European Journal of Molecular Biotechnology

160-168

Authors: Khadidja Bouzid, Fouzia Toumi Benali, Rabah Chadli, Mohamed Bouzouina, Aman Bouzid, Amal Benchohra, Mustapha Mahmoud Dif

The aim of the current study was to evaluate the total phenolic, flavonoid content and to investigate the antioxidant capacities of the fruit extracts of Arbutus unedo L. that grows in Tiaret area (Western Algeria). First we have extracted the fruit by some non-polar solvent (chloroform, ethyl acetate, 1-butanol). Total phenolic content and total flavonoid content were evaluated according to the Folin-Ciocalteu procedure, and a colorimetric method, respectively. Extracts content was determined by using a high-performance liquid chromatography (HPLC)-UV method. The total phenolic contents of A.unedo L. varied between 12.75±0.06 to 34.17±1.36 mg gallic acid equivalent/g of dry weight of extract. The total flavonoid varied from 2.18±0.10 to 6.54±1.14 mg catechin equivalent/g. The antioxidant potential of all extracts was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, the IC50 of acetate ethyl was the best by 0,009 mg/ml may due to the phenolic compound, in the second was the chloroform extract by IC50=0,015mg/ml, in the third was butanol extract by IC50= 0,022 mg/ml and in the last was water extract by IC50= 0,048mg/ml. the antioxidant activity of all extracts was better than ascorbic acid. The extract obtained under optimum conditions was analyzed by HPLC and five flavonoid compounds were identified; they are catechin, apiginin, silybin, fisetine and naringin.

180-209

Authors: Oleg Mosin, Ignat Ignatov, Dmitry Skladnev, Vitaly Shvets

Biological influence of deuterium on cells of various taxonomic groups of prokaryotic and eucaryotic microorganisms realizing methylotrophic, chemoheterotrophic, photo-organotrophic, and photosynthetic ways of assimilation of carbon substrates (methylotrophic bacteria Brevibacterium methylicum, chemoheterotrophic bacteria Bacillus subtilis, photo-organotrophic halobacteria Halobacterium halobium, and green micro algae Chlorella vulgaris) was investigated at the growth on media with heavy water (2H2O). For investigated microorganisms are submitted the data on growth and adaptation on the growth media containing as sources of deuterated substrates 2H2O, [2H]methanol and hydrolisates of deutero-biomass of methylotrophic bacteria B. methylicum, obtained after multistage adaptation to 2H2O. The qualitative and quantitative composition of intra- and endocellular amino acids, proteins, carbohydrates and fatty acids in conditions of adaptation to 2H2O is investigated. It is shown, that the effects observed at adaptation to 2H2O, possess a complex multifactorial character and connected to cytological, morphological and physiological changes – the magnitude of the lag- period, time of cellular generation, output of biomass, a parity ratio of synthesized amino acids, proteins, carbohydrates and lipids, and also with an evolutionary level of the organization of the investigated object and the pathways of assimilation of carbon substrates as well.

223-228

Authors: Ajay Tomer, Ramji Singh, Manoj Kumar Maurya

Deoiled cakes of Neem and Jatropha served as source of diversified nutrition for Pseudomonas flourescens when used as substrate for mass culturing of antagonist. The present investigation were undertaken to test the suitability of neem and Jatropha cakes for liquid Concentration and longevity of Pseudomonas flourescens in vitro. Increasing the dilution of stock of initial inoculums resulted in decrease of total viable counts of Pseudomonas flourescens was comparatively highest population (64.67×105) and (201.67×105) lesser after 15 days than after 7 days of inoculation on Neem and Jatropha cake concentration respectively.

104-119

Authors: Ignat Ignatov, Oleg Mosin

This review article views predominately the structure and function of animal and bacterial photoreceptor pigments (rhodopsin, iodopsin, bacteriorhodopsin) and their aspects of nano- and biotechnological usage. On an example of bacteriorhodopsin is described the method of its isolation from purple membranes of photo-organotrophic halobacterium Halobacterium halobium by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, the solubilization with 0,5 % (w/v) SDS-Na and subsequent fractionation by methanol and gel filtration chromatography on Sephadex G-200 Column balanced with 0.09 M Tris-borate buffer (pH = 8,35) with 0,1 % (w/v) SDS-Na and 2,5 mM EDTA. Within the framework of the research the mechanism of color perception by the visual analyzer having the ability to analyze certain ranges of the optical spectrum, as colors was studied along with an analysis of the additive mixing of two colors. It was shown that at the mixing of electromagnetic waves with different wavelengths, the visual analyzer perceive them as separate or average wave length corresponding to mix color.

