Biotechnologia Acta

23-35

Authors: T. P. Pirog, D. I. Khomyak, N. A. Grytsenko, A. P. Sofilkanych, А. D. Konon, K. A. Pokora

The literature and own experimental data, concerning biotechnological potential of bacteria of Nocаrdia genus are given. The use of these microorganisms as destructors of aliphatic (octane, pentadecanol, eicosane, octacosane, hexatriacontane, pristane), aromatic (phenol, octylbenzene, phenanthrene, anthracene), nitroaromatic (4-nitrophenol), heterocyclic (pyridine, ?-picoline) hydrocarbons is described. The prospects of use of Nocаrdia in processes of substances bio-transformation (production of daidzein, ibuprofen, nicotinic acid) and synthesis of some valuable metabolites, in particular antimicrobial and cytotoxic substances (ayamycin, transvalencin А, nocathiacin, brasilibactin A, nocaracins etc.) as well as substances with surface-active and emulsifying properties are discussed. The own experimental data concerning optimization of cultivation conditions and intensification of surfactant synthesis on glycerol (byproduct of biodiesel production) by oil oxidizing bacteria strain Nocardia vaccinii K-8, that was isolated from oil polluted samples of soil are presented. The ability of strain K-8 to assimilate some aromatic compounds (phenol, benzene, toluene, naphthalene, hexachlorbenzene, sulfanilic acid and N-phenylanthranilic acid, 0.3–0.5%) was determined. It was shown that the highest oil destruction degree (94–98%) in polluted water (2.6 g/L) was achieved in the case of treatment with suspension of N. vaccinii K-8 cells (9.8 x 107 CFU/mL) after 30 days, while surfactant preparation of post fermentative cultural liquid (100–300 mL/kg) was more effective for remediation (destruction of 74–83% of oil) of oil polluted soil (20 g/kg). It was determined that surfactants (0.085–0.85 mg/mL) and other exocellular metabolites of strain К-8 possess antimicrobial activity against some phytopathogen bacteria of Pseudomonas and Xanthomonas strains. In this connection the quantity of living cells decreased by 80–100% after the treatment with the investigated preparations.

46-52

Authors: O. Y. Dudchenko, V. N. Pyeshkova, O. O. Soldatkin, O. P. Soldatkin, 2, S. V. Dzyadevych

The conductometric biosensor for fructose determination based on fructose dehydrogenase and potassium ferricyanide mediator as electron acceptor has been developed. The enzyme was immobilized on a surface of the conductometric transducer together with bovine serum albumin using crosslinking with glutaraldehyde. Working conditions of the discribed fructose biosensor were optimized. The results concerning influence of the buffer solution concentration and potassium ferricyanide concentration on the biosensor performance are given. The fructose biosensor is characterized by high signal reproducibility and selectivity to fructose. The developed conductometric biosensor can be successfully used for fructose monitoring in the procedures of food and clinical diagnostic.

63-68

Authors: A. N. Sukach, T. D. Liashenko, M. V. Shevchenko

It is shown that isolated brain cells of newborn rats are heterogeneous suspension consisted of differentiated cells, stem and progenitor cells, which can proliferate and differentiate in conditions of in vitro cultivation. During cultivation in presence of serum the neural cells of the newborn rats form aggregates, where reconstitution of cellular microenvironment lost in isolation takes place followed by cell reparation. After attachment to a substrate the aggregate cells migrate and spread, form a monolayer of astroglia, which the neuroblasts and colonies of stem/progenitor cells are formed on. The findings show feasible application of newborn rat neural cells as model of postnatal neural tissue for studying the influence of cellular environment on process of development and recovery of differentiated neural cells, as well as proliferation and differentiation of stem/progenitor cells.

