3/2016

Author

page: - pp
DOI: 10.11134/btp.3.2016.1
Starovoitova S.A.
National University of Food Technologies
68, Vladimirskaya str., Kiev, 01601, Ukraine
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Abstract

Types of tannins and their sources for selection, and characteristics such as the structure and properties of the tannins, especially those of bacterial origin, were considered. The positive and negative properties of various tannins, mainly those of vegetable origin, were reviewed. The methods of biodegradation of the main types of tannins and enzymes involved in the processes of their decomposition were analysed. The possibility of using tannins and their properties in various industries, especiallyin food and pharmaceutical industries, was discussed. The data of many known researchers with respect to bacterial tannase activity and methods of determination and isolation of bacteria with tannase activity were summarized. The main tannase-producing bacteria, especially those with potential probiotic properties and use in creating bacterial drugs and functional foods (namely, bacteria that belong to the genera such asLactobacillus, Bacillus, Enterococcus,andPentococcus), were shown. Application perspectives of the tannase activity of probiotic microorganisms as a basis for pharmaceutical products-probiotics and functional foods enriched with probiotic microorganisms that have relevant antioxidant and anti-tumour properties—were discussed. The practical uses of tannase are still quite limited because of insufficient information on its properties and the complexity of its production and purification.

Keywords: tannase activity, gut microbiota, probiotics, antioxidant properties

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Author

page: - pp
DOI: 10.11134/btp.3.2016.2
Ryabushkina N., Abugalieva S., Turuspekov Y.
Institute of Plant Biology and biotechnology
Timiryazev street 45, Almaty 050040, Kazakhstan
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Abstract

Plants are vitally important part of biodiversity that determines a global stability of ecosystems. According to the International Convention on Biological   Diversity,   two thirds of plants in the world are underextinction due to anthropogenic activities and global climate change. The strategies of plant protection on regional and global levels must be focused on evaluation of current conditions of bioresources, the development of systematic statuses of all living plant species, including those under the extinction. There is a necessity to fullydescribe floristically rich regions with purpose to develop a proper strategies for conservation of ecosystems, rare, endemic, and industrially important wild species both in situ and ex situ. Based on international experience, the achievement of this goal should be achieved through creation ofprotected natural territories in Kazakhstan, including National Reserves and National Parks. One of the modern approaches for the integrated study of the plant genetic resources is molecular systematics, which is based on combination of research studies in botany and molecular genetics. 

Keywords: flora, biodiversity, inventory, preservation, protected natural territories, genbank, DNA

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Author

page: - pp
DOI: 10.11134/btp.3.2016.3
Aхambayeva A.S.1, Shagyrova Zh.S.1, Nurgozhin T.S.2, Zhienbay E.2, Shustov A.V.1
1 National Center for Biotechnology
Korgalzhyn hwy, 13/5, Astana, 010000, Kazakhstan
2 Nazarbayev University
Kabanbai Batyr avenue, 53, Astana, 010000, Kazakhstan

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Abstract

Mussel adhesive proteins (MFPs) help mussels to attach to various surfaces. Natural MFPs present in the adhesive discs of the byssal threads of the mussels exhibit a number of good adhesiveproperties thatcan be used in medicine, orthodontics, and cell and tissue engineering. MFPs can be used to glue a variety of materials, natural and artificial. For use in medicine, it is important that the MFPs are waterproof adhesives, effectively bonding surfaces submerged in water. Natural MFPs are already being used as adhesives, but their utilization is limited because of high costs.

Recombinant MFPs are an attractive alternative because they can be produced in largequantities. For the recombinant MFPs produced by Escherichia coli to possess adhesive properties, post-translational modification of tyrosine residues to 3,4-dihydroxyphenylalanine (DOPA) is required.

This paper describes an expression system in which the recombinant adhesive protein Fp-131 is produced, and the newly synthesized polypeptide undergoes modification of the tyrosine residues to DOPA. Hydroxylation of tyrosine occurredin vivo in the bacterial cells because of the activity of tyrosinase, which is coexpressed with Fp-131. Coexpression of Fp-131 and tyrosinase was achieved through the expression of the proteins from two plasmids with different origins of replication and resistance markers.

