Biotechnologia Acta

Number 2, 2018

5-22
ELECTRONIC INFORMATION SYSTEMS IN BIOTECHNOLOGY

Authors: O. M. KLYUCHKO

Abstract

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The aim of the work was to generalize and analyze the use of electronic information systems in biotechnology in order to create new versions of these systems. The publications concerning the systems of different types for solving the problems in biotechnology were studied. Similar systems which traditionally belong to biology and medicine were classified. The prospects of their application for development of more advanced electronic systems were considered.

23-29
SYNTHESIS AND CHARACTERIZATION OF POLY(D,L-LACTIC-CO-GLYCOLIC)ACID MICROPARTICLES LOADED BY DIPHTHERIA TOXOID

Authors: T. O. Chudina, V. A. Skoryk, E. I. Legach

Abstract

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Number of views: 338

The aim of the study was to synthesize poly(D,L-lactide-co-glycolide) particles — PLGA and obtain their complexes with a non-toxic diphtheria toxin recombinant fragment subunit B fused with enhanced green fluorescent protein EGFP-SubB; to characterize the main physical and chemical properties of the resulting complexes. Two types of the diptheria toxoid loaded PLGA microparticles were obtained: particles with immobilized diphtheria toxoid (PLGA 1) and particles with encapsulated (PLGA) diphtheria toxoid (PLGA 2). Micropartcles obtaines were characterized for antigen loading, particle size, polydispersity index and in vitro antigen release.

30-39
RECOGNITION OF Mycobacterium tuberculosis ANTIGENS MPT63 AND MPT83 WITH MURINE POLYCLONAL AND scFv ANTIBODIES

Authors: A. A. Siromolot, O. S. Oliinyk, D. V. Kolybo

Abstract

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The goal of this study was to characterize serum immunoglobulin G (IgG) antibody responses during experimental immunization of laboratory mice by purified recombinant proteins MPT63, MPT83 of M. tuberculosis and artificial fusion protein MPT83-MPT63 and obtain the recombinant single chain variable fragments of antibodies (scFv) against these antigens. This study demonstrates that the humoral immune response to MPT63, MPT83, MPT83-MPT63 fusion protein and equimolar set of MPT63 and MPT83 was highly different. For each antigen, serum antibody levels were evaluated by a cutoff value based on optical density index. A crucial role of MPT83 for immunogenicity of chimeric protein and/or cocktail of individual antigens under conditions of immunization of laboratory animals. We obtained also specific scFv antibodies against MPT63 and MPT83. These antibodies can be used for the development of the system for quantitative determination of antigens as well as for their biological properties investigation. Thereby, based on the results of the immune response and mycobacterial proteins antigenicity we showed highly immunogenicity properties of N-terminal part of MPT83 antigen for enhencement of ELISA sensitivity. We suggest MPT83-MPT63 fusion protein as a potential candidate on the role of antigenic substance for the serological diagnosis of tuberculosis.

40-46
MATHEMATICAL MODELLING OF OPTIMIZATION NUTRIENT MEDIUM COMPOSITION FOR ENTOMOPATHOGENIC BACTERIA STRAIN Bacillus thuringiensis 87/3 CULTIVATION

Authors: М. V. Boiko, N. V. Patyka, S. М. Shulga, O. O. Tigunova, H. S. Andriiash

Abstract

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Number of views: 361

The aim of the research was to develop an optimal liquid medium composition for bacteria strain B. thuringiensis 87/3 which is the most favorable for the identification the potential production of biologically active components. In this work we have used new entomopathogenic bacteria strain B. thuringiensis var. thuringiensis (Bt H1) № 87/3 selected in vitro, isolated from the larvae of leaf-eating insects natural populations of Leptinotarsa decemlineata Say (L4). Nutrient medium for cultivation of Bt 87/3 was optimized on base a full-fledged xperiment, according to the Box-Behnken (33) plan. Optimumal concentrations were investigated in the medium of carbon source (15 g/l of glucose), nitrogen (10 g/l of corn extract) and phosphorus-containing inorganic salts (1.5 g/l of diammonium phosphate). When this strain grown in periodic conditions at 30 ºC, the number of viable cells reached 4.4.109 cells/ml. Nutrient medium that was optimized can be recommended for cultivation of Bacillus thuringiensis 87/3 in production conditions.

47-56
POLYMORPHISM OF SOME TRANSCRIPTION FACTOR GENES RELATED TO DROUGHT TOLERANCE IN WHEAT

Authors: Lakhneko O. R., Stepanenko A. I., Kuzminskiy Ye. V., Morgun B. V.

