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 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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The interest in calcium phosphates arises from the fact that bones contain a high percentage of mineralized calcium phosphate . In this study, pure and biocompatible hydroxyapatite (HAP) powder was successfully synthesized using hen’s egg shell as calcium source and phosphoric acid by precipitation method.The precipitate obtained was subjected to ripenning process for 24 hours, filtered, air dried, and calcined at temperatures of 400,800,900,and 1000 ºC.
X-Ray diffraction(XRD) technique was used to investigate the formation of HAP powder, XRD results revealed the HAP formation and also indicate no occurrence of secondary phases. Fourier Transform Infrared(FT-IR) spectrum shows the characterstic peaks for phosphate and hydroxyl grou
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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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 physica
... Show MorePorosity 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 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
Basrah crude oil Vacuum residue 773+ K with specific gravity 1.107 and 4.87wt. % sulfur, was treated with hexane commercial fraction provided from Al-Taji Gas Company for preparing deasphaltened oil(DAO)suitable for hydrotreating process. Deasphaltening was carried out with 1h mixing time, 10ml:1g solvent to oil ratio and at room temperature. Hexane deasphaltened oil was hydrotreated on presulfied commercial Co-Mo/γ-Al2O3 catalyst in a trickle bed reactor. The hydrotreating process was carried out at temperature 660 K,LHSV 1.3 h^ –1, H2/oil ratio 300 l/l and constant pressure of 4MPa. The hydrotreated product was distillated under vacuum distillation unit. It is found that the mixture of 75% of vacuum residue with 25% anthracene satisfie
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