[Specific structural features and immunomodulatory properties of the lipopolysaccharides of Pseudomonas bacteria].

The results of in vitro studies of the immunomodulatory action of the lipopolysaccharides (LPS) of the Pseudomonas bacteria--P. fluorescens biovar I strains IMV 4125 = ATCC 13525, IMV 7769, and IMV 1152; P. fluorescens biovar IV strain IMV 2111; P. syringae pv. syringae IMV 281 = CPPB 281 = ATCC 19310 and IMV 467; and P. wieringae IMV 7923--on the mouse spleenocytes and human peripheral blood mononuclear cells (PBMC), B lymphocytes, and T lymphocytes are described. The proliferative activity of mouse spleenocytes correlated with the degree of LPS toxicity. The PBMC mitogenic activity induced by the P. fluorescens IMV 7769 LPS preparation exceeded the activity of E. coli 026:B6 LPS. The immunomodulatory effect of LPS on T cells was strain and dose dependent. The LPS of P. syringae pv. syringae INV 467 displayed a comparatively pronounced immunomodulatory effect on human blood B lymphocytes.

[Characterization of lipopolysaccharides from Pseudomonas fluorescens IMB 2108 (biovar II) and IMB 2111 (biovar IV) with O-chains represented by alpha-glucan].

Results of studies of the structurally unique O-chains of lipopolysaccharides, which were isolated from the dry biomass of Pseudomonas fluorescens IMB 2108 (biovar II) and IMB 2111 (biovar IV) by the Westphal technique and purified by repeated ultracentrifugation, are reported. The bulk of the lipopolysaccharide preparations contained S- and R-molecules at an average molar ratio of 1: 2. The main components of the hydrophobic moiety of lipid A were 3-hydroxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, dodecanoic, hexadecanoic, and octadecanoic acids, as well as hexadecenoic and octadecenoic acids. Glucosamine and phosphoethanolamine were identified as components of the hydrophilic moiety of lipid A. The degree of lipid A phosphorylation amounted to 3-4%. Fractions of the core oligosaccharide contained glucose, galactose, mannose, rhamnose, arabinose, glucosamine (only in strain IMB 2108), alanine, phosphoethanolamine, phosphorus, and 2-keto-3-deoxyoctulosonic acid (KDO). Heptose was present in trace amounts. O-specific polysaccharide chains were represented by a linear polymer of D-glucose units, which were linked together via alpha-(1,4) glycoside bonds. The existence of P. fluorescens strains that have alpha-1,4-glucan as the O-chain of their lipopolysaccharides has not been described before.

[A comparative analysis of the ice nucleation activity of pseudomonad cells and lipopolysaccharides]. / Sravnitel'nyi analiz l'doobrazuiushchikh svoistv kletok i lipopolisakharidov bakterii roda Pseudomonas.

The paper deals with the study of the ice nucleation activity of the cells, extracellular lipopolysaccharides (ELPSs), lipopolysaccharides (LPSs), and their structural components (lipid A, core oligosaccharide, and O-specific polysaccharide) of Pseudomonas fluorescens, P. syringae, P.fragi, and P. pseudoalcaligenes. The aqueous suspensions of the intact cells of P. syringae IMV 1951 and IMV 185 began to freeze at -1 and -4 degrees C, respectively. This suggests that these cells possess ice nucleation activity. The aqueous cell suspensions of two other strains, P. fluorescens IMV 1433 and IMV 2125, began to freeze at lower temperatures than did distilled water (-9 degrees C), which suggests that the cells of these strains possess antifreeze activity. The ice nucleation activity of the bacterial strains studied did not show any correlation with their taxonomic status. The ice nucleation activity of ELPSs depended little on their concentration (within a concentration range of 0.2-0.4%). In most cases, the ice nucleation activity of ELPSs, LPSs, and their structural components differed from that of the intact cells from which these biopolymers were obtained. This may indicate that the biopolymers under study play a role in ice nucleation, but this role is not crucial. The relationship between the structure of LPSs and their effect on ice nucleation is discussed.