Abstract

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Number of views: 446

The aim of the research was to study polymorphism of preselected gene loci of three transcription factors (TaNAC2a, TaWRKY2, and TaWRKY19) and the Late Embryogenesis Abundant (LEA) proteins dehydrin (Td29b) related to wheat drought tolerance. The genes structure and chromosome location were established via bioinformatics tools. It is stated that TaWRKY2 and TaWRKY19 genes were comprised of 4 exons and 3 introns located on 2BS and 1DS chromosome arms, respectively; TaNAC2a — 2 exons and 1 intron 7AS; Td29b — single exon gene 3AS. Using polymerase chain reaction, no polymorphism was observed. Polymorphic bands were detected for TaWRKY2 locus. The screening of the distribution of the revealed polymorphic loci was carried out for a set of wheat and rye varieties, old landraces and interspecific hybrids. The polymorphism of TaWRKY2 locus indicated the presence of some other possible alleles of the gene. The obtained data are important for further investigations of wheat drought tolerance.

57-63
APPLICATION OF AERATION-OXIDATIVE JET-LOOPED SETUP FOR BIOLOGICAL WASTEWATER TREATMENT

Authors: O. M. Obodovych, L. A. Sablii, V. V. Sydorenko, M. S. Korenchuk

Abstract

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Number of views: 361

The purpose of the study was the design of an aerator-oxidizer and optimal parameters of its work to ensure the conditions of mixing and dissolution of oxygen, in which there is no disturbance of the state of activated sludge. The testing of the work of the aeration-oxidation setup of rotor type with the use of a sludge mixture with different design of aerator-oxidizers and in different operating modes was performed. The assessment of the effect of processing on the state of the activated sludge was carried out according to standard parameters: sludge index, chemical oxygen demand. In addition, the number and state of activated sludge organisms were evaluated. The results of qualitative and quantitative analysis of activated sludge before and after processing in the setup are presented. The parameters in which the activated sludge functions in satisfactory mode are revealed.

64-71
NECESSITY OF TRANSLOCATION DOMAIN FOR REALISATION OF CYTOSTATIC EFFECT OF NON-TOXIC DERIVATIVES OF DIPHTHERIA TOXIN

Authors: K. Y. Manoilov, O. I. Krynina, A. Ju. Labyntsev, S. I. Romaniuk, D. V. Kolybo

Abstract

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Number of views: 367

The aim of the work was to evaluate in vitro the cytostatic effect of recombinant fragments of the non-toxic point mutant of diphtheria toxin — CRM197, which was suggested as a potent medication for treatment of triple negative breast cancer. For this purpose, non-toxic recombinant derivatives of diphtheria toxin — CRM197, subunit B SbB and receptor domain Rd had been isolated by Ni-NTA agarose affinity chromatography and their effect on the growth of individual colonies of triple negative breast cancer MDA–MB–231 cells were characterized by such parameters as average colony area, perimeter and circularity index. According to the obtained results, CRM197 and SbB, whose molecules contain the translocation domain Td, exhibited the same cytostatic effect against MDA–MB–231 cells, reducing the area and perimeter of individual colonies. Rd protein did not affect the last two parameters that characterize the size of the colonies, but changed the form of the margin of colonies, as evidenced by an increase in the circularity index. It is supposed that Td may be involved in the implementation of cytostatic action due to its inherent pore-forming activity in relation to lipid membranes. It is concluded that Rd and Td, unlike the catalytic domain of diphtheria toxin, play important roles in the implementation of the cytotoxic properties of CRM197, while SbB consisting of Rd and Td is the structural DT fragment of smallest molecular weight that can be used as the analog of CRM197.

72-78
PECULIARITIES OF ANTIBIOTIC BATUMIN ACTION ON BIOFILM FORMATION BY Staphylococcus aureus AND Pseudomonas batumici

Authors: L. N. Churkina, V. V. Klochko, S. .D Zagorodnya, L. V. Yaroshenko, O. B. Luitko

Abstract

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Number of views: 337

The aim of the work was the research of antibiotic batumin action on the biofilm formed by staphylococci and on the process of the biofilm formation by strain-producer Pseudomonas batumici. The minimal inhibiting concentration (MIC) of batumin was studied according to The Clinical and Laboratory Standards Institute standards — CLSI. Biofilm formation was studied using photometric O’Toole method with flatbed photometer at wavelength 540 nm. The batumin penetrated into biofilms of staphylococci and reduced the biomass of the formed biofilm of Staphylococcus aureus and S. epidermidis at the antibiotic concentration of 0.5 μg/ml, which was only twice higher than MIC of batumin for planktonic cells. The degree of biofilm changes in the presence of batumin differed for different staphylococcus strains. However, these differences did not depend on strains sensitivity or their resistance to the antibiotic. The ability of batumin strain-producer to form the biofilm and the ability of antibiotic to stimulate this process in concentrations of 1 and 10 μg/ml was established for the first time. In addition, it was found that in the Pseudomonas batumici B-321 planktonic culture in the presence of 10 μg/ml of batumin, the increase in length of producer cells on average by 15% was noted. The obtained data shed the light on batumin antimicrobic action on the staphylococci biofilm. It can be argued that is a perspective for treatment of staphylococcal infections. It enabled to consider batumin as a promising drug for the staphylococcal infections treatment.