[Department of Biochemistry of Microorganisms--start of the path (1951-1973)]. / Otdel biokhimii mikroorganizmov--nachalo puti (1951-1973).

Creation of the Department of Biochemistry of Microorganisms at the Institute of Microbiology and Virology of the Academy of Sciences of Ukrainian SSR in the 30's of the last century was determined by a necessity of profound investigation of vital activity biochemism of microorganisms from various systematic groups which were studied in microbiological department of the Institute. Such complexity can explain certain diversity of the Department research at initial stages of its existence. The research of saccharose transformation into dextran Leuconostoc mesenteroides, when production solutions become slingy at sugar-refinaries, was one of the first most significant works of the Department. The enzyme saccharose-glycosyl-transferase performing this process was described for the first time. A cycle of works on the study of enzymes splitting lactose in milk under the effect of Streptococcus lactis has been carried out. Complex investigation of a number of proteins, polysaccharides, enzymes in enterobacteria has shown that the blocking of the enzyme aldolase is one of the reasons of alkali formation. A method has been developed for isolation of arenarin, antibiotic of plant origin, from sandy everlasting, the nature of its acting basis has been established. Nufarin, an active antibiotic, was isolated from the roots of white water lily when studying nitrogen fixation processes, special attention was given to interaction of hydrogenase and enzymes, taking part in nitrogen fixation, to the effect of ATP on these processes, ways of its synthesis, localization of ATPase in the cell membranes. Works on the study of lypopolysaccharides and polysaccharides of Gram-negative enterobacteria, bacteria of Pseudomonas genus were started with the purpose to use the obtained data to specify systematic propositions of the investigated microorganisms. Further on these works became the basis of thematic department. There are numerous reviews dedicated to their development.

[Functional groups in the alpha-galactosidase active site in Cladosporium cladosporioides]. / Issledovanie funktsional'nykh grupp v aktivnom tsentre alpha-galaktozidazy Cladosporium cladosporioides.

The activity of alpha-galactosidase isolated from culture fluid of micromycete Cladosporium cladosporioides (Fres.) de Vries 16,038 has been studied as affected by cations, anions and specific chemical reagents (p-chlormercurybenzoate (p-ChMB), iodacetamide, N-ethylmaleimide, L-cysteine, dithiotreitol, beta-mercaptoethanol, EDTA, o-phenanthroline, sodium azide). It has been established that Ag+ ions inhibited competitively alpha-galactosidase at pH 4.0 and 6.0, the inhibition constants (Ki) made 3.6 x 10(-5) M and 4.3 x 10(-6) M, respectively. Galactose in concentration of 1 mM to 5 mM preserved the enzyme from the negative effect of Ag+ ions, while L-cysteine did not manifest the protective effect. Ions of Hg2+ p-ChMB inhibited noncompetitively the activity of alpha-galactosidase, Ki for Hg2+ and p-ChMB made 5.7 x 10(-7) M and 4.7 x 10(-6) M, respectively. Preincubation with galactose does not preserve alpha-galactosidase from the inhibiting effect of Ag+ and p-ChMB, but th[not readable: see text] compounds (L-cysteine, dithiotreitol, beta-mercaptoethanol) restore the enzyme activity. Participation of histidine imidazole group in the catalytic action is supposed on the basis of the inhibitory and kinetic analysis. Sulphydryl groups do not take part in the catalysis but play an important role in supporting the active conformation of the protein molecule. The groups containing the atoms of metals are absent in the alpha-galactosidase molecule.

[The molecular biological mechanisms of the functioning of microbial glycopolymers and carbohydrases]. / Molekuliarno-biologicheskie mekhanizmy funktsionirovaniia mikrobnykh glikopolimerov i karbogidraz.

Special attention has been paid to glycopolymers which perform various functions in the microbial cell, determine its interaction with the environment, possess a broad range of biological activity (antigenicity, cytotoxicity, immunomodulating properties, participation in the bean-rhizobial symbiosis). Composition and structure of the following polysaccharides, lipopolysaccharides and their components: O-polysaccharide, core oligosaccharide and lipid A from bacteria of genera Clavibacter, Ralstonia, Pseudomonas and Rhizobium, have been studied. Phenotypization of the studied strains has been carried out and chemotaxonomic criteria for creation of serological classification schemes have been proposed. Serogrouping of strains is determined by the availability and chemical nature of lateral substituents of the basic chain of O-specific polysaccharide. Interrelation between physico-chemical characteristics of the polysaccharide complex of rhizobia and formation of bean-rhizobial symbiotic structures has been shown. It is established that polysaccharides, lipo- and exopolysaccharides are efficient immunomodulators, manifest high antileucosis and antimetastasis activities. Physico-chemical properties, specificity, kinetics and action mechanism, structure of active and substrate-binding centres of bacterial and fungal enzymes: alpha-amylase, alpha-galactosidase, galactose oxidase. Data obtained permitted developing recommendations on enzymes application to some fields of the national economy.

Characterization of lipopolysaccharides from Pseudomonas syringae (serogroup II).

