Bacterial Polysaccharide Structures Research Articles

Structure elucidation and gene cluster of the O-antigen of Shewanella xiamenensis strain DCB-2-1 containing an amide of d-glucuronic acid with d-alanine and its bonding with U, Cr and V

The aim of this work was to examine the structure and gene cluster of O-OPS of S. xiamenensis strain DCB-2-1 and survey its conceivability for chelating uranyl, chromate and vanadate ions from solution. O-polysaccharide (OPS, O-antigen) was isolated from the lipopolysaccharide of Shewanella xiamenensis DCB-2-1 and studied by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy and sugar analysis. The following structure of the brunched pentasaccharide was established: [Display omitted] where d-β-GlcpA(d-Ala) is d-glucuronic acid acylated with NH group of d-Ala. The OPS structure established is unique among known bacterial polysaccharide structures. Interestingly, that dN-(d-glucuronoyl)-d-alanine derivative is not found in bacterial polysaccharides early.The O-antigen gene cluster of Shewanella xiamenensis strain DCB-2-1 has been sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in agreement with the OPS structure. Based on the analysis of the IR spectra of the isolated polysaccharide DCB-2-1 and the products of its interaction with UO2(NO3)2 ∗ 6H2O, NH4VO3 and K2Cr2O7, a method of binding them can be proposed. Laboratory experiments show that the use of polysaccharide can be effective in removing uranyl, chromate and vanadate from solution.

Structure and gene cluster of the O-antigen of Escherichia coli strain SDLZB008

The O-polysaccharide (O-antigen) of Escherichia coli SDLZB008 was isolated from the lipopolysaccharide and studied by sugar analyses along with 1H and 13C NMR spectroscopy. The following structure of the branched pentasaccharide repeating unit was established, which is unique among the known structures of bacterial polysaccharides:▪The O-antigen gene cluster of E. coli SDLZB008 has been sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the O-polysaccharide structure.

The unique structure of bacterial polysaccharides - Immunochemical studies on the O-antigen of Proteus penneri 4034-85 clinical strain classified into a new O83 Proteus serogroup

The serological classification scheme of the opportunistic Proteus bacilli includes a number of Proteus penneri strains. The tested P. penneri 4034-85 strain turned out to be serologically distinguished in ELISA and Western blotting. The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of this strain and studied by sugar and methylation analyses and dephosphorylation along with 1H and 13C NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HSQC, HMBC, and HSQC-TOCSY experiments, The O-polysaccharide was found to have a linear repeating unit containing glycerol 1-phosphate and two residues each of Gal and GlcNAc. The following O-polysaccharide structure was established, which, to our knowledge, is unique among known bacterial polysaccharide structures:→4)-β-d-GlcpNAc-(1→3)-α-d-Galp-(1→3)-β-d-GlcpNAc-(1→2)-β-d-Galp-(1→3)-Gro-1-P-(O→.

Structural and genetic characterization of the colitose-containing O-specific polysaccharide from the lipopolysaccharide of Herbaspirillum frisingense GSF30T

The lipopolysaccharide (LPS) of Herbaspirillum frisingense GSF30T (HfGSF30), a non-pathogenic diazotrophic endobiont, was isolated by phenol-water extraction from bacterial cells and was characterized by chemical analyses and SDS PAGE. The O-specific polysaccharide (OPS, O-antigen), obtained by mild acid hydrolysis of the LPS, was examined by sugar and methylation analysis, along with 1H and 13C NMR spectroscopy, including 2D 1H,1H COSY, 1H,1H TOCSY, 1H,1H ROESY, 1H,13C HSQC, and 1H,13C HMBC experiments. The OPS was found to consist of branched tetrasaccharide repeating units of the following structure:→3)−α−L−Rhap−1→3−β−D−Galp−1→3−β−D−GlcpNAc−(1→2↑1α−ColpThis structure is unique among the known bacterial polysaccharide structures. Analysis of the HfGSF30 genome showed that it contained a set of sequentially arranged operons (presumably a cluster of genes) associated with the O-antigen. Amino acid sequence analysis using the BLAST program demonstrated the specificity of this putative cluster for Herbaspirillum spp. The genes responsible for the biosynthesis of the OPS of HfGSF30 were dispersed in the genome, constituting small operons. A putative O-antigen gene cluster of HfGSF30 was identified and found to be consistent with the OPS structure.

