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Vol 59, No 2 (2023)

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Articles

Extracellular Vesicles of Bacteria Mediate Intercellular Communication: Practical Applications and Biosafety (Review)

Chernov V.M., Mouzykantov A.A., Baranova N.B., Chernova O.A.

Abstract

Extracellular vesicles, secreted by bacterial cells, are the focus of close attention of researchers. They are enriched with bioactive molecules, mediate the intercellular communication of micro- and macroorganisms, participate in the adaptation of bacteria to stressful conditions, reprogramming target cells, modulating immunoreactivity in higher organisms, changing the structure of microbial communities and ecosystems. The unique properties of bacterial extracellular vesicles (BEVs) open up broad prospects for their practical application – in clinical medicine, agriculture, biotechnology and ecology as diagnostic markers, vaccines, new biological products and means of their delivery. However, to implement the practical applications, a number of problems need to be solved. This review focuses on the ambiguous role of BEVs in the regulation of living systems, the problem of assessing the safety of BEVs and approaches to its solution related to innovative technologies.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):107-119
pages 107-119 views

Probiotic Properties of Saccharomycetes (Review)

Ryabtsevа S.A., Khramtsov A.G., Sazanova S.N., Budkevich R.O., Fedortsov N.M., Veziryan A.A.

Abstract

The purpose of the review is to summarize and analyze information on the molecular genetic basis and methods for studying the probiotic activity of Saccharomycetes fungi, the mechanisms of their physiological action, and their application in biotechnology. The relevance of research in this area is confirmed by the dynamics of the growth of publications. The effectiveness of Saccharomyces boulardii in the treatment and prevention of diarrhea of various etiologies, relapses of C. difficile infection, side effects of H. pylori infection therapy has been established with a high level of evidence. Genetic, cytological, cultural and biochemical features of S. boulardii determine their probiotic activity. Other Saccharomyces strains with probiotic potential are most often isolated from national fermented plant and dairy products. A unified methodology for studying the probiotic properties of yeast has not yet been created; clinical trials involving people are needed to confirm their status. Promising probiotics are strains of the species S. cerevisiae and K. marxianus, which have an international safety status. Possible mechanisms of physiological action of Saccharomycetes include antimicrobial and antitoxic, trophic, antisecretory and anti-inflammatory effects. Some of the mechanisms of yeast probiotic action differ from those of bacteria, and not all of them are yet understood. Saccharomycetes probiotics can be used to improve the biological value, quality and safety of food products.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):120-132
pages 120-132 views

Application of Sulfate-reducing Bacteria in Bioremediation from Heavy Metals and Metalloids (Review)

Bryukhanov A.L., Khijniak T.V.

Abstract

Pollution of the environment with heavy metals, metalloids and radionuclides is a global problem that seriously affects the state of the biosphere. In particular, chromium compounds have a toxic, mutagenic and carcinogenic effect. The main principle of purification of anthropogenic and natural ecosystems from chromates is the reduction of Cr(VI) to Cr(III), the salts of which are significantly less toxic and insoluble. However, currently used electrochemical and ion-exchange cleaning methods are quite expensive and require the use of special reagents. At the same time, sulfate-reducing bacteria (SRB) are of particular interest for bioremediation of this kind, since many of them are very resistant to high concentrations of heavy metals and are able to effectively reduce them in the presence of hydrogen as an electron donor. The review summarizes known data on the interaction of heavy metals, metalloids and radionuclides with SRB. The features of the metabolism of these microorganisms, leading to intracellular accumulation of heavy metals and metalloids, are considered. Complex and finely regulated enzymatic mechanisms for the reduction of toxic metals (using various cytochromes, hydrogenases, oxidoreductases, highly specific metal reductases, and thioredoxin/thioredoxin-reductase systems), as well as the possibility of using immobilized cells and biofilms of SRB in the effective bioremediation of natural waters, soils, and industrial effluents, are described.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):133-149
pages 133-149 views

The Use of Phage Antibodies for Microbial Cells Detection (Review)

Guliy O.I., Evstigneeva S.S., Dykman L.A.

