Teichoic acids of three type strains of the Bacillus subtilis group, Bacillus mojavensis VKM B-2650, Bacillus amyloliquefaciens subsp. amyloliquefaciens VKM B-2582, and Bacillus sonorensis VKM B-2652

Cell walls of three type strains of the Bacillus subtilis group, Bacillus mojavensis VKM B-2650, Bacillus amyloliquefaciens subsp. amyloliquefaciens VKM B-2582, and Bacillus sonorensis VKM B-2652, are characterized by the individual set of teichoic acids. All strains contained 1,3-poly(glycerol phosphates), unsubstituted, acylated with D-alanine, and glycosylated. The latter differ in the nature of the monosaccharide residue. Teichoic acids of B. mojavensis VKM B-2650^T and B. amyloliquefaciens subsp. amyloliquefaciens VKM B-2582^T contained α-glucopyranose, while those of B. sonorensis VKM B-2652^T contained β-glucopyranose and N-acetyl-α-D-glucosamine. Moreover, cell walls of B. mojavensis VKM B-2650^T contained a teichoic acid of poly(glycosylglycerol phosphate) nature with the following structure of the repeating unit: -4)-α-D-α-D-GlcpNAc-(1 → 3)]-Glcp-(1 → 2)-sn-Gro-(3-P-. The type strains have been characterized according to the composition of cell wall sugars and polyols. Application of teichoic acids (set and structure) as chemotaxonomic characteristics is discussed for six type strains of the Bacillus subtilis group. Polymer structures were determined by chemical and NMR spectroscopic techniques.     

Disaccharide 1-phosphate polymers of some representatives of the Bacillus subtilis group

Disaccharide 1-phosphate polymers as well as teichoic acids of various structures have been found in the cell walls of the representatives of the Bacillus subtilis group, namely Bacillus subtilis subsp. spizizenii VKM B-720 and VKM B-916, B. subtilis VKM B-517, and Bacillus vallismortis VKM B-2653^T. Disaccharide 1-phosphate polymers are composed of repeating units of the following structure: -P-4)-β-D-GlcpNAc-(1→6)-α-D-Galp-(1-, the N-acetylglucosamine residues are partially acetylated at positions O3 and O6 (VKM B-720 and VKM B-916); -P-4)-β-D-Glcp-(1→6)-α-D-GlcpNAc-(1-, the glucopyranose residues are partially acetylated at positions O2 or O3 (VKM B-517); -P-6)-α-D-GlcpNH ₃ ⁺ /α-D-GlcpNAc-(1→2)-α-D-Glcp-(1-, the N-acetylglucosamine residues are partially deacetylated (VKM B-2653^T). The structures of the two last disaccharide 1-phosphate polymers have not been reported so far for Gram-positive bacteria. The teichoic acids in the studied strains are O-D-alanyl-1,5-poly(ribitol phosphates) substituted with β-D-glucopyranose (VKM B-517, VKM B-720, VKM B-916) or 2-acetamido-2-deoxy-β-D-glucopyranose (VKM B-2653^T). The structures of the phosphate-containing polymers have been studied by chemical methods and by NMR spectroscopy.     

Cell wall glycopolymers of type strains from three species of the genus <Emphasis Type="Italic">Actinoplanes</Emphasis>

The structures of cell wall glycopolymers from the type strains of three Actinoplanes species were investigated using chemical methods, NMR spectroscopy, and mass spectrometry. Actinoplanes digitatis VKM Ac-649^T contains two phosphate-containing glycopolymers: poly(diglycosyl-1-phosphate) →6)-α-D-GlcpNAc-(1-P-6)-α-D-GlcpN-(1→ and teichoic acid →1)-sn-Gro-(3-P-3)-β-[β-D-GlcpNAc-(1→2]-D-Galp-(1→. Two glycopolymers were identified in A. auranticolor VKM Ac-648^T and A. cyaneus VKM Ac-1095^T: minor polymer–unsubstituted 2,3-poly(glycerol phosphate), widely abundant in actinobacteria (Ac-648^T), and mannan with trisaccharide repeating unit →2)-α-D-Manp-(1→2)-α-D-Manp(1→6)-α-D-Manp-(1→(Ac-1095^T). In addition, both microorganisms contain a teichuronic acid of unique structure containing a pentasaccharide repeating unit with two residues of glucopyranose and three residues of diaminouronic acids in D-manno- and/or D-gluco-configuration. Each of the strains demonstrates peculiarities in the structure of teichuronic acid with respect to the ratio of diaminouronic acids and availability and location of O-methyl groups in glucopyranose residues. All investigated strains contain a unique set of glycopolymers in their cell walls with structures not described earlier for prokaryotes.     

O-Specific Polysaccharides (O-Antigens) of a New Species of Enteric Bacteria Escherichia albertii Closely Related to Escherichia coli

Russian Journal of Bioorganic Chemistry - The data on the structure of O-specific polysaccharides (O-antigens) of all nine known molecular types (potential O-serotypes) of a new type of...     

Structure and serological characterization of the O-antigen of Proteus mirabilis O18 with a phosphocholine-containing oligosaccharide phosphate repeating unit

A phosphorylated, choline-containing polysaccharide was obtained by O-deacylation of the lipopolysaccharide (LPS) of Proteus mirabilis O18 by treatment with aqueous 12% ammonia, whereas hydrolysis with dilute acetic acid resulted in depolymerisation of the polysaccharide chain by the glycosyl phosphate linkage. Treatment of the O-deacylated LPS with aqueous 48% hydrofluoric acid cleaved the glycosyl phosphate group but, unexpectedly, did not affect the choline phosphate group. The polysaccharide and the derived oligosaccharides were studied by NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HMQC and HMQC-TOSCY experiments, along with chemical methods, and the following structure of the pentasaccharide phosphate repeating unit was established: [carbohydrate structure in text] Where ChoP=Phosphocoline Immunochemical studies of the LPS, O-deacylated LPS and partially dephosphorylated pentasaccharide using rabbit polyclonal anti-P. mirabilis O18 serum showed the importance of the glycosyl phosphate group in manifesting the serological specificity of the O18-antigen.     

