysenteriae O-specific polysaccharides are discussed.     

Characterization of the lipopolysaccharide from Rahnella aquatilis 1-95

The lipopolysaccharide from the freshwater bacterium Rahnella aquatilis 1-95 has been isolated and investigated for the first time. The structural components of the lipopolysaccharide molecule: lipid A, core oligosaccharide, and O-specific polysaccharide were isolated by mild acidic hydrolysis. In lipid A, 3-hydroxytetradecanoic and tetradecanoic acids were found to be the predominant fatty acids. In the core oligosaccharide, galactose, arabinose, fucose, and an unidentified component were shown to be the major monosaccharides. The O-specific polysaccharide consists of a regularly repeating trisaccharide unit with the acyl and phosphate following structure: [structure: see text] groups have been shown to be responsible for the toxic and pyrogenic properties of the lipopolysaccharide of R. aquatilis.     

Structural studies of the side chain of outer membrane lipopolysaccharide from Pseudomonas syringae pv. coriandricola W-43.

The lipopolysaccharide (LPS) was isolated from Pseudomonas syringae pv. coriandricola W-43 by hot phenol-water extraction. Rhamnose and 3-N-acetyl-3-deoxyfucose were found to be the major sugar constituents of the LPS together with N-acetylglucosamine, N-acetylgalactosamine, heptose, and 3-deoxy-D-manno-octulosonic acid (Kdo). The main fatty acids of lipid A of the LPS were 3-OH-C:10, C12:0, 2-OH-C12:0, and 3-OH-C12:0. The O-specific polysaccharide liberated from the LPS by mild-acid hydrolysis was purified by gel permeation chromatography. The compositional analysis of the O-specific polysaccharide revealed the presence of L-rhamnose and 3-N-acetyl-3-deoxy-D-fucose in a molar ratio of 4:1. The primary structure of the O-specific polysaccharide was established by methylation analysis together with 1H and 13C nuclear magnetic resonance spectroscopy, including two-dimensional shift-correlated and one-dimensional nuclear Overhauser effect spectroscopy. The polysaccharide moiety was found to consist of a tetrasaccharide rhamnan backbone, and 3-N-acetyl-3-deoxy-D-fucose constitutes the side chain of the branched pentasaccharide repeating unit of the polysaccharide.     

Structure and serological analysis of the Hafnia alvei 481-L O-specific polysaccharide containing phosphate in the backbone chain

The lipopolysaccharide was extracted from cells of Hafnia alvei 481-L bacterial strain and, after mild acid hydrolysis, the O-specific polysaccharide was isolated and characterised. On the basis of chemical analyses and NMR spectroscopic studies of the polysaccharide and oligosaccharides obtained after Smith degradation, or hydrogen fluoride treatment, it was found that the repeating unit of the O-specific polysaccharide is a phosphorylated hexasaccharide: [see text]. The biological repeating unit of the H. alvei 481-L O-antigen has galactose phosphate at the nonreducing terminus. Serological tests indicate that this strain represents an individual serotype in the H. alvei genus.     

Structure of the O-specific polysaccharide from a marine bacterium Oceanisphaeralitoralis KMM 3654(T) containing ManNAcA

The O-specific polysaccharide was isolated from the lipopolysaccharide of a marine bacterium Oceanisphaeralitoralis KMM 3654(T) and studied by chemical methods along with (1)H and (13)C NMR spectroscopy. The following new structure of the O-specific polysaccharide of O. litoralis containing D-glucose and two residues of 2-acetamido-2-deoxy-D-mannuronic acid was established: →4)-α-D-Glcp-(1→4)-β-D-ManpNAcA-(1→4)-β-D-ManpNAcA-(1→.     

Antigenic bacterial polysaccharides. 36. A study of the structure of the O-specific polysaccharide chain of the lipopolysaccharide of Pseudomonas fluorescens IMV 2763 (biostrain G)

Chemical and serological characterization of the Pseudomonas fluorescens IMV 2763 (biovar G) lipopolysaccharide was carried out. The O-specific polysaccharide chain of the lipopolysaccharide is composed of D-mannose, 6-deoxy-L-talose, N-acetyl-D-galactosamine and O-acetyl groups in the ratio of approximately 2:1:1:1. The polysaccharide is branched and a half of residues of 6-deoxytalose and monosubstituted mannose carry O-acetyl groups. On the basis of methylation, partial acid hydrolysis and 13C NMR analysis it was concluded that the repeating unit of the polysaccharide has the following structure: (formula; see text)     

Structure of O-specific polysaccharide from Yersinia pseudotuberculosis of serotype VI lipopolysaccharide

Using methylation studies, partial hydrolysis and 13C NMR spectroscopy data, the following structure of O-specific polysaccharide from lipopolysaccharide of Yersinia pseudotuberculosis VI serovar has been proposed: (Formula: see text).     

Structural studies of the O-specific polysaccharide of Hafnia alvei strain PCM 1207 lipopolysaccharide.