Structure and gene cluster of the O antigen of Escherichia coli F17, a candidate for a new O-serogroup

Escherichia coli F17 isolated from horse feces was studied in respect to the O antigen (O polysaccharide) structure and genetics. The lipopolysaccharide was isolated by phenol-water extraction of bacterial cells and cleaved by mild acid hydrolysis to yield the O polysaccharide, which was studied by sugar analysis and selective solvolysis with CF3CO2H along with one- and two-dimensional 1H and 13C NMR spectroscopy. The O polysaccharide was found to have a branched pentasaccharide repeat (O-unit) containing one residue each of d-galactose, d-mannose, l-rhamnose, d-glucuronic acid, and N-acetyl-d-glucosamine; about 2/3 units bear a side-chain glucose residue. To our knowledge, the F17 O-polysaccharide structure established is unique among known bacterial polysaccharide structures. The O-antigen gene cluster of E. coli F17 between the conserved genes galF and gnd was sequenced and found to be 99% identical to that of E. coli 102,755 assigned to a novel OgN8 genotype (A. Iguchi, S. Iyoda, K. Seto, H. Nishii, M. Ohnishi, H. Mekata, Y. Ogura, T. Hayashi, Front. Microbiol. 7 (2016) 765). Genes in the cluster were annotated taking into account the F17 O-polysaccharide structure. The data obtained confirm that E. coli F17 and E. coli strains belonging to the OgN8 genotype can be considered as a candidate to a new E. coli O-serogroup. The O antigen of this novel type was demonstrated to make for an effective shield protecting the intimate outer membrane surface of bacteria from direct interaction with bacteriophages.

Structure of the O-polysaccharide of Escherichia coli O60

Structure of the O-polysaccharide (O-antigen) of Escherichia coli O60 was studied by sugar analysis, partial solvolysis with CF3CO2H, and 1D and 2D 1H and 13C NMR spectroscopy. The O-polysaccharide was found to consist of D-galactose and L-rhamnose. The structure of its branched tetrasaccharide repeating unit was established, which is unique among known bacterial polysaccharide structures.

Biosynthesis of lipopolysaccharides with different length of the O-specific region as a virulence factor of Gram-negative bacteria

The outer membrane of Gram-negative bacteria is a biological structure with a unique composition that significantly contributes to the survival of bacteria in the unfavorable conditions of the host organism. The lipopolysaccharide constitutes about 70% of the external part of the outer membrane. The LPS molecule is composed of three different parts: lipid A, core oligosaccharide and O antigen. Despite the O-specific antigen being one of the most intensely studied surface structures of bacterial polysaccharides, a number of questions regarding the mechanism of the O antigen biosynthesis and its transport to the cell surface are still unanswered. The paper describes the biosynthesis of the lipopolysaccharide molecule, with particular emphasis on the O-specific chain biosynthesis, the mechanism of lipopolysaccharide length regulation and the influence of the type of synthesized O-specific chains on bacterial survival in adverse host organisms.

Structural studies on the O-polysaccharide of Escherichia coli O57

Mild acid hydrolysis of the lipopolysaccharide of Escherichia coli O57 afforded an O-polysaccharide, which was isolated by gel permeation chromatography (GPC) and studied by sugar analysis, Smith degradation and solvolysis with trifluoroacetic acid, along with 2D 1H and 13C NMR spectroscopy. The O-polysaccharide was found to contain d-Glc, d-Gal, d-GalA, d-GlcNAc, and l-FucNAc, as well as O-acetyl groups. Smith degradation of the O-deacetylated polysaccharide destroyed side-branch β-Glсp and α-GalpA to give a modified linear polysaccharide. Solvolysis cleaved selectively the linkage of α-l-FucpNAc to give a pentasaccharide corresponding to the O-polysaccharide repeat. A comparison of the NMR spectra of the initial and O-deacetylated polysaccharides showed that α-GalpA is non-stoichiometrically O-acetylated at position either 2 (∼30%) or 3 (∼40%). The following structure of the O-polysaccharide was established, which is unique among known bacterial polysaccharide structures: [Display omitted]