Lipopolysaccharides (LPS) from the strains of Pseudomonas syringae pv.syringae 90a, 435, pv. atrofaciens K-1025, pv. morsprunorum CF-4 referred by Pastushenko and Simonovich (1979) to serogroup II have been studied. The strains were shown to be heterogeneous by chemical composition of core and lipid A and structure of O-specific polysaccharide. The preparations heterogeneity in serological cross reactions were also detected. O-specific polysaccharides of the strains having similar structures were not identical in serological tests. The assumption on the lipid A role in serogrouping of P. syringae strains has been advanced.

[The characteristics of the lipid A of the lipopolysaccharides in Pseudomonas syringae strains]. / Kharakteristika lipida A lipopolisakharidov shtammov Pseudomonas syringae.

Component composition of lipid A of Pseudomonas syringae pv. syringae 218 and P-55, pv. syringae (holci) 8299, pv. phaseolicola 120a, pv. atrofaciens 2399 has been studied. The lipid A composition of all the strains studied includes 3-hydroxydecanoic fatty acid (3-OH-C10:0), 2-hydroxydodecanoic (2-OH-C12:0), 3-hydroxydodecanoic (3-OH-C12:0), dodecanoic (C12:0), hexadecanoic (C16:0), octadecanoioc (C18:0), hexadecenic (C16:1), octadecenic (C18:1) fatty acids. The carbohydrate part of the lipid A macromolecule of all strains after acid hydrolysis contains ethanolamine, phosphoethanolamine, glucosamine.

[Antigenic bacterial polysaccharides. Structure of O-specific polysaccharides from Pseudomonas cepacia serogroups C, I, O1, and O4]. / Antigennye polisakharidy bakterii. Stroenie O-spetsificheskikh polisakharidov Pseudomonas cepacia serogrupp C, I, O1 i O4.

Like some Pseudomonas cepacia serogroups studied earlier, serogroups C, I (Nakamura), O1 and O4 (Heidt) are characterized by the presence of at least two structurally different O-antigenic polysaccharide chains in cell-wall lipopolysaccharides. On the basis of acid hydrolysis, methylation, 1H- and 13C-NMR spectroscopy, including computer-assisted 13C-NMR-based analysis, the complete structures of the predominant polysaccharides of serogroups I (I), C and O4 (III) and the minor polysaccharides of serogroups I (II) and O1 (V) were established, and the structure of the predominant polysaccharide of serogroup O1 (IV) established earlier (Cox A. D., Wilkinson S. G.@Carbohydr. Res. 1990. V. 195. No 2. P. 295-301) was confirmed.

[The specificity of an immune serum to the exocellular lipopolysaccharide of Pseudomonas wieringae]. / Spetsifichnost' immunnoi syvorotki k ékzotselliuliarnomu lipopolisakharidu Pseumonas wieringae.

The serum obtained to exocellular lipopolysaccharide (ELPS) of Pseudomonas wieringae selectively agglutinated strains of pathovar of P. syringae and did not agglutinated strains of P. cichorii, P. solanacearum, P. gladioli pv. allicola, P. fluoroviolaceus, strains of nonphytopathogenic pseudomonads as well as bacteria of the genera Erwinia, Bacillus, Xanthomonas, Klebsiella. Consequently, the antigen determinant common with antigen of the species Pseudomonas syringae is present in the composition of ELPS.

[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)

[Polysaccharide--a stimulant of DNA reparative synthesis]. / Polisakharid--stimuliator reparativnogo sinteza DNK.

It is stated that regulation of the repair synthesis of DNA underlies the antimutagenic action of the polysaccharide isolated from bacteria. Protective effect of polysaccharides is also displayed.

[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.

[Immunologic and structural studies of lipopolysaccharides from Pseudomonas cepacia]. / Immunologicheskie i strukturnye issledovaniia lipopolisakharidov Pseudomonas cepacia.

O-serotyping of 30 Pseudomonas cepacia strains isolated from the soil and rhizosphere of different plant species in the territory of the USSR has been performed using 15 O-typing antisera according to the Heidt and Nakamura schemes. It is suggested to introduce two new O-serogroups (serogroups K and L) into the available P. cepacia classification scheme. They are most often met among the P. cepacia strains in different geographical areas of the USSR simultaneously with serogroups 2 (G) and 1 (D). To elucidate the molecular principles of serological inhomogeneity of the species the immunochemical studies of lipopolysaccharides of a number of P. cepacia strains have been conducted and the structure has been determined for repeating links of O-specific polysaccharides of P. cepacia strains attributed to 4 Nakamura serogroups, 3 Heidt serogroups, to serogroups K and L, as well as for certain strains from the collection of the Institute of Microbiology and Virology of the Ukr. SSR Academy of Sciences.

[Antigenic polysaccharides of bacteria. 35. Establishment of the structure of polysaccharide chains of lipopolysaccharides of Pseudomonas cepacia IMV 4176 and IMV 4202 (Serotype 3)]. / Antigennye polisakharidy bakterii. 35. Ustanovlenie struktury polisakharidnykh tsepei lipopolisakkharidov Pseudomonas cepacia IMV 4176 i IMV 4202 (serotip 3).