[Structure and properties of the lipopolysaccharide of Pseudomonas fluorescens IMV 2366 (biovar III)]. / Stroenie i svoistva lipopolisakkharida Pseudomonas fluorescens IMV 2366 (biovar III).

The lipopolysaccharide (LPS) preparation isolated from the bacterial mass of Pseudomonas fluorescens IMV 2366 (biovar III) by Westphal's method and purified by repeated ultracentrifugation was characterized by the presence of the S- and R-forms of molecules. The following structural portions of the LPS molecule were obtained in the individual state and characterized: lipid A, core oligosaccharide, and O-specific polysaccharide. The main components of the lipid A hydrophobic moiety were 3-hydoxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, dodecanoic, and hexadecanoic fatty acids. Glucosamine, phosphoethanolamine, and phosphorus were identified as the components of the lipid A hydrophilic moiety. Rhamnose, glucose, galactose, glucosamine, galactosamine, alanine, phosphoethanolamine, phosphorus, 2-keto-3-desoxyoctulosonic acid (KDO), as well as 2-amino-2,6-didesoxygalactose (FucN) and 3-amino-3,6-didesoxyglucose (Qui3N), were revealed in the composition of the core oligosaccharide fractions. O-specific polysaccharide chains were established to be composed of repeating trisaccharide units consisting of residues of L-rhamnose (L-Rha), 2-acetamido-2,6-didesoxy-D-galactose (D-FucNAc), and 3-acylamido-3,6-didesoxy-D-glucose (D-Qui3NAcyl), where Acyl = 3-hydroxy-2,3-dimethyl-5-hydroxyprolyl. Neither double immunodiffusion in agar not the immunoenzyme assay revealed serological relations between the strain studied and the P. fluorescens strains studied earlier.

[Comparative characteristics of lipopolysaccharides of various Pseudomonas fluorescens strains (Biovar I)]. / Sravnitel'naia kharakteristika lipopolisakharidov razlichnykh shtammov Pseudomonas fluorescenns (biovar I).

From the biomass of five Pseudomonas fluorescens biovar I strains, including the P. fluorescens type strain IMV 4125 (ATCC 13525), lipopolysaccharides (LPS) were isolated (by extraction with a phenol-water mixture followed by repeated ultracentrifugation), as well as individual structural components of the LPS macromolecule: lipid A, the core oligosaccharide, and O-specific polysaccharide (O-PS). 3-Hydroxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, dodecanoic, hexadecanoic, octadecanoic, hexadecenoic, and octadecenoic fatty acids were present in lipid A of the LPS of all the strains studied. Glucosamine, ethanolamine, and phosphoethanolamine were revealed in the lipid A hydrophilic part of all of KDO, a trace amount of heptoses, ethanolamine, phosphoethanolamine, alanine, and phosphorus were identified as the main core components. Interstrain differences in the core oligosaccharide composition were revealed. Structural analysis showed that the O-PS of the type strain, as distinct from that of other strains, is heterogeneous and contains two types of repetitive units, including (1) three L-rhamnose residues (L-Rha), one 3-acetamide-3,6-dideoxy-D-galactose residue (D-Fuc3NAc) as a branching substitute of the L-rhamnan chain and (2) three L-Rha residues and two branching D-Fuc3NAc residues. The type strain is also serologically distinct from other biovar I strains due to the LPS O-chain structure, which is similar to those of the strains of the species Pseudomonas syringae, including the type strain. The data of structural analysis agree well with the results of immunochemical studies of LPS.

[The distinctive features of the structure of the Pseudomonas fluorescens IMV 247 (biovar II) lipopolysaccharide]. / Osobennosti stroeniia lipopolisakharida Pseudomonas fluorescens IMB 247 (biovar II).