Fp-131 was purified using metal affinity chromatography under denaturing conditions. After dialysis and freeze-drying, a product was obtained with an yield of 25 mg from 1 L of the induced culture. Presence of DOPA was demonstrated in Fp-131 by using a colour reaction with NBT. The adhesion strength of Fp-131 was measured using the lap-shear test. For this test, Fp-131 was used to glue two flat adherends, and a shear load was attached to the glued joint. The adhesive strength was 1.1 MPa, which is comparable to that of natural MFPs.

Keywords: adhesive protein, mussels, tyrosinase, DOPA, adhesion strength, biocompatible glue

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Author

page: - pp
DOI: 10.11134/btp.3.2016.4
Karalnik B.V.1, Denisova T.G.1, Omarova M.N.1, Zhunussova G.B.1, Tugambayev T.I.2, Zakaryan S.B.2, Fedosov Y.S.1
1H.Zhumatov Scientific Hygiene and Epidemiology Center
34,Makataev str., Almaty, 050002, Kazakhstan
2M. Aykimbayev Kazakh Scientific Center of Quarantine and Zoonotic Diseases
14,Kapalskaya str., Almaty, 050054,Kazakhstan

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Abstract

The aim of this study was to develop immunologic reagents from a synthetic sorbent for early brucellosis diagnostics and compare them with previously developed erythrocyte immunologic reagents by using animal immunization experiments and examination of patients. Rabbits; veterinary brucellosis vaccines for sheep and cattle; lipopolysaccharides (LPS) of different species of Brucella and Yersinia chemically activated (with -NH2or –COOH groups) with polystyrene beads(diameter,2 µm) and tagged or not tagged with a fluorochrome (natriumisothiocyanate); previously developed Brucella erythrocyte immunologic reagent for revealing lymphocytes with receptors for the antigens; and mononuclear fractions of rabbits and patients suspected to have brucellosis were used. Methods of adhesion of cells/corpuscles and their blocking with LPS and statistical comparisons were used. A Brucella immunologic reagent was developed from the synthetic sorbent for determining LfRLPS specificity. Advantages of the developed immunologic reagent were as follows: absence of non-specific adhesion due to binding of the sorbent rather than LPS and higher accuracy of determination of LfR; non-obligation of parallel determination of sorbent binding per se by lymphocytes; and simplification and decrease of spending time for the LfR count. Features of the sorbent did not influence the detection of LfR specific to taxonomically close bacteria. The results substantiate the expediency of use of the immunologic reagent from a synthetic sorbentfor the determination of LfR.

Keywords: sorbed immunologic reagents, early diagnostics, brucellosis

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Author

page: - pp
DOI: 10.11134/btp.3.2016.5
Nadirova L.T., Stanbekova G.E., Beisenov D.K., Iskakov B.K.
M. Aitkhozhin Institute of Molecular Biology and Biochemistry
86 Dosmukhamedov str., Almaty, 050012, Kazakhstan
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Abstract

Potato is one of the most consumed crop products. It is grown inan area of 190000 hectares in Kazakhstan. The potato spindle tuber viroid (PSTVd) is the causative agent of the so-called ‘gothic’ disease, and it causesthe degradation of potato tubers anda decrease in yield of up to 65%. PSTVd has been found in every continent;in Kazakhstan, molecular diagnostics for PSTVd have not yet been performed.

The first survey of PSTVd in Kazakhstan was performed, and spindle- and pear-shaped tubers collected from private farms in the Almaty region were grown under greenhouse conditions. Total RNAwas isolated from the leaves and analysed for the presence of PSTVd. Reverse transcription-polymerase chain reaction (RT-PCR) with specific primers revealed a 360-bp DNA fragment in a number of samples. The correspondence to PSTVd was confirmed using sequencing. Subsequently, the isolated PSTVd clone was used as a labelled probe for detecting the viroid in other plants. RT-PCR and nucleic acid hybridization assay revealed the presence of PSTVd in 26% of the selected spindle tubers.

Currently, there are no effective methods to prevent the infection of potatoes by PSTVd; therefore, diagnostic methods are crucial for preventing the spread of PSTVd through seed material.