Structure and genetics of the O-specific polysaccharide of Escherichia coli O27

The O-specific polysaccharide (O-antigen) is a part of the lipopolysaccharide on the cell surface of Gram-negative bacteria. The O-polysaccharide was obtained by mild acid hydrolysis of the lipopolysaccharide of Escherichia coli O27 and studied by sugar analysis and Smith degradation along with 1H and 13C NMR spectroscopy. The following structure of the branched hexasaccharide repeating unit was established, which is unique among known structures of bacterial polysaccharides: [Display omitted] where GlcA is non-stoichiometrically O-acetylated at position 3 (∼22%) or 4 (∼37%). Functions of genes in the O-antigen gene cluster of E. coli O27 were tentatively assigned by comparison with sequences in the available databases and found to be consistent with the O-polysaccharide structure.

Structures and gene clusters of the O-antigens of Escherichia albertii O3, O4, O6, and O7

The O-specific polysaccharides (OPSs) called O-antigens were obtained by mild acid degradation of the lipopolysaccharides of Escherichia albertii serotypes O3, O4, O6, and O7 and studied by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy. The following structure was established for the OPS of E. albertii O4, which, to our knowledge, is unique among known bacterial polysaccharide structures:→2)-α-l-Rhap-(1 → 2)-α-l-Fucp-(1 → 2)-β-d-Galp-(1 → 3)-α-d-GalpNAc-(1 → 3)-β-d-GlcpNAc-(1→The OPS structure of the strain of E. albertii O7 studied was identical to that of strain LMG 20973 (= Albert 10457), whose structure has been reported earlier (R. Eserstam et al. Eur. J. Biochem. 269 (2002) 3289–3295). E. albertii O3 and O6 shared the OPS structures with Escherichia coli O181 and O3, respectively, except for the lack of O-acetylation in E. albertii O3, which is present in E. coli O181. The gene clusters driving the O-antigen biosynthesis of the E. albertii strains were sequenced, the genes were annotated by comparison with sequences in the available databases, and the predicted functions of the encoded proteins were found to be consistent with the OPS structures established. In accordance with the relatedness of the OPS structures, the O-antigen gene clusters of E. albertii O3 and O6 contain the same genes and have the same organization as those of E. coli O181 and O3, the entire gene clusters being 83% and 98% identical, respectively.

Influence of Feed Composition on the Monomeric Structure of Free Bacterial Extracellular Polysaccharides in Anaerobic Digestion.

Six 5.0-L fill-and-draw batch reactors were used with different feed compositions containing a range of carbohydrates (glucose, sucrose, fructose) and nitrogen sources (urea, NH4Cl) at various concentrations to investigate free extracellular polysaccharide (EPS) production during anaerobic digestion (AD). This work analyzed not only their monosaccharide components, but also their specific linkage patterns and the change associated with different chemical nature in carbon substrates or nitrogen sources; all of these parameters can have profound biological consequences, and were correlated to macronutrients present in the feed. It is believed that feed composition is a major factor which determines the physicochemical characteristics of the free EPS. Our findings also suggest that the differences associated with the digestion of various carbon substrates and/or nitrogen sources could alter monomeric saccharide composition and concentrations of the free EPS. Such insights demonstrate that previous studies on feed C/N ratios tended to overestimate EPS production, while variations in the chemical nature of the nitrogen source were overlooked. Our results also link the physiochemical properties of free EPS with underlying biofouling mechanisms, and demonstrate that membrane fouling is to some extent dependent upon the prevailing nutritional environment and feed composition.