75-82

Authors: O. V. Sytar, A. M. Gabr, N. Yu. Taran, I. M. Smetanska

The growth parameters of transformed root cultures, total phenolic content and phenolic acids composition has been studied in root cultures, which were obtained from various explants of buckwheat by Agrobacterium rhizogenes strains A4. The methods of obtaining of the transformed root cultures, total phenol estimation, gas-liquid chromatography and polymerase chain reaction has been used. Elevated levels of total phenols in transformed roots of buckwheat from different sources of explants have been found. The high content of chlorogenic, p-hydroxybenzoic, p-anisic and caffeic acids has been discovered in the root cultures, which can be used for their industrial production. Maximal root growth was equal 21.2 g/l of dry weight in the roots as source for root culture, 17.7 g/l with leaves and 14.6 g/l with stems at 3 week after placement. Molecular analysis by polymerase chain reaction amplification was confirmed that the rol B gene (652 bp) which transferred info hairy roots from Ri-plasmid in Agrobacterium rhizogenes is responsible for induction of root from plant species.

85-99

Authors: I. V. Goraichuk, V. I. Bolotin, A. P. Gerilovych, O. S. Solodyankin, R. O. Kucheryavenko, V. V. Kucheryavenko

Bovine viral diarrhea is a widespread infection of cattle that has a wide range of clinical symptoms in domestic and wild ruminants. It is a major problem in cattle and causes significant economic losses in the cattle industry. The virus infects bovines of all ages and causes both immunosuppression and reproductive, respiratory and digestive disorders. Persistently infected cattle are the main factor in transmission of the disease between and among herds. Comparative results of antibodies presence received by two methods of enzymoimmunoassay and virus neutralization test are given in the paper. During the work, 1010 samples of blood serum of cattle from three farms in the Kharkiv region were selected and analyzed. Bovine viral diarrhea virus concerning antibodies were found by enzymoimmunoassay in 704 samples (69.7%) using commercial kit and in 690 samples (68.3%) using in house method. After results clarification by virus neutralization test, bovine viral diarrhea antibodies were found in 712 samples (70.5%). Immunoenzyme analysis is recommended for mass screening of cattle for viral diarrhea occurrence. The results confirm that the sensitivity immunoenzyme analysis satisfies the requirements of the diagnostic methods. Using the neutralization reaction of viruses as the «gold standard» of serological methods, it is appropriate to clarify the results of immunoenzyme analysis. Since the results contain a signi ficant number of false positive results, it is necessary to carry out comprehensive studies using both serological and molecular genetics methods.

100-109

Authors: O. B. Mykchaylova, N. L. Poyedinok

Results of the study on growth and morphological peculiarities of valuable medicinal mushroom Cordyceps militaris from the culture collection of mushrooms of Kholodny Institute of Botany of the National Academy of Sciences of Ukraine are given. Using the method of scanning electron microscopy, the micro-morphological structures specific to this species were studied. This allows identifying this species in pure culture. Culture-morphological studies were performed on four agar nutrient media. Glucosepeptone-yeast agar medium, malt agar extract and incubation temperature of 16 °C were the most favorable for the vegetative growth of all the tested strains. Temperature of 36 °C is critical. Maximal growth of C. militaris was observed at pH 6.0?6.5. The optimal sources of carbon for vegetative mycelium growth were glucose, lactose and sucrose, whereas peptone and yeast extract were the best sources of nitrogen. Light irradiation of different nature stimulated the radial growth in agar media and accumulation of their biomass in liquid ones.

116-120

Authors: S. M. Boiko, M. E. Ryazanova

Activity of cellulolytic enzymes cultures Stereum hirsutum (Wild.) Gray, different nuclear status, depending on the substrate have been studied. The most productive by synthesis endoglucanases monokaryotic cultures of Sh1-4 and Sh7-2 were determined. It is found that nutrient medium containing filtering paper is optimal for synthesis of cellulase (culture Sh1-4, maximum of cellulolytic activity is 319 ?М/mg, culture Sh7-2 — 101 ?М/mg). Maximum of cellulolytic activity at cultivation with sawdust of trees for culture Sh1-4 observed at the 10th day was 142 ?М/mg (oak sawdust), for culture Sh7-2 at the 20th days it was 34 ?М/mg (ash-tree sawdust). Study of quality cellulolytic enzymes enabled to establishe presence of four exoenzymes of Cx complex with a relative electrophoretic mobility of 0.22; 0.24; 0.47; 0.74. Enzyme with relative electrophoretic mobility 0.47 showed high concentration and activity.