Abstract

Phage antibody display technology has revolutionized the field of bacterial immunodetection. This technology allows the expression of an antibody fused to the coat protein of a filamentous bacteriophage. The use of phage display makes it possible to obtain high-affinity antibodies by passing the stage of animal immunization, reducing the time for obtaining stable antibody-producing clones from several months to several weeks, significantly reducing the cost of the process. These advantages make phage antibodies an important tool for bacterial detection. The paper presents a brief description of the technological methods for obtaining phage antibodies to microbial cells. The possibilities and prospects for using phage antibodies as a selective agent in analytical systems, including biosensors, are discussed.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):150-166
pages 150-166 views

Production of Soluble Human Gamma Interferon in the Escherichia coli Expression System with a Decrease in Cultivation Temperature

Volosnikova E.A., Esina T.I., Shcherbakov D.N., Volkova N.V., Gogina Y.S., Tereshchenko T.A., Danilenko E.D.

Abstract

A recombinant strain producing human gamma interferon (IFN-γ) E. coli BL 21/pET-IFN-γ was constructed, providing a high level of its expression. A method has been developed for obtaining a soluble form of recombinant IFN-γ, consisting of the processes of producing a biomass of a producer strain containing a target protein in an amount of 32–37% of the total content of cellular proteins, protein isolation and purification. The purification process included the stages of disintegration, clarification of the cell lysate, chromatographic purification and dialysis. The developed method makes it possible to obtain from 1 g of wet biomass up to 5 mg of the drug with a purity of at least 95% and high specific (antiviral) activity.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):167-173
pages 167-173 views

Formation of Various Antimicrobial Peptide Emericellipsin Isoforms in Emericellopsis alkalina under Different Cultivation Conditions

Kuvarina A.E., Sukonnikov M.A., Rogozhin E.A., Serebryakova M.V., Timofeeva A.V., Georgieva M.L., Sadykova V.S.

Abstract

A microbiological screening of the target component of emericellipsin A of the Emericellopsis alkalina E101 strain was carried out in various biotechnological systems at various pH. The content of emericellipsin A was quantified under these conditions.It has been established that the new approved membrane-liquid cultivation method at pH 10 contributes to an increase in the yield of the main component of emericellipsin A. It was shown that the new method of cultivating the strain E. alkalina E101 also promotes the synthesis of various isoforms of the main component of emericellipsin A. Some comparative analysis of them was carried out.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):174-181
pages 174-181 views

Synergistic Interaction of Arabinases of Different Types of Action in the Bioconversion of Sugar Beet Pulp and Apple Pomace

Semenova M.V., Kuryshkina M.S., Sinitsyn A.P.

Abstract

The interaction of endoarabinase (endoA) with exo-type enzymes was studied during their joint hydrolysis of branched arabinane (BAra), sugar beet pulp (SBP) and apple pomace (AP). It was shown that mixtures of endoA with arabinofuranosidase (AF) or arabinoxylan-arabinofuranhydrolase (AXH) with endoA content of 20 and 40%, respectively, were the most effective in the hydrolysis of BAra. As a result of the optimization of the complex of arabinases, cellulases and pectinase, almost complete conversion of AP into monosaccharides (arabinose, glucose, fructose) was carried out. During the hydrolysis of SBP, the conversion rate of hemicellulose (arabinane) was more than 50%, cellulose – 75%.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):182-190
pages 182-190 views

Decolorization of Dyes in the Bioelectrochemical System Depending on the Immobilization of Shewanella oneidensis MR-1 Cells on the Anode Surface and Electrical Stimulation of the External Circuit

Samkov A.A., Chugunova Y.A., Kruglova M.N., Moiseeva E.V., Volchenko N.N., Khudokormov A.A., Samkova S.M., Karaseva E.V.

