Structure of O-Polysaccharide and Lipid A of Pantoea Agglomerans 8488.

The Pantoea agglomerans 8488 lipopolysaccharide (LPS) was isolated, purified and characterized by monosaccharide and fatty acid analysis. The O-polysaccharide and lipid A components of the LPS were separated by mild acid degradation. Lipid A was studied by electrospray ionization mass spectrometry (ESI-MS) and found to consist of hexa-, penta-, tetra- and tri-acylated species. Two-dimensional nuclear magnetic resonance (NMR) spectroscopy revealed the following structure of the O-polysaccharide repeating unit →3)-α-L-Rhap-(1→6)-α-D-Manp-(1→3)-α-L-Fucp-(1→3)-ß-D-GlcNAcp-(1→. The LPS showed a low level of toxicity, was not pyrogenic, and reduced the adhesiveness index of microorganisms to 2.12, which was twofold less than the control. LPS modified by complex compounds of germanium (IV) and tin (IV) were obtained. It was found that six LPS samples modified by Sn compounds and two LPS samples modified by Ge compounds lost their toxic activity when administered to mice in a dose of LD50 (105 µg/mice or 5 mg/kg). However, none of the modified LPS samples changed their serological activity in an Ouchterlony double immunodiffusion test in agar.

Development and characterization of chitosan/polyvinyl alcohol polymer material with elastolytic and collagenolytic activities.

The biologically active polymeric material with entrapped peptidase Bacillus thuringiensis var. israelensis with caseinolytic, collagenase and elastase activities was developed as a promising product for medical use. We have evaluated and reported here the following physical-chemical and biochemical characteristics of entrapped enzyme: peptidase/polymer interaction and morphology analyses, film thickness, water content, time of dissolution in water and physiological saline, kinetic of casein hydrolysis and pH- and thermoprofiles of proteolytic activity. Scanning electron microscopy images shows the relative uniformity of the film surface with entrapped peptidase. The released peptidase was characterized by increased proteolytic activity in the acidic (14%-35%) and alkaline (7%-32%) regions. After nine months of storage, peptidase in chitosan/polyvinyl alcohol films retains more than 95% of its initial proteolytic activity. We consider this film as a perspective biotechnological agent in medicine.

Pantoea agglomerans P1a lipopolysaccharide: Structure of the O-specific polysaccharide and lipid A and biological activity.

O-specific polysaccharide and lipid A were obtained from the lipopolysaccharide from new strain of Рantoea agglomerans P1a by mild acid hydrolysis. It was found that the major form of lipid A presented by tetraacylated derivative containing biphosphorylated GlcN disaccharide, three 14:0 (3-OH) and 12:0 residues. The structure of the O-specific polysaccharide was established by chemical, NMR and computational methods: →3)-α-D-Manp-(1 → 4)-ß-D-Fucp-(1 → 4)-ɑ-D-Fucp-(1→The LPS of Р. agglomerans P1a showed low level of toxicity and pyrogenicity to compare with LPS of E. coli O55:B5 and pyrogenal (respectively).

Lipopolysaccharides of Pantoea agglomerans 7604 and 8674 with structurally related O-polysaccharide chains: Chemical identification and biological properties.

Structurally related O-specific polysaccharide (O-antigen) and lipid A components were obtained by mild acid degradation of the lipopolysaccharides (LPSs) of two strains of bacteria Pantoea agglomerans, 7604 and 8674. Studies by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy enabled elucidation of the following structures of the O-polysaccharides, which differ only in the linkage configuration of a side-chain glucose residue: R=α-d-Glcp in strain 7604 or ß-d-Glcp in strain 8674 Lipid A samples were studied by GC-MS and high-resolution ESI-MS and found to be represented by penta- and tetra-acyl species; lipid A of strain 8674 also included hexaacyl species. A peculiar feature of lipid A of both strains is the presence of the major cis-9-hexadecenoic (palmitoleic) acid, which has not been found in P. agglomerans strains studied earlier. The LPSs of both strains were pyrogenic, reduced the average adhesion and the index of adhesiveness and showed a relatively low level of lethal toxicity. O-antiserum against strain 7604 showed one-way cross-reactivity with the LPS of strain 8674, and O-antisera against both strains cross-reacted with LPSs of some other Р. agglomerans strains but more strains were serologically unrelated. These structural and serological data indicate immunochemical heterogeneity of Р. agglomerans strains and will find demand in classification of Р. agglomerans by O-antigens.

Lipopolysaccharide of Pantoea agglomerans 7969: Chemical identification, function and biological activity.

Lipopolysaccharide (LPS) of Pantoea agglomerans 7969 isolated from apple tree was purified and characterized chemically by sugar and fatty acid analysis. Lipid A was analysed by negative-ion mode ESI MS and found to consist mainly of hexa- and tetra-acyl species typical of E. coli lipid A. The O-specific polysaccharide of the LPS was studied by sugar analysis, Smith degradation, and one- and two-dimensional 1H and 13C NMR spectroscopy. The polysaccharide is built up of linear tetrasaccharide repeating units, and about ∼25% repeats contain glycerol 1-phosphate on the GlcNAc residue: →3)-α-l-Rha p-(1→6)-α-d-Man p-(1→3)-α-d-Fuc p-(1→3)-ß-d-Glc pNAc-(1→∼25% Gro-1-P-(O→6)⌋ The LPS showed low levels of toxic and pyrogenic activities and reduced the average adhesion and the index of adhesiveness.

Isolation and structure elucidation of two different polysaccharides from the lipopolysaccharide of Rahnella aquatilis 33071T.

Two different polysaccharides were obtained by mild acid degradation of the lipopolysaccharide of Rahnella aquatilis 33071(T). These were studied by sugar and methylation analyses along with 1D and 2D (1)H and (13)C NMR spectroscopy. The following structures were established for the polysaccharides: -->4)-alpha-D-Rhap-(1-->3)-alpha-D-Rhap-(1-->3)beta-D-Galp-(1-->3)-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1--> beta-D-Glcp-(1-->3)-alpha-D-galp-(1-->4)-alpha-D-GlcpA-(1-->2). The former structure is new, whereas the latter has been reported earlier as the structure of the O-specific polysaccharide of R. aquatilis 95 U003 (Zdorovenko, E. L.; Varbanets, L. D.; Zatonsky, G. V.; Kachala, V. V.; Zdorovenko, G. M.; Shashkov, A. S.; Knirel, Y. A. Carbohydr. Res.2008, 343, 2494-2497).

Structure of the O-polysaccharide of the lipopolysaccharide of Rahnella aquatilis 1-95.

The O-polysaccharide was isolated by mild acid hydrolysis of the lipopolysaccharide of Rahnella aquatilis 1-95 and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including NOESY and 1H,13C HSQC experiments for linkage and sequence analysis. The following structure of the branched trisaccharide repeating unit of the O-polysaccharide was established: [carbohydrate structure: see text].