European Journal of Molecular Biotechnology

Number 1 (7), 2015

4-11
Bactericidal Effect of Aqueous Extracts of the Bark of the Pomegranate (Punica granatum L.) on Bacteria

Authors: Rabah Chadli, Aman Bouzid, Khadidja Bouzid, Hamida Nader

Abstract

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This research concerns the study of antibacterial properties of different aqueous extracts of the bark of the pomegranate (Punica granatum L.). Three bacterial strains were used in this test: Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella. Very interesting bactericidal properties of aqueous extracts of the bark of the pomegranate were found on bacteria. The inhibition zones have a very large diameter up to 20 mm and the MIC and MBC are low, of the order of 0.78 mg/ml. This work has shown antibacterial activity against three bacteria may contribute to the fight against infectious diseases, and possibly offer the possibility of using pomegranate peel pharmaceutical and food industries.

12-26
Electrochemically Activited Water: Biophysical and Biological Effects of Anolyte and Catholyte Types of Water

Authors: Georgi Gluhchev, Ignat Ignatov, Stoil Karadzhov, Georgi Miloshev, Nikolay Ivanov, Oleg Mosin

Abstract

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This article outlines the results on the antimicrobial action of electrochemically activated water solutions (anolyte/catholyte), produced in the anode and cathode chamber of the electrolitic cell. Under laboratory conditions the cell culture and suspensions of classical swine fever (CSF) virus were treated with the anolyte. After inoculating them with cell cultures, the viral presence (the presence of viral antigen) was measured using the immunoperoxidase technique. It was found that anolyte did not affect the growth of the cell culture PK-15; viral growth during the infection of a cell monolayer with a cell culture virus was affected in the greatest degree by the anolyte in 1:1 dilution and less in other dilutions; whereas the viral growth at the infection of a cell suspension with the CSF virus was affected by the anolyte in dilution 1:1 in the greatest degree, and less by other dilutions; viral growth at the infection with a virus in suspension of the cell monolayer was affected by the anolyte in all dilutions. Unexpectedly, the stronger biocidal effect of the catholyte was observed when a strain of E. coli DH5 was treated by the anolyte and catholyte, respectively. In order to provide additional data about the antiviral activity of the electrochemically activated water and the distribution of H2O molecules according to the energies of hydrogen bonds, the non-equilibrium energy spectrum (NES) and differential non-equilibrium energy spectrum (DNES) of the anolyte and catholyte were measured.

27-36
Uses Semi–quantitative and Relative Quantity Methods to Analysis Gene Expression of DGAT1 Gene Responsible for the Olive Diacylglcerol Acyltransferases in 10 Cultivars of Olive (Olea europaea. L)

Authors: Ali Saeed Atiyah AL-Janabi

Abstract

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In this study gene expression for DGAT1 gene was analyzed. Diacylglycerol acyltransferases (DGATs) catalyze the final step of the triacylglycerol (TAG) biosynthesis of the Kennedy pathway. Two major gene families have been shown to encode DGATs, DGAT1 (type-1) and DGAT2 (type-2). Gene expression were analyzed for 10 Olive cultivars (Olea europaea L.) (Khaderi, Qaysi, Manzenillo, Baashiqi, Arabqween, Nabali, Labeeb, Dahkan, Shami and Sorani). Different plant organs as plant materials (mature leaves, mesocarp and seeds for drups) used for analysis. Two methods for analysis gene expression were used, first method was called semi – quantitative and second method was called relative – quantitative, used in relative method (Real time PCR and Actine gene as Housekeeping gene). On the other hand chemical analysis was used on fruits like moisture % and oil % of dry and fresh weigh. The results revealed the following: DGAT1 gene expression in leaves, mesocarp and seeds by two methods (semi- quantitative and relative quantity) were the convergent results and clear, also if this results compared with chemical analysis shows that the best cultivars were Arabqween, Khaderi, Qaysi and Labeeb. The cultivars Shami and Khaderi then were contain in fruits desirable qualities of olive oil, low moisture and high oil percentages ratios. While Nabali, Manzanello, and Sorani cultivars middle desirable quantity, and Baashiqi and Dahkan cultivars had undesirable because of low oil quantity and high moisture in contain fruits. Some cultivars have low intensity in semi- quantitative and little fold in relative quantity but it have high oil in contain fruits that may be indicate that these cultivars were complete gene expression and begin to accumulation and save oil in tissue. Therefore particular emphasis was given to the temporal regulation of olive DGATs during drupe development. In olive fruit, TAGs are formed and stored in both the mesocarp and the seed .Two drupe compartments that have different physiological functions and roles and also display difference in the mode of TAG accumulation. DGATI share an overlapping expression pattern after 28 WAF, suggesting that they probably function at those stages. However, following maximal mRNA levels at 22 WAF, DGAT1 transcription declined substantially.

37-52
The Biosynthesis of Deuterium Labeled Amino Acids Using a Strain of Facultative Methylotrophic Bacterium Вrevibacterium Methylicum 5662 With RuMP Cycle of Carbon Assimilation

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

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We used Gram-positive aerobic facultative methylotrophic bacterium, Brevibacterium methylicum, L-phenylalanine producer with ribulose-5-monophosphate (RuMP) cycle for carbon assimilation for microbiological preparation of [2H]phenylalanine via conversion of low molecular weight substrates ([U-2H]MeOH and 2H2O). For this purpose, the cells of the methylotroph with improved growth characteristics were used on minimal salt media M9 supplemented with 2 % (v/v) [U-2H]MeOH and increasing gradient of 2Н2O concentration from 0; 24,5; 49,0; 73,5 up to 98 % (v/v) 2Н2O. L-phenylalanine was isolated from the growth medium after adding 5 M 2HCl (in 2Н2О), pH = 2,0 by extraction with isopropanol and subsequent crystallization in ethanol (output 0,65 g/l). Alanine, valine, and leucine/isoleucine were produced and accumulated exogenously in amounts of 5–6 mol in addition to the main product of biosynthesis. The method allows to obtain [2Н]amino acids with different levels of deuterium enrichment, depending on 2Н2O concentration in growth media, from 17 atom% 2Н (2 deuterium atoms) (on the growth medium with 24,5 % (v/v) 2Н2О) up to 75 atom% 2Н (6 deuterium atoms) (on the growth medium with 98 % (v/v) 2Н2О) with introduction of deuterium to benzyl С6Н5СН2-fragment of molecule that is confirmed with the data of electron impact (EI) mass spectrometry analysis of methyl ethers of N-5-dimethylamino(naphthalene)-1-sulfochloride [2H]amino acids after the separation by reverse-phase HPLC.