Keywords: potato spindle tuber viroid, diagnostic, reverse transcription, PCR, northern blotting

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Author

page: - pp
DOI: 10.11134/btp.3.2016.6
Tagimanova D.S., Khapilina O.N., Amenov A.A., Kalendar R.N.
National Center for Biotechnology
13/5, Kurgalzhyn road, Astana, 010000, Kazakhstan
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Abstract

High-quality DNA is necessary for molecular genetic studies. It is difficult to extract quality DNA from plant tissues, especially herbarium specimens, since they contain significant amounts of polysaccharides, phenols, and pigments that reduce the efficiency of PCR. An effective method for extracting genomic DNA from herbarium specimens is required. Nucleic acid purification methods can be classified into two general categories: liquid phase and columns containing sorbents.In this paper, wedescribed various methods for extracting DNA from herbarium specimens of Rhodiola rosea. The results showed low efficiency of methods based on the use of chaotropic salts such as guanidin tiotsionata, PVP and β-mercaptoethanol.The use of SDS-extraction buffer reduced the quantitative parameters of the DNA samples. For the extraction of DNA from herbarium material, the most effective protocol was based on acid cetyltrimethylammonium bromide buffer with hot chloroform. The buffer increased the yield of high-quality DNA because it prevented oxidative processes and the formation of DNA chemical components with pigments and polysaccharides. Use of this method resulted in 100% recovery of DNA from all the samples.

Keywords: Rhodiola rosea, cenopopulation, DNA, herbarium, molecular genetic analysis

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Author

page: - pp
DOI: 10.11134/btp.3.2016.7
Mukantayev K., Shustov A., Tursunov K., Ingirbay B., Adish Zh., Ramanculov E., Mukanov K.
National Center for Biotechnology
13/5, Korgalzhyn hwy, Astana, 010000, Kazakhstan
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Abstract

Matrix proteins of RNA viruses are the main structural components and are required for virus assembly. The  assembly of these virus particles is based on the ability of binding of the matrix protein to the nucleocapsid. This property suggests a promising role for the matrix protein, as it acts as a bridge between the nucleocapsid and plasma membrane. Analysis of literature on the biological functions of basic components of the rabies virus andthe spread of rabies in the Republic of Kazakhstan showed the importance of research on the recombinant matrix antigen of the virus. In this study, we used genetic engineering to show the results of receipt ofthe recombinant matrix protein of the rabies virus. As a result of research carried out design of the expression genetic construction containing gene of matrix protein of the rabies virus, and was performed synthesis of this gene with length 654 nucleotides under de novo conditions. The nucleotide sequence of the synthesized gene showed the correct assembly. Using expression plasmids, anEscherichia coli strain that produced the recombinant matrix antigen of the rabies virus was obtained. Electrophoretic analysis showed the expression of the protein with a molecular weight of 41 kDa. We developed protocols for the cultivation of the obtained strain and purification of the recombinant matrix antigen. Immunization of mice and analysis of sera from the immunized mice with the enzyme-linked immunosorbent assay were used to study the immunogenic properties of the recombinant antigen.

Keywords: rabies, virus, matrix protein, recombinant antigen, genetic construction, production strain

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Author

page: - pp
DOI: 10.11134/btp.3.2016.8
Mukantayev K.N., Shustov A.V., Tursunov K., Іnіrbay B., Adish J., Rаmanculov E.M., Mukanov K.K.
National center of biotechnology
13/5, Korgaldjin st., Astana, 010000, Kazakhstan
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Abstract

The foot-and-mouth disease (FMD) is a highly contagious disease found in both agricultural and wild cloven-hoofed animals. The basic and effective method for preventing the emergence and spread of the infection is vaccination of susceptible livestock and creation of a buffer zone for the immunized animals. Identification of the infected animals ina herd of vaccinated animals is important for preventingFMD. In this paper, we have presented materials associated with obtaining 2C, 3A, 3B, and 3D recombinant non-structural proteins of the FMD virus (FMDV) and detecting their diagnostic properties by usingthe enzyme-linked immunosorbent assay (ELISA). On the basis of amino acid sequencing of the 2C, 3A, 3B, and 3D non-structural proteins of different types of FMDV,genes of the most immunogenic sites of the antigenswere selected. We designed genetic constructions containing genes of the 2C, 3A, 3B, and 3D non-structural proteins of FMDV. The pET32 vector was used as the expression plasmid.

For ELISA, recombinant 2C, 3A, 3B, and 3D non-structural antigens of FMDV were obtained. The molecular mass of the 2C, 3A, 3B, and 3D recombinant non-structural antigens was 40, 45, 38, and 34 kDa, respectively.

Keywords: biotechnology, virus, ELISA, FMD, antigen, non-structural antigen

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