- Details
- Category: 6_2011(en)
- Hits: 176
ISSN 1995-5537
Р. 9-17, Bibliography 47, Russian.
Universal Decimal classification: 602.6:582.5/.9
Yu. S. Luchakivs’ka, O. M. Kishchenko, M. Yu. Vasylenko, Yu. V. Simonenko, N. V. Kuchuk
Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, Kiyv
Recent studies have shown high immunogenicity of secretory Mycobacterium tuberculosis Ag85B and ESAT6 proteins. That's why it was suggested to use them as components of a new tuberculosis vaccine instead of the widely used BCG nowadays.
Production of recombinant proteins, especially vaccines, in transgenic plants has many economic and safety advantages as compared to animal and bacterial production systems, thus transgenic plants with raw-edible leaves, fruits and taproots could be discussed as so-called «edible vaccines». In this study we report the obtaining of carrot plants of Nantskaya, Karotel, Red Giant, Perfektziya and Amsterdamskaya varietes expressing the genes of secretory Mycobacterium tuberculosis proteins. The genes were transferred into plants via Agrobacterium-mediated transformation using Agrobacterium tumefaciens GV3101 and Agrobacterium rhizogenes A4 strains. The binary vectors contained genes of Ag85B, Ag85B(ΔTMD), ESAT6 and fused Ag85B(ΔTMD)-ESAT6 proteins driven by constitutive 35S CaMV promoter and root-specific Mll sugar beet promoter. PCR and RT-PCR analysis proved the presence and transcription for nptII selective gene and fbpB and esxA genes for all the analyzed carrot plants in case of nptII and for 92–95% of them in case of target genes. Western-blot analysis proved presence of the recombinant proteins for several plant lines.
Key words: carrot, recombinant protein, secretory proteins, Mycobacterium tuberculosis, Agrobacterium-mediated transformation.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008
- Details
- Category: 6_2011(en)
- Hits: 199
ISSN 1995-5537
"Biotechnology" journal V. 4, No. 6, 2011
Р. 18-30, Bibliography 37, Ukrainian.
Universal Decimal classification: 575.24+615.222:581.143.6
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine, Kyiv
A growth and development as a complete organism as a plant tissue culture have determined by different parameters oscillation of the cell system that formed culture lines, tissue or an organism as a whole not only at level processes with circadian rhythms (circadian organization level) but at level processes with passage rhythms during passage organization. However this basic processes have not been adequately explored.
Dynamic of cell population systems in vitro has been studed by the example of Rauwolfia serpentina Benth culture tissues at passage level, specifically temporal organization and robustness of this system. The phase trajectories of productivity indexes dynamics (biomass and indoline alkaloids accumulation), morphogenesis index dynamic (traheid elements dynamic), as well as dynamics of cell part with different DNA content in the nucleus and different nucleoli square at passage level under culturing condition variation for the high-productive Rauwolfia serpentina K-27 strain (ajmaline producer) were researched. The obtained results have indicated point of view that applied biotechnological methods of productivity increasing in the researched cell culture are based on dynamics hereditary memory.
Key words: cell population in vitro, dynamic of the cell systens, phase trajectories, Rauwolfia serpentina.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008
- Details
- Category: 6_2011(en)
- Hits: 180
ISSN 1995-5537
"Biotechnology" journal V. 4, No. 6, 2011
Р. 31-35, Bibliography 14, Ukrainian.
Universal Decimal classification: 577.151:544.023.223
PHENOL OXIDATION USING COVALENTLY IMMOBILIZED HORSERADISH PEROXIDASE
I. I. Romanovska,O. V. Oseychuk,S. S. Dekina, Y. A. Shesterenko,
O. V. Sevastyanov, T. Yu. Gromovoy
Bogatsky’s Physico-Chemical Institute of National Academy of Sciences of Ukraine, Odesa
Chuiko Institute of Surface Chemistry of National Academy of Sciences of Ukraine, Kyiv
Odesa National Medical University
According to modified Bakh’s method, partially purified horseradish peroxidase preparation (RZ = 1,0; activity 100 U/mg of protein) was isolated. Molecular mass of obtained HPR (42 972 Da) was confirmed by mass-spectrometry: matrix activated laser desorption/ ionization. The enzyme was covalently immobilized by periodate oxidation of polysaccharide membrane «Diacell», followed by sodium borohydride reduction, at mass ratio enzyme: support 0.2:1. The biocatalyst with quantitative protein coupling, 88% of activity’s pH- and thermooptima, increased termostability, stable at storage (8 months) was obtained. Kinetic studies had shown the increasing of hydrogen peroxide and pyrogallol Kм values and decreasing of Vmax values during immobilized HPR-catalyzed reactions. In the batch reactor the biocatalyst developed promoted the quantitative phenol oxidation (1 mmol/dm3) during 7 cycles of usage and high level of it’s bioconversion (95–50%) during the next 15 cycles. Molecular weight (about 2021 Da) of polymer product, insoluble in water and most organic solvent, of peroxidase oxidation of phenol — polioxyfenilene (particle size is from 15 to 75 mm) consisting of 21 units was first determined by laser desorption/ionization.
