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The aim of the work was to determine the influence of a number of coordination compounds of germanium, in particular cation-anionic type, on the activity of α-L-rhamnosidases of three producers: Cryptococcus albidus, Eupenicillium erubescens and Penicillium tardum ІМВ F-100074. Activity of α-L-rhamnosidases was determined according to Davis method, using naringin as substrate. The specific α-L-rhamnosidase activity of preparations was 12 units/mg of protein for C. аlbidus, 120 units/mg for E. еrubescens and 27 units/mg for P. tardum. Protein concentration in preparations was 0.01 mg/ml. Gernanium compounds were synthesized in alcohol-water solution according to standardized methods. In all compounds, the same bis (citrato) germanate anion is realized; however, the composition of the cation changes: the protonated form of phenanthroline and bipyridine or the complex cations of d-metals. It was established that none of the complex compounds exhibited an inhibitory effect on the activity of the enzymes under study. Maximum activating effect was observed with compound tris(bi-pyridine)nickel(II) bis(citrate)germanate monohydrate in concentration of 0.1% on α-L-rhamnosidases of C. albidus (10-fold), E. еrubescens (2.5-fold) and P. tardum (5-fold). Notably, enzymatic activity increased by 45, 47 and 60% respectively in presences of compound bi-pyridine bis(citrate)germanate dihydrate in same concentration. Hence, it was shown that these two compounds can be used as effectors of studied enzymes.

The purpose of the research was to investigate the influence of stanum (IV) and germanium (IV) coordination compounds on peptidases 1 and 2 of Bacillus thuringiensis var. israelensis ІМV В-7465 activity. The study of coordination compounds effect on peptidase activity was carried out by complexes with the enzymes incubation and residual activity to the collagen, elastin and fibrin determination. It was revealed the regularity in the influence of different structure complex compounds on peptidases of B. thuringiensis var. israelensis ІМV В-7465 activity. The stanum (IV) complexes with salicyloylhydrazones of aromatic aldehydes increase collagenase and elastase activities. Substituents replacement in aldehyde fragment of stanum (IV) with isonicotinoylhydrazones of aromatic aldehydes complexes by the less polar ones contributed to increase of elastase activity of both enzymes while an absence of substituents enables to increase peptidase 2 fibrinolytic activity. The complexes of germanіum (IV) with isonicotinoylhydrazone of salicylic aldehyde which contain Zn and Co, increased collagenase activity of the peptidase 1 as well as elastase and fibrinolytic activities of the peptidase 2. In general, all tested complexes may be considered as peptidase effectors of B. thuringiensis var. israelensis ІМV В-7465. A difference of complexes effect on activities of the both enzymes is due to the characteristics of the structure of coordination compounds.

The cultivation conditions of Bacillus thuringiensis ІМV В-7324 for synthesis of the elastolytic peptidase were studied. By mono- and two-factorial experiments it was optimized the nutrient medium and conditions of growth for the synthesis of B. thuringiensis ІМV В-7324 elastolytic peptidase. It was established that maximal synthesis of the enzyme (5.15 U/mg of the protein) occurs at the exponential phase of growth on 18 h in submerged cultivation. As a result of screening experiments it was shown that all components of the basic medium except gelatin are significant for the enzyme biosynthesis. The elimination of gelatin leads in 9.8-fold increase of the elastolytic activity (50.55 U/mg of the protein). The influence of the nitrogen and carbon sources on the enzyme synthesis was studied. It was established that the optimal sources are the ammonium sulfate and arabinose. Their usage allows us to increase in 17.4 and 4.6 times the elastolytic activity (90 and 24 U/mg of the protein). The optimal concentrations of the ammonium sulfate and arabinose in the medium which allow to increase the elastolytic activity in 24.7 times (127.45 U/mg of the protein) was determined by the bifactorial experiment on three levels. The optimized nutrient medium contains (g/l): arabinose — 13.0; (NH4)2SO4 — 14.0; КН2РО4 — 1.6; (CH3COO)2Zn — 0.25; MgSO4∙7H2O — 0.75. The elastolytic activity of B. thuringiensis ІМV В-7324 was 132.5 U/mg of protein during the growth by submerged cultivation for 18 h in 200 ml of the optimized nutrient medium at initial pH 7.0, on a shaker at 220 rpm at 37 °C.

The purpose of this work was to investigate α-L-rhamnosidase that hydrolytically cleaves the terminal unreduced α-1,2-, α-1,4- and α-1,6-linked rhamnose residues in both synthetic and natural glycosides, oligo-, and polysaccharides, various glycoconjugates: flavonoid derivatives such es rutin, neohesperidin, hesperidin, naringin, quercitrin, saponins, terpene glycosides. These properties of the enzyme could be used for the needs of food industry, pharmaceutical and chemical industry: to improve the quality of beverages (reduction of bitterness, flavor enhancing wines), for production of food additives, medicine preparations and rhamnose. As a result of screening conducted among 9 strains of micromycetes, ability to synthesize α–Lrhamnosidase was revealed only in Penicillium sp. 2918. Complex enzyme preparation was obtained from culture supernatant of this micromycete by fractionation with ammonium sulfate (90% saturation) and its physico-chemical properties such as pH- and thermooptimum, pH- and thermal stability and substrate specificity were studied as well. It is shown that enzyme has pH optimum is about 6.0 and thermooptimum is about 60 оC. Preparation of Penicillium sp. 2918 with α-L-rhamnosidase reveals α-D-glucosidase, α-D-galactosidase and α D-glucosaminidase activity.

