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 present study aims to detection optimal conditions of production of amylase enzyme from isolate of B. subtillis A4. Nine carbonic sources were represented by starch, maltose, fructose, sucrose, glucose, arabinose, xylose, sorbitol and mannitol) at concentration of 1% for each source. It was found that the best was represented by starch carbonic, which showed higher activity and qualitative activity of 7.647 Unit/ ml and 461.56 Unit/ mg. Ten nitrogen sources were selected, including yeast extract, peptone, trypton, gelatin, urea and meat extract as organic sources Ammonium sulphate, Sodium nitrate, Potassium nitrate and Ammonium chloride as inorganic sources. These sources were added at aconcentration of 0.5% to the production medium. Th
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A new bio-electrochemical system was proposed for simultaneous removal of organic matters and salinity from actual domestic wastewater and synthetically prepared saline water, respectively. The performance of a three-chambered microbial osmotic fuel cell (MOFC) provided with forward osmosis (FO) membrane and cation exchange membrane (CEM) was evaluated with respect to the chemical oxygen demand (COD) removal from wastewater, electricity generation, and desalination of saline water. The MOFC wasinoculated with activated sludge and fueled with actual domestic wastewater. Results revealed that maximum removal efficiency of COD from wastewater, TDS removal efficiency from saline water, power density, and current density were
... Show MoreThis work deals with preparation of zeolite 5A from Dewekhala kaolin clay in Al-Anbar region for drying and desulphurization of liquefied petroleum gas. The preparation of zeolite 5A includes treating kaolin clay with dilute hydrochloric acid 1N, treating metakaolin with NaOH solution to prepare 4A zeolite, ion exchange, and formation. For preparation of zeolite 4A, metakaolin treated at different temperatures (40, 60, 80, 90, and 100 °C) with different concentrations of sodium hydroxide solution (1, 2, 3, and 4 N) for 2 hours. The zeolite samples give the best relative crystallinity of zeolite prepared at 80 °C with NaOH concentration 3N (199%), and at 90 and 100°C with NaOH concentration solution 2N (184% and 189%, respectively). Ze
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This paper deals with a method called Statistical Energy Analysis that can be applied to the mechanical and acoustical systems like buildings, bridges and aircrafts …etc. S.E.A as a tool can be applied to the resonant systems in the circumstances of high frequency or/and complex structure». The parameters of S.E.A such as coupling loss factor, internal loss factor, modal density and input power are clarified in this work ; coupled plate sub-systems and explanations are presented for these parameters. The developed system is assumed to be resonant, conservative, linear and there is an equipartition of energy between all the resonant modes within a given frequency band in a given sub-system. The aim of th
... Show MoreAims: The aim of this study was to evaluate the value and accuracy of longitudinal strain in detection of coronary artery disease compared to coronary angiography. Results: The left ventricular longitudinal strain-speckle tracking showed evidence of stenosis of left anterior descending artery, circumflex artery and right coronary artery in (86.1%), (76.4%), and (84.7%) respectively. For the stenosis in left anterior descending artery, the current study showed that the longitudinal strain was a good predictor for presence of significant stenosis with a sensitivity of (93.8%), specificity (75%) and accuracy (91.7%) compared with coronary angiography. For the stenosis in right coronary artery, the left ventricular longitudinal strain had
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In this article, Pb2Ba1.7Sr0.3Ca2Cu3O10+δ superconductor material was synthesized using conventional solid-state reaction method. X-ray diffraction (XRD) analysis demonstrated one dominant phase 2223 and some impurities in the product powder. The strongest peaks in the XRD pattern were successfully indexed assuming a pseudo-tetragonal cell with lattice constants of a = 3.732, b = 3.733 and c = 14.75 Å for a Pb-Based compound. The crystallite size and lattice strain between the layers of the studied compound were estimated using several methods, namely the Scherrer, Williamson-Hall (W.H), sizestrain plot (SSP) and Halder Wagner (H.W) approach. The values of crystallite size, calculated by Scherrer, W.H, SSP and H.W methods, were 89.454077
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The present study attempts to shed light on some biological aspects and characteristics of Piaractus brachypomus, including some biometrics, phenotypic and feeding pattern that characterizes this species. Besides, the study touches upon the body shape and the Otolith. These fish species have recently been seen frozen in the Iraqi local market. The standard length of fish specimens in this study reach 26.55cm it exceeded the specimens of Pacu fish collected from other studies from other countries, As well the specimens weight was 632gm it exceeded other studies mentioned in this manuscript. As the irregularity in the distribution of teeth rows, especially in the lower jaw was clear in our specimens. The average weight of some skull b
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The present investigation is concerned for the purification of impure zinc oxide (80-85 wt %) by using petroleum coke
(carbon content is 76 wt %) as reducing agent for the impure zinc oxide to provide pure zinc vapor, which will be
oxidized later by air to the pure zinc oxide.
The operating conditions of the reaction were studied in detail which are, reaction time within the range (10 to 30 min),
reaction temperature (900 to 1100 oC), air flow rate (0.2 to 1 l/min) and weight percentage of the reducing agent
(petroleum coke) in the feed (14 to 30 wt %).
The best operating conditions were (30 min) for the reaction time, (1100 oC) for the reaction temperature, (1 l/min) for
the air flow rate, and (30 wt %) of reducing