[Comparative analysis of tobacco and Arabidopsis insertional mutants, transformed with equal vector constructions].

We have analyzed genetic effects of heterologous plant genes insertion on genome functioning of higher plants, belonging to different systematic groups (tobacco, Arabidopsis). Plants of different species were responding differently to the insertion of the same transgene, which is likely to be associated with the location of alien DNA insertion and could manifest in morphological changes spectrum and target gene expression level.

[Extracellular proteolytic activity of bacteria from soda-salt lakes of Transbaikalia].

Influence of nitrogen source on proteinases synthesis in aerobic alkalotolerant and halotolerant bacteria from soda-salt lakes of Transbaikalia was studied. Maximal accumulation of proteinases was revealed on medium with peptones. Introduction of various sources of nitrogen in the medium did not result in increase of enzyme activity in cultural liquid.

[The study of two alkaliphilic thermophile bacteria of the anoxybacillus genus as producers of extracellular proteinase].

Two strains of alkaliphilic thermophile bacteria of the genus Anoxybacillus from hydrothermal vents of Lake Baikal were detected and characterized. It was demonstrated that proteinases secreted by these bacteria had wide substrate specificity, hydrolyzed proteins and n-nitroanilide substrates, and showed maximal activity at pyroglutamyl-alanine-alanine-leucine n-nitroanilide hydrolysis. We determined maximal activity of the proteinases at alkaline pH values (10.

[Proteolytic enzymes of the fungi: extracellular proteases of xylotrophic basidiomycetes].

Proteolytic enzymes of the fungi attract attention of investigators due to many reasons among which their large diversity, wide substrate specificity, stability under extreme conditions (pH, temperature) are most important. Their functional significance, including various processes, from hydrolysis of macromolecular substrates under deficiency of nitrogen compounds to initiation and maintenance of pathogenesis, is also very interesting. The present review deals with classification and biochemical properies of extracellular fungal proteinases, their physiological role as well as fields of practical application.

[Extracellular proteases of mycelial fungi as participants of pathogenic processes].

The interest in proteases secreted by mycelial fungi is due to several reasons of which one of the most important is their involvement in the initiation and development of the pathogenic process. A comparison of saprophytic and phytopathogenic mycelial fungi revealed one characteristic feature, namely, the appearance of a new trypsin-like activity in phytopathogens that is absent in saprophytes. To clear up the question of whether the degree of pathogenicity of a fungus is related to the activity of secreted trypsin-like protease, several species of Fusarium of various pathogenicity were compared.

[Properties of post-proline cleaving enzymes from Tenebrio molitor].

Two post-proline cleaving enzymes PRE1 and PRE2 with molecular masses of 101 and 62 kDa, respectively, capable of hydrolyzing Z-AlaAlaPro-pNA were isolated for the first time from the midgut of the flour beetle Tenebrio molitor and characterized. PRE1 is active only in acidic media, with a maximum at pH 5.6, whereas PRE2, both in acidic and alkaline media with a maximum at pH 7.

[Isolation and properties of cysteine protease from Serratia proteamaculans 94].

A new cysteine protease (SpCP) with a molecular mass of about 50 kDa and optimal functioning at pH 8.0 was isolated from the culture medium of a Serratia proteamaculans 94 psychrotolerant strain using affinity and gel permeation chromatography. The enzyme N terminal amino acid sequence (SPVEEAEGDGIVLDV-) exhibits a reliable similarity to N terminal sequences of gingipains R, cysteine proteases from Polphyromonas gingivalis.

[Extracellular proteinases of filamentous fungi as potential markers of phytopathogenesis].

The presence of proteins in the culture liquid of filamentous fungi under study was found to induce the secretion of proteinases. The inhibitory analysis of the major extracellular proteinases of the saprotrophic fungus Trichoderma harzianum and the phytopathogenic fungus Alternaria alternata showed that they both belong to the group of serine proteinases. The substrate specificity of these proteinases and their sensitivity to inhibitors suggest that the enzyme of T.

[Extracellular proteolytic eyzymes of microscopic fungi from thermal springs of the Barguzin Valley (Northern Baikal region)].

Production of extracellular proteolytic enzymes was studied in thermophilic fungi Paecelomyces variotii and Aspergillus carneus, isolated from thermal springs of the Barguzin Valley. Protease synthesis in these fungi requires protein in the ambient medium. The composition of the enzymes secreted by A.

[Degradation of proteinaceous substrates by xylotrophic basidiomycetes].

The ability of various xylotrophs to produce extracellular proteolytic enzymes has been studied, with emphasis on medium-related factors regulating their secretion. Direct measurement of proteolytic activity in the culture liquid and postelectrophoresis determination of protease activity in polyacrylamide gel copolymerized with gelatin demonstrated that the secreted enzymes are quantitatively and qualitatively diverse. Activity levels of extracellular proteolytic enzymes strongly depend on pH and contents of protein and carbohydrate in the medium.

[Proteinase inhibitors as antistress proteins in higher plants].

Physicochemical and functional characteristics of plant protein proteinase inhibitors as antistress biopolymers were studied to determine the mechanisms for plant resistance to phytopathogens and to obtain disease-resistant cereal and leguminous cultures. The activity of trypsin, chymotrypsin, and subtilisin inhibitors varied in monocotyledonous and dicotyledonous cultures. Study varieties of leguminous and cereal cultures were shown to contain endogenous inhibitors specific to proteinases of phytopathogenic fungi Fusarium, Colletotrichum, Helminthosporium, and Botrytis.

