Structural studies of the lipopolysaccharide from Haemophilus parainfluenzae strain 20

Haemophilus parainfluenzae is a Gram-negative bacterium that colonizes the upper respiratory tract of humans and is a part of normal flora. In this study, we investigated the lipopolysaccharide (LPS) expressed by H. parainfluenzae strain 20. Using NMR and MS techniques on LPS, oligosaccharide samples and lipid A, the structures for O-antigen, core oligosaccharide and lipid A could be established. It was found that the biological repeating unit of the O-antigen is →4)-α-d-GalpNAc-(1→P→6)-β-d-Glcp-(1→3)-α-d-FucpNAc4N-(1→, in which d-FucpNAc4N is 2-acetamido-4-amino-2,4,6-trideoxy-d-galactose. This sugar is in β-configuration when linked to O-4 of the glucose residue of β-d-Galp-(1→2)-l-α-d-Hepp-(1→2)-[PEtn→6]-l-α-d-Hepp-(1→3)-[β-d-Glcp-(1→4)]-l-α-d-Hepp-(1→5)-[PPEtn→4]-α-Kdo-(2→6)-lipid A. LPS from a wbaP mutant of H. parainfluenzae strain 20 did not contain an O-antigen, consistent with the wbaP gene product being required for expression of O-antigen in fully extended LPS.     

Structure of the Bordetella trematum LPS O-chain subunit

Analysis of the O-chain subunit of the lipopolysaccharide (LPS, endotoxin) isolated from Bordetella trematum, a recently identified human pathogen, was undertaken. The polysaccharide (PS) moiety was shown to contain only two O-chain subunits, which differed in the anomeric bond of their first sugar. A trisaccharide fragment resulting from the cleavage of a FucNAc glycosidic bond was isolated after treatment of the PS with anhydrous HF. Nitrous deamination of the LPS led to the release of the following heptasaccharide corresponding to two trisaccharide subunits linked to an anhydromannitol residue. beta-ManNAc3NAmA-(1-4)-beta-ManNAc3NAmA-(1-3)-alpha-FucNAc-(1-4)-beta-ManNAc3NAmA-(1-4)-beta-ManNAc3NAmA-(1-3)-beta-FucNAc-(1-6)-2,5-anhManol.     

The structure of the O-polysaccharide from the lipopolysaccharide of Providencia alcalifaciens O30

The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Providencia alcalifaciens O30. Studies by sugar and methylation analyses along with (1)H and (13)C NMR spectroscopy, including two-dimensional (1)H,(1)H COSY, TOCSY, ROESY, and H-detected (1)H,(13)C HSQC, HMBC, and HMQC-TOCSY experiments, showed that the polysaccharide has a linear pentasaccharide repeating unit of the following structure:     

Structure of the O-polysaccharide leads to classification of Proteus penneri 31 in Proteus serogroup O19

O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide (LPS) of Proteus penneri strain 31. Sugar and methylation analyses along with NMR spectroscopic studies, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C and 1H,31P HMQC experiments, demonstrated the following structure of the polysaccharide: [carbohydrate structure: see text] where FucNAc is 2-acetamido-2,6-dideoxygalactose and EtnP is 2-aminoethyl phosphate. The polysaccharide studied has the same carbohydrate backbone as the O-polysaccharide of Proteus vulgaris O19. Based on this finding and close serological relatedness of the LPS of the two strains, it is proposed to classify P. penneri 31 in Proteus serogroup O19 as an additional subgroup. In contrast, D-GlcNAc6PEtn and alpha-L-FucNAc-(1-->3)-D-GlcNAc shared with a number of other Proteus O-polysaccharides could not provide any significant cross-reactivity of the corresponding LPS with rabbit polyclonal O-antiserum against P. penneri 31.     

Structural elucidation of the capsular polysaccharide isolated from Kaistella flava

The Gram-negative bacterium under study belongs to the genus Kaistella. It was isolated from a soil sample of the Haian Island in China, and it produces a lipophilic polysaccharide characterised by a branched hexasaccharide repeating unit, counting four 6-deoxy-alpha-l-mannose (Rha) residues, one 2-acetamido-2-deoxy-beta-d-glucose (GlcNAc) and a 2-acetamido-2,6-dideoxy-beta-d-galactose (FucNAc) unit. The structure of the repeating unit, assigned through 2D-NMR spectroscopy, is herein reported for the first time: [carbohydrate structure: see text]     

The elucidation of the structure of the core part of the LPS from Plesiomonas shigelloides serotype O17 expressing O-polysaccharide chain identical to the Shigella sonnei O-chain

Plesiomonas shigelloides O17 LPS contains the same O-antigenic polysaccharide chain as a causative agent of dysentery, Shigella sonnei. This polysaccharide can be used as a component of a vaccine against dysentery. Core part of the P. shigelloides O17 LPS was studied using NMR and mass spectrometry and the following structure was proposed: Significant similarity of the P. shigelloides O17 LPS core with the structure of the P. shigelloides O54 core was observed.