131-148

Authors: Oleg Mosin, Ignat Ignatov

Methylotrophs – aerobic chemoheterotrophic microorganisms submitted by cocci and bacilli mobile forms, are inhabitants of reservoirs and soils of various type, where there are going on various processes of decomposition of organic substances with formation of the one-carbon С1-compounds and some С2-, and С3-compounds, capable to be assimilated by methylotrophs. These microorganisms assimilating carbon on ribuloso-5-monophospate and serine pathways, are allocated from soil ground, the sewage containing decomposing vegetative remainss, from ruminant paunch and other sources. Methylotrophic bacteria recently draw the increasing attention of biotechnology as feasible sources of natural biologically active compounds – fodder fibers and irreplaceable amino acids, carotenoid pigments, lipids and policarbohydrates. For preparation of these compounds are used genetically marked strains of methylotrophic bacteria, obtained via genetic engineering approaches and selection. Recently developed gene-engineering methods of manipulation with the methylotrophic genom allow create on the basis of microbic DNA of methylotrophs expression vectors of eukaryotic proteins for medical and veterinary purposes, as human insulins. In this review article there are submitted data including the results of the authors’ own research on evolution of methylotrophic bacteria, the metabolism and their biotechnological usage.

52-71

Authors: Ignat Ignatov, Oleg Mosin, Borislav Velikov, Enrico Bauer, Georg Tyminski

In this paper are submitted data on longevity factors and mountain water in factorial research of phenomenon of longevity in mountainious and field areas of Bulgaria. The dependence was established among various internal and external factors on a phenomenon of longevity – residence area, health status, gender and heredity. It was shown that water is among the most important factors for longevity. Natural waters derived from various Bulgarian water springs were investigated by non-equilibrium energy (NES) and differential non-equilibrium energy spectrum of water (DNES) method. The biological effects of water with varrying content of deuterium are also discussed. It was shown, that the increased content of deuterium leads to physiological, morphological and cytology alterations of the cell, and also renders negative influence on cellular metabolism, while deuterium depleted water with reduced deuterium content on 20–30 % has beneficial effects on health. By using IR-spectroscopy were investigated various samples of water with varying contents of deuterium, received from Bulgarian water springs and blood serum of cancer patients as well. As estimation factor was measured the values of the average energy of hydrogen bonds (∆EH...O) among H2O molecules, as well as local maxima in the IR-spectra of various samples of water and human blood serum at -0,1387 eV and wavelength 8,95 μm. For a group of people in critical condition of life and patients with malignant tumors the greatest values of local maxima in IR-spectra are shifted to lower energies relative to the control group. This testifies to the structural changes of water. The obtained results testify to necessity of consumption of clean natural water which quality satisfies mountain water from Bulgarian water springs.

86-94

Authors: Alexander A. Shiroky, Alexander V. Volkov, Valery V. Novochadov

The article is an attempt to explain the renewal of the articular cartilage in normalcy and osteoarthritis development by principles of mathematical modeling. Such models help to develop advanced methods of prevention, detection and treatment of osteoarthritis including molecular biotechnologies based on tissue engineering conception. We used histological images to perform structural analysis to discover the signs of active system and its states. Received data are useful to develop research protocols in cartilage tissue engineering.

4-10

Authors: Pavel A. Krylov

To give the empirical space for development of molecular biotechnologies in regenerative medicine the article presents a systematic review of new data for the last 5 years, dedicated to chondrocyte receptor apparatus, connected to physiological and reparative regeneration in articular cartilage. The main attention is given to the receptors of chondrocytes involved in synthesis and degradation extracellular matrix, differentiation and apoptosis of chondrocytes.

25-40

Authors: Oleg Mosin, Ignat Ignatov, Dmitry Skladnev, Vitaly Shvets

With using of a strain of of L-phenylalanine secreted Gram-positive aerobic facultative methylotrophic bacteria Brevibacterium methylicum В-5652, assimilating methanol via ribulose-5-monophosphate (RMP) cycle of carbon assimilation it was carried out the preparative microbiological synthesis of phenylalanine and metabolically related amino acids (alanine, valine, leucine/isoleucine in the amount of 5–6 mmol/l), labelled with deuterium (2H). The data on adaptation of L-phenylalanine secreted methylotrophic bacterium В. methylicum to the maximal concentration of deuterium in minimal growth medium M9 with 98 % 2Н2O and 2 % [2Н]methanol, and data on biosynthesis of deuterium labelled L-phenylalanine with different levels of enrichment are submitted. The developed method for biosynthesis allows to obtain [2Н]phenylalanine with different levels of isotopic enrichment, depending on 2Н2O concentration in growth media M9, from 17 % (2 deuterium atoms) (on growth medium with 24,5 % 2Н2О) right up to 75 % (6 deuterium atoms) (on growth medium with 98 % 2Н2О) with introduction of deuterium to benzyl С6Н5СН2-fragment of molecule that is confirmed with the data of the electron impact (EI) mass-spectrometry analysis of methyl ethers of N-dimethylamino(naphthalene)-5-sulfochloride (dansyl) derivatives of [2Н]phenylalanine, isolated from the liquid culture after its separation by RP HPLC.