Biotechnologia Acta

9-24

Authors: Kanyuk M. I.

The purpose of the work is to summarize the literature data concerning ultrafine diamonds, namely their industrial production, as well as considerable photostability and biocompatibility that promote their use in modern visualization techniques. It is shown that due to the unique physical properties, they are promising materials for using in nanotechnology in the near future. Possibility of diverse surface modification, small size and large absorption surface are the basis for their use in different approaches for drug and gene delivery into a cell. The changes in the properties of nanodiamond surface modification methods of their creation, stabilization and applications are described. It can be said that fluorescent surface-modified nanodiamonds are a promising target in various research methods that would be widely used for labeling of living cells, as well as in the processes of genes and drugs delivery into a cell.

35-42

Authors: Kapustianenko L. G., Iatsenko T. A., Iusova O. I., Grinenko T. V.

Our aim was to develop a method for isolation of human plasminogen kringle 5 possessing functional activity. The proposed method includes the following steps: hydrolysis of plasminogen with elastase, separation of mini-plasminogen from kringle fragments 1–3 and 4 on Lys-Sepharose, mini-plasminogen hydrolysis with pepsin, affinity chromatography on AH-Sepharose and polyacrilamide gel electrophoresis. We obtained the electrophoretically pure fragment of human plasminogen kringle 5 showing functional activity towards the ligands with high and low molecular mass. Weight yield was 3.8% that corresponds to 25.3% of the theoretically possible. It was established that affinity chromatography on AH-Sepharose was the sufficient step to isolate kringle 5 from mini-plasminogen hydrolysate with pepsin. This approach does not require additional purification steps while the ability of kringle 5 to bind specifically to AH-Sepharose demonstrates the functional activity of the kringle.

49-53

Authors: Matselyukh B. P., Matselyukh D. Ya., Golembiovska S. L., Gural S. V.

The aim of the paper was the selection of the improved strains of Phaffia rhodozyma using chemical mutagenesis and the identification of individual carotenoids synthesized by isolated more pigmented mutants. Hyperpigmented mutants of P. rhodozyma NRRL Y-17268-1 and IMB Y-5021-15 were isolated from initial strains NRRL Y-17268 and IMB Y-5021 by nitrosoguanidine mutagenesis. Pigments were purified by TLC and identified using HPLC and liquid chromatography-mass spectrometry. It was shown that initial strains P. rhodozyma NRRL Y-17268 and IMB Y-5021 and obtained from them mutants NRRL Y-17268-1 and IMB Y-5021-15 produced torulene and torularhodin without illumination in shaking flasks at 20 оC. The content of torularhodin produced by the mutant strains Y-17268-1 (18.2 μg) and Y-5021-15 (16.5 μg) per 1.0 g of dry biomass was increased to 33.8 and 18.4%, respectively, in comparison with the content of this pigment in the initial parental strains. The obtained strains present interest for further selection of more active producers of carotenoids and examination of the action of reactive oxygen species as stimulators of carotenoid production in yeasts.

61-70

Authors: Kuchma M. D., Shablii V. A., Kyryk V. M.3, Svitina A. N.2, Shablii Yu. N., Prokopets Yu. K., Indichenko T. M., Lukash L. L., Lobyntseva G. S.

The aim of the work was the comparative studing of the character of differentiation of hematopoietic progenitor cells of the placenta and umbilical cord blood in vivo and their multipotent properties in vitro. The proposed methods were used for mononuclear cells isolation from umbilical cord blood, placental tissue and mature fetal chorion, of flow cytometry and of analysis of the potential for differentiation. We found that majority of hematopoietic progenitor cells both in mature placenta and umbilical cord blood remains uncommitted, however in placental tissue we found more amount of differentiated cells that include myeloid progenitor with a phenotype CD34+CD45lowCD33+SSClow, later myeloid progenitors with a phenotype CD34+CD45lowCD14+SSClow (their content is significantly higher than in cord blood), erythroid progenitors with a phenotype CD34+CD45lowCD235+SSClow (their number significantly above than that in cord blood), B-lymphoid progenitors with a phenotype CD34+CD45lowCD19+SSClow, T-lymphoid progenitors and Natural Killer Cells-progenitors with a phenotype CD34+CD45lowCD7+SSClow, and also T-lymphocytes at the different stages of maturation with a phenotypes CD7+CD45+ and CD7+CD45RA+CD45+ respectively. Placental hematopoietic progenitor cells have similar potential for differentiation in vitro in comparison with cord blood ones. Presence of hematopoietic cells in placental tissue at different stages and lines of differentiation suggests that the placental hematopoiesis last during all term of gestation.

