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.
Used 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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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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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 aim of this research is to benefit from recycl the aircraft waste oils which is discarded in sewage network, to be used in preparation of greases for industrial purposes and to reduce the environmental pollution. In this research synthetic greases were prepared with special specifications by mixing the waste oils after treating with (silica gel as adsorbent agent, and filtration to precipitate impurities then heated to 110 C? to get rid of water) bentonite produced in Iraq which is available and cheap with existence of high density polyethylene at specific conditions of ( heating and mixing) . The best weight proportion were reached, then paraffin wax and additives were added to improve the properties of grease and give the
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The ability of pulverized walnut-shell to remove oil from aqueous solutions has been studied. It involves two-phase process which consists of using walnut-shell as a filtering bed for the accumulation and adsorption of oil onto its surface. Up to 96% oil removal from synthetic wastewater samples was achieved while tests results showed that 75% of oil can be removed from the actual wastewater discharged from Al- Duara refinery in the south of Baghdad.
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Used 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
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The study included the determination of pollen grains features for 8 genera and 13 taxa of Mimosoideae subfamily grown in Baghdad/ Iraq by using each of light and scanning electron microscope. The samples of taxa were collected from various sites in Baghdad province in central Iraq located on 32 45° 0-33 45 0 N and 44 0 0- 44° 45 0 E. the results from this study revealed different pollen types as monad in each of Leucaena, Prosopis, and Neltuma, tetrad in Mimosa and polyads in Acacia, Albizia, Calliandra, Pithecellobium and Vachellia. Each taxa of these genera characterized by special palynological features as shape, size, number of polyads grain and conplateuration as well as other parameters included other dimensions, and these
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Crude soybean peroxidase (SBP), isolated from soybean seed coats (hulls) at unusually low concentrations, catalyses the oxidative polymerisation of hazardous aqueous benzidine and its 3,3′-dichloro, 3,3′-dimethyl and 3,3′-dimethoxy derivatives in the presence of hydrogen peroxide. The optimum operating conditions for oxidation of 0·10 mM benzidine were investigated. At pH 5, the hydrogen peroxide-to-substrate concentration ratio was 1·5 and the minimum SBP concentration required to achieve at least 95% conversion of the benzidine in synthetic wastewater was 0·43 mU/ml. Progress curves were established for the conversion of the four substrates, and apparent first-order rate constants were derived. Enzyme-catalysed polym
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