By ammonium sulfate fractionation and chromatography on TSK-gels Toyopearl HW- 60 and Fractogel TSK DEAE-650-s from supernantant of cultural liquid of Penicillium tardum 1MB F-100074 was isolated and purified in 23 times preparation of enzyme with α-L-rhamnosidase activity with yield of 3.12 %. The specific activity was 27.7 U/mg of protein, molecular mass 67 kDa, thermo- and pH optimum - 60 °C and pH 5.0 respec- tively. It was shown high pH- and thermostability of purified P tardum a-L-rhamnosidase which may be dued to presence of 12 % of carbohydrate component.

From Achromobacter sp. 7a, that was isolated with Black sea, aquatoria of island Zmi- inyi, was isolated enzyme with α-amylase activity, that able also to split the synthetic substrates: p-nitrophenyl-α-D-glucopyranoside and p-nitrophenyl-α, -β-D-xylopyranoside. Methods of isolation and purification of enzyme were selected which included: ammonium sulfate precipitation and affinity sorption on starch, that improved enzyme activity in 7 times in comparison with activity in the supernatant of cultural liquid. a-Amylase showed maximal activity at pH 7.0 and 11.0 and to the temperature 50 °C. Enzyme remained fully stable during 24 hours in the range of pH from 7.0 to 12.0, during 3 hours at a temperature 37 °C and 50 °C at pHopt 7.0, and also 87.5 % and 75 % of initial activity saved during 3 h of incubation at a temperature 37 °C and 50 °C at pHopt 11.0 respectively. It is shown that addition of antihunt agents (ions of calcium, chloride of natrium) did not protect an enzyme from thermoinactivation (60 °C, 70 °C).

The serological and phytotoxic properties of lipopolysaccharide (LPS) of plant pathogens--Pantoea agglomerans were studied. It is known that the thin variations in the structure of the O-specific polysaccharides determining serological specificity of gram- negative bacteria and used as a molecular basis of serological classification schemes. For P. agglomerans still does not exist a classification scheme based on serology specificity of their LPS. The results of cross serological tests demonstrate immunochemical heterogeneity of species P agglomerans. Only three strains of the 8488, 8490 and 7969 according to the agglutination of O-antigens and direct hemagglutination and inhibition direct hemagglutination can be attributed to a single serogroup. Other strains--each separate group, although some have a relationship. Compared with control plants under the influence of seed treatment of LPS in plants may be reduced, and in some cases increased root length, height and weight sprout, depending on the strain from which the selected LPS. Dive seedlings of tomatoes in the solutions of the studied preparations FSC caused the loss, and after some time, restore turgor.

The effect of cations and anions on the activity of Aspergillus flavus var. oryzae and Bacillus subtilis α-amylases showed that the tested enzymes are sensitive to most of cations and resistant to anions. The most significant inhibitory effects on the activity of A. flavus var. oryzae α-amylase have been demonstrated by Al3+ and Fe3+ ions, while on the activity of B. subtilis α-amylase - Hg2+, Cu2+ and Fe3+ ions. Inactivation of A. flavus var. oryzae and B. subtilis α-amylases in the presence of EGTA is indicated on the presence within their structure of metal ions. An important role in the enzymatic catalysis of both enzymes play carboxyl groups as evidenced by their inhibition of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methiodide. Inhibition of B. subtilis α-amylase by p-chloromercuribenzoate, N-ethylmaleimide and sodium sulfite is indicated on the probable involvement of the sulfhydryl groups in the functioning of the enzyme. Unlike most studied glycosidases the tested enzymes do not contain histidine imidazole group in the active center.

The influence of some technological cultivation parameters of Penicillium tardum to synthesize of the extracellular α.-L-rhamnosidase were studied. It was shown that rhamnose (0.8%), yeasts autolysate (0.2%), temperature of the cultivation 25 degrees C, pH 5.0 are necessary for maximal α-L-rhamnosidase production. The enzyme reaches the maximal activity level in 96 hours with sulphitic number equal 0.44. At cultivation of P. tardum in the picked up conditions the α-L-rhamnosidase synthesis has raised in 4 times.