80-84

Authors: Zimina O. V., Sytnyk K. S., Parii M. F., Alkhimova O. G.

The goal of the work was, using the database «The Arabidopsis Information Resource» TAIR, to select 12 SSLP-markers distributed along the Arabidopsis chromosomes and chromosome arms, to optimize the conditions of amplification of each fragment and for simultaneous amplification of several fragments. For identification of A. thaliana ecotypes and their hybrid, the SSLP sequences were used. These DNA markers are highly polymorphic in Arabidopsis and easy to use. Using this database, the primers were selected for 12 SSLP-markers distributed along all chromosomes and their arms. A. thaliana ecotypes Columbia and Landsberg erecta were used. The experiments revealed that two-stage PCR using two annealing temperatures of primers in each cycle allows efficient amplification of all the fragments considered. The conditions for carrying out two multiplex PCR, each of which allows the two fragments were amplified and a single multiplex PCR allowing three markers for amplification were defined. The developed system of DNA markers can be used to study the behavior and inheritance of each chromosome of maternal and paternal genomes of Arabidopsis hybrids and enables quick and efficient genetic analysis.

92-99

Authors: Krupodorova T. A., Barsteyn V. Yu., Peshuk L. V., Haschuk O. I., Kostenko E. E.

The aim of this work was the study of influence of cultivation substrates (agriculture wasters) on biomass accumulation, amino acid composition, polysaccharide content and sorption ability towards heavy metals for the obtained biomass of edible myshroom P.ostreatus. The intensity of P. ostreatus biomass accumulation (18–24,1 g/L) and high conversion of substrates (33,3–44,6%) have shown prospects for P. ostreatus cultivation on new substrates such as wheat germ oil meal, CO2-extraction waste — amaranth flour and rapeseed meal. The optimum concentration of selected substrates were 70 g in 1 liter of distilled water for wheat germ oil meal and amaranth flour, 60 g/l — for rapeseed mea. It was foundl 17 amino acids, including 9 essential ones in fungi biomass hydrolyzate. Significant influence of cultivation substrate on quantitative composition of amino acids has been established. To all biomass samples the prevalence of glutamic and aspartic acids, arginine among the nonessential amino-acids, leucine, lysine and cystine among the essential amino-acids were common. Endopolysaccharides content in mushroom biomass and exopolysaccharides in culture liquid were slightly different depending on the selected substrates. Sorption of heavy metals by P. ostreatus biomass was increased in series Hg2+ < Pb2+ < Cd2+. High biological activity of the biomass as a source of important essential amino acids and endopolysaccharides as well as sorption capacity towards toxic ions of Pb2+, Cd2+, Hg2+ were determined. It could be a good purpose for usage of P. ostreatus biomass as an ingredient in the composition of functional food or food for special purpose to enhance both: its nutritional value and excretion of heavy metals from the human body.

107-117

Authors: Danylenko S. G., Kigel N. Ph., Burtseva G. V.

Principal criteria for the selection of microorganisms with a wide range of biological and technological properties for fermentation of raw meats are considered. Attention is paid to the main groups of microorganisms such as Micrococсus, Staphylococcus, Lactobacillus, Bifidobacterium and Propionibacterium which are promising for creation of bacterial preparations. To create bacterial preparations, the basic criteria of selection for microorganisms were determined as follows: the ability of microorganisms to be developed within the specific ecological niche (raw meat materials) and their influence on flavor characteristics of the final product under the conditions of intensification of production technologies of meat products. Methods used for search and retrieval of technologically promising strains from different natural sources (fresh meats, minced meats, meat, dairy and sour-milk products, vegetables, fruit, brines and mixtures for salting) are considered.