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.
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The removal of heavy metal ions from wastewater by sorptive flotation using Amberlite IR120 as a resin, and flotation column, was investigated. A combined two-stage process is proposed as an alternative of the heavy metals removal from aqueous solutions. The first stage is the sorption of heavy metals onto Amberlite IR120 followed by dispersed-air flotation. The sorption of metal ions on the resin, depending on contact time, pH, resin dosage, and initial metal concentration was studied in batch method .Various parameters such as pH, air flow rate, and surfactant concentration were investigated in the flotation stage. Sodium lauryl sulfate (SLS) and Hexadecyltrimethyl ammonium bromide (HTAB) were used as anionic and cationic surfactant re
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Calcareous nannofossils were documented from the upper part of the Cretaceous Balambo Formation in northern Iraq with the aim of determining an evidence for the Oceanic Anoxic Event. A detailed investigation of the calcareous nannofossils led to the identification of twenty-four species. Regarding these data, Discolithus litterarius (Górka, 1957) was identified at the studied interval with the age of Early Aptian.
Early Aptian assemblages are dominated by nannoconids that drop sharply within the D. litterarius nannofossil zone, which may be related to the nannoconid crisis recorded in the Early Aptian in the other parts of the world. This event is coincided by a decrease in CaCO3<
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Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen and a model bacterium for studying virulence and bacterial social traits. While it can be isolated in low numbers from a wide variety of environments including soil and water, it can readily be found in almost any human/animal-impacted environment. It is a major cause of illness and death in humans with immunosuppressive and chronic conditions, and infections in these patients are difficult to treat due to a number of antibiotic resistance mechanisms and the organism’s propensity to form multicellular biofilms. One hundred twenty clinical samples and forty hospital environmental samples (various sources) were collected from hospitals in Baghdad city during the period from Oc
... Show MoreThe degradation of Toluidine Blue dye in aqueous solution under UV irradiation is investigated by using photo-Fenton oxidation (UV/H2O2/Fe+). The effect of initial dye concentration, initial ferrous ion concentration, pH, initial hydrogen peroxide dosage, and irradiation time are studied. It is found put that the removal rate increases as the initial concentration of H2O2 and ferrous ion increase to optimum value ,where in we get more than 99% removal efficiency of dye at pH = 4 when the [H2O2] = 500mg / L, [Fe + 2 = 150mg / L]. Complete degradation was achieved in the relatively short time of 75 minutes. Faster decolonization is achieved at low pH, with the optimal value at pH 4 .The concentrations of degradation dye are detected by spectr
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Excessive intake of fluoride, mainly through drinking water is a serious health hazard affecting humans worldwide. In this study, the defluoridation capacities of locally available raw waste beef bones have been estimated. Several experimental parameters including contact time, pH, bone dose, fluoride initial concentration, bone grains size, agitation rate, and the effect of co-existence of anions in actual samples of wastewater were studied for fluoride removal from aqueous solutions. Results indicated excellent fluoride removal effeciency up to 99.7% at fluoride initial concentration of 10 mg F/L and 120 min contact time. Maximum fluoride uptake was obtained at neutral pH range 6-7. Fluoride removal kinetic was well described by the ps
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Nanotechnology is a continually expanding field for its uses and applications in multiple areas i.e. medicine, science, and engineering. Biosynthesis is straightforward, less-toxicity, and cost-effective technology. TiO2 NPs biosynthesis has attained consideration in recent decades. In this study, probiotic bacteria were isolated from cow’s raw milk samples, and then were identified by using the Vitek2 system; as Leuconostoc spp. included Leuconostoc mesenteroides subsp. mesenteroides (Leu.1), Leuconostoc mesenteroides subsp. cremoris (Leu.4), and Leuconostoc pseudomesenteroides (Leu.14). All Leuconostoc spp. isolates showed an ability for TiO2 NPs bio-production, after being incubated at anaerobic conditions (30 o C/ 24 h) in DeM
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An experimental study was conducted with low cost natural waste adsorbent materials, barley husks and eggshells, for the removal of Levofloxacine (LEVX) antibacterial from synthetic waste water. Batch sorption tests were conducted to study their isothermal adsorption capacity and compared with conventional activated carbon which were, activated carbon > barley husks > eggshells with removal efficiencies 74, 71 and 42 % with adsorbents doses of 5, 5 and 50 g/L of activated carbon, barley husks, and eggshells respectively. The equilibrium sorption isotherms had been analyzed by Langmuir, Freundlich, and Sips models, and their parameters were evaluated. The experimental data were correlated well with the Langmuir model which gives the
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