[Formation of Ag/AgCl Nanoparticles in the Matrix of the Exopolysaccharide of a Diazotrophic Strain Azotobacter chroococcum XU1].

A complex of Ag/AgCl nanoparticles was synthesized on the basis of the extracellular polysaccharide of Azotobacter chroococcum XU1 and 10 mM AgNO3 solution. The complex was characterized by UV spectroscopy, X-ray diffraction, and scanning electron microscopy. Colloidal solutions of the complex had absorption peaks at 260 and 420 nm, indicating the formation Ag/AgCl nanoparticles. The size of the nanoparticles varied from 6 to 50 nm. The nanobiocomposite consisting of the exopolysaccharide matrix and Ag/AgCl nanoparticles exhibited a fungicidal effect against such plant pathogens as Fusarium oxysporum f. sp. vasinfectum and Verticillium dahliae.

[Plasmids of Salt-tolerant Rhizosphere Microorganisms].

Five plasmid DNAs were detected in three of microorganisms, which formed microbial composition «Ð—амин-М¼. One plasmid (23.1 kb) was found in Bacillus megaterium СКБ-310 cells and another one (55.0 kb) was found in cells of Pseudomonas stutzeri СКБ-308, but cells of Bacillus subtilis СКБ-309 contained 3 plasmids (48.5 kb, 30.0 kb and 13.3 kb). It was assumed that these plasmids may carry genes of resistance to adverse environmental conditions, including the high content (10 %) of ions Cl- and SO4(2-).

[State of Fungal Lipases of Rhizopus microsporus, Penicillium sp. and Oospora lactis in Border Layers Water-Solid Phase and Factors Affecting Catalytic Properties of Enzymes].

We demonstrated that a change in the catalytic activity of fungal lipases synthesized by Rhizopus microsporus, Penicillium sp. and Oospora lactis and their ability to absorb on different sorbents depended on the nature of groups on the solid phase surface in the model systems water: lipid and water: solid phase. Thus, the stability of Penicillium sp. lipases increased 85% in the presence ofsorsilen or DEAE-cellulose, and 55% of their initial activity respectively was preserved. In the presence of silica gel and CM-cellulose, a decreased rate of lipid hydrolysis by Pseudomonas sp. enzymes was observed in water medium, and the hydrolysis rate increased by 2.4 and 1.5 times respectively in the presence of aminoaerosil and polykefamid. In an aqueous-alcohol medium, aminoaerosil and polykefamid decreased the rate of substrate hydrolysis by more than 30 times. The addition of aerosil to aqueous and aqueous-alcohol media resulted in an increase in the hydrolysis rate by 1.2-1.3 times. Sorsilen stabilized Penicillium sp. lipase activity at 40, 45, 50 and 55 degrees C. Either stabilization or inactivation of lipases was observed depending on the pH of the medium and the nature of chemical groups localized on the surface of solid phase. The synthetizing activity of lipases also changed depending on the conditions.

[Purification of extracellular proteinases from B. subtilis SKB 256 by biospecific chromatography].

Abstract-A simple and efficient method of preparing highly purified extracellular proteinases of B. subtilis B-1 (SKB 256) has been developed. A sorbent based on sorsilen impregnated with hemoglobin or cytochrome c has been synthesized for this purpose. A significant difference between the efficiency of hemoglobin and cytochrome c as biospecific ligands has been observed: the enzyme yield amounted to 40.6 and 65.6% of the total amount of enzyme adsorbed, respectively. The culture was shown to contain two major proteinase forms with different molecular masses that could be separated by chromatography on a Sephadex G-50 but gave only one band with MW 27 kDa upon denaturing electrophoresis in 12.5% PAG in the presence of 0.1% SDS. The influence of eluent pH, ionic strength and ethanol concentration on the sorption of the proteinases on the biospecific sorbent, as well as on the desorption from it, has been investigated. Positive influence of 20% ethanol on proteinase desorption has been demonstrated.

[The effect of acid proteinases on the activity and stability of glucoamylase preparations].

It was demonstrated that the presence of acid proteinases in the preparation isolated from Aspergillus awamori decreased the activity and stability of glucoamylase. Patterns of changes in the enzymatic activity and stability of glucoamylase at increased temperature and various pH values were studied over a long-term storage. A biospecific sorbent for removal of acid proteinases was synthesized, and glucoamylase preparations free of proteolytic activity were produced.

