Comparative structural study of the lipopolysaccharides of Y. enterocolitica serovars 0:7.8 and 0:19.8

The lipopolysaccharides ofYersinia enterocolitica, serovars 0:7.8 (strain 106) and 0:19.8 (strain 842), isolated from the microbial mass by phenol-water extraction, contained residues of L-fucose, 6-deoxy-D-gulose, D-mannose, D-galactose, D-glucose, D- and L-glycero-D-mannoheptoses, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and 2-keto-3-deoxyoctonic acid (KDO). The polysaccharides obtained by mild acid hydrolysis of the lipopolysaccharides followed by gel filtration on Sephadex G-50 were a mixture of the O-specific polysaccharide and the core, which could not be separated even by repeated rechromatography because of the comparability of their molecular masses. On the basis of the results of monosaccharide analysis, methylation, Smith degradation, and partial hydrolysis, a structure has been suggested for the repeating unit of the O-specific polysaccharides of the lipopolysaccharides ofY. enterocolitica of the serovars studied.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Branch, USSR Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 763–770, November–December, 1989.     

Comparative structural study of the lipopolysaccharides of Y. enterocolitica serovars 0:7.8 and 0:19.8

The lipopolysaccharides ofYersinia enterocolitica, serovars 0:7.8 (strain 106) and 0:19.8 (strain 842), isolated from the microbial mass by phenol-water extraction, contained residues of L-fucose, 6-deoxy-D-gulose, D-mannose, D-galactose, D-glucose, D- and L-glycero-D-mannoheptoses, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and 2-keto-3-deoxyoctonic acid (KDO). The polysaccharides obtained by mild acid hydrolysis of the lipopolysaccharides followed by gel filtration on Sephadex G-50 were a mixture of the O-specific polysaccharide and the core, which could not be separated even by repeated rechromatography because of the comparability of their molecular masses. On the basis of the results of monosaccharide analysis, methylation, Smith degradation, and partial hydrolysis, a structure has been suggested for the repeating unit of the O-specific polysaccharides of the lipopolysaccharides ofY. enterocolitica of the serovars studied. Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Branch, USSR Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 763–770, November–December, 1989.     

Investigation of the protein component of the lipopolysaccharide-protein complex ofYersinia pseudotuberculosis

It has been established by electrophoresis in polyacrylamide gel that the protein component of the lipopolysaccharide-protein complex ofYersinia pseudotuberculosis consists of two poly-peptides with molecular weights of 42,000–45,000 and 18,000–20,000. These polypeptides have been detected in the cell wall ofY. pseudotuberculosis as the main components. Protein fractions B-2 and B-3, differing with respect to solubility in phenol, were isolated from the LPPC ofY. pseudotuberculosis by extraction with hot 45% phenol. These protein fractions had the same polypeptide composition, which is characteristic for membrane proteins. B-3 included the LPS component and B-2 contained lipid A or fragments of it as impurity. Fractions B-3 and B-2 precipitated with antiserum to the LPPC.Pacific Ocean Institute of Bioorganic Chemistry of the Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 92–97, January–February, 1980.     

Exoglycans of Yersina pseudotuberculosis

Summary 1. From a culture medium of strain 12 ofYersinia pseudotuberculosis of subtype IB a mixture of exoglycans has been isolated which consists of acidic heteropolysaccharides bound ionically to a protein component.2. The yield and monosaccharide composition of the exoglycans produced depend on the conditions of growth and the form of the colonies.3. The exoglycans of the local strain No. 12 and of a standard strain (No. 159) ofY. pseudotuberculosis of subtype IB contain monosaccharides (galactose, glucose, mannose, xylose, arabinose, fucose, glucuronic acid, glucosamine) and differ only by the ratio of the monosaccharide residues.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Scientific Center of the Academy of Sciences of the USSR. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 315–321, May–June, 1975.     

