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2017
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MODERN BIOTECHNOLOGICAL APPROACHES TO LIFESPAN EXTENSION OF ANIMALS AND HUMANS E. L. Levitsky

Biotechnologia Acta V. 10, No 2, 2017, Р. 7-21, Bibliography 49, English
https://doi.org/10.15407/biotech10.02.007

Abstract:
The purpose of the research was to analyze current data concerning the problem of extending the life of multicellular animals and humans. The modern views about the processes of aging and prolongation of life are presented. The analysis focused on the genetic mechanisms of aging and mainly biotechnological approaches (genetic engineering, gene therapy, the use of stem cells, and the reprogramming of the genome) to prolong the life of multicellular organisms. For comparison, some traditional methods of prolonging life are described (drug therapy, exercise training, calorically restricted nutrition). This analysis allows   to postulate the perspectives and advantages of using biotechnological methods for prolonging life in comparison with traditional ones.

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INDUSTRIAL WASTE BIOCONVERSION INTO SURFACTANTS  BY   Rhodococcus erythropolis ІMV Ас-5017, Acinetobacter calcoaceticus ІMV В-7241 and  Nocardia vaccinii ІMV В-7405 T. P. Pirog, M. O. Shulyakova, L. V. Nikituk, S. I. Antonuk, I. V. Elperin

Biotechnologia Acta V. 10, No 2, 2017, Р. 22-33, Bibliography 35, English
https://doi.org/10.15407/biotech10.02.022

Abstract:
The aim of the work is to realize an alternative processing of toxic industrial waste into surfactants by strains Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241 and Nocardia vaccinii IMV B-7405 for remediation of environment.The studied strains were grown in liquid media containing such sources of carbon as waste (fried) sunflower oil, technical glycerol (by-product of biodiesel production), and aromatic compounds. The synthesis of surfactants was evaluated by emulsification index, conditional concentration of surfactants and concentration of extracellular surfactants, which was determined gravimetrically after their extraction from supernatant by the mixture of methanol and chloroform. The concentration of oil in water and soil was analyzed by gravimetric method after extraction with hexane.
It was shown that with increasing concentration of the inoculum up to 10−15% and two times increase of nitrogen source content in medium containing 7−8% (v/v) of crude glycerol, concentration of surfactants synthesized by R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B 7241 and N. vaccinii IMV B-7405 was 3.4; 5.0 and 5.3 g/l, respectively, that is 1.6−1.7 times higher as compared with values on basal medium with the same content of substrate. The maximum concentration (3.9−4.3 g/l) of surfactants synthesized by A. calcoaceticus IMV B-7241 on fried sunflower oil (4%) was achieved by using the inoculum grown on refined oil. The ability of R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B-7241 and N. vaccinii IMV B-7405 to decompose aromatic compounds (phenol, naphthalene, toluene, hexachlorobenzene, benzoic and N-phenylanthranilic acid) with simultaneous synthesis of extracellular metabolites with surface-active and emulsifying properties was established. In the presence of surfactants in the form of culture liquid (5−10%), the degree of degradation of complex oil with heavy metal (Cu2+, Cd2+, Pb2+, 0.01−0.5 mmol) pollution in water (3−6 g/l ) and soil (20 g/kg) after 20 days was 82−92%.
Bioconversion of industrial waste into surfactants for environmental technology allows recycling toxic waste, reducing costs of microbial surfactants and provides double effect of environmental purification, which is achieved in the production and use of microbial surfactants.

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ANTIVIRAL ACTIVITY OF LIPOPOLYSACCHARIDES OF Pseudomonas chlororaphis subsp. aureofaciens L .D. Varbanets1, S. L. Rybalko2, D. B. Starosyla

Biotechnologia Acta, V. 10, No 2, 2017, Р. 34-39, Bibliography 9, English
https://doi.org/10.15407/biotech10.02.034

Abstract:
The aim of the study was to investigate the ability of lipopolysaccharides  of two strains of Pseudomonas chlororaphis subsp. aureofaciens to inhibit in vitro the reproduction of human viruses: influenza  A/FM/1/47 (H1N1), herpes simplex type 2 and bovine diarrhea, which is used as a model of hepatitis C virus, as well as to suppress hepatitis C virus production in model system of cells transfected with cDNA of this virus. It has been established that for both lipopolysaccharides in three types of cultures (MDCK, Vero and MDBK) the toxicity is not manifested even in a concentration of 100.0 μg/ml, and decreasing in infectious virus titer more than by 2.0 lg TCD50 (ED99) was already achieved at concentrations of 1.55 mg/ml. Selectivity indexes determination of lipopolysaccharides preparations against the influenza A/FM/1/47 (H1N1) virus, herpes simplex virus type 2 and bovine diarrhea virus shows that lipopolysaccharides of P. chlororaphis subsp. aureofaciens UCM B-306 and UCM B-111 are effective inhibitors of investigated viruses reproduction: selectivity index is at least 64. In the model of Jurkat cells transfected with human hepatitis C virus cDNA, viral RNA loading was determined in cells treated with lipopolysaccharides of P. chlororaphis subsp. aureofaciens. The results of the studies indicate that when lipopolysaccharides of both strains are administered, the production of the hepatitis C virus is completely inhibited.

