The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtration chromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the already used in published researches, which depend on the costly affinity chromatography and other expensive methods of purification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR) .The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated on Yeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C˚ for 24 h .Loop fully of the yeast culture was transferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C˚for 24h , after that it was stored at 4C˚ ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae was identified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cell of S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negatively charged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtration chromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fraction was measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISA Kit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weight of GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plotted between log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR that extracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ion exchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient (0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration of sodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured in the fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M of NaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity 71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don't give any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step of purification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical with the peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed in these fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and 39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S. cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.
In this study, sawdust as a cheap method and abundant raw material was utilized to produce active carbon (SDAC). Physiochemical activation was utilized where potassium hydroxide used as a chemical activating agent and carbon dioxide was used as a physical activating agent. Taguchi method of experimental design was used to find the optimum conditions of SDAC production. The produced SDAC was characterized using SEM to investigate surface morphology and BET to estimate the specific surface area. SDAC was used in aqueous lead ions adsorption. Adsorption process was modeled statistically and represented by an empirical model. The highest specific surface area of SDAC was 688.3 m2/gm. Langmuir and Freundlich isotherms were used to
... Show MoreThis study was aimed to evaluate atotal phenolic content, antibacterial activity, and antioxidant activity of M. communis callus extracts were evaluated. Callus induction in general Murashige and Skoog (MS) media is completed by the Benzil adenine's unique knowledge of callus formation. A well diffusion experiment was used to examine antibacterial interest in Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. The DPPH radical scavenging activity test was used to measure antioxidant activity. FTIR and HPLC have been used to pinpoint the presence of polyphenol compounds in calluses. The total phenol content of plant leaves extract (0.1, 0.5, and 1) mg/ml was 42.12, 94.08, and 189 mg of Gallic ac
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Five different bacterial isolates [ Vibrio cholera (Ogawa) , Vibrio cholera (Inaba) , Salmonella typhi , Salmonella paratyphi and ? Salmonella typhimurium ] were obtained from the Central Health Laboratory . Both sensitivity tests (MIC , MBC and wells method ) against these bacteria were performed by using the aqueous of leaves extract of Marjoram plant. The results cleared that the values of MIC for Vibrio cholera serotypes Ogawa and Inaba were 100 mg/ml , while the value of MBC was 200 mg/ml. The value of the Inhibition zone at 100 mg /ml concentration for both Ogawa and Inaba were 13 mm and 9 mm respectively. Our results showed that the three types of Salmonella didn’t show any inhibition zone at 200 mg/ml .
The removal of boron from aqueous solution was carried out by electrocoagulation (EC) using magnesium electrodes as anode and stainless steel electrodes as cathode. Several operating parameters on the removal efficiency of boron were investigated, such as initial pH, current density, initial boron ion concentration, NaCl concentration, spacing between electrodes, electrode material, and presence of carbonate concentration. The optimum removal efficiency of 91. 5 % was achieved at a current density of 3 mA/cm² and pH = 7 using (Mg/St. St. ) electrodes, within 45 min of operating time. The concentration of NaCl was o. 1 g/l with a 0.5cm spacing between the electrodes. First and second order rate equation were applied to study adsorp
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This study focused on treating wastewater to remove phosphorus by adsorption onto naturaland local materials. Burned kaolin, porcelinite, bauxite and limestone were selected to be testedas adsorption materials.The adsorption isotherms were evaluated by batch experiments, studyingthe effects of pH, temperature and initial phosphorus concentration. The results showed that at pH6, temperature 20°C and 300 mg/l initial phosphorus concentration; the sorption capacity was0.61, 9, 10 and 13 mg/g at 10 h contact time, for burned kaolin, porcelanite, limestone and bauxiterespectively. As the pH increased from 2 to 10 the removal efficiency for the materials differs inbehaviour. The removal efficiency increased from 40 to 90 % for limestone, and dec
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The performance of a batch undivided electrochemical reactor with a rotating cylinder electrode of woven-wire (60 mesh size), stainless steel 316, is examined for the removal of copper from synthetic solution of o.5 M sodium chloride containing 125 ppm at pH ≈ 3.5. The effect of total applied current, rotation speed on the figures of merit of the reactor is analyzed. For an applied current of 300 mA at 100 rpm, the copper concentration decreased from 125 to mg l-1 after 60 min of electrolysis with a specific energy consumption of 1.75 kWh kg-1 and a normalized space velocity of 1.62 h-1. The change in concentration was higher when the total applied currents were increased because of the turbulence
... Show MoreNicotine was separated from eggplant and green pepper seeds (Solanaceous) by High Performance Liquid Chromatography (HPLC).The concentration of nicotine in the eggplant extract (0.871-0.877 μg/ml) was determined by injecting standard material with 0.5 and 1.5 μg/ml, while the concentrations of nicotine in green pepper extract (0.613-0.618 μg/ml) was determined when the standard material was injected with 0.5 and 1.5 μg/ml. The qualitative chemical data was calculated from derivations of the standard material. Nicotine concentration was measured qualitatively in both extracts through the calibration curve and method of the standard addition. This technique has high accuracy and compatibility, bringing the proportion of relati
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The degradation and mineralization of 4-chlorophenol (4-CP) by advanced oxidation processes (AOPs) was investigated in this work, using both of UV/H2O2 and photo-Fenton UV/H2O2/Fe+3 systems.The reaction was influenced by the input concentration of H2O2, the amount of the iron catalyst, the type of iron salt, the pH and the concentration of 4-CP. A colored solution of benzoquinon can be observed through the first 5 minutes of irradiation time for UV/H2O2 system when low concentration (0.01mol/L) of H2O2 was used. The colored solution of benzoquinon could also be observed through the first 5 minutes for the UV/H2O2/Fe+3 system at high
concentration (100ppm) of 4-CP. The results have shown that adding Fe+3 to the UV/H2O2 system enhanced
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In this study miconazole nitrate was formulated as topically applied emulgel; different formulas were prepared using sodium carboxymethylcellulose (SCMC) and carboxypolymethylene (carbomer 941) as gelling agents. The influence of type of gelling agent and concentration of both oil phase and emulsifying agent on drug release was studied and compared with commercially available miconazole nitrate cream (Mecozalen®). The results of in vitro release showed that SCMC emulgel bases gave better release than carbomer 941 bases and the release of drug increase from both bases as a function of increasing the concentration of emulisifying agent. The oil phase had retardation effect when
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Room temperature ionic liquids show potential as an alternative to conventional organic membrane solvents mainly due to their properties of low vapour pressure, low volatility and they are often stable. In the present work, the technical feasibilities of room temperature ionic liquids as bulk liquid membranes for phenol removal were investigated experimentally. In this research several hydrophobic ionic liquids were synthesized at laboratory. These ionic liquids include (1-butyl-3-methylimidazolium bis (trifluoromethylsulfonyl) imide[Bmim][NTf2], 1-Hexyl-3-methylimidazolium bis (trifluoromethylsulfonyl) imide[Hmim][NTf2], 1-octyl-3-methylimidazolium bis (trifluoromethylsulfonyl)imide[Omim][NTf2],1‐butyl
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