Investigation of the core of the lipopolysaccharides ofYersinia pseudotuberculosis

Lipopolysaccharides have been isolated from two R mutants of the pseudotuberculosis microbeYersinia tuberculosis, serovar VA. Mild acid hydrolysis followed by gel filtration on Sephadex G-25 gave the core oligosaccharides OS I and OS II. Complete acid hydrolysis showed that OS I consisted of residues of D-galactose, G-glucose, two heptoses (D-glycero-D-mannoheptose and L-glycero-D-mannoheptose), and D-glucosamine in a ratio of 1:2.5:4:1, while OS II consisted of residues of D-glucose, heptoses, and D-glucosamine in a ratio of 1:2.5:0.2. On the basis of the results of monosaccharide analysis, methylation, Smith degradation, etc., a partial structure of the core oligosaccharide of the LPS ofY. pseudotuberculosis has been put forward.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 751–755, November–December, 1985.     

Chemical composition of the low-molecular-mass fraction of Yersinia pseudotuberculosis as a function of the temperature of cultivation

The dependence of the chemical composition of the low-molecular-mass fraction of the microbial cells ofYersinia pseudotuberculosis (str. 2781) on the temperature of cultivation (6 and 37°C) has been investigated.Scientific-Research Institute of Epidemiology and Microbiology, Siberian Branch, Russian Academy of Medical Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, Nos. 3,4, pp. 312–316, May–August, 1992.     

Structural investigation of the lipopolysaccharide ofYersinia pseudotuberculosis of type IB

Summary Using the chromato-mass spectrometry of the methylated sugars formed in the hydrolysis of the fully methylated lipopolysaccharide fromYersinia pseudotuberculosis of type IB the nature of the glycosidic bonds between the monosaccharide residues has been established. The hapten formed in the partial hydrolysis of the lipopolysaccharide has also been studied by methylation. The results obtained have confirmed the results of an investigation of the methylated lipopolysaccharide; they have enabled an idea to be put forward of the composition of the repeating unit of the O-specific side chains of the lipopolysaccharide.Pacific-Ocean Institute of Bioorganic Chemistry of the Far-Eastern Scientific Center of the Academy of Sciences of the USSR. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 563–570, September–October, 1975.     

An investigation of a lipopolysaccharide-protein complex fromYersinia pseudotuberculosis by the light-scattering method

Summary An estimate has been made of the molecular weights and characteristic viscosities of aggregates of the LPPC fromYersinia pseudotuberculosis in water, and also in aqueous solutions of sodium chloride, urea, guanidine hydrochloride, and sodium dodecyl sulfate. It has been shown that a high ionic strength of the solution and the presence of disaggregating agents lead to a considerable decrease in the molecular weight of the LPPC.Pacific Ocean Institute of Bioorganic Chemistry of the Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 51–53, January–February, 1979.     

Specific polysaccharides of the lipopolysaccharides of Gram-negative bacteria

This review generalizes the result of structural investigations of the polysaccharides of the O-specific side chains of the lipopolysaccharides (LPSs) of the most studied representations of the family Enterobacteriaceae. The O-specific polysaccharides are the serologically dominating part of the molecule that is responsible for the O-antigenic specificity of the LPSs. The structures of the specific polysacchrides from various Gram-positive bacteria are given. The serological specificity of the O-antigens is discussed and a connection is shown between the chemical structures of the polysaccharides and their serological affinity.Pacific Institute of Bioorganic Chemistry of the Far-Eastern Scientific Center of the USSR Academy of Sciences, Vladivostok. Translated from khimiya Prirodnykh Soedinenii, No. 5, pp. 532–547, September–October, 1986.     

Investigation of the core of the lipopolysaccharides ofYersinia pseudotuberculosis

Lipopolysaccharides have been isolated from two R mutants of the pseudotuberculosis microbeYersinia tuberculosis, serovar VA. Mild acid hydrolysis followed by gel filtration on Sephadex G-25 gave the core oligosaccharides OS I and OS II. Complete acid hydrolysis showed that OS I consisted of residues of D-galactose, G-glucose, two heptoses (D-glycero-D-mannoheptose and L-glycero-D-mannoheptose), and D-glucosamine in a ratio of 1:2.5:4:1, while OS II consisted of residues of D-glucose, heptoses, and D-glucosamine in a ratio of 1:2.5:0.2. On the basis of the results of monosaccharide analysis, methylation, Smith degradation, etc., a partial structure of the core oligosaccharide of the LPS ofY. pseudotuberculosis has been put forward.     