The results of the study of the Pseudomonas fluorescens IMV 247 (biovar II) lipopolysaccharide (LPS) isolated from the dry bacterial mass by Westphal's method and purified by repeated ultracentrifugation are presented. The macromolecular organization of the LPS is characterized by the presence of S and R forms of LPS molecules in a 1:1 ratio. The structural components of the LPS molecule--lipid A, the core oligosaccharide, and the O-specific polysaccharide--were isolated and characterized. 3-Hydroxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, and dodecanoic acids proved to be the main lipid A fatty acids. Glucosamine, phosphoethanolamine, and phosphorus were identified as the components of the lipid A hydrophilic portion. Glucose, galactose, arabinose, rhamnose, glucosamine, alanine, phosphoethanolamine, phosphorus, and 2-keto-3-deoxyoctulonate (KDO) were revealed in the heterogeneous fraction of the core oligosaccharide. The O-specific polysaccharide chain was composed of repeating tetrasaccharide units consisting of L-rhamnose (L-Rha), 3,6-dideoxy-3-[(S)-3-hydroxybutyramido]-D-glucose (D-Qui3NHb), 2-acetamido-2,4,6-trideoxy-4[(S)-3-hydroxybutyramido-D-glucose (D-QuiNAc4NHb), and 2-acetamido-2-deoxy-D-galacturonic acid (D-GalNAcA) residues. A peculiarity of the O-specific polysaccharide was that it released, upon partial acid hydrolysis, the nonreducing disaccharide GalNAcA-->QuiNAc4NHb with a 3-hydroxybutyryl group glycosylated intramolecularly with a QuiN4N residue. Double immunodiffusion in agar and lipopolysaccharide precipitation reactions revealed no serological interrelationship between the strain studied and the P. fluorescens strains studied earlier.

[Characteristics of Pseudomonas fluorescens IMV 1433 (Biovar I) lipopolysaccharide]. / Kharakteristika lipopolisakharida Pseudomonas fluorescens IMV 1433 (biovar I).

The lipopolysaccharide of strain Pseudomonas fluorescens IMV 1433 (biovar I) was isolated and investigated. 3-Hydroxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, dodecanoic, hexadecanoic, hexadecenoic, and octadecenoic acids were identified in the composition of lipid A. The total amount of hydroxy acids makes up 60%. In the hydrolysate of lipid A, components of the were revealed hydrophilic skeleton--glucosamine and phosphoethanolamine--were revealed. Glucose, galactose, rhamnose, glucosamine, galactosamine, alanine, phosphoethanolamine, phosphorus, and 2-keto-3-deoxyoctulosonic acid were identified in the core of the lipopolysaccharide molecule. O-specific polysaccharide chains of the lipopolysaccharide are built from repeating trisaccharide units consisting of the residues of 2-acetamido-2,6-dideoxy-L-glucose, 2-acetamido-2,6-dideoxy-D-glucose and 4-acetamido-4,6-dideoxy-D-galactose. The lipopolysaccharide studied is similar to the lipopolysaccharide of P. fluorescens IMV 1152 (biovar I) in the structure of lipid A and core oligosaccharide, and identical to it in the structure of the O-chain (this identity correlates with the high level of the serological relationship of the strains).

[Characterization of the lipopolysaccharide from the Pseudomonas fluorescens strain IMV 472 (biovar I)]. / Kharakteristika lipopolisakharida Pseudomonas fluorescens IMV 472 (biovar I).