Screening of protease producers with specificity to insoluble and hard soluble protein substrates of animal origin (collagen, fibrin, elastin and keratin) was carried out. It was studied the bacterial cultures (24 strains) isolated from water and periphyton of enclosures with dolphins, and also from exhalations, oral cavity and skin of dolphins. Some bacterial strains isolated from water and periphyton of enclosures hydrolyzed collagen (5-23 U/ml) and elastin (20-32 U/ml). Thus all tested cultures did not possess the property of extracellular keratinases synthesis. The streptomycetes (48 strains) were isolated from the soil of Black Sea coastal strip near Odessa and Saky, from parkland and the shores of freshwater lake in Saky and from the soil of Atlantic Ocean coastal strip near Albufena (Portugal). Several streptomycetes have been found to appeare the perspective producers of extracellular keratinase and collagenase. The strains isolated from the soil of the coastal strip area both sea and freshwater lake in Saky possessed the highest activity (up to 5 U/mg).

Influence of coordinative compounds of germanium (IV) and stanum (IV) (complexes of germanium (IV) with nicotinamide (Nad) [GeCl2(Nad)4]Cl2 (1) and complexes of stanum (IV) with 2-hydroxybenzoilhydrazone 4-dimetylaminobenzaldehide (2-OH-HBdb) [SnCl4(2-OH-Bdb-H)] (2), 3-hydroxy-2-naphtoilhydrazone 2-hydroxynaphtaldehide (3-OH-H2Lnf) [SnCl3(3-OH-HLnf)] (3) and izonicotinoilhydrazone 2-hydroxyibenzaldehide [SnCl3 (Is·H)] (4) on activity of peptidases 1 and 2 Bacillus thuringiensis, α-L-rhamnosidase Cryptococcus albidus, Eupenicillium erubescens and α-amylase Aspergillus flavus var. oryzae. Results testify that all studied compounds differ on their influence on activity of the enzymes tested: significantly don't change elastolytic activity of peptidases 1 and 2 B. thuringiensis, completely inhibit A. flavus var. oryzae amylase, activate or oppress of α-L-rhamnosidase C. albidus and E. erubescens. Considerable differences in compounds (3, 4) on activity observed in case of the last. It's possible that peculiarity of influence (1) in compare with (2-4) is connected with existence of different central atoms of complexants: germanium (IV) (1) and stanum (IV) (2-4). A certain analogy in oppression of C. albidus α-L-rhamnosidase by compounds (1) and (4) can explain with presence of a pyridinic ring at molecules of their ligands. The less activsty displayed compound (2) with coordinative knot {SnCl4ON}. Nature of compounds (3, 4) activity was absolutely different: essential increase of activity of C. albidus α-L-rhamnosidase and full oppression of E. erubescens α-L-rhamnosidase by compound (3), while the action of compound (4) was feed back. Taking into account identical coordination knot {SnCl3O2N} the major role in this case play change of a hydrazide fragment in molecules of their ligands.

The component composition of Cryptococcus albidus and Eupenicillium erubescers alpha-L-rhamnosidases have studied. It was shown that enzymes have a monomeric structure. Enzyme preparations of C. albidus and E. erubescens have similar qualitative but differ in quantitative amino acid composition. alpha-L-rhamnosidase of C. albidus characterised by high amount of histidine, proline, cysteine, methionine in compared with alpha-L-rhamnosidase of E. erubescens. alpha-L-Rhamnosidase of E. erubescens, in contrast to the alpha-L-rhamnnosidase of C. albidus, contained higher levels of lysine, arginine, threonine, alanine, isoleucine, leucine, tyrosine, phenylalanine. It is shown that purified preparations of alpha-L-rhamnosidase C. albidus and E. erubescens contained 5 and 1% carbohydrates respectively. Enzyme preparations differ in quantitative monosaccharide composition, which represented by rhanmose, xylose, mannose, galactose and glucose. Furthermore, alpha-L-rhannosidase C. albidus contained fuicose, whereas alpha-L-rhamnosidase E. erubescens--ribose and arabinose. A significant percentage of hydrophobic amino acids, which is 31 and 34% of the total content, and the presence of the carbohydrate component are essential in stabilization of enzymes molecule.

The study is dedicated to research of phytotoxic properties of Ralstonia solanacearum lipopolysaccharides. This causative agent is one of the most dangerous among potato bacterial diseases. It is revealed that the inhibitory effect of LPS solution on seedlings germination is more noticeable on crops susceptible to brown rot. Maximal total phytotoxic properties have been shown by LPS from strains 35, 52b, TX1 and TS3, which were characterized by relatively low rhamnose content. Relative to the control plants LPS may diminish and some ones--increase the root length, height and weight of seedlings, subject to particular strain. But the stimulation revealed is minor.

All strains tested are characterized by proteolytic (caseinolytic) activity, while elastase one was revealed only in two Gracilibacillus strains 6T2 and 7Tl. The activity was high enough (23.1 and 34.7 E/Ml, respectively). These values are at the level of bacterial producers which are described in literature: Bacillus mesentericus 316 M (6 E/Ml), Bacillus thuringiensis IMB B-7324 (50-55 E/Ml). The ability of two strains tested to synthesize enzyme, active against elastine, is important, so far as microbial enzyme may be perspective for using in medicine: elastases are able to dissociation of elastin fibres of connective tissues. These two strains display also fibrinolytic activity, however it was insignificant. Six of eight strains studied manifested alpha-amylase activity (0.01 - 1.173 E/Ml). It was shown that no strains, isolated from the Dead Sea costal ecosystems are able to manifest alpha-L-rhamnosidase activity.