The significance of enterobacterial mutants for the chemical investigation of their cell-wall polysaccharides

The outer cell surface of Enterobacteriaceae, i.e. the cell wall, consist of a rigid structure (murein) on which additional proteins, lipids, and polysaccharides are deposited. In the bacterial wild types (S forms) the polysaccharide is species-specific and carries the serologically determinant groups of the respective O antigen. These specific polysaccharides often consist of a large number of monosaccharides; up to eight different monosaccharides may be involved. Bound to lipid A, the cell-wall lipopolysaccharides represent the endotoxins of the bacteria. During the past few years the structures of the enterobacterial cell-wall polysaccharides have been elucidated by chemical, immunochemical, biochemical, and genetic investigations, particularly in the Salmonella. Here the polysaccharides consists of a basal polysaccharide common to all species, to which (in the S form) are bound longer species-specific side chains, consisting of repeating oligosachharide units. Spontaneous S → R mutation leads to R forms which are deficient mutants of the wild types in regard to the biosynthesis of the complete cell-wall polysaccharide. In contrast to the multiplicity of the serological specificities of the somatic antigens of the S forms, only a few serological types were found among the R forms (R I, R II, etc.). These R polysaccharides correspond to intermediates in the biosynthesis of the wile-type polysaccharides. The S → R mutation leads to the loss (or block) of an enzymes (transferase, synthetase) participating in the synthesis of the S polysaccharides. Recently many deficient mutants have been isolated and analysed, and in this way numerous stages in the biosynthesis of the cell-wall polysaccharides, for example, of Salmonella minnesota, have been made accessible to direct analysis. According to these investigations, the cell-wall polysaccharides of Salmonella consist of a basic polyheptose phosphate core which is bound to lipid A via ketodeoxyoctonic acid. To the basic core are linked pentasaccharide units of (R II structure). All other R forms are structurally deficient mutants of R II. In the complete polysaccharides of Salmonella S forms (wild types), the repeating oligosaccharide units of the specific side, chain are bound to the terminal N-acetylglucosamine of the R II structure.     

Mass‐Spectrometric Studies of Providencia SR‐Form Lipopolysaccharides and Elucidation of the Biological Repeating Unit Structure of Providencia rustigianii O14‐Polysaccharide

Enterobacteria Providencia are opportunistic human pathogens causing multiple types of infections. Earlier we have studied the S‐ and R‐form lipopolysaccharides (LPSs) of Providencia strains of various O‐serogroups and established the structures of the O‐polysaccharides (O‐antigens) and core‐region oligosaccharides, respectively. Now we report on mass spectrometric studies of oligosaccharides consisting of the core moiety with one O‐polysaccharide repeating unit attached, which were derived from the SR‐form LPSs of Providencia strains. The site of attachment of the O‐polysaccharide to the core and the structure of the O‐polysaccharide biological repeating unit were elucidated in Providencia rustigianii O14 using NMR spectroscopy.     

Full structure of the carbohydrate chain of the lipopolysaccharide of Providencia rustigianii O34

A lipopolysaccharide isolated from an opportunistic pathogen of the Enterobacteriaceae family Providencia rustigianii O34 was found to be a mixture of R-, SR-, and S-forms consisting of a lipid moiety (lipid A) that bears a core oligosaccharide, a core with one O-polysaccharide repeating unit attached, and a long-chain O-polysaccharide, respectively. The corresponding carbohydrate moieties were released from the lipopolysaccharide by mild acid hydrolysis and studied by sugar and methylation analyses along with one- and two-dimensional NMR spectroscopy and high-resolution electrospray ionization mass spectrometry. As a result, the structures of the core and the O-polysaccharide were established, including the structure of the biological repeating unit (an oligosaccharide that is preassembled and polymerized in biosynthesis of the O-polysaccharide), as well as the mode of the linkage between the O-polysaccharide and the core. Combining the structure of the carbohydrate moiety thus determined and the known structure of lipid A enabled determination of the full lipopolysaccharide structure of P. rustigianii O34.     

Specific polysaccharides of the lipopolysaccharides of Gram-negative bacteria

This review generalizes the result of structural investigations of the polysaccharides of the O-specific side chains of the lipopolysaccharides (LPSs) of the most studied representations of the family Enterobacteriaceae. The O-specific polysaccharides are the serologically dominating part of the molecule that is responsible for the O-antigenic specificity of the LPSs. The structures of the specific polysacchrides from various Gram-positive bacteria are given. The serological specificity of the O-antigens is discussed and a connection is shown between the chemical structures of the polysaccharides and their serological affinity.     

