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.
Polyphenol oxidase (PPO) is an enzyme containing copper, presents in various fruits and vegetables. It is responsible for the browning reactions when the cells are damaged during handling. The best conditions for extraction of polyphenol oxidase from banana peel was by using an extraction buffer containing phosphate buffer (0.05 M, pH 7), 0.01 M ascorbic acid and 0.5% polyethylene glycol, with extraction ratio 1:4 (w:v) for one minute by using blender. The enzyme activity was measured spectrophotometrically at 425 nm. PPO was studied to prevent the browning of banana peel which results in the loss of their marketability. The aim of this study was to determine the optimum conditions for polyphenol oxidase extraction from banana peel.
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Porosity and permeability are the most difficult properties to determine in subsurface reservoir characterization. The difficulty of estimating them arising from the fact that porosity and permeability may vary significantly over the reservoir volume, and can only be sampled at well location. Secondly, the porosity values are commonly evaluated from the well log data, which are usually available from most wells in the reservoir, but permeability values, which are generally determined from core analysis, are not usually available. The aim of this study is: First, to develop correlations between the core and the well log data which can be used to estimate permeability in uncored wells, these correlations enable to estimate reservoir permeabil
... Show MoreUsed vegetable oil was introduced to transesterfication reaction to produce Biodiesel fuel suitable for diesel engines. Method of production was consisted of filtration, transesterfication, separation and washing. Transesterfication was studied extensively with different operating conditions, temperature range (35-80oC), catalyst concentration (0.5-2 wt. % based on oil), mixing time (30-120 min.) with constant oil/methanol weight ratio 5:1 and mixing speed 1300 rpm. The concentration of Fatty acid methyl esters (Biodiesel) was determined for the transesterficated oil samples, besides of some important physical properties such as specific gravity, viscosity, pour point and flash point. The behavior of methyl esters production and the phys
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Purpose/objective:
1 - To explain the financial impact of the activities and areas of human resources management and the adoption of the methodology for estimating costs on the basis of conduct and statement of how to assess costs and benefits of human resource activities.
2 - Measuring human capital, and its impact on the financial statements.
Design/methodology/approach:
Concentrated dimensions of the research paper's lack of financial statements prepared by the organizations for information mandated human resource its components of the three (attraction - development
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The purpose of this research is to explain the effect of the dropping of three zeroes from Iraqi dinar on the basic role of the dinar and derivative and dynamic in transaction of Iraqi economic. the importance of this research comes from the effect of this operation on the evaluation of the exchange between the individuals of Iraqi society and the other between the Iraqi individuals and the foreign whom use the Iraqi dinar as a tool for exchange also the effect of the operation on the reserves of the Iraqi dinar the results of this research are as the following:-
1-the operation of dropping of zeroes must be associated with increase of the sharing of economic sectors wi
... Show MoreElectronic Alattarh been studied long flexible factors forming the nucleus of boron in the shell model framework multipolar been identified factors was introduced into the effects of polarization heart in the first place accounts
The optimum conditions for production of fibrinolytic protease from an edible mushroom Pleurotus ostreatus grown on the solid medium , Sus medium, composed of Sus wastes (produced from extracted medicinal plant Glycyrrhiza glabra) were determined. Addition of 5% of Soya bean seeds meal in Sus medium recorded a maximum fibrinolytic protease activity resulting in 7.7 units / ml. The optimum moisture content of Sus medium supplemented with 5% Soya bean seeds meal was 60% resulting in 7.2 units / ml.Pleurotus ostreatus produced a maximum fibrinolytic protease activity when the spawn rate,pH of medium and incubation temperature were 2,6 and 30°C, respectively. The maximum fibrinolytic protease activity was 7.6 units / ml when incubat
... Show MoreIn this study, a novel application of lab-scale dual chambered air-cathode microbial fuel cell (MFC) has been developed for simultaneous bio-treatment of real pharmaceutical wastewater and renewable electricity generation. The microbial fuel cell (MFC) was provided with zeolite-packed anodic compartment and a cation exchange membrane (CEM) to separate the anode and cathode. The performance of the proposed MFC was evaluated in terms of COD removal and power generation based on the activity of the bacterial consortium in the biofilm mobilized on zeolite bearer. The MFC was fueled with real pharmaceutical wastewater having an initial COD concentration equal to 800 mg/L and inoculated with anaerobic aged sludge. Results demo
... Show MoreThis study concluded detection of Toxoplasma gondii in milk, immunologically by using Elisa and nested PCR)nPCR (based on B1 gene, also to investigate the effect of toxoplasmosis, parity, breed and flock on some milk composition in the Iraqi local and Shami goats in the middle of Iraq. A total of 80 milk samples of the lactating goats were collected. Results of this study showed the prevalence of Toxoplasmosis was 21.25% and 28.75% by Elisa and nPCR respectively without significant differences. The sensitivity of Elisa was a low (30.43%) whereas the specificity was a high (82.45%). The degree of agreement estimated by Kappa coefficient revealed a slight agreement (0.14) between two methods. The results indicated that goats infected
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