The substrate specificity of Cryptococcus albidus and Eupenicillium erubescens alpha-L-rhamnosidases has been investigated. It is shown that the enzymes are able to act on synthetic and natural substrates, such as naringin, neohesperidin. alpha-L-Rhamnosidases hydrolysed the latter ones very efficiently, in this case E. erubescens enzyme was characterized by higher values of V(max) in comparison with the enzyme of C. albidus. However the C. albidus alpha-L-rhamnosidase showed greater affinity for naringin and neohesperidin than the enzyme of E. erubescens (K(m) 0.77 and 3.3 mM and 5.0 and 3.0 mM, respectively). As regards the synthetic derivatives of monosaccharides, both enzymes exhibited narrow specificity for glycon: E. erubescens alpha-L-rhamnosidase--only to the p-nitrophenyl-alpha-L-rhamnopiranoside (K(m) 1.0 mM, V(max) 120 micromol/min/mg protein), and C. albidus--to p-nitrophenyl-alpha-D-glucopyranoside (K(m) 10 mM, V(max) 5 micromol/min/mg protein). Thus, it was found that the enzyme preparations of E. erubescens and C. albidus are differed by their substrate specificity. The ability of E. erubescens and C. albidus alpha-L-rhamnosidases to hydrolyse natural substrates: naringin and neohesperidin, evidences for their specificity for alpha-1,2-linked L-rhamnose. Based on these data, we can predict the use of E. erubescens and C. albidus alpha-L-rhamnosidases in various industries, food industry in particular. This is also confirmed by the fact that the investigated alpha-L-rhamnosidases were stable at 20% concentration of ethanol and 500 mM glucose in the reaction mixture.

Investigations of physico-chemical properties of the Bacillus thuringiensis IMV B-7324 fibrinolytic peptidase showed that optimal activity of enzyme displayed at pH 10.0 and temperature 60 degrees C. Stability of peptidase retained in the range of pH from 6.0 to 11.0 and temperature from 20 to 50 degrees C over 1 h. Inhibition of fibrinolytic peptidase activity by ethylene glycol tetraacetic acid (EGTA) and tetrasodium salt of ethylenediaminetetraacetic acid (trilon B) indicates the belonging of this enzyme to the group of metallopeptidases. It was established that cations Ag+, Mg2+ and Ba2+ increased the fibrinolytic activity by 40 %, 25 % and 30 %, respectively, but Ca2+, Zn2+, Cu2+, Pb2+ and Hg2+ reduced it by 30-60%. Several of studied anions (F-, Br-, SO2-, S2O(2-)3, AsO(3-)4, NO-(3) and NO2(-) inhibit the activity of B. thuringiensis IMV B-7324 fibrinolytic peptidase by 25-100%.

The current literature data on producers, physico-chemical properties and substrate specificity of a-amylases produced by microbes from different taxonomic groups such as bacteria, fungi and yeasts are discussed in the survey. Synthesis of alpha-amylase majority is an inducible process which is stimulated in the presence of starch or products of its hydrolysis. It is possible to increase enzymes activity level by optimization of cultivation conditions of strains-producers. alpha-Amylases, isolated from different sources are distinguished in their physico-chemical properties, particularly in their molecular weights, pH- and thermooptimums, inhibitors and activators. The enzymes hydrolyse soluble starch, amylose, amylopectin, glycogen, maltodextrins, alpha- and beta3-cyclodextrins and other carbohydrate substrates. It is well known that alpha-amylases belong to GH-13 family of glycosyl-hydrolases, which contain the catalytic domain A as (beta/alpha)8-barrel. In addition to domain A, alpha-amylases contain two other domains: B and C, which are localized approximately on opposite sides of (beta/alpha)8-barrel. Most of the known alpha-amylases contain calcium ion, which is located on the surface between domains A and B and plays an important role in stability and activity of the enzyme.

The fibrinolytic enzymes are peptidases, which belong to the class of hydrolytic enzymes. They are able to dissolve the insoluble fibrin. This review presents the fibrinolytic peptidases of Bacillus. The methods of increasing biosynthesis of these enzymes, determination of their activity, isolation and purification have been observed. The fibrinolytic peptidases of Bacillus are serine- and metallopeptidases. Its can hydrolyze both the natural substrates (fibrin, fibrinogen and hemoglobin) and synthetic substrates. The high stability of these enzymes in a wide range of pH (5-12) and temperature (10-75 degrees C) permits using them in pharmacology, medicine for the prevention and treatment of cardiovascular diseases.