Key words: horseradish peroxidase, Diacell membrane, covalent immobilization, phenol oxidation, polyhydroxyphenylene.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008
- Details
- Category: 6_2011(en)
- Hits: 175
ISSN 1995-5537
"Biotechnology" journal V. 4, No. 6, 2011
Р. 42-50, Bibliography 33, Russian.
Universal Decimal classification: 579.22:581.557
SOYBEAN LECTIN EFFECT ON METABOLISM AND SYMBIOTIC PROPERTIES OF Bradyrhizobium japonicum STRAINS
D. M. Sytnikov, О. D. Krugova, N. M. Mandrovskaya
Kholodnyi Institute of Botany of National Academy of Sciences of Ukraine, Kyiv;
Institute of Plant Physiology and Genetics of National Academy of Sciences of Ukraine, Kyiv
The effects of soybean seed lectin (SBA) on growth, metabolism and symbiotic properties expression of soybean nodule bacteria (Bradyrhizobium japonicum) strains with different activity (634b, 646, 21110 and 604k) were studied in laboratory and vegetative experiments. It was shown that preliminary incubation of rhizobia with host-plant lectin stimulated their growth and increased the peroxidase activity of strains independently of the potential symbiotic activity. Soybean lectin initiated the appearance of new proteins in bacteria cells of rhizobial active strain 646. It was established that specific plant lectin was able to stimulate or inhibit the nodulation and nitrogen-fixing of strains with different activity under «soybean - nodule bacteria» symbiosis condition. Thus, homologous lectin through metabolism and character of symbiotic properties of rhizobia is multidirectional in its action on the genetically determinated microsymbiont potential.
Key words: Bradyrhizobium japonicum, soybean lectin, soybean-rhizobial symbiosis, nitrogenase activity, antioxidant enzymes.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008
- Details
- Category: 6_2011(en)
- Hits: 169
ISSN 1995-5537
"Biotechnology" journal V. 4, No. 6, 2011
Р. 51-59, Bibliography 33, Russian.
Universal Decimal classification: 621.014.46(546.57+546.56:66-911.38).001.5
STRUCTURE, PROPERTIES AND TOXICITY OF NANOPARTICLES
OF SILVER AND COPPER OXIDES
Institute of Occupational Health of National Academy of Medical Sciences of Ukrainу, Kyiv;
Paton Institute of Electro-welding of National Academy of Sciences of Ukraine, Kyiv;
Chuyko Institute of Surface Chemistry of National Academy of Sciences of Ukraine, Kyiv;
Palladin Institute of Biochemisry of National Academy of Sciences of Ukraine, Kyiv
Currently, a number of general methods for nanoparticles synthesis have been created. In terms of opportunities for industrial production of nanomaterials in Ukraine today, the most advanced is nanotechnology based on physical methods of their obtaining. Therefore today finding stable organic colloidal dispersions stabilized by surfactants and standardization of the experiments techniques in vitro and in vivo, development of safety criteria, and acceptable thresholds for toxicity of metal nanoparticles is an urgent problem. Physicochemical properties of nanoparticles of silver and copper oxides in colloidal solutions of albumin, polyvinylpyrrolidone, and dextran have been studied. Cytotoxicity of nanoparticles of silver and copper oxides in polyvinylpyrrolidone and dextran was evaluated. Toxicity of silver oxide nanoparticles was assessed during per os subacute experiment on Wistar rats. High toxicity of 46.6 mg/l concentration of silver oxide nanoparticles was detected. Biochemical study of enzyme system activity, measurement of trace elements content in whole blood and morphological study of blood cellular composition enabled to reveal minor changes in the composition of blood cells and amount of trace elements, as well as a significant reduction in ceruloplasmin activity and increased activity of ATP and zincprotoporphirine in response to per os subacute exposure of silver oxide nanoparticles with a size of 32 nm and 2.5 mg/l concentration.
Key words: nanoparticles of silver and copper oxides, albumin, polyvinylpyrrolidone, dextran, microelements, enzymes.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008
- GROWTH OF Ganoderma applanatum (Pers.) Pat. AND G. lucidum (Curtis) P. Karst. STRAINS AND SYNTHESIS OF POLYSACCHARIDES IN SUBMERGED CULTURE T. A. Krupodorova
- NUTRIENT MEDIA FOR in vitro CULTIVATION OF Ungernia victoris Vved. ex Artjushenko TISSUES O. M. Bublyk, I. O. Andreev, К. V. Spiridonova, L. P. Mozhylevska, V. A. Kunakh
- EFFECT OF DIESEL FUEL ON BIOSYNTHESIS OF PROTEINS, CARBOHYDRATES AND LIPIDS IN Chlorella vulgaris Beijer. А. I. Gorda, V. V. Grubinko
- COMPARATIVE STUDY OF ANTITUMOR EFFECT OF PRISTINE C60 FULLERENES AND DOXORUBICIN S. V. Prylutska, А. P. Burlaka, Yu. I. Prylutskyy, U. Ritter, P. Scharff