Structure of the O-polysaccharide of Aeromonas hydrophila O:34; a case of random O-acetylation of 6-deoxy-L-talose

The O-polysaccharide of Aeromonas hydrophila O:34 was obtained by mild-acid degradation of the lipopolysaccharide and studied by chemical methods and NMR spectroscopy before and after O-deacetylation. The polysaccharide was found to contain D-Man, D-GalNAc and 6-deoxy-L-talose (L-6dTal), and the following structure of the tetrasaccharide repeating unit was established [carbohydrate structure see text] where 6dTal(I) is O-acetylated stoichiometrically at position-2 and 6dTal(II) carries no, one or two O-acetyl groups at any positions.     

Structure of an O-acetylated acidic O-specific polysaccharide of Proteus vulgaris O46

An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O46 and studied by chemical methods (O-deacetylation, sugar and methylation analyses, partial solvolysis) and 1H and 13C NMR spectroscopy. Solvolysis of the O-deacetylated polysaccharide with trifluoromethanesulfonic acid resulted in a alpha-D-GlcpNAc-(1 --> 3)-D-GlcA disaccharide that demonstrated the usefulness of this reagent for selective cleavage of heteropolysaccharides. The following structure for the polysaccharide was established: --> 4)-alpha-D-Glcp6Ac(1 --> 3)-beta-D-GlcpA4Ac-(1 --> 3)-alpha-D-GlcpNAc-(1 --> 3)-beta-D-GlcpA4Ac-(1 --> where the degree of O-acetylation is approximately 65% at position 6 of Glc and 80-95% at position 4 of GlcA residues.     

Structure of the O-specific polysaccharide of the bacterium Proteus mirabilis O29

An O-specific polysaccharide was obtained by mild acid degradation of P. mirabilis O29 lipopolysaccharide (LPS) and found to contain 2-acetamido-2-deoxy-D-galactose and D-glucuronic acid (D-GlcA) in the ratio 3:1. Studies of the polysaccharide by 1H- and 13C-NMR spectroscopy including two-dimensional correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), nuclear Overhauser effect spectroscopy (NOESY), and H-detected 1H,13C-heteronuclear multiple-quantum coherence (HMQC) experiments demonstrated the following structure of the branched tetrasaccharide repeating unit:     

Anionic polymers of the cell wall of Streptomyces sp. VKM An-2534

The cell wall of Streptomyces sp. VKM An-2534, the causative agent of common scab in potato tubers, which does not synthesize thaxtomin and is phylogenetically close to phytopathogen Streptomyces setonii sp. ATCC 25497, contains two anionic carbohydrate-containing polymers. The major polymer is teichuronic acid, whose repeating unit is disaccharide --> 4)-beta-D-ManpNAc3NAcyA-(1 --> 3)-alpha-D-GalpNAc-(1-->, where Acy is a residue of acetic or L-glutamic acid. The polymer of such structure has been found in Gram-positive bacteria for the first time. The minor polymer is teichoic acid [1,5-poly(ribitol phosphate)], in which a part of the ribitol residues are glycosylated at C4 with beta-D-Glcp and, probably, with beta-D-GlcpNAc and some residues are O-acylated with Lys residues. The structures were proved by chemical and NMR spectroscopic methods. It is likely that the presence of acidic polysaccharides on the surface of the phytopathogenic streptomycete is necessary for its attachment to the host plant.     

Temperature-dependent variations and intraspecies diversity of the structure of the lipopolysaccharide of Yersinia pestis

Yersinia pestis spread throughout the Americas in the early 20th century, and it occurs predominantly as a single clone within this part of the world. However, within Eurasia and parts of Africa there is significant diversity among Y. pestis strains, which can be classified into different biovars (bv.) and/or subspecies (ssp.), with bv. orientalis/ssp. pestis most closely related to the American clone. To determine one aspect of the relatedness of these different Y. pestis isolates, the structure of the lipopolysaccharide (LPS) of four wild-type and one LPS-mutant Eurasian/African strains of Y. pestis was determined, evaluating effects of growth at mammalian (37 degrees C) or flea (25 degrees C) temperatures on the structure and composition of the core oligosaccharide and lipid A. In the wild-type clones of ssp. pestis, a single major core glycoform was synthesized at 37 degrees C whereas multiple core oligosaccharide glycoforms were produced at 25 degrees C. Structural differences occurred primarily in the terminal monosaccharides. Only tetraacyl lipid A was made at 37 degrees C, whereas at 25 degrees C additional pentaacyl and hexaacyl lipid A structures were produced. 4-Amino-4-deoxyarabinose levels in lipid A increased with lower growth temperatures or when bacteria were cultured in the presence of polymyxin B. In Y. pestis ssp. caucasica, the LPS core lacked D-glycero-D-manno-heptose and the content of 4-amino-4-deoxyarabinose showed no dependence on growth temperature, whereas the degree of acylation of the lipid A and the structure of the oligosaccharide core were temperature dependent. A spontaneous deep-rough LPS mutant strain possessed only a disaccharide core and a slightly variant lipid A. The diversity and differences in the structure of the Y. pestis LPS suggest important contributions of these variations to the pathogenesis of this organism, potentially related to innate and acquired immune recognition of Y. pestis and epidemiologic means to detect, classify, control and respond to Y. pestis infections.