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Anionic phosphate-containing cell wall polymers of bacilli are represented by teichoic acids and poly(glycosyl 1-phosphates). Different locations of phosphodiester bonds in the main chain of teichoic acids as well as the nature and combination of the constituent structural elements underlie their structural diversity. Currently, the structures of teichoic acids of bacilli can be classified into three types, viz. poly(polyol phosphates) with glycerol or ribitol as the polyol; poly(glycosylpolyol phosphates), mainly glycerol-containing polymers; and poly(acylglycosylglycerol phosphate), in which the components are covalently linked through glycosidic, phosphodiester, and amide bonds. In addition to teichoic acids, poly(glycosyl 1-phosphates) with mono- and disaccharide residues in the repeating units have been detected in cell walls of several Bacillus subtilis and Bacillus pumilus strains. The known structures of teichoic acids and poly(glycosyl 1-phosphates) of B. subtilis, B. atrophaeus, B. licheniformis, B. pumilus, B. stearothermophilus, B. coagulans, B. cereus as well as oligomers that link the polymers to peptidoglycan are surveyed. The reported data on the structures of phosphate-containing polymers of different strains of B. subtilis suggest heterogeneity of the species and may be of interest for the taxonomy of bacilli to allow differentiation of closely related organisms according to the 'structures and composition of cell wall polymers' criterion.

cell wall, teichoic acids, Bacillus, poly(glycosyl 1-phosphates)

NCBI PubMed ID: 21999535
Journal NLM ID: 0376536
Publisher: Nauka/Interperiodica
Correspondence: potekchina@hotbox.ru
Institutions: Lomonosov Moscow State University, Russia

The teichoic acids (TAs) of type strains, viz. Bacillus licheniformis VKM B-511T and Bacillus pumilus VKM B-508T, as well as phylogenetically close bacteria VKM B-424, VKM B-1554, and VKM B-711 previously assigned to Bacillus pumilus on the basis of morphological, physiological, and biochemical properties, were investigated. Three polymers were found in the cell wall of each of the 5 strains under study. Strains VKM B-508T, VKM B-424, and VKM B-1554 contained polymers of the same core: unsubstituted 1,3-poly(glycerol phosphate) (TA I) and 1,3-poly(glycerol phosphate) with O-D-Ala and N-acetyl-a-D-glucosamine substituents (TA II and TA III', respectively). The cell walls of two remaining strains contained TA I, TA II, and a poly(glycosylpolyol phosphate) with the following structure of repeating units: -6)-a-D-GlcpNAc(1-1)-snGro-(3-P-(TA III'') in 'Bacillus pumilus' VKM B-711 (100% 16S rRNA gene similarity with the type strain of Bacillus safensis) and -6)-a-D-Galp-(1-2)-snGro-(3-P-(TA III''') in Bacillus licheniformis VKM B-511T. The simultaneous presence of three different TAs in the cell walls was confirmed by the NMR spectroscopic DOSY methods. The structure of the polymers and localization of O-D-Ala residues were investigated by the chemical and NMR spectroscopic methods.

NMR spectroscopy, teichoic acid, Bacillus licheniformis, taxonomy, Bacillus pumilus

Publication DOI: 10.1134/S0026261712030125
Journal NLM ID: 0376652
Publisher: Moskva: Izdatelstvo Nauka
Correspondence: streshinskaya@mail.ru
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, All-Russian Collection of Microorganisms (VKM), Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Russia, Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia
Methods: 13C NMR, 1H NMR, NMR-2D, 31P NMR, paper chromatography, composition analysis, genetic methods, paper electrophoresis

The structure of the capsular polysaccharide antigen of A. pleuropneumoniae serotype 7 was determined using 1D- and 2D-n.m.r. methods. Dephosphorylation, methylation, and g.l.c.-mass spectrometry methods were used to confirm the analysis. The type-7 specific polysaccharide was a high-molecular-weight teichoic acid-type polymer of D-galactose, glycerol, and phosphate (2:1:1), composed of glycosylglycerol repeating units joined through monophosphate diester linkages and with the structure: [formula: see text].

structure, serotype, capsular polysaccharide, Actinobacillus pleuropneumoniae

NCBI PubMed ID: 2036652
Publication DOI: 10.1016/0008-6215(91)80158-J
Journal NLM ID: 0043535
Publisher: Elsevier
Institutions: Division of Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6 Canada
Methods: 13C NMR, 1H NMR, methylation, GLC-MS, NMR-2D, HF solvolysis, sugar analysis, 31P NMR, acid hydrolysis, NMR-1D

Information on the prevalence, compositions, and structures of anionic carbohydrate-containing polymers of cell walls of Gram-positive bacteria is summarized. The data suggest that these polymers are important for normal functioning of bacterial cells and require further studies. Structural data on teichoic acids found in the literature published over the last few years are discussed. This is a very diverse class of polymers whose structure-specific pathways of degradation were studied and NMR spectra were examined. Unique comprehensive tables of 13C-NMR spectroscopic data (mainly obtained by the authors) on these polymers are given in the Appendix. Other tables summarize data on teichuronic acids, sugar-phosphate polymers, acid polysaccharides, and structural variants of bonds between acid polysaccharides and peptidoglycans known from the literature. Functions of anionic polymers and their possible chemotaxonomic applications are discussed