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WATER BINDING IN COMPOSITE SYSTEMS BASED ON MILLED MEDICINAL PLANTS AND NANOSILICA V. V. Turov, T. V. Krupska, А. P. Golovan, L. S. Аndriyko, М. Т. Кartel

Biotechnologia Acta V. 10, No 2, 2017, Р. 40-56, Bibliography 22, English
https://doi.org/10.15407/biotech10.02.040

Abstract:
The aim of the study was to identify the influence of hydrodensified nanosilica particles on the binding of water by milled plant raw materials in neutral and acidic media. Flowers of Hibiscus sabdariffa and Calendula officinalis were used as the model materials. According to the microphotographs and low temperature 1H NMR spectroscopy data, the silica film forms on the surface of the milled plant particles, and it can significantly influence their hydration. According to the suggested scheme, some of the water from the inner cavities of plant raw materials moves (as evidenced by the decreasing radius of water-filled pores) to the zone of contact of the composite components (the radius of clusters of adsorbed water increases). In studies of desorption of active substances from milled medicinal herbs and their composites by the initial and hydrodensified nanosilica, it has been shown that the formation of a composite significantly reduces the rate of desorption. Minimal desorption is observed in composites containing hydrodensified nanosilica. The studied composite systems are promising for biomedical researches.

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IDENTIFICATION OF Psy1 GENE ALLELES RESPONSIBLE FOR CAROTENOID ACCUMULATION IN WHEAT GRAINS O. V. Stepanenko, A. І. Stepanenko, Ye. V. Kuzminskiy, B. V. Morgun

Biotechnologia Acta, V. 10, No 2, 2017, Р.  57-66, Bibliography 21, English
https://doi.org/10.15407/biotech10.02.057

Abstract:
The aim of the research was to select and optimize marker systems for identification of Psy1 genes alleles, which are responsible for different levels of carotenoid pigments accumulation in wheat grains, as well as to screen varieties for the selection of valuable genotypes. 162 wheat samples were analyzed by the polymerase chain reaction method. Among them, varieties and lines with different allelic states of Psy-A1 and Psy-B1 genes were identified. The Psy-D1 gene did not show any polymorphism. As a result, samples with valuable alleles of Psy1 genes, which potentially contain increased carotenoids content in grains, were selected.

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PROBIT ANALYSIS FOR Cd, Pb, Cu, Zn  PHYTOTOXICITY ASSESSMENT N. O. Ryzhenko, V. M. Kavetsky

Biotechnologia Acta, V. 10, Np 2, 2017, Р.  67-74, Bibliography 21, English
DOI: 10.15407/biotech10.02.067

Abstract:
The aim of investigation was to develop a new approach in Cd, Zn, Cu, Pb phytotoxicity assessing. This approach provided the probit analysis using. Methods included probit analysis, thin layer chromatography, statistical methods (calculation of the least significant difference, correlation analysis). We applied «doze-effect» model to obtain the ranking of the metals according to their phytotoxicity in spring barley field. We offered to estimate the phytotoxicity by PhLD50 index. Research results were: graphic formalization of "dose-effect" dependence and calculation of phytotoxic doses (PhLD50 and PhLD95) for Cd, Cu, Pb, Zn in polluted soil. According to PhLD50 value we conducted a comparative assessment of Cd, Cu, Pb, Zn phytotoxicity relatively to spring barley.   According to PhLD50 value metals could be ranked: Cd> Cu> Pb> Zn.   The most toxic metal was Cd.   PhLD  values, on which the estimation of metals phytotoxicity, were: Cd –  50, Cu –129, Pb –537, Zn –603 mg / kg mobile forms  in turf-podzol sandy loam soil. Our findings are relevant to estimating the metal hazard and controlling the condition of the crop growth.

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ANTIMICROBIAL, ENTOMOPATHOGENIC AND ANTIVIRAL ACTIVITY OF GAUPSIN BIOPREPARATION CREATED ON THE BASIS OF Pseudomonas chlororaphis STRAINS E. A. Kiprianova, L. A. Safronova, A. O. Prosyanov

Biotechnologia Acta, V. 10, No 1, 2017, Р. 7-16, Bibliography 25, English
https://doi.org/10.15407/biotech10.01.007

Abstract:
The aim of this review was to present the results of more than ten-year study of gaupsin biopreparation created on the basis of two strains Pseudomonas chlororaphis subsp. aureofaciens UCM В-111 and UCM В-306 with antifungal, entomopathogenic and antiviral activities. Data about antibiotic substances produced by these strains — phenazine and phenylpyrrole derivatives — are presented. Entomocidal properties against the wide spectrum of insect pests have been found out in the strains-producers. Antiviral activity of gaupsin due to the production of thermostable exopolymers containing neutral monosaccharides has been shown using the tobacco mosaic virus as a model. Lipopolysaccharides of the strains В-111 and В-306 also appeared to be highly active antiviral agents. Structure of their O-specific polysaccharides has been established. The last one are structurally heterogenic, presented by linear tri-and tetrasaccharide repeated links and have specific structure that has not been described previously.

