Biotechnologia Acta

9-21

Authors: E. A. Shesterenko, I. I. Romanovska O. V. Sevastyanov,S. A. Andronati

The classification, structure, and mechanism of catalytic action of carboxylesterase of different origin are presented in the review. The prospects of carboxylesterases application for metabolism and both several drugs and prodrugs activation investigation in vitro are shown. The enzyme usage as biocatalyst of stereoselective hydrolysis and synthesis of a wide range of acyclic, carbocyclic and heterocyclic compounds — esters are also urgent. It was established that enantiomers obtainable with the help of carboxylesterase are characterized by high chemical yields and optical purity; immobilization on different supports stabilizes the enzyme and allows the repeated usage of obtained biocatalysts. The own studies conducted and the enzymatic hydrolysis features of news 3-acylhydroxy-1,4-benzodiasepin-2-ones — potential anxiolytic and hypnotic means, with a help of pig liver microsomal fraction carboxylesterase have been established. For the first time the enantioselective hydrolysis of 3-acetoxy-7-bromo-1-methyl-5-phenyl-1,2-dihydro-3H-1,4-benzdiazepine-2-one was accomplished using free and immobilized in phyllophorine and alginate, stabilized by Ca2+ microsomal fraction. The S-enantiomer of substrate was isolated, which suggests the increased specificity of pig liver microsomal fraction carboxylesterase to its R-enantiomer.

34-45

Authors: S. А. Starovoitova, O. I. Skrotska

Modern tendencies of recombinant microorganisms creation for obtaining on their basis a new effective biopreparations (probiotics) with wider spectrum of biological and therapeutic properties were considered. A lot of attention was focused on the main genera of perspective bacteria for creation of recombinant probiotics particularly: Lactococcus, Bifidobac terium,Bacillus, Escherichia. The main created Ukrainian and foreign gene-modified strains, that are widely used today in creation of effective recombinant biopreparations were characterized. Some fundamental directions and methods of gene-modified strains obtaining, which are used in getting effective biopreparations that used for therapy and prophylactic illness were reported, under which this group of pharmaceutical drugs were not used earlier. The safety matters of probiotics using on basis of genemodified strains were examined. Medical and veterinary biopreparations on basis of recombinant microorganisms could be used directly and effectively for therapy and prophylaxis of different illness, beginning from disbacteriosis up to cardiovascular diseases. It is related with some probiotic microorganisms ability for lowering of serum cholesterol at the host organism.

73-80

Authors: L. D. Varbanets, О. V. Matselyukh, N. А. Nidyalkova, Е. V. Аvdiyuk, А. V. Gudzenko, I. I. Seifullina, G. N. Маsаnоvets, N. V. Khitrich

Chloride, bromide and isothiocyanate complexes of cobalt(II) with N-substituted thiocarbamoyl-N?-pentamethylenesulfenamides (1)–(12), and also complexes of cobalt(II, Ш) with derivatives of morpholine-4-carbodithioic acid (13)–(18) have been used as modificators of enzymes of hydrolytic action — Bacillus thurin-giensis ІМВ В-7324 peptidases, Bacillus subtilis 147 and Aspergillus flavus var. oryzae 80428 amylases, Eupenicillium erubescens 248 and Cryptococcus albidus 1001 rhamnosidases. It was shown that cobalt (II, Ш) compounds influence differently on the activity of enzymes tested, exerted both inhibitory and stimulatory action. It gives a possibility to expect that manifestation of activity by complex molecule depends on ligand and anion presence — Cl–, Br– or NCS–. The high activating action of cobalt(II) complexes with N-substituted thiocarbamoyl-N?-pentamethylenesulphenamides (1)–(12) on elastase and fibrinolytic activity of peptidases compared to tris(4-morpholinecarbodithioato)cobalt(ІІІ) (14) and products of its interaction with halogens (15)–(17), causes inhibitory effect that is probably due to presence of a weekly S–N link, which is easy subjected to homolytic breaking. The studies of influences of cobalt(II) complexes on activity of C. аlbidus and E. еrubescens ?-Lrhamnosidases showed, that majority of compounds inhibits of its activity, at that the most inhibitory effect exerts to C. аlbidus enzyme.To sum up, it is possible to state that character of influence of cobalt(II) complexes with N-substituted thiocarbamoyl-N?-pentamethylenesulphenamides, and also cobalt(II, Ш) complexes with derivatives of morpholine-4-carbodithioic acid varies depending on both strain producer and enzyme tested. The difference in complex effects on enzymes tested are due to peculiarities of building and functional groups of their active centers, which are also responsible for binding with modificators.

86-96

Authors: A. A. Tykhomyrov, E. I. Yusova, S. I. Diordieva, V. V. Corsa, T. V. Grinenko

Components of plasminogen/plasmin system play crucial role in fibrinolytic rocesses. They are also involved in regulation of cell activity in normalcy and various pathological conditions. In particular, kringle-containing proteolityc plasminogen fragments, which are denoted as angiostatins, participate in processes during neovascularization, metastasis, tumor growth, inflammation. Angiostatins are considered to be potential markers of diseases associated with vascular pathologies. Therefore, elaboration of specific and sensitive methods for their detection is still important area of inquiry. In the present study, approaches for obtaining of plasminogen fragments through its limited proteolysis by pancreatic elastase and further purification of proteolytic derivates by means of gel filtration and affinity chromatography on Lys-Sepharose are described. Polyclonal antibodies raised to fragment K1-3 have been produced, and their principal immunochemical properties have been studied. It has been found that antibodies purified on immunoaffine sorbent demonstrate different affinity toward plasminogen and its fragment, as follows: Glu-Pg > Lys-Pg > К1-3 > mini-Pg > К4. Based on the data obtained, immunological features of plasminogen kringle domains are discussed. Antibodies against fragment K1-3 could be applied in immunochemical analysis, in particular, Western blot, for detection of angiostatins as markers of tumor growth, metastasis, cardiovascular diseases and inflammation processes, and used as molecular tool for investigation of plasminogen/plasmin system functioning in health and disease.

115-118

Authors: Sergeeva L. E., Komisarenko A. G., Bronnikova L. I., Mikhalska S. I., Tishchenko O. M.

Low frequency of in vitro plant regeneration limits significantly wide expansion of biotechnology methods. The screening of explants with stable and high level of the morphogenesis is an open problem. It is known, that free proline fluctuations correlate with processes of plant growth and development in vivo and in vitro. Sunflower mature seeds were germinated in vitro. The contents of free proline at somatic tissues with different regenerational abilities were measured. Following explants were estimated: cotyledons from 1- or 4-days cultivating shoots; primary explants prepared for the regeneration procedure (segments of 4-day cultivating shoots); primary explants with and without initial morphogenesis, 6 days on the regeneration medium; shoots and certain explants at the moment of their mutual separation; explants, 10 days after shoots release. The top levels of free proline were observed in tissues with fulfiled organogenesis. The hypothesis about proline involvement to the process of sunflower morphogenesis in vitro is discussed. During primary screenings of explants the level of free proline can be appropriate index of in vitro sunflower organogenesis.