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.
A study of the Torymid collection of Iraq. resulted in undescribed species of the genus
Liodontonierus Gah. L. longicorpus sp. n. with 2 figures.
In the present work, 9-fluorenone-2-carboxylic acid methyl ester (1) was prepared from 9-fluorenone-2-carboxylic acid and then converted into the acid hydrazide (2). Compound (2), is the key intermediate for the synthesis of several series of new compounds such as substituted 1,3,4-oxadiazole derivatives (3-6) were synthesized from the condensation of different substituted benzoic acids with compound (2) using POCl3 as condensing agent. Treatment of compound (2) with formic acid gave the N-formyl hydrazide (7), which upon refluxing with phosphorous pentoxide in benzene yielded the corresponding 5-(9-fluorenone-2-yl)-1,3,4-oxadiazole (8). Reaction of hydrazide (2) with phenyl isocyanate to give N-phenyl semicarbazide derivative (9), then thi
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In this research, the efficiency of low-cost unmodified wool fibers were used to remove zinc ion from industrial wastewater. Removal of zinc ion was achieved at 99.52% by using simple wool column. The experiment was carried out under varying conditions of (2h) contact time, metal ion concentration (50mg/l), wool fibers quantity to treated water (70g/l), pH(7) & acid concentration (0.05M). The aim of this method is to use a high sensitive, available & cheep natural material which applied successfully for industrial wastewater& synthetic water, where zinc ion concentration was reduced from (14.6mg/l) to (0.07mg/l) & consequently the hazardous effect of contamination was minimized.
(1)
Various industrial applications include the dyeing of textiles, paper, leather, and food products, as well as the cosmetics industry. Physic-chemical methods are required to breakdown dyes because they are known to be harmful and persistent in the environment. Many companies' treated effluents contain small amounts of dyes. When it comes to removing dye from wastewater, adsorption has verified to be aneconomical alternative to more traditional treatment procedures. It's important to degrade color impurities in industrial effluents since they constitute a serious health and environmental concern. One way that's been tried is using clay minerals as an adsorbent. Using adsorption for removing
... Show MoreJudo has witnessed tremendous developments since its inception until the present day. It has been distinguished by its adaptation to the various challenges it has faced throughout the ages. Judo is one of the sports that have been affected by social, technological and cultural changes. These changes reflect its transformation from the traditional Japanese martial art to a global sport practiced. All over the world, therefore, studying the historical development of judo is important, as it provides valuable insights into the development of martial arts over a century, by studying the origins, principles and techniques of judo for the period (1880 - 1980), and also enables us to gain an understanding A deeper understanding of how the art form
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This study aimed to extraction of essential oil from peppermint leaves by using hydro distillation methods. In the peppermint oil extraction with hydro distillation method is studied the effect of the extraction temperature to the yield of peppermint oil. Besides it also studied the kinetics during the extraction process. Then, 2nd -order mechanism was adopted in the model of hydro distillation for estimation many parameters such as the initial extraction rate, capacity of extraction and the constant rat of extraction with various temperature. The same model was also used to estimate the activation energy. The results showed a spontaneous process, since the Gibbs free energy had a value negative sign.
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