Studies on the O-specific polysaccharide of the lipopolysaccharide from the Pseudomonas mediterranea strain C5P1rad1, a bacterium pathogenic of tomato and chrysanthemum

An O-specific polysaccharide (OPS) was isolated from the lipopolysaccharide of Pseudomonas mediterranea strain C5P1rad1, the causal agents of tomato pith necrosis and Chrysanthemum stem rot, and studied by one- and two-dimensional 1H and 13C NMR spectroscopy. The following structure of the trisaccharide repeating unit of the OPS was established, which, to our knowledge, is unique among the known bacterial polysaccharide structures:→4)-β-d-ManpNAc3NAcA-(1 → 4)-β-d-ManpNAc3NAcA-(1 → 3)-α-d-QuipNAc4NAc-(1→where QuiNAc4NAc and ManNAc3NAcA indicate 2,4-diacetamido-2,4,6-trideoxyglucose and 2,3-diacetamido-2,3-dideoxymannuronic acid, respectively. Pre-treatment of leaves with LPS or OPS preparations at 250 and 50 μg mL−1 did not inhibit development of a hypersensitivity reaction induced by P. mediterranea C5P1rad1 on tobacco, tomato and chrysanthemum plants. The same preparations at 250 μg mL−1 partially prevented elicitation of the hypersensitivity reaction by Pseudomonas syringae KVPT7RC on chrysanthemum but not tobacco and tomato.

Structure and gene cluster of the O-antigen of Escherichia albertii O1 resembling the O-antigen of Pseudomonas aeruginosa O5

The O-specific polysaccharide (O-antigen) was obtained by mild acid degradation of the lipopolysaccharide of Escherichia albertii serotype O1 strain SP20140089 and studied by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy. The following structure was established for the trisaccharide repeating unit of the O-polysaccharide:→4)-β-d-ManpNAc3NAcA-(1 → 4)-β-d-GlcpNAm3NAcA-(1 → 3)-α-d-GlcpNAc-(1→where ManNAc3NAcA and GlcNAm3NAcA indicate 2,3-diacetamido-2,3-dideoxymannuronic acid and 2-acetimidoylamino-3-acetamido-2,3-dideoxyglucuronic acid, respectively. While showing some similarity with O-polysaccharide structures of a group of Pseudomonas aeruginosa serotypes (O2, O5, O16, O18, and O20), that of E. albertii O1 is unique among known bacterial polysaccharide structures. The gene cluster for biosynthesis of the O1-antigen was sequenced and functions of the genes were predicted by comparison with sequences in the available databases, including those involved in the synthesis of nucleotide precursors of 2,3-diamino-2,3-dideoxyhexuronic acid derivatives in P. aeruginosa O5.

Identification serologically, chemically and genetically of two Escherichia coli strains as candidates for new O serogroups.

Escherichia coli strains are normally identified by the combination of their O and H (and sometimes K) antigens, and serotyping based on the antigens is believed to be crucial for clinical detection and epidemiological investigation. Two E. coli strains, G5413 and G5287, were isolated from faecal samples of female patients with diarrhoea and were not agglutinated with any antisera that cover the well-known O serogroups of E. coli. We elucidated the O-polysaccharide (OPS) structures and analysed the O-antigen gene clusters of these bacteria. The OPS structure of G5413 established by monosaccharide analysis and NMR spectroscopy was found to be unique amongst known bacterial polysaccharide structures. The O-antigen gene cluster of this strain was sequenced and did not match sequence data with any of the 184 O serogroups that have been recognized internationally. Gene functions were tentatively assigned and were appropriate to the OPS structure. Based on these data, we suggest G5413 as a candidate for a new E. coli O serogroup. Both the OPS structure and O-antigen gene cluster of G5287 were identical to those of E. coli L-19, a candidate for another new O serogroup characterized by us recently. Recognition of these two provisional O serogroups increases the number of known O-antigen forms of E. coli to 186.

Structural and genetic studies of the O-antigen of Escherichia coli O163

An acidic O-polysaccharide (O-antigen) of Escherichia coli O163 was obtained by mild acid hydrolysis of the lipopolysaccharide and studied by sugar analysis and Smith degradation along with 1D and 2D 1H and 13C NMR spectroscopy. The following structure of the linear tetrasaccharide repeating unit was established, which is unique among known structures of bacterial polysaccharides: [Display omitted] Functions of genes in the O-antigen gene cluster of E. coli O163 were tentatively assigned by comparison with sequences in the available databases and found to be in agreement with the O-polysaccharide structure. Relationships between O-antigen structures and gene clusters of E. coli O163 and Salmonella enterica O41 are discussed.