On mild acid degradation of the Pseudomonas cepacia strain IMV 4176 lipopolysaccharide, two polysaccharides were obtained, one of which is a homopolymer of N-acetyl-D-galactosamine and the other is composed of equal amounts of N-acetyl-D-galactosamine and D-ribose. Partial hydrolysis with aqueous oxalic acid caused depolymerization of the heteropolysaccharide, and the homopolysaccharide was isolated in the individual state. On the basis of methylation and 13C NMR analysis, it was concluded that both polysaccharides are built up of disaccharide repeating units having the following structures: ----4)-alpha-D-GalpNAc-(1----4)-beta-D-GalpNAc-(1---- and ----4)-alpha-D-GalpNAc-(1----2)-beta-D-Ribf-(1----. The heteropolysaccharide from P. cepacia strain 4176 is identical by the structure of the repeating unit to the O-specific polysaccharide of P. cepacia strain IMV 4202 (serotype 3), Pseudomonas aeruginosa O12 and Serratia marcescens O14.

[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.

[Antigenic polysaccharides of bacteria. 33. The structure of O-specific polysaccharide chain of lipopolysaccharide from Pseudomonas cepacia serotype 6]. / Antigennye polisakharidy bakterii. 33. Struktura O-spetsificheskoi polisakharidnoi tsepi lipopolisakharida Pseudomonas cepacia, serotip 6.

On mild acid degradation of the Pseudomonas cepacia serotype 6 lipopolysaccharide, the O-specific polysaccharide was obtained, which contains D-mannose and D-galactose residues in the ratio approximately 1:1, as well as O-acetyl groups. On the basis of 1H and 13C NMR analysis, calculation of specific optical rotation, and methylation, it was concluded that the polysaccharide possesses the following structure: (formula; see text) Regularities in glycosidation effects in 13C NMR spectra of 1,3-linked disaccharides containing furanoside residues are discussed.

[Antigenic polysaccharides of bacteria. 32. The structure of O-specific polysaccharide chains of Pseudomonas cepacia serotype B and E lipopolysaccharides containing D-fucose]. / Antigennye polisakharidy bakterii. 32. Struktura O-spetsificheskikh polisakharidnykh tsepei lipopolisakharidov Pseudomonas cepacia serotipov B i E, soderzhashchikh D-fukozu.

On the basis of acid hydrolysis, methylation, 1H and 13C NMR analysis, and calculation of specific optical rotation, the following structures were established for O-specific polysaccharides of Pseudomonas cepacia serotypes B and E: ----3)-beta-D-Galf-(1----3)-alpha-D-Fucp-(1----serotype B ----3)-beta-D-GlcpNAc-(1----3)-alpha-D-Fucp-(1----serotype E A characteristic feature of the polysaccharides is the presence of D-fucose, rather rare for bacterial antigens.

[Serine proteinase with lytic properties]. / Serinovaia proteinaza s liticheskimi svoistvami.

A thermophilic Bacillus licheniformis strain can synthesize extracellular serine proteinase, which is similar to subtilisin of the Carlsberg type in its amino acid composition, the specificity of its action, the character of its inhibition by certain compounds and immunochemical properties, but differs from the latter in its greater thermostability and in the ability to cause lysis of Gram-negative bacteria and yeast living cells.

[Antigenic polysaccharides of bacteria. 25. Structure of the O-specific polysaccharide chain of Pseudomonas cepacia 673/2 lipopolysaccharide containing L-glycero-D-manno-heptose]. / Antigennye polisakharidy bakterii. 25. Struktura O-spetsificheskoi polisakharidnoi tsepi lipopolisakharida Pseudomonas cepacia 673/2, soderzhashchei L-glitsero-D-manno-geptozu.

O-Specific polysaccharide, consisting of D-rhamnose and L-glycero-D-manno-heptose (LD-Hep) in a 2 : 1 ratio, was obtained on the mild acid degradation of the Pseudomonas cepacia IMV 673/2 lipopolysaccharide; monosaccharide LD-Hep has not previously been found in O-specific chains of lipopolysaccharides. On the basis of methylation and 13C-NMR data, it was concluded that the polysaccharide is composed of trisaccharide repeating units having the following structure: ----3)-alpha-D-Rha-(1----3)-alpha-D-Rha-(1----2)-alpha-LD-Hep-(1----

[Purification and study of the properties of a desulfonating enzyme in Pseudomonas alcaligenes]. / Ochistka i izuchenie svoistv desul'foniruiushchego fermenta Pseudomonas alcaligenes.

An enzyme desulfonating anionic alkyl benzene sulfonate (ABS) surfactants was isolated from Pseudomonas alcaligenes TR and purified. The physicochemical and catalytic characteristics of the enzyme were studied. The kinetic constants of ABS desulfonation were determined and shown to depend on the length of a hydrocarbon radical. The molecular mass of the enzyme was found to be close to 60,000.