Features of nucleic acids sequences of genes of lipid A deacylases of R. solanacearum, which are presented in GenBank database, were analyzed. Primers to this gene were selected, and using them 12 strains of R. solanacearum were analyzed for the presence of this gene in their genome by means of PCR method. PagL gene was identified in strain 749, and dependence of its transcription on some environmental conditions was established.

Results of monosaccharide analysis of Ralstonia solanacearum lipopolysaccharides of the strains of different geographical origin are shown. Such monosaccharides as glucose, rhamnose and glucosamine prevail. Following the results of analysis, strains with supposedly truncated O-polysaccharide are revealed. As to their quantitative and monosaccharide composition of LPS the strains have demonstrated high level of heterogeneity.

The scheme of isolation and purification of Bacillus thuringiensis IMV B-7324 peptidase has been developed. This scheme includes ammonium sulfate precipitation and chromatography on neutral and charged TSK-gels. It was found that the enzyme hydrolyzes elastin and fibrin. The molecular weight is 26 kDa. It was shown that the enzyme is an alkaline serine peptidase. The optimal pH of hydrolysis of elastin and fibrin were 9.0 and 10.0, respectively. The optimal temperature of elastin and fibrin hydrolysis are 40 and 50 degrees C, respectively. The high stability of the purified preparation in the studied range of pH and temperature was shown. The stabilizing effect of zinc at a concentration of 1 mM on the elastase activity, and the inhibitory effect of other divalent cations under study have been established. The investigated chloride and acetate anions reduced activity by 20%, while phosphate anions increased activity by 15-30%.

Fibrinolytic peptidase of Bacillus thuringiensis IMV B-7324 was isolated by ammonium sulfate fractionation, gel-filtration and ion exchange chromatography on TSK-gels--Toyopearl HW-55 and DEAE 650 (M). Fibrinolytic activity of the purified enzyme was 87.9 U/mg of protein that was 19.9 times higher compared with the supernatant cultural liquid, the yield on its activity reached 31%. The gel-filtration on Sepharose 6B and by SDS-PAGE electrophoresis demonstrated the homogeneity of the purified fibrinolytic peptidase, which molecular weight was approximately 24 kDa.

By immunochemical investigations of eight strains of Ralstonia solanacearum six strains were attributed to four serogroups. Two of them are formed by pairs of R. solanacearum strains 4 and 526; 758 and 7954; two others are represented by single strains--TX1 Ta TS3, correspondingly. Antigenic structure of R. solanacearum 7954 O-polysaccharide unites antigenic epitopes of R. solanacearum strains 4, 35, 526, 749, however the absence of cross-reactivity does not permit uniting them into the same group. The latter, and also the fact that the antiserum to R. solanacearum 749 in the reaction with LPS of R solanacearum 526 forms two precipitation lines (while in the homological system it forms only one line) may be explained by differences in the component composition of heat-stable immunogens (which were used for antiserum obtaining), and also purified LPS which were utilized as antigens in immunochemical reactions.

The effect of cations, anions and specific chemical reagents: 1-[3-(dimethylamino)propyl]-3-ethylcarbodimide methiodide, EDTA, o-phenantroline, dithiotreitol, L-cysteine, beta-mercaptoethanol, p-chlormercurybenzoate (p-ChMB), N-ethylmaleimide on the alpha-L-rhamnosidase activity of Cryptococcus albidus has been investigated. The essential role of Ag+ which inhibits the alpha-L-rhamnosidase activity by 72.5% was shown. Rhamnose at 1-5 mM protect the enzyme from the negative effect of Ag(+). It was expected that carboxyl group of C-terminal aminoacid and imidazole group of histidine would participate in the catalytic action of alpha-L-rhamnosidase on the basis of inhibition and kinetic analysis.

The results of the comparative toxicity studies of native lipopolysaccharide (LPS) of Rahnella aquatilis 96U037 and that modified by tin complexes indicates that, due to the modification of LPS by tin complex with benzoylhydrazone of 4-dimethylaminobenzaldehyde, a decrease of its toxicity was observed that led to disappearance of the pyrogenic effect. All obtained derivatives lost completely the antigenic activity both in homologous and heterologous systems which may indicate to the interaction of modifying complexes with certain groups being the components of antigenic determinant. The paper is presented in Ukrainian.

Influence ofsome technological parameters of cultivation ofproducers Cryptococcus albidus, Eupenicillinum erubescens, Bacillus sp. on the process of synthesis of extracellular enzyme alpha-L-rhamnosidase has been studied. The authors have determined optimal sources of carbon (0.2-0.3% rhamnose) and nitrogen (0.2% sodium nitrate for C. albidus and E. erubescens and ammonium sulphate for Bacillus sp.) (the ratio 1:2), cultivation temperature (28 degrees C, 25 degrees C, 42 degrees C, respectively) for maximum synthesis of alpha-L-rhamnosidase. Use of the medium with initial pH value from 4 to 8 was most efficient for all the studied strains. The maximum level of alpha-L-rhamnosidase activity of E. erubescens and Bacillus sp. was established at the value of sulphite number of 0.44, while for C. albidus--it was 0.56. Maximum alpha-L-rhamnosidase activity of C. albidus, E. erubescens, Bacillus sp. is achieved at 4, 8 days and 27 hours of cultivation, respectively. The cultures being grown in selected conditions, the alpha-L-rhamnosidase synthesis has increased by 30, 50 and 20%, respectively.