Synthesis of antigenic bacterial polysaccharides and their fragments

Conclusions The free trisaccharide-D-mannopyranosyl(1-4)--L-rhamnopyranosyl(1 3)-D-galactopyranose was obtained, which is the repeating unit of the backbone chain of the O-specific polysaccharides ofSalmonella of serological groups A, B, and D₁.Translated from Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, No.1, pp.165–167, January, 1976.     

Synthese von tri- und tetrasaccharid-einheiten des O-antigens aus Aeromonas salmonicida

The following oligosaccharide sequences containing the repeating unit of the O-specific chain of lipopolysaccharides from aeromonas salmonicida have been synthesized: α-D-Glcp-(1→3)-α-L-Rhap-(1 →3)-β-D-ManpNAcO(CH₂)₈CO₂Me, α-D-Glcp-(1 →4)-α-D-Glcp-(1→3)-α-L-Rhap-(1→3)-β-D-ManpNAcO(CH₂)₈CO₂Me and α-D-Glcp(1→4)-α-D-Glcp-(1→3)-[β-D-ManpNAc(1→4)]-L-Rha.     

Chemical and immunochemical characterization of the lipopolysaccharides of Yersinia kristensenii serovars 0:12,25; 0:12,26; and 0:25,35

A comparative and immunochemical characterization of the lipopolysaccharides of three serovars ofY. kristensenii has been performed and the ratios of the monosaccharides have been established. The results of the immunochemical investigations confirmed the serotyping of the microorganisms.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Branch, Academy of Sciences of the USSR, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 335–338, May–June, 1988.     

Pyrolysis–desorption chemical ionization and high-energy tandem mass spectrometry of the O-specific antigen of Yersinia ruckeri

Application of pyrolysis-desorption chemical ionization mass spectrometry (Py-DCI-MS) is demonstrated for the structural characterization of the native O-specific antigen of Yersinia ruckeri. Under proper pyrolytic and chemical ionization conditions this antigenic polysaccharide yields anhydrohexosamine fragments which facilitate identification of the trimeric repeating unit sequence. Tandem mass spectrometric analyses of the trimeric repeating unit fragments provided additional evidence for the proposed structure and permitted rationalization of the observed fragment ions. Furthermore, Py-DCI-MS of smaller hexosamine subunits isolated during off-line pyrolysis experiments showed fragmentation characteristics almost identical with those observed in the mass spectra of the whole antigen, hence confirming the stability of the pyrolysis fragments.     

Unusual monosaccharides in the O-factors of lipopolysaccharides of Gram-negative bacteria

Information is generalized on the structure of the O-specific polysaccharides forming components of the lipopolysaccharides of Gram-negative bacteria in which residues of unusual monosacchrides rarely found in nature have been identified. A chemical substantiation of the immunological specificity of the O-factors of specific chains is given. The role of the unusual monosaccharides in the manifestation of O-factor specificity is discussed.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Branch, USSR Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 743–763, November–December, 1989.     

Structural investigation of the lipopolysaccharide ofYersinia enterocolitica serovar 0:8

The lipopolysaccharide ofYersinia enterocolitica serovar 0:8 (strain 161) isolated from the microbial mass by aqueous-phenol extraction contains residues of L-fucose-6-deoxy-D-gulose, D-mannose, D-galactose, D-glucose, D- and L-glycero-D-mannoheptoses, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and 2-keto-3-deoxyoctonic acid (KDO). The polysaccharide was obtained by mild acid hydrolysis of the lipopolysaccharide followed by gel filtration on Sephadex G-50. On the basis of the results of monosaccharide analysis, methylation, Smith degradation, and partial hydrolysis the following structure is suggested for the repeating unit of the O-specific polysaccharide of the LPS ofYersinia enterocolitica, serovar 0:8:Pacific Ocean Institute of Bioorganic Chemistry of the Far-Eastern Scientific Center of the USSR Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedenenii, No. 5, pp. 657–664, September–October, 1987.