Lipopolysaccharide was isolated from dry biomass of the Pseudomonas fluorescens strain IMV 472 (biovar I) by the Westphal procedure and purified by ultracentrifugation. Fractions of the structural components of the lipopolysaccharide macromolecule-lipid A, the core oligosaccharide, and the O-chain-were isolated and characterized. 3-Hydroxydecanoic, 2-hydroxydodecanoic, 3-hydroxydodecanoic, hexadecenoic, octadecenoic, and hexadecanoic acids were identified as the main fatty acids of lipid A. The hydrophilic moiety of lipid A was found to include glucosamine, phosphoethanolamine, and phosphate. Glucose, galactose, rhamnose, glucosamine, galactosamine, alanine, phosphoethanolamine, and 2-keto-3-deoxyoctulosonic were identified in the core region of lipid A. The O-specific polysaccharide chain consisted of a repeating tetrasaccharide fragment that included two L-rhamnose residues, a D-fucose residue, and an N-acetyl-D-glucosamine residue as a side substituent. Double immunodiffusion in agar and the lipopolysaccharide precipitation reaction revealed no serological interrelation between strain IMV 472 and P. fluorescens strains studied previously.

[The determination of phosphoethanolamine and other amino components in the lipopolysaccharides of gram-negative bacteria by ion-exchange chromatography]. / Opredeleneie fosfoétanolamina i drugikh aminokomponentov v lipopolisakharidakh gramotritsatel'nykh bakterii metodom ionoobmennoi khromatografii.

A method for analysis of phosphoethanolamine in hydrolysates of the fractions structural parts of lipopolysaccharide macromolecule on the column with cation-exchange resin Ostion LG ANB (CSFR) in the system of step gradient of sodium-citrate buffers is suggested, optimal conditions for acidic hydrolysis of polysaccharide-standard were chosen to determine phosphoethanolamine and other amino compounds contained in it. The conditions for chromatography analysis of phosphoethanolamine together with aminosugars and amino acids have been developed. Using the suggested procedure the presence of phosphoethanolamine was found in preparations of lipid A hydrolysates and core oligosaccharide isolated from some lipopolysaccharide strains of Pseudomonas fluorescens.

[Characteristics of Pseudomonas fluorescens lipopolysaccharide]. / Kharakteristika lipopolisakharida Pseudomonas fluorescens.

A lipopolysaccharide was isolated from Pseudomonas fluorescens IMV 1152 (biovar I). The fractions of the structural moieties of lipopolysaccharide macromolecule were extracted and studied separately. 3-hydroxydecanoic, 3-hydroxydodecanoic and 2-hydroxydodecanoic fatty acids were identified in lipid A, total quantity of those was more than 80%. In acid hydrolysate of lipid A the glucosamine and phosphoethanolamine were found. The rhamnose, arabinose, glucose, glucosamine and phosphorus were identified as the components of core oligosaccharide fractions. A trisaccharide repeating unit of the O-cpecific polysaccharide chain consists of the residues of aminosugars: 4-acet-amido-4,6-dideoxy-D-galactose, 2-acetamido-2,6-dideoxy-D-glucose and 2-acetamido-2,6-dideoxy-L-glucose. The studied lipopolysaccharide resembled the structure of lipid A and core oligosaccharide with lipopolysaccharide from type strain of Pseudomonas fluorescens IMV 4125 (ATCC 13525), whereas their O-chains were different.

[Antigenic bacterial polysaccharides. 36. A study of the structure of the O-specific polysaccharide chain of the lipopolysaccharide of Pseudomonas fluorescens IMV 2763 (biostrain G)]. / Antigennye polisakharidy bakterii. 36. Issledovanie struktury O-spetsificheskoi polisakharidnoi tsepi lipopolisakharida Pseudomonas fluorescens IMV 2763 (viobar G).

Chemical and serological characterization of the Pseudomonas fluorescens IMV 2763 (biovar G) lipopolysaccharide was carried out. The O-specific polysaccharide chain of the lipopolysaccharide is composed of D-mannose, 6-deoxy-L-talose, N-acetyl-D-galactosamine and O-acetyl groups in the ratio of approximately 2:1:1:1. The polysaccharide is branched and a half of residues of 6-deoxytalose and monosubstituted mannose carry O-acetyl groups. On the basis of methylation, partial acid hydrolysis and 13C NMR analysis it was concluded that the repeating unit of the polysaccharide has the following structure: (formula; see text)

[Fatty acid composition of lipid A in Pseudomonas fluorescens]. / Zhirno-kislotnyi sostav lipida A u Pseudomonas fluorescens.