For the first time lipopolysaccharides (LPS) of 6 Budvicia aquatica strains--representatives of new Enterobacteriaceae species have been isolated. It was shown that the yield of LPS ranged from 0.9 to 7.0 % of cells dry weight. On the basis of monosaccharide composition LPS of tested strains may be referred to 3 groups. The serological studies indicated the immunochemical heterogeneity of B. aquatica species: LPS interacted only in homological system and showed no cross-reactivity with heterological antisera.

The influence of content of cultivation media and fermentation conditions on synthesis of alpha-L-rhamnosidase by Cryptococcus albidus 1001 and 1003 were studied. It was shown that L-rhamnose and NaNO, are the optimum sources of carbon and nitrogen, respectively. The temperature of 25 degrees C, pH 6.0, growing in 50 ml of the medium at rotation rate 220 rev/min during 6 days are optimum parameters for both strains.

Screening of producers of alpha-L-rhamnosidase among 331 yeast strains (representatives of 12 genera and 17 species) has been performed. A capacity to synthesize the enzyme was revealed in 45% of the studied strains with activity 0.01-0.6 un/ml. Complex enzymatic preparations of alpha-L-rhamnosidase were obtained from cultural liquid of two the most active producers by fractionation with ammonium sulfate (90% saturation). The influence of pH and temperature on enzymes activities and also specificity towards natural and synthetic substrates were studied.

Microbial proteolytic enzymes are able to hydrolyze various protein substrates. Data concerning the ability to degrade proteins amongst various groups of microorganisms, such as bacteria, fungi, actinomycetes and yeasts, are collected in this article. The physical and chemical properties of proteolytic enzyme preparations and methods of their separation and purification have been examined. The possible ways of these enzymes application in industry and medicine have been discussed. The paper is presented in Ukrainian.

The studies of lipopolysaccharides (LPS) of eight Rahnella aquatilis strains, isolated from different sources have shown that they contain both S- and R-types of molecules. This fact is evidenced by the presence of high-molecular fraction of O-specific polysaccharides (O-PS) and low-molecular fraction of core oligosaccharides. The predominant monosaccharides of core oligosaccharides were glucose, galactose. The presence of only one high-molecular fraction O-PS was a characteristic feature of all investigated strains. The predominant monosaccharides of O-PS were galactose, glucose, mannose, rhamnose and fucose. 3-hydroxytetradecanoic (48.9-93.1%), dodecanoic (2.9-12.1%), tetradecanoic (4.1-25.3%) and hexadecanoic (2.8-15.3%) acids have been obtained in lipids A of LPS depending on the strain. The presence in lipid A of R. aquatilis of only 3-hydroxytetradecanoic acid which is characteristic of Enterobacteriaceae proves the correct ascribing of the isolated strains to this family.

Lipopolysaccharides (LPS) of eight strains of R. aquatilis isolated from different sources have been studied. The studies of neutral monosaccharide composition evidence that all of LPS contain galactose (13.4-68.5%), glucose (5.7-29.8%) and heptose (2.6-8.3%) (depending on strains). Some monosaccharides, such as ribose (95U007), rhamnose (95U011, 95U012, 96U036), fucose (95U003, 95U004, 95U007) and mannose (95U012, 96U035, 96U036, 96U037) were absent in LPS. Arabinose was present in two strains--95U003 and 95U007. On the basis of monosaccharide composition all investigated LPS can be divided into six groups. It was shown by double immunodiffusion in agar that all R. aquatilis LPS displayed antigenic activity in homological systems. The results of serological cross reactions indicate the immunochemical heterogeneity of R. aquatilis species.

The influence of different factors on biosynthesis of extracellular alpha-amylase of Aspergillus sp. 55 grown at submerged cultivation has been studied. The optimal composition of nutrient medium (1 g of starch and 0.5 g of NaNO3 per 1 l) was chosen. The enzyme preparation has a wide pH optimum of activity (4.5-9.0), thermooptimum at pH 6.5 was 60 degrees C, at pH 4.5 and 9.0--50 degrees C. The inhibitory effect of coordinative germanium compounds on amylolytic activity of the preparation was shown. The paper is presented in Ukrainian.

The current data on microbial mannanases are summarised in the work. The questions concerning the spreading of mannanases substrates--mannans in nature, their structural variations are regarded. The production of mannanases by micro-organisms of different taxonomic groups, their physico-chemical properties and substrate specificity are described. The ways of mannans hydrolysis by mannanase are estimated. The questions of cloning and expression of mannanase genes have been discussed. These results are very important for producing highly effective microbial biosynthetics.