The structure of the O-specific side-chain of the Hafnia alvei strain PCM 1207 lipopolysaccharide (LPS) has been investigated. Methylation analysis, partial acid hydrolysis, matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) MS, fast atom bombardment (FAB)-MS/MS and 1H- and 13C-NMR spectroscopy were the principal methods used. Glycerol phosphate was identified as a constituent in the polysaccharide and the following structure of a pentasaccharide repeating unit was established: The polysaccharide is partially (approximately 10%) substituted with O-acetyl groups. The lipopolysaccharide was also subjected to high resolution magic angle spinning (HR-MAS) NMR analysis, which showed both the signals of the O-specific polysaccharide as well as several signals from unsubstituted core oligosaccharides. This confirmed the presence of the described structure in the native LPS.     

Antigenic polysaccharides of bacteria of the genus Shigella. Determination of the structure of polysaccharide chains of Shigella boydii type 9 lipopolysaccharide and detection of unusually high molecular weight glycolipid

Specific acidic polysaccharide has been isolated from the Shigella boydii type 9 antigenic lipopolysaccharide after mild hydrolysis followed by chromatography on Sephadex G-50. The polysaccharide consists of D-glucose, D-glucuronic acid, 2-acetamido-2-deoxy-D-glucose, and L-rhamnose. From the results of methylation analysis, partial acid hydrolysis and 13C NMR data the structure of the repeating unit of the polysaccharide was deduced as follows: [----4)DGlcp(alpha 1----4)DGlcAp(beta 1----3)DGlcNAcp(alpha 1----3)LRhap(alpha 1----]n. The lipopolysaccharide from Sh. boydii 9 was fractionated by gel chromatography on the Sephadex G-200 column in a buffer containing sodium deoxycholate into three fractions. PAGE-SDS of the fractions obtained, 13C NMR- and chromato-mass-spectrometry data indicated that the three fractions contained the O-specific polysaccharide as the only carbohydrate component. The substance from the most high-molecular weight fraction contained unusually long O-specific chains (60,000 dalton). In the fat acid composition this fraction differed from other lipopolysaccharides by absence of beta-hydroxymyristic acid.     

Structure of O-specific polysaccharide from Pseudoalteromonas nigrifaciens strain KMM 161

On mild acid degradation of the lipopolysaccharide of the marine microorganism Pseudoalteromonas nigrifaciens KMM 161 an O-specific polysaccharide containing D-galactose, 2-acetamido-2-deoxy-D-glucose, 3,6-dideoxy-3-(4-hydroxybutyramido)-D-galactose, and 2-acetamido-2-deoxy-L-guluronic acid residues was obtained. From the results of Smith degradation, O-deacetylation of the polysaccharide, and NMR spectroscopy the following structure of the tetrasaccharide repeating unit of the O-specific polysaccharide was established [see reaction]. It should be noted that the same structure occurs in the antigenic polysaccharide of Pseudoalteromonas nigrifaciens KMM 158 described earlier as Alteromonas macleodii 2MM6.     

Antigenic bacterial polysaccharides. 24. The structure of the O-specific polysaccharide chain of Salmonella arizonae 063 (Arizona 08) lipopolysaccharide

The O-specific polysaccharide chain of the Salmonella arizonae O63 lipopolysaccharide is composed of D-glucose, D-galactose, N-acetyl-D-galactosamine, and 3-acetamido-3,6-dideoxy-D-galactose (Fuc3NAc) residues in the ratio 1:1:2:1. On the basis of methylation analysis and calculations of 13C-NMR-spectra of the polysaccharide and of the product of its selective cleavage with anhydrous hydrogen fluoride, the linear polymer lacking 3-acetamido-3,6-dideoxygalactose, it was concluded that the polysaccharide has the following structure: (Formula: see text).     

Inhibition of prokaryote and eukaryote cells adhesion by sea proteobacteria Pseudoalteromonas nigrifaciens KMM 156 lipopolysaccharide and its fragments]

The impact of Pseudoalteromonas nigrifaciens KMM 156 lipopolysaccharide and its fragments on adhesion of prokaryote and eukaryote cells was studied. The lipopolysaccharide and O-specific polysaccharide identical by its structure to capsular polysaccharide lowered the number of the Yersinia pseudotuberculosis bacteria attached to sheep erythrocytes and adhesion of mouse neutrophils to plastic. The kora oligosaccharide had no effect on the processes.     

Structure of the O-specific polysaccharide of Proteus vulgaris O4 containing a new component of bacterial polysaccharides, 4,6-dideoxy-4

A high-molecular-mass O-specific polysaccharide was obtained by mild acid degradation of Proteus vulgaris O4 lipopolysaccharide followed by GPC. The polysaccharide was studied by chemical methods along with 1H and 13C NMR spectroscopy, including two-dimensional COSY, TOCSY, NOESY, H-detected 1H,13C HMQC, and 1H,13C HMBC experiments. Solvolysis of the polysaccharide with trifluoromethanesulfonic (triflic) acid resulted in a GlcpA-(1 --> 3)-GlcNAc disaccharide and a novel amino sugar derivative, 4,6-dideoxy-4-[N-[(R)-3-hydroxybutyryl]-L-alanyl]amino-D-glucose [Qui4N(HbAla)]. On the basis of the data obtained, the following structure of the tetrasaccharide repeating unit of the O-specific polysaccharide was established: --> 4)-beta-D-GlcpA-(1 --> 3)-beta-D-GlcpNAc-(1 --> 2)-beta-D-Quip4N(HbAla)-(1 --> 3)-alpha-D-Galp-(1 -->. This structure is unique among the O-specific polysaccharides, which is in accordance with classification of the strain studied in a separate Proteus serogroup.