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MOLECULAR MECHANISMS OF PLURIPOTENCY INDUCTION AND REPROGRAMMING OF SOMATIC CELLS T. O. Deputatova

Biotechnologia Acta, V. 10, No 1, 2017, Р. 17-25, Bibliography 41, English
https://doi.org/10.15407/biotech10.01.017

Abstract:
To analyze the recent studies elucidating the molecular mechanisms of pluripotency induction and crucial stages of the reprogramming process was the aim of the review. The key focus is on the factors enabling switch between the reprogramming stages. It is concluded that one of the key barriers for iPSC applications is the multi-stage nature of somatic cells reprogramming that features both stochastic early phases and deterministic establishment of pluripotent regulatory network. Despite thousands of scientific studies, various reprogramming protocols restrict effective research analysis and identification of molecular reprogramming mechanisms. In order to specify accurate reprogramming algorithms and develop more effective protocols of patient specific reprogrammed cells cultivation, the future researches require focus on the phase transition switchers.

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YEAST β-MANNANASE ACTIVITY N. V. Borzova, L. D. Varbanets, V. S. Pidgorskyi, O. D. Ianieva

Biotechnologia Acta, V. 10, No 1, 2017, P. 26-33, Bibliography 22, English
https://doi.org/10.15407/biotech10.01.026

Abstract:
The aim of the research was to determine the mannan-degrading activity of yeasts cultures isolated from various sources and select strains with high β-mannanase activity. As a result of screening of 245 yeast strains, which are the representatives of 7 genera and 14 species, the active producers of extracellular β-mannanase were identified. To increase β-mannanase activity, the cultures were grown under submerged conditions using guar gum galactomannan as a carbon source and an inducer. β-Mannanase activity was determined by dinitrosalicylic method. The most active biosynthetic species were Cryptococcus albidus, C. gastricus, C. magnus, C. terreus, C. laurentii, Saccharomyces cerevisiae, Williopsis californica, Metschnikowia pulcherrima, Pichia anomala and P. guilliermondii. The activity in culture supernatant was ranged from 0.2 to 75 U/ml. α-Galactosidase activity was found in two strains (Debaryomyces polymorphus UCM Y-152 and Debaryomyces hansenii var. fabryi UCM Y-2400). None of the tested cultures demonstrated both β-mannanase and α-galactosidase activity, that is, they are unable to attack both the main and side chains of galactomannan.

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DOMESTIC BUTANOL-PRODUCING STRAINS OF THE Clostridium GENUS O. O. Tigunova, N. Ye. Beiko, H. S. Andriiash, S. H. Pryiomov, S. M. Shulga

Biotechnologia Acta, V. 10, No 1, 2017, P. 34-42, Bibliography 54, English
https://doi.org/10.15407/biotech10.01.034

Abstract:
The aim of the work was to summarize the results of own research concerning obtaining butanol producing strains of Clostridium genus, to identify them by physiological, morphological and genetic methods. Further study of characteristics and biological features of the strains, and various approaches in biotechnological process of butanol production are discussed. The work includes methods to increase butanol accumulation by producer strains. Perspectives of using chemical mutagenesis in Clostridia as a method of increasing butanol production are considered. The feasibility of using non-food raw material as a substrate for fermentation is discussed. Different methods of pretreatment and their impact on the accumulation of butanol in the liquid medium are compared. Butanol accumulation is shown to increase significantly if the synthesis precursors are added as components of enzymatic medium, and the “reverse bard” is used to reduce waste production without affecting the level of butanol synthesis. The problem of conservation of producing strains is given, and protective medium for microorganisms during the freeze-drying is defined.

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THE CONTENTS OF NEUTRAL AND POLAR LIPIDS IN CLOSTRIDIA CELLS UNDER CULTIVATION IN THE PRESENCE OF BUTANOL S. I. Voychuk

Biotechnologia Acta, V. 10, No 1, 2017, P. 41-51, Bibliorg. 16, English
https://doi.org/10.15407/biotech10.01.043

Abstract:
The aim of the study was to evaluate changes in the portion of polar and neutral lipids in the cells of Clostridium during their cultivation in the presence of butanol. Four natural isolates of Clostridium genus were studied with flow cytometry approaches. Under the optimal culture conditions, the polar lipids prevailed over neutral ones in bacterial cells; the content of neutral lipids doubled in spores of these microorganisms, while the content of polar ones was reduced. Strains No 1 and No 2 were able to grow at 1% butanol in the medium, and the strain No 4 was at 1.5%. When cultivated in the presence of different concentrations of butanol, the bacterial strains did not differ in such cytomorphological features as granularity and cell size. The quantitative content of polar and neutral lipids in the presence of butanol varied depending on the content of butanol in the medium, however this effect had a strain-specific character and did not show a correlation with the resistance of these bacteria to butanol. So, the content of polar and neutral lipids varied depending on butanol content in the medium. However this effect was strain-specific independently of resistance of these bacteria to butanol. The use of bacterial biomass as a source of lipids for the production of biofuels requires further optimization of the process to increase the content of the neutral lipid fraction in bacterial cells.

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