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.
This study expands the state of the art in studies that assess torsional retrofit of reinforced concrete (RC) multi-cell box girders with carbon fiber reinforced polymer (CFRP) strips. The torsional behavior of non-damaged and pre-damaged RC multi-cell box girder specimens externally retrofitted by CFRP strips was investigated through a series of laboratory experiments. It was found that retrofitting the pre-damaged specimens with CFRP strips increased the ultimate torsional capacity by more than 50% as compared to the un-damaged specimens subjected to equivalent retrofitting. This indicated that the retrofit has been less effective for the girder specimen that did not develop distortion beforehand as a result of pre-loading. From
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In this research, production of ethanol from waste potatoes fermentation was studied using Saccharmyses cerevisiae. Potato Flour was prepared from potato tubers after cooking and drying at 85°C. Homogenous slurry of potato flour was prepared in water at solid liquid ratio 1:10. Liquefaction of potato flour slurry with α-amylase at 80°C for 40 min followed by saccharification with glucoamylase at 65°C for 2 hr .Fermentation of hydrolysate with Saccharomyces cerevisiae at 35°C for two days resulted in production of 33 g/l ethanol.
The parameters studied were; temperature, time of fermentation and pH. It was found that Saccharification process is affected by enzyme Amylo 300 conc
... Show MoreThe extraction process of chlorophyll from dehydrated and pulverized alfalfa plant were studied by percolation method. Two solvent systems were used for the extraction namely; Ethanol-water and Hexane-Toluene systems . The effect of circulation rate, solvent concentration, and solvent volume to solid weight ratio were studied. In both ethanol water, and Hexane-Toluene systems it appears that solvent concentration is the most effective variable.
Objective: The goal of this research was to evaluate where selenium nanoparticles impact the activity of antibodies in immunized lambs with foot and mouth vaccines by modulating the immune system. Materials and Methods: Two groups of lambs of 3–4 months of age were injected with 1 ml of ARRIAH-VAC vaccine intramuscularly in the neck, five Lambs were given selenium nanoparticles (size 100 nm) oral administration of selenium nano dose of 0.1 mg/kg of body mass once every day for sixty days considered as group one (G1) while the other five used as control Group 2 (G2). Results: This resulted in the establishment of an immune response, as evidenced by a rise in antibody titer in the blood using the ELISA test for three serotypes A,
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Bauxite residue (red mud) is a waste material from alumina refineries in the Bayer process, containing significant quantities of valuable metals, notably scandium (Sc). The objective of this study is to recover Sc (III) from Hungarian bauxite residue by using hydrometallurgical processes, including solvent extraction and leaching. Red mud directly leached with hydrochloric acid to generate the leachate solution. The significant iron content (~38 %) in red mud makes it hard to recover scandium selectively due to comparable physicochemical characteristics. According to the findings, Fe (III) could be effectively extracted from hydrochloric acid leachate as HFeC14 using diethyl ether before Sc extraction. Protocol B demonstrated superior recov
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Bauxite residue (red mud) is a waste material from alumina refineries in the Bayer process, containing significant quantities of valuable metals, notably scandium (Sc). The objective of this study is to recover Sc (III) from Hungarian bauxite residue by using hydrometallurgical processes, including solvent extraction and leaching. Red mud directly leached with hydrochloric acid to generate the leachate solution. The significant iron content (~38 %) in red mud makes it hard to recover scandium selectively due to comparable physicochemical characteristics. According to the findings, Fe (III) could be effectively extracted from hydrochloric acid leachate as HFeC14 using diethyl ether before Sc extraction. Protocol B demonstrated superior recov
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