Structure and gene cluster of the O antigen of Escherichia coli L-19, a candidate for a new O-serogroup.

Escherichia coli L-19 isolated from a healthy individual did not agglutinate with any of 21 polyvalent antisera that cover 174 E. coli O-serogroups. The strain was studied in respect to the O-antigen (O-specific polysaccharide, OPS) structure and genetics. The LPS was isolated by phenol-water extraction of bacterial cells and cleaved by mild acid hydrolysis to yield the OPS. The OPS was studied by sugar and methylation analyses, along with 1D and 2D (1)H and (13)C NMR spectroscopy. The established structure of the linear tetrasaccharide repeating unit was found to be unique among known bacterial polysaccharide structures. A peculiar component of the L-19 OPS was an amide of glucuronic acid with 2-amino-1,3-propanediol (2-amino-2-deoxyglycerol) (GroN). The O-antigen gene cluster of L-19 between the conserved genes galF and gnd was sequenced, and gene functions were tentatively assigned by a comparison with sequences in the available databases and found to be in agreement with the OPS structure. Except for putative genes for synthesis and transfer of GroN, the sequences in the L-19 O-antigen gene cluster were little related to those of reference strains of the 174 known E. coli O-serogroups. The data obtained suggest that L-19 can be considered as a candidate for a new E. coli O-serogroup.

Structure and gene cluster of the O-antigen of Escherichia coli O30

The acidic O-polysaccharide (O-antigen) of Escherichia coli O30 was isolated from the lipopolysaccharide and studied by sugar analysis and NMR spectroscopy. The following structure of the branched tetrasaccharide repeating unit was established, which is unique among known structures of bacterial polysaccharides: [Display omitted] The O-antigen gene cluster of E. coli O30 was sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the O-polysaccharide structure.

Structure and gene cluster of the O-antigen of Escherichia coli O36

The O-polysaccharide (O-antigen) of Escherichia coli O36 was isolated from the lipopolysaccharide and studied by sugar analyses and Smith degradation along with 1H and 13C NMR spectroscopy. The following structure of the branched pentasaccharide repeating unit was established, which is unique among the known structures of bacterial polysaccharides: [Display omitted] The O-antigen gene cluster of E. coli O36 has been sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the O-polysaccharide structure.

Structure, biosynthesis, and function of bacterial capsular polysaccharides synthesized by ABC transporter-dependent pathways

Bacterial capsules are formed primarily from long-chain polysaccharides with repeat-unit structures. A given bacterial species can produce a range of capsular polysaccharides (CPSs) with different structures and these help distinguish isolates by serotyping, as is the case with Escherichia coli K antigens. Capsules are important virulence factors for many pathogens and this review focuses on CPSs synthesized via ATP-binding cassette (ABC) transporter-dependent processes in Gram-negative bacteria. Bacteria utilizing this pathway are often associated with urinary tract infections, septicemia, and meningitis, and E. coli and Neisseria meningitidis provide well-studied examples. CPSs from ABC transporter-dependent pathways are synthesized at the cytoplasmic face of the inner membrane through the concerted action of glycosyltransferases before being exported across the inner membrane and translocated to the cell surface. A hallmark of these CPSs is a conserved reducing terminal glycolipid composed of phosphatidylglycerol and a poly-3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) linker. Recent discovery of the structure of this conserved lipid terminus provides new insights into the early steps in CPS biosynthesis.

Structure and gene cluster of the O-antigen of Escherichia coli O120

The acidic O-polysaccharide (O-antigen) of Escherichia coli O120 was isolated from the lipopolysaccharide and studied by sugar analysis and NMR spectroscopy. The following structure of the branched hexasaccharide repeating unit was established, which is unique among the known structures of bacterial polysaccharides:▪The O-antigen gene cluster of E. coli O120 was sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the O-polysaccharide structure.