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.
For this research, the utilisation of electrocoagulation (EC) toremove theciprofloxacin (CIP) and levofloxacin (LVX) from aqueous solutions was examined. The effective removal efficiencies are 93.47% for CIP and 88.00% for LVX, under optimum conditions. The adsorption isotherm models with suitable mechanisms were applied to determine the elimination of CIP and LVX utilizingtheEC method. Thefindingsshowed the adsorption of CIP and LVX on iron hydroxide flocs followed the Sips isotherm, with correlation coefficient values (R2) of 0.939 and 0.937. Threekinetic models were reviewed to determine the accurate CIP and LVX elimination methods using the EC method. The results showed that itfittedfor the second-order model, which indicated that the c
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A hierarchically porous structured zeolite composite was synthesized from NaX zeolite supported on carbonaceous porous material produced by thermal treatment for plum stones which is an agro-waste. This kind of inorganic-organic composite has an improved performance because bulky molecules can easily access the micropores due to the short diffusion path to the active sites which means a higher diffusion rate. The composite was prepared using a green synthesis method, including an eco-friendly polymer to attach NaX zeolite on the carbon surface by phase inversion. The synthesized composite was characterized using X-ray diffraction spectrometry, Fourier transforms infrared spectroscopy, field emission scanning electron microscopy, energy d
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From a large number of bacterial samples collected from different hospital in Iraq in central health laboratory ,only ten isolates were identified primary as Vibrio. A number of morphology and biochemical test were carried out to complete this identification that showed all bacterial isolates were related to Vibrio cholerae .In this study all Vibrio isolates were investigated for Bio typing and the result showed that all (10) isolate were related to (Eltor biotypes) .Also, the susceptibility test towards eight antibiotics were carried out .
Results shows that ciprofloxacin , Norfloxacin, Erythromycin, Ampicillin, ceftriaxone and Amikacin were the most effective
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Titanium oxide nanoparticles-modified smectite (SMC-nTiO2) as a low-cost adsorbent was investigated for the removal of Rhodamine B (RhB) from aqueous solutions. The adsorbents (SMC and SMC-nTiO2) were characterized by scanning electron microscopy, Fourier transforms infrared spectroscopy, and energy-dispersive X-ray spectroscopy. The effects of various parameters like contact time, adsorbent weight, pH, and temperatures were examined. Three kinetic equations (pseudo-first-order (PFO), pseudo-second-order (PSO), and intra-particle diffusion) were used to evaluate the experimental kinetic of the data and the results showed that the adsorption process is in line with the PSO kinetic model. Adsorption equilibrium isotherms were modeled using La
... Show MoreCitrus fruit contain variety of flavonoids such as Hesperidin (the principal flavonoid in oranges and grapefruit). Hesperidin is found in high concentration in fruit peel of oranges and in substantially lower concentration in juice of these fruits. Hesperidin was extracted from oranges peel by treating the peels with calcium hydroxide. HPLC technique was used to determine hesperidin. Hesperidin was saperated and purified in a purity of about 90.1-95.7% and yield about 1.5 %w/w from oranges peel dry powder. Both hesperidin and oranges peel extract showed significan antibacterial activity. Sensitivity to hesperidin and oranges peel extracts were not similar for the chosen bacteriaCrude orange peel extract gave a various antimicro
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This article reviews the technical applicability of nanofiltration membrane process for the removal of nickel, lead, and copper ions from industrial wastewater.
Synthetic industrial wastewater samples containing Ni(II), Pb(II), and Cu(II) ions at various concentrations (50, 100, 150 and 200 ppm), under different pressures (1, 2, 3 and 4 bar), temperatures (10, 20, 30 and 40 oC), pH (2, 3, 4, 5 and 5.5), and flow rates (1, 2, 3 and 4 L/hr), were prepared and subjected treated by NF systems in the laboratory. Suitable NF membrane was chosen after testing a number of NF membranes (University of Technology-Baghdad), in terms of production and removal. NF system was capable of removing more than (85%, 78%, and 66% for Ni(II
... Show MoreThe present paper deal with the issue of the beginning of the culturally
renaissance in emirates of Arab Gulf from 1914-1945 between tow world war
has been attracting the attention of academic about the developments in many
fields in the Arab Gulf at this time.
The paper is divided into five sections. First section, deals with the
geographic importance for the Arab Gulf region. Second section, the economic
situations in the region before and after oil. The third section, talk for social
situations, like population, tribe and tribes in society, and immigration. The
fourth section, deals with the factors of rise the culture and political in the Arab
Gulf before discovery of oil period. The five section, the cultu
The uptake of Cd(II) ions from simulated wastewater onto olive pips was modeled using artificial neural network (ANN) which consisted of three layers. Based on 112 batch experiments, the effect of contact time (10-240 min), initial pH (2-6), initial concentration (25-250 mg/l), biosorbent dosage (0.05-2 g/100 ml), agitation speed (0-250 rpm) and temperature (20-60ºC) were studied. The maximum uptake (=92 %) of Cd(II) was achieved at optimum parameters of 60 min, 6, 50 mg/l, 1 g/100 ml, 250 rpm and 25ºC respectively.
Tangent sigmoid and linear transfer functions of ANN for hidden and output layers respectively with 7 neurons were sufficient to present good predictions for cadmium removal efficiency with coefficient of correlatio
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