[Effects of nutrient medium composition and temperature on the germination of conidia and the entomopathogenic activity of the fungi].

The effects of nutrient medium composition and temperature on the germination of conidia of the fungi Beauveria bassiana (strain AlG) and Metarhizium anisopliae (strain M-99) and their entomopathogenic activity have been studied. It was demonstrated that the presence of carbohydrates alone was sufficient for the spores of M. anisopliae M-99 to germinate, whereas the germination of B. bassiana AlG spores was inhibited by carbohydrates. Addition of KJ, ZnSO4, or KBr into the Czapek medium increased the entomopathogenic activity of B. bassiana. The optimum temperature for spore germination was 20-35 degrees C in both fungal species.

[Conditions for cultivation of the fungus Penicillium melinii UzLM-4 and its biosynthesis of lipases]. / Usloviia kul'tivirovaniia i biosintez lipaz gribom Penicillium melinii UzLM-4.

Cultivation of the fungus Penicillium melinii UzLM-4 on a Raistrick's medium of our own modification made it possible to increase the biosynthesis of lipases three to four times. The following conditions ensured a high rate of synthesis of the extracellular lipase: age of the inoculum, 15 days; concentration of the inoculum, 15 x 10(6) conidia per 100 ml medium; initial pH of the nutrient medium, 8.0; and cultivation in a shaker at 150 rpm (25 degrees C).

[Effect of culture medium composition on peroxidase biosynthesis by the fungus Pleurotus ostreatus UzBI-I105]. / Vliianie sostava pitatel'noi sredy na biosintez peroksidazy gribom Pleurotus ostreatus, shtamm UzBI-I105.

Production of extracellular peroxidase during submerged cultivation of the xylotrophic bazidiomycete Pleurotus ostreatus UZBI-I105 in nutrient media with lignocellulosic wastes, exhausted cottonseed oil cake, cotton stalks, rice husks, or ambary hemp was studied. The enzyme production increased threefold to fivefold in the presence of exhausted cottonseed oil cake extract in the nutrient medium. The dynamics of peroxidase production in various media was investigated.

[Insecticidal activity of Bacillus thuringiensis cells]. / Entomotsidnaia aktivnost' bakterial'nykh kletok Bacillus thuringiensis.

The influence of sixteen different nutrient media on the entomopathogenic activity of three Bacillus thuringiensis strains was studied. The medium composition based on potato, yeast extract, and molasses was optimized. B. thuringiensis No 1 grown on the media No 7 and 9 displayed the highest entomopathogenic activity (94.3 and 90.6%, respectively).

Bioreactors based on immobilized cells.

For purification of oil-containing sewage water of oil-proceeding enterprises it has been elaborated by authors the laboratory bioreactor on the basis of immobilized mycelial Mucor miehei fungus cells with periodical and constant supply of purified solution. The purification process is tested on model and real oil-containing liquid wastes. It has been established that under true bioreactor exploitation, 1 g of Mucor miehei immobilized cells enables to purify more than 100 L of sewage water, containing 0.1-0.15% of lipid substances.

[Enzymatic utilization of cotton soap stock]. / Frmentativnyi sposob utilizatsii khlopkovogo soapstoka.

Enzymatic hydrolysis of neutral fat of cotton oil soap stock with a nonspecific lipase produced by Oospora lactis F-500 was designed. The culture liquid and a preparation of enzyme obtained by precipitation with isopropanol from a filtrate of the culture liquid were used. Utilization of cotton oil soap stock as the only source of carbon during cultivation of the fungus was studied. The rate of hydrolysis of soap stock fat strongly depended on the way of biological conversion of cotton oil soap stock. The most effective utilization was observed during cultivation of the fungus in the medium containing soap stock.

Purification and characterization of a Penicillium sp. lipase which discriminates against diglycerides.

A lipase was isolated from Penicillium sp. strain UZLM-4 and characterized. This lipase has a molecular weight of 27,344 (determined by mass spectrometry) and hydrolyzes triglycerides in preference to mono- and diglyceride substrates. Among various triglyceride substrates, tributyrin is hydrolyzed about four times faster than any other tested. The lipase has a preference for hydrolysis at the 1,3 positions of the lipids and shows a weak stereoselectivity for the S enantiomer. Unlike most other lipases, this lipase is stable and has a high activity at low surface pressures (5-10 mN/m).