Effect of lipopolysaccharides (LPS) and their structure components (lipid A, oligosaccharide core, and O-specific polysaccharides) from 8 strains of P. fontium on the adhesion process have been investigated for the first time. It was demonstrated, that LPS and their structural components may compete with E. coli adhesins and phytohemagglutinin under binding with receptors on the surface of the rabbit erythrocytes. An analysis of different structural components influence indicated, that in comparison with O-specific polysaccharides and core oligosaccharide lipids A of all investigated strains of P. fontium displayed the highest inhibitory action on the adhesion process. It was shown that there existed certain dependence between LPS composition and their effects on the adhesion process. LPS, obtained from P. fontium strains grown at different temperatures (36, 25, 4 degrees C) and characterized by various quantitative and qualitative monosaccharide and fatty acid composition, demonstrated different adhesion properties.

Modified lipopolysaccharides (LPS) of Pragia fontium were obtained with germanium complexes (IV) of 2-aminobenzoylhydrazon of salicylic aldehyde (2-NH2-H2Bs), 2-hydroxybenzoylhydrazon salicylic aldehyde (2-OH-H2Bs) and nicotinoylhydrazon of salicylic aldehyde (H2Ns). The modification of LPS was confirmed by IR spectroscopy. Comparative investigations of pyrogenic activity of native and modified LPS showed, that only P. fontium 20125 LPS, modified by germanium complexes with 2-hydroxybenzoylhydrazon of salicylic aldehyde (2-OH-H2Bs) has lost the pyrogenic activity. In the homological reactions of double immunodiffusion in agar it was shown that all modified LPS unlike the native ones lose completely antigenic activity.

The schemes of isolation and purification of proteolytic complex of Bacillus sp. have been developed, which include fractionation by ammonium sulphate and separation on TSK-gels: ion-exchange chromatography on Toyopearl DEAE-650 (M) and gel-filtration on Toyopearl HW-50. It has been established that the proteolytic complex Bacillus sp. is presented by neutral and alkali proteases (pH optimum at 6.0-8.0 and 10.0), which hydrolyse a number of protein substrates: casein, gelatin, hemoglobin, fibrin, and elastin. Two proteolytic complexes with dominating fibrinolytic and elastase activity and molecular weight 28.7 and 22.7 kDd were isolated as a result of purification. It was found out that the obtained enzymatic preparations belong to the group of serine protease which need metal ions for development of their activity.

On the basis of the data of metal ions influence it was established that Penicillium commune 266 alpha-L-rhamnosidases are dependent of metals. The cations of Cu2+, Mn2+, Ag+, Hg2+ essentially influenced the activity decreasing its up to 45-55%. Ba2+ and Mn2+ ions completely inhibited alpha-L-rhamnosidase 2 activity in concentration of 10(-3) M. The role of carboxylic group and histidine imidazole group in hydrolysis of rhamnoglycosides have been estimated by the Dixon, Lineweaver-Burke and photooxidation methods. It was also shown that sulfhydrilic groups which do not take part in catalysis, but are necessary for displaying the enzymatic activity are located at a certain distance from P. commune 266 alpha-L-rhamnosidase active centre.

Microbial keratinolytic enzymes hydrolyze such a hard dissoluble protein substrate as keratin. Data conserning ability to degrade keratin by various groups of microorganisms and their potential biological role are discussed in the paper. The physical and chemical properties of enzyme preparations of keratinases and methods of their separation and purification have been examined. The available and possible ways of these enzymes application in industry and medicine have been discussed.

Structure components (lipid A. oligosaccharide core, and O-specific polysaccharide) of the lipopolysaccharide from 8 strains of Pragia fontium were obtained as a result of mild acidic hydrolysis. The macromolecular organization of lipopolysaccharides of studied strains was heterogeneous and presented both by S-forms, and R- or SR-forms. It was established that as to fatty acid composition lipids A of tested strains were represented by one chemotype with predominant 3-hydroxytetradecanoic fatty acid. An analysis ofmonosaccharide composition of core oligosaccharides has detected 5 chemotypes, and glucose, galactose, rhamnose, ribose, arabinose, xylose and mannose were identified in them. It was shown that O-specific polysaccharides consisted of glucose, galactose, rhamnose, and mannose that demonstrated their belonging to 4 chemotypes.

The paper deals with the capacity of the coordination germanium compounds with citric acid in a complex with some aminoacids (aspartic acid, methionine, cysteine, threonine, phenylalanine, L-glutamic acid) as bioligands to affect synthesis and activity of the proteolytic enzyme complex of Bacillus sp. 27 and Yarrowia lipolytica 2061. Thus, biscitrategermanium acid H2[Ge(H4Citr)2] and its complex with aspartic acid (HAsp)2 [Ge(H4Citr)2] can be used with success for inducing biosynthesis ofcaseinolytic (182.1 and 197.4%) and elastase (182.6 and 383.3%) activities of Bacillus sp. 27, respectively. Biscitrategermanium acid can be used to induce biosynthesis of caseinolytic (182.1 and 197.4%) and elastase (182.6 and 383.3%) activities (respectively) of Bacillus sp. 27. Biscitrategermanium acid with methionine induced biosynthesis of caseinolytic (127.6%) and haemoglobinolytic (110.0%) activities of Y. lipolytica 2061. The tested coordinative compounds could not induce activity ofproteolytic complexes of both microorganisms. The capacity of complexes of citrategermanium acid with aspartic acid, methionine, cysteine, threonine, phenylalanine in concentration of 0.1% to inhibit activity of proteolytic complex of Y. lipolytica 2061 can be used in further studies of the catalytic centre of enzymes.

