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.
The existence of the tax Coupled with the existence of the power of the political Sociality and the evolution of its concept differes according to the concept of the role of the state and its evolution in the economical life and it represents on of the main sources of funding one hand and an effective means that enables the state to intervene in various aspects of the economic life on the other hand. Tax also can be considered as a tool used to re-distribue the in class. Perhaps Iraq suffers from and in the marginalization to the role of taxes in funding and in turn the adoption of the Iraqi budget on oil revenues to finance the aspects of expenitvre which can pose a risk on the total Iraq
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The analysis and efficiency of phenol extraction from the industrial water using different solvents, were investigated. To our knowledge, the experimental information available in the literature for liquid-liquid equilibria of ternary mixtures containing the pair phenol-water is limited. Therefore the purpose of the present investigation is to generate the data for the water-phenol with different solvents to aid the correlation of liquid-liquid equilibria, including phase diagrams, distribution coefficients of phenol, tie-lines data and selectivity of the solvents for the aqueous phenol system.
The ternary equilibrium diagrams and tie-lines
... Show MoreIn the present study, advanced oxidation process / heterogeneous photocatalytic process (UV/TiO2/Fenton) system was investigated to the treatment of oily wastewater. The present study was conducted to evaluate the effect of hydrogen peroxide concentration H2O2, initial amount of the iron catalyst Fe+2, pH, temperature, amount of TiO2 and the concentration of oil in the wastewater. The removal efficiency for the system UV/TiO2/Fenton at optimal conditions and dosage (H2O2 = 400mg/L, Fe+2 = 40mg/L, pH=5, temperature =30oC, TiO2=75mg/L) for 1000mg/L load was found to be 77%.
Aluminum foil cover around the re
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Cadmium is one of the heavy metal found in the wastewater of many industries. The electrocoagulation offers many advantages for the removal of cadmium over other methods. So the removal of cadmium from wastewater by using electrocoagulation was studied to investigate the effect of operating parameters on the removal efficiency. The studied parameters were the initial pH, initial concentration, and applied voltage. The study experiments were conducted in a batch reactor with with two pairs of aluminum electrodes with dimension and 2mm in thick with 1.5 cm space between them. The optimum removal was obtained at pH =7, initial concentration = 50 mg/L, and applied voltage = 20 V and it was 90%.
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Activated carbon was Produced from coconut shell and was used for removing sulfate from industrial waste water in batch Processes. The influence of various parameter were studied such as pH (4.5 – 9.) , agitation time (0 – 120)min and adsorbent dose (2 – 10) gm.
The Langmuir and frandlich adsorption capacity models were been investigated where showed there are fitting with langmmuir model with squre regression value ( 0.76). The percent of removal of sulfate (22% - 38%) at (PH=7) in the isotherm experiment increased with adsorbent mass increasing. The maximum removal value of sulfate at different pH experiments is (43%) at pH=7.
In this study, measuring effectiveness Alauriz in some seeds powder and found that the seeds of-sophistication sex had the highest effective enzymatic reach 353 units / gProtein and Alkabbatah study enzyme extract under different storage conditions and Altaj showed that the enzyme loses Thbatih and whether Mrkbhetwan adding Alklasrin concentration of 10% for the systematic solution
In this study, the adsorption of Zn (NO3)2 is carried out by using surfaces of malvaparviflora. The validity of the adsorption is evaluated by using atomic absorption Spectrophotometry through determination the amount of adsorbed Zn (NO3)2. Various parameters such as PH, adsorbent weight and contact time are studied in terms of their effect on the reaction progress. Furthermore, Lagergren’s equation is used to determine adsorption kinetics. It is observed that high removal of Zn (NO3)2 is obtained at PH=2. High removal of Zn (NO3)2 is at the time equivalent of 60 min and reaches equilibrium,where 0.25gm is the best weight of adsorbant . For kinetics the reaction onto malvaparviflora follows pseudo first order Lagergren’s equation.
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Experimental activity coefficients at infinite dilution are particularly useful for calculating the parameters needed in an expression for the excess Gibbs energy. If reliable values of γ∞1 and γ∞2 are available, either from direct experiment or from a correlation, it is possible to predict the composition of the azeotrope and vapor-liquid equilibrium over the entire range of composition. These can be used to evaluate two adjustable constants in any desired expression for G E. In this study MOSCED model and SPACE model are two different methods were used to calculate γ∞1 and γ∞2
n this work, a series of new nucleoside analogues (β-glucose liked to pyrazoline moiety) was synthesized. In the beginning, chalcone [1-3] was formed from the reaction of acetophenone and benzaldehyde derivatives in the presence of sodium hydroxide. Pyrazolines [4-6] were obtained from the reaction of the prepared chalcones and hydrazine hydrate in the presence of ethanol absolute. These pyrazolines were treated with β-glucose pentaacetate to afford a series of desirable protected nucleoside analogues [8-10]. After that hydrolysis of protected nuclioside analogues in sodium methoxide gave free nucleoside analogues [11-13]. These new formed compounds were diagnosed by 13C-NMR and 1H- NMR for some of them and FT-IR spectroscopy.
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