The fatty acid composition of lipid A was studied using gas-liquid chromatography (GLC) and GLC-mass spectrometry in Pseudomonas fluorescens strains of biovars A, B, C, i, F and G, the type strain ATCC 13525 (biovar A) inclusive. The following fatty acids were identified as predominant in the composition of lipid A in the strains representing biovars A, B, C, i, F and G: 3-hydroxydecanoic (3-OH C10:0), 2-hydroxydodecanoic (2-OH C12:0), 3-hydroxydodecanoic (3-OH C12:0), dodecanoic (C12:0), hexadecanoic (C16:0), octadecanoic (C18:0), hexadecenoic (C16:1) and octadecenoic (C18:1) acids. Lipid A of a biovar G strain differed noticeably from other strains in its fatty acid composition. Its main components were as follows: 3-hydroxytetradecanoic (3-OH C14:0), 3-hydroxypentadecanoic (3-OH C15:0) and dodecanoic (C12:0) fatty acids. The coefficients of similarity were determined for lipid A specimens isolated from the studied strains of P. fluorescens by calculating their fatty acid composition with a computer.

[Immunochemical characteristics of a lipopolysaccharide from Pseudomonas aurantiaca]. / Immunokhimicheskaia kharakteristika lipopolisakharida Pseudomonas aurantiaca.

A lipopolysaccharide was isolated from Pseudomonas aurantiaca IMB 31 by extraction with aqueous phenol and purified by ultracentrifugation. The lipopolysaccharide was confined to the phenol phase. Fucosamine (2-amino-2,6-dideoxygalactose) (36%) and bacillosamine (2,4-diamino-3,4,6-trideoxyglucose) (23%) were identified as hypothetic components of the O-chain in the carbohydrate moiety of the macromolecule using the techniques of paper chromatography, gas-liquid chromatography and ion-exchange chromatography on an amino acid analyser. Rhamnose, glucose, galactose, glucosamine and galactosamine were detected as hypothetical components of the core in the lipopolysaccharide composition, as well as 2-keto-3-deoxyoctonic acid, heptose, alpha-alanine and phosphorus, usual components of the core in Pseudomonas. The following predominant fatty acids were identified in the composition of lipid A using the techniques of gas-liquid chromatography with standard compounds and gas-liquid mass spectrometry: 3-OH C10:0 (14.4%), C12:0 (30.5%), 2-OH C12:0 (14.9%), 3-OH C12:0 (17.4%), C16:0 (9.9%). The serological relationship between P. aurantiaca strains was studied, and their phylogenetic relationship with P. fluorescens is discussed.

[Comparative biochemical studies of polyhedral proteins of nuclear polyhedrosis viruses]. / Sravnitel'noe binokhimicheskoe issledovanie poliedrennykh belkov virusov iadernogo poliedroza.

Using disc polyacrylamide gel electrophoresis, the molecular weights of polyhedral proteins of nuclear polyhedrosis viruses (NPV) of Porthetria dispar, Mamestra brassicae, and Aporia crataegi were found to be 28000 +/- 3000. It was shown that NPV polyhedra of Bombyx mori, Galleria mellonella, P. dispar, and M. brassicae contain a protease. During dissolution of the polyhedra at pH 10,5 this protease specifically cleaves the matrix protein into 2--5 fragments. The amino acid compositions of NPV polyhedral proteins of P. dispar, M. brassicae, A. crataegi, Hyphantria cunae were shown to be very similar. It was found that tyrosine is a C-terminal amino acid of NPV polyhedral proteins of P. dispar, M. brassicae, and A. crataegi.