The influence of different factors on the biosynthesis of extracellular proteolytic complexes by strains-producers Bacillus circulans 693, Bacillus sp. 27 and Yarrowia lipolytica 2061 at submerged cultivation has been investigated. It has been shown that ammonium hydrocarbonate and gelatin with glucose were optimum carbon and nitrogen sources for synthesis of proteolytic activity of B. circulans strain 693, gelatin with arabinose--for Bacillus sp. 27, gelatin and glycine with sorbose--for Y. lipolytica 2061. It has been established that the cultivation of producers on optimal sources of carbon and nitrogen increased protease activity of cultural liquid of B. circulans 693 3.8 times, Bacillus sp. 27--2.7 times, Y. lipolytica 2061--3.4 times. It has been found that the usage of different protein substrates in cultural medium permitted to obtain the proteolytic enzymes with various specificity with respect to globular and fibrillar proteins.

The results of the Department of Biochemistry investigations during the recent 5 years are presented. For the first time the structures of O-specific polysaccharides of lipopolysaccharides of representatives of Enterobacteriaceae new genus--Rahnella aquatilis have been established. The structures are represented by linear or branched mono-, tri- and hexasaccharides. As well as in other Enterobacteriaceae representatives only one hydrohylated acid--3-hydroxytetradecanoic has been established in lipids A of R. aquatilis. It was shown that toxicity and pyrogeneity of lipopolysaccharides R. aquatilis and Ralstonia solanacearum are due to the acyl-, phosphoryl- and free hydroxylic groups of lipids A. Physico-chemical properties and substrated specificity of enzymes-glycoproteins (alpha-L-rhmanosidase, a-N-acetylgalactosaminidase, collagenase and keratinase) have been studied. On the basis of the data concerning substrate specificity of the above enzymes the possible trends of their practical usage are considered.

Screening of producers of proteases among 367 strains of microorganisms has been carried out. A capacity to synthesize the enzymes was revealed in 25% of the studied strains of Bacillus genus (from 0.0054 to 0.892 un./mg of protein) and in 35% of the yeasts of Yarrowia genus (0.95 to 1.349 un./mg of protein). Elastase, gelatinase, caseinolytic, fibrinolytic and haemoglobinolytic activities were found in culture liquid of 7 the most active strains, their values depended on time of producer cultivation.

The influence of some technological parameters of cultivation of the producer Bacillus subtilis 147 on the process of extracellular enzymes alpha-amylase synthesis was investigated. The optimum sources of carbon (0.1% insoluble starch) and nitrogen (0.2% sodium nitrate) for maximum production of alpha-amylase are established. It was shown, that the temperature 42 degrees C, the carbon nitrogen ratio 1:2, pH 7.0, volume of a nutritious medium 100 ml, rotation rate 220 rev/min during 4-6 days are optimum parameters of the producer cultivation. As a result the enzyme activity was increased 14 times.

Lipopolisaccharides (LPS) of 8 strains of Pragia fontium--representatives of a new genus of Enterobacteriaceae family, was isolated for the first time. It was demonstrated, that lipopolysaccharides yields were 10-20% of dry cells weight. Investigations of monosaccharide composition have determined the presence of glucose, galactose, rhamnose and mannose in lipopolysaccharides, that permitted to establish 4 chemotypes ofpolysaccharide part of LPS. Serological investigations showed immunochemical heterogeneity of Pfontium: 8 investigated strains were divided by the O-antigenicity into 7 serogroups.

The spot sweetflag virus (SSV) as to its morphology and structural components in comparison with phytorhabdovirus of curly potato dwarf virus (CPDV) and rhabdoviruses of vesicular stomatitis virus (VSV), pathogenic for people and animals, corresponds to the definition of rhabdovirus and belongs to Rhabdoviridae family. Virions of SSV have a bacillus-like form and dimensions 110-130 x 45 nm. SSV contains structural proteins 130, 66, 43-39, 32-30 and 25 kDa. In the virion structure the fatty acids have been identified: palmitic (47 %), linolic (4.2%), oleic (14.9%), stearic (3.94%), holesterol (23%), and also carbohydrates: glucose (25.3%), galactose (18.3%), arabinose (16%), rhamnose (3.1%) and mannose (2.32%). Aminosaccharides: glucosamine and galactosamine, with correlation 1:7.2, were also found out. The paper is presented in Ukrainian.