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 work was conducted to study the recovery of catalyst and desirable components from tar formed in phenol production unit and more particularly relates to such a method whereby better recovery of copper salts, phenol, benzoic acid and benzoate salts from tar by aqueous acid solution was accomplished.
The effect of solvent type, solvent concentration (5, 10, 15, 20, 25 and 30 wt%), agitation speed (100, 200, 300 and 400 rpm), agitation time (5, 10, 15, 20 and 25 min), temperature (90, 100, 110, 120, 130 and 140 oC) , phase ratio (1/1, 2/1, 3/1, 4/1 and 5/1) and number of extraction (1, 2, 3, 4, and 5) were examined in order to increase the catalyst and desirable components extraction.
Four types of solvent were used; hydrochloric
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Seventy of Klebsiella pneumoniae isolates had been collected from some Hospitals in Baghdad city from October to December 2017. The 70 isolates were taken from diverse clinical specimens. All K. pneumoniae isolates were identified based on API 20 E and Vitek2 compact system. Antibiotics sensitivity test was carried out toward 10 antibiotics using discs diffusion method. The level of antibiotics resistance was 81.42% for Ceftriaxone, whereas the low level of antibiotics resistance was 37.14% for Piperacillin. K. pneumoniae isolates were typed genotypically by using two different methods of amplification, multiplex-PCR and enterobacterial repetitive intergenic consensus (ERIC)-PCR typing methods. Results showed that out of 70 isolates, there
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A microbubble air flotation technique was used to remove chromium ions from simulated wastewater (e.g. water used for electroplating, textiles, paints and pigments, and tanning leather). Experimental parameters were investigated to analyze the flotation process and determine the removal efficiency. These parameters included the location of the sampling port from the bottom of the column, where the diffuser is located to the top of flotation column (30, 60, and 90 cm), the type of surfactant (anionic, SDS, or cationic, CTAB) and its concentration (5, 10, 15, and 20 mg/L), the pH of the initial solution (3, 5, 7, 9, and 11), the initial contaminant concentration (10, 20, 30, and 40 mg/L), the gas flow rate (0.1, 0.2, 0.3, and 0.5 L/mi
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New 1,2,4-triazole derivatives of 2-mercaptobenzimidazole (MB) are reported. Ethyl (benzimidazole-2-yl thio) acetate (1) has been prepared by condensing 2-mercaptobenzimidazole with ethylchloroacetate. The ester (1) on reacting with hydrazine hydrate gave the corresponding acetohydrazide(2)which was reacted separately with phenylisocyanate and phenylisothiocyanate, followed by ring closure in an alkaline medium giving 3-[(benzimidazole-2-yl thio) methyl]-4-phenyl-1,2,4-triazole-5-ol and 3-[(benzimidazole-2-yl thio) methyl]-4-phenyl-1,2,4-triazole-5-thiol respectively (6,7). Reaction of acetohydrazide (2) with CS2 and ethanol/KOH, gave dithiocarbazate salt (8). Cyclization of (8) with hydrazine hydrate gave 3-[(benzimi
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The presence of alkaloids in Crassula ovata is a topic that is still unexplored, as there are no published studies on the matter. This study demonstrates the presence of an alkaloid compound (and its class) for the first time in Crassula ovata. The plant material was defatted with n-hexane, and a Soxhlet apparatus was used for the extraction process, while the acid-base method was used for the isolation of alkaloids from the chloroform fractions. The quaternary alkaloid was precipitated from the aqueous layer spontaneously, in high quantity. By using standard spectroscopic methods (including liquid chromatography - mass spectroscopy) we were able to clarify the structure of the precipi¬tated compound as a tetrahydroprotoberberine a
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The aim of this work was to prepare zeolite type 13X from locally available kaolin and to study the effects of using some binding materials through the process of agglomeration of this zeolite. This study was focused on using kaolin binder in different weight percents (10,15,25,35 and 45%).Physical and mechanical properties of the agglomerates such as porosity , apparent density , pore volume, crushing strength , loss on attrition , surface area and finally the adsorption capacity had been measured and evaluated .The preparation step was achieved by mixing the reactants consisting of metakaolin , source of silica as ( sodium trisilicate ) and sodium hydroxide . The conditions was temperature of 70° C and time of mixing as 8, 10,24,34,50
... Show MoreIn this work 2-hydrazino pyrimidine (1) was prepared from 2-mercapto pyrimidine with hydrazine hydrate. Treatment of (1) with active methylene compounds gave 2-(3,5-dimethyl -1 H – Pyrazole-1-yl) pyrimidine , whereas the reaction of (1) with carboxylic anhydride namely maleic anhydride or 1,2,3,6-tetra hydro phthalic anhydride yielded 1-Pyrimidine-2-yl-1,2-dihydro pyridazine-3,6-dione (3) and 2 – Pyrimidin -2-yl -2,3,4 a ,5,8 a – hexahydro phthalazine 1,4 – dione (4) . Reaction of (1) with phenyl isothiocyanate and ethyl chloro acetate afforded 3-Phenyl-1,3-thiazolidine-2,4-dione-2( pyrimidine -2- yl hydrazone (6) Azomethine (7-10) were prepared through condensation of (1) with aromatic aldehydes or ketones, then comp
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