[Various lipases for producing products enriched with polyenic acids in fish fat hydrolysis]. / Issledovanie razlichnykh lipaz dlia polucheniia produktov, obogashchennykh polienovymi kislotami pri gidrolize rybikh zhirov.

The enzymic hydrolysis of fish with lipases from various sources was studied. The lipase from the fungus Rhizopus microsporus preferentially removes saturated fatty acids, while lipase from the pyloric caeca of salmon unsaturated fatty acids upon hydrolysis of fish fats. The enzymes can be used to obtain fatty products enriched with eicosanopentaenoic acid, mono- and diacylglycerols by enzymic hydrolysis of the ivasi fat.

[Properties of lipases from Oospora lactis]. / Svoistva lipaz Oospora lactis.

The procedure of isolation and purification of lipases from the fungus Oospora lactis have been developed. The existence of two different lipases has been demonstrated, one of which (Mr = 43000) is localized in the periplasmic space and can be liberated into the external medium in an unchanged form, while the other (Mr = 40000) is tightly bound to the membranes and can be solubilized by detergent treatment. The most essential properties of the lipases are discussed and a detailed analysis of the functional and physicochemical properties of extracellular lipase is given.

[Multiplicity of lipase forms from Rhizopus microsporus]. / Priroda mnozhestvennosti form lipazy Rhizopus microsporus.

The lipase from Rhizopus microsporus of glycoprotein origin is represented by five forms differing in their molecular weights and localization in the cell. The lipase forms are distinguished by their relative content of A and B subunits (AB, 2A + B, A + 2B, 4A + B, 3A + 2B). Only three isoenzymes containing the bulk of the non-protein moieties were detected in the culture fluid. The other two forms can exist only as intracellular proteins.

[Lipase activity of Oospora lactis protoplasts]. / Lipaznaia aktivnost' protoplastov Oospora lactis.

The paper describes how to prepare osmotically sensitive forms from the fungus Oospora lactis. The lytic enzyme from actinomycetes at a concentration of 25 mg/ml causes abundant formation of protoplasts at 36 degrees C and pH 7.5 within 10--15 min. The best stabilizing agent is 1 M NaCl solution or a mixture of 1 M NaCl with 1 M mannitol solution (1:1). In order to induce lysis of cell walls by the enzyme from Helix pomatia, the cells must be pretreated with a solution containing 40 mM tris (hydroxymethyl)-aminomethane, 5 mM EDTA and 0.2 M cysteine. The production of Oospora lactis protoplasts was accompanied with the liberation of lipase from the periplasmic space. The activity of lipase was 90% of the overall intracellular activity as was confirmed by the results of differential centrifugation of a homogenate prepared by mechanical disintegration of the cells. Since the Rf in disc electrophoresis, the pH optimum and the substrate specificity of exocellular lipase were identical with those of periplasmic lipase, the enzyme must be liberated without a change in the molecular weight.

[Extracellular lipases of the fungus Rhizopus microsporus UzLT-1]. / Izuchenie vnekletochnykh lipaz griba Rhizopus microsporus, UzLT-1.

Lipase biosynthesis by the fungus Rhizopus microsporus UzLT-1 was investigated. Under certain conditions the fungus could produce two forms of extracellular lipases. The paper describes the process of purification of the enzyme L-I,N- and C-terminal amino acids of the lipases have been determined. Identity of terminal amino acids of both enzymes has been demonstrated. It has been shown that both forms of lipase from Rhizopus microporus hydrolyze predominantly alpha-ester bonds.

[Rhizopus microsporus strain UzLT-I--a thermotolerant producer of lipase]. / Rhizopus microsporus shtamm UzLT-1-termotolerantnyi produtsent lipazy

The properties of the thermotolerant fungus Rhizopus microsporus strain UzLT-1--producer of lipolyptic enzymes are described. Optimal cultivation conditions--40 degrees, C, pH 4.5--Are determined. The lipolytic activity of the culture on the medium consisting of corn extract (2%), cotton-seed oil (1%) and water is 850 ml 0.1 n KOH per 100 ml culture liquid. The enzymic preparations of lipase have been precipitated by isopropanol and ammonium sulphate. The preparation precipitated by isopropanol shows its macimum activity at pH 4.2 and 7.8 and a temperature of 40--50 degrees C.