13C-NMR, NMR, structure, cell wall, composition, teichoic acid, teichuronic acid, Gram-positive, gram-positive bacteria, taxonomy

NCBI PubMed ID: 9360295
Journal NLM ID: 0376536
Publisher: Nauka/Interperiodica
Institutions: School of Biology, Lomonosov Moscow State University, Russia

Sugar nucleotide-dependent glycosyltransferases (GTs) are key enzymes that catalyze the formation of glycosidic bonds in nature. They have been increasingly applied in the synthesis of complex carbohydrates and glycoconjugates with or without in situ generation of sugar nucleotides. Human GTs are becoming more accessible and new bacterial GTs have been identified and characterized. An increasing number of crystal structures elucidated for GTs from mammalian and bacterial sources facilitate structure-based design of mutants as improved catalysts for synthesis. Automated platforms have also been developed for chemoenzymatic synthesis of carbohydrates. Recent progress in applying sugar nucleotide-dependent GTs in enzymatic and chemoenzymatic synthesis of mammalian glycans and glycoconjugates, bacterial surface glycans, and glycosylated natural products from bacteria and plants are reviewed.

carbohydrate, glycosyltransferase, enzymatic synthesis, sugar nucleotide, chemoenzymatic synthesis, biocatalyst

NCBI PubMed ID: 33310623
Publication DOI: 10.1016/j.cbpa.2020.10.007
Journal NLM ID: 9811312
Publisher: London: Elsevier
Correspondence: Chen Xi
Institutions: Department of Chemistry, University of California Davis, Davis, CA, US

Capsule polymers are crucial virulence factors of pathogenic bacteria and are used as antigens in glycoconjugate vaccine formulations. Some Gram-negative pathogens express poly(glycosylglycerol phosphate) capsule polymers that resemble Gram-positive wall teichoic acids and are synthesized by TagF-like capsule polymerases. So far, the biotechnological use of these enzymes for vaccine developmental studies was restricted by the unavailability of enantiopure CDP-glycerol, one of the donor substrates required for polymer assembly. Here, we use CTP:glycerol-phosphate cytidylyltransferases (GCTs) and TagF-like polymerases to synthesize the poly(glycosylglycerol phosphate) capsule polymer backbones of the porcine pathogen Actinobacillus pleuropneumoniae, serotypes 3 and 7 (App3 and App7). GCT activity was confirmed by high-performance liquid chromatography, and polymers were analyzed using comprehensive nuclear magnetic resonance studies. Solid-phase synthesis protocols were established to allow potential scale-up of polymer production. In addition, one-pot reactions exploiting glycerol-kinase allowed us to start the reaction from inexpensive, widely available substrates. Finally, this study highlights that multidomain TagF-like polymerases can be transformed by mutagenesis of active site residues into single-action transferases, which in turn can act in trans to build-up structurally new polymers. Overall, our protocols provide enantiopure, nature-identical capsule polymer backbones from App2, App3, App7, App9, and App11, Neisseria meningitidis serogroup H, and Bibersteinia trehalosi serotypes T3 and T15. IMPORTANCE Economic synthesis platforms for the production of animal vaccines could help reduce the overuse and misuse of antibiotics in animal husbandry, which contributes greatly to the increase of antibiotic resistance. Here, we describe a highly versatile, easy-to-use mix-and-match toolbox for the generation of glycerol-phosphate-containing capsule polymers that can serve as antigens in glycoconjugate vaccines against Actinobacillus pleuropneumoniae and Bibersteinia trehalosi, two pathogens causing considerable economic loss in the swine, sheep, and cattle industries. We have established scalable protocols for the exploitation of a versatile enzymatic cascade with modular architecture, starting with the preparative-scale production of enantiopure CDP-glycerol, a precursor for a multitude of bacterial surface structures. Thereby, our approach not only allows the synthesis of capsule polymers but might also be exploitable for the (chemo)enzymatic synthesis of other glycerol-phosphate-containing structures such as Gram-positive wall teichoic acids or lipoteichoic acids.

NMR, Neisseria meningitidis, polysaccharides, capsule, glycosyltransferase, teichoic acid, enzymatic synthesis, glycerol, glycoconjugate vaccine, antibiotic resistance, Recombinant Protein Expression, animal health, TagF-like polymerases, veterinary vaccine development

NCBI PubMed ID: 34076489
Publication DOI: 10.1128/mBio.00897-21
Journal NLM ID: 101519231
Publisher: Washington, DC: American Society for Microbiology
Correspondence: Timm Fiebig
Institutions: Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany, Fraunhofer International Consortium for Anti-Infective Research (iCAIR), Hannover, Germany, Department of Biosciences, University of Salzburg, Salzburg, Austria, Max Planck Institute for Dynamics of Complex Technical Systems, Bioprocess Engineering, Magdeburg, Germany
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, 31P NMR, Western blotting, cloning, enzymatic synthesis, HPLC-AEC, solid-phase synthesis, BLASTp