[Protease inhibitors: use to increase plant tolerance to insects and pathogens].

The review deals with analysis of the possibility of the use of genes of inhibitors of proteolytic enzymes of plants to increase plant tolerance to insect pests and phytopathogens. The idea of using protease inhibitors for plant defense is strongly supported, first, by their wide distribution in plant tissues and high activity towards various proteolytic enzymes of insects, bacteria and fungi. The results obtained for the last years indicate that the genetic engineering approach is perspective for solving of this kind of problems.

[Effect of the antioxidant ionol on proteolytic apparatus of aging coleoptiles of wheat seedlings grown in light].

The dynamics of changes in total proteolytic activity and activities of various groups of proteases in the coleoptiles of 3- to 12-day-old wheat seedlings grown in light with and without antioxidant BHT (2,6-di-tert-butyl-4-methylphenol) was studied. It was established that the specialized proteases that easily hydrolyze specific synthetic substrates and the enzymes actively hydrolyzing histone H1 dominate in young coleoptiles of 3- to 4-day-old seedlings. Proteases that degrade equally well the majority of the studied substrates are accumulated in the cells of old coleoptiles of 11- to 12-day-old seedlings.

[Isolation and primary structure of trypsin from the red king crab Paralithodes camtschaticus].

Trypsin from hepatopancreas of the Paralithodes camtschaticus crab was isolated in homogeneous state by successive ion-exchange chromatography on DEAE-Sephadex, affinity chromatography on Agarose modified with peptide ligands from trypsin hydrolysate of salmin, and ion-exchange chromatography on a Mono Q column. The total yield of the protein was 64%. Its N-terminal amino acid sequence was determined (IVGGTEVTPG-).

[Manganese-dependent ribonucleotide reductase of Propionibacterium freudenreichii subsp. shermanii: partial purification, characterization, and role in DNA biosynthesis].

Like Lactobacillus leichmanii, Rhizobium meliloti, and Euglena gracilis, P. freudenreichii implicates cobalamin in DNA anabolism via adenosylcobalamin-dependent ribonucleotide reductase. However, in the absence of corrinoids, P.

[Amino acid sequence of anionic protease inhibitors from buckwheat seeds].

Complete amino acid sequence of IT1 protease inhibitor and partial amino acid sequences of IT2 and IT4 protease inhibitors from buckwheat Fagopyrum esculentum Moench seeds were determined by automatic Edman degradation and mass spectrometry. IT1 inhibitor comprises 69 amino acid residues and its molecular mass is 7743.8 D.

[Isolation and properties of serine proteinase PC from the Kamchatka crab, Paralithodes camtschatica--a proteolytic enzyme with broad specificity].

A homogeneous serine proteinase PC has been isolated from the Camchatka crab (Paralithodes camtschatica) hepatopancreas using affinity chromatography on arginine-Sepharose, protamine tryptic peptide-agarose and ion-exchange chromatography on Mono-Q, with a 68% yield. The enzyme is completely inhibited by diisopropylfluorophosphate, a typical inhibitor for serine proteinases. The molecular mass of the proteinase is 29 kDa, pI is 3.

[Inhibition of exogenous serine proteinases by a trypsin inhibitor from the buckwheat IT-1 seeds].

The possibility of inhibition of exogenous trypsin- and chymotrypsin-like proteinases by a proteinase inhibitor from buckwheat (IT-1) seeds has been studied. The inhibition constants for bovine trypsin and alpha-chymotrypsin and human granulocyte cathepsin G by IT-1 are equal to 1.1, 67 and 200 nm, respectively.

[Isolation and properties of carboxypeptidase from the Kamchatka crab Paralithodes camtshatica].

Homogeneous carboxypeptidase PC from a hematopancreas of kamchatka crab Paralithodes camtshatica was obtained by means of an affinity chromatography on sorbents containing arginine, protamine hydrolysate, and phenylalanine as ligands with an yield 23% and purification degree 37.4. The isolated enzyme has a molecular mass 34 kDa, as evidenced by an SDS-PAGE; pI 3.

[Changes in activity of proteolytic enzymes (BAPAases) in germinating buckwheat and rye seeds].

The interrelationship between the activity of proteolytic enzymes (BAPAases) from buckwheat and rye seeds hydrolyzing Nalpha-benzoyl-DL-arginine-p-nitroanilide (BAPA) and the amount of the antiserum to these enzymes necessary to obtain a certain inhibition level has been studied at different stages of seed germination. The data obtained show that the increase of the BAPAase activity in germinating rye seeds is due to de novo synthesis of this enzyme. During this process antigenically identical enzyme molecules are synthesized in roots and shoots of the developing plant.

[Immunochemical characterization of proteolytic enzymes (BAPAase) from buckwheat and rye seeds].

A comparative study of the immunochemical properties of two serine proteolytic enzymes (BAPases) from buckwheat and rye seeds hydrolyzing N alpha-benzoyl-DL-arginine-p-nitroanilide (BAPA) has been performed. It has been shown that buckwheat and rye seed BAPAases were partially antigenically identical. This was demonstrated using double immunodiffusion in agar and inhibition of one enzyme with an antiserum to the other.