Abstract

The effect of the polarity of the electrical stimulation of the external circuit of the bioelectrochemical systems, as well as the immobilization of Shewanella oneidensis MR-1 cells containing the DyP peroxidase gene on the rate of discoloration of dyes of different types was found. For the crystal violet triphenylmethane dye, the maximum decolorization rate by suspended S. oneidensis MR-1 cells 2.05 ± 0.07 μM/h was noted in the case of connecting a 1.2 V direct polarity DC voltage source. One of the minimum rates was observed in the case of reverse polarity of the connection. In the case of cells immobilized on the anode, the rate was higher, reaching 2.91 ± 0.09 μM/h and did not decrease with increasing substrate concentration. The lowest values were also noted for the reverse connection of the voltage source. In case of the azo dye congo red, the maximum rate was found for a source with direct connection and an open circuit (0.26 ± 0.01 and 0.29 ± ± 0.02 μM/h, respectively), the minimum value is 0.11 ± 0.02 μM/h for reverse connection. For the crystal violet decolorization products, a significant decrease in the intensity of the main absorption peak at 590 nm band was found, with no notable hypsochromic shift. The qualitative changes in the decolorization products composition are indicated by the appearance, in case of a direct polarity of the ionistor connection, of a new absorption maximum in the region of 360 nm. The results may be of interest for the development of new methods of bioelectrochemical cleaning.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):191-199
pages 191-199 views

Influence of Bacterial Mutualists and Phytopatogenes on Changes in Concentrations of cAMP and H2O2 in Pea Seedles of Rondo Varieties and its Clutterless and Superclub Mutants

Lomovatskaya L.A., Zakharova O.V., Goncharova A.M., Romanenko A.S.

Abstract

Changes in the concentrations of hydrogen peroxide and cyclic adenosine monophosphate (cAMP) in the roots of seedlings of pea cv. Rondo and its supernodulating mutant Nod3 and anodulating K14 were studied during infection with Rhizobium leguminosarum bv. vicea (strain RCAM 1022) or Pseudomonas syringae pv. pisi (strain 1845). It was shown that 360 min after infection of pea seedlings of the Rondo variety, the level of endogenous hydrogen peroxide slightly differed from the control. In the roots of Nod3 seedlings, this level significantly decreased, and in the roots of K14 it significantly increased when infected with the 1845 strain, but remained unchanged when exposed to bacteria of the RCAM 1022 strain. and young root hairs of Rondo seedlings, while strain 1845 had no effect on this parameter. Both types of bacteria had no effect on the concentration of cAMP in the roots of seedlings of the Nod3 mutant, whereas in K14, under the influence of RCAM 1022, the cAMP level almost doubled, and under the influence of 1845, it decreased. It is assumed that hydrogen peroxide and cAMP may be involved in the formation of supernodulating and nodulating phenotypes of mutants, as well as in the formation of resistance to a specific pathogen, Pseudomonas syringae pv. pisi. It is possible that this phenomenon can be used to diagnose the resistance of newly created mutants and pea varieties to the blight pathogen.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):200-207
pages 200-207 views

Immobilized Microorganisms-based Biopreparation as a Major Component for New Generation High-effeciency Organic Fertilizers

Yudina N.Y., Kozlova T.N., Abramova T.N., Arlyapov V.A., Asulyan L.D., Alferov V.A., Reshetilov A.N.

Abstract

Two biopreparations made of the bacteria Bacillus megaterium and Azotobacter chroococcum immobilized in modified polyvinyl alcohol hydrogel have been created. It has been shown that after application of the constructed biopreparations, an increase in soil moisture content (by a factor of two), germination (by up 100%), the length of the cress stem and roots (15–30%) was achieved. The addition of Azotobacter chroococcum-based biopreparation led to a 2-time increase of nitrogen in soil. The survival period for bacteria immobilized in hydrogel based on polyvinyl alcohol modified with succinic acid was longer than 30 days. It is concluded that the studied biopreparationsexhibited high potential as a major material for efficient biofertilizer of new generation.

Prikladnaâ biohimiâ i mikrobiologiâ. 2023;59(2):208-216
pages 208-216 views