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.
KE Sharquie, AA Noaimi, ZN Al-Khafaji…, Journal of Cosmetics, Dermatological Sciences and Applications, 2016 - Cited by 2
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Anadara granosa is a species of the class bivalve commonly found on the east coast of South Sumatra as a fishery commodity. This species has not been widely studied as a source of new bioactive compounds that have antioxidant abilities. This study aims to analyze the antioxidant ability of A. granosa against DPPH radicals and its phytochemical profile qualitatively. Samples were taken at the fishing port of Sungsang Village, South Sumatra, Indonesia. Furthermore, the samples were extracted using ethanol as a solvent and tested for antioxidants against DPPH radicals, total phenol analysis, and preliminary phytochemical test. Based on the antioxidant test results, the IC50 value of the ethanolic extract of
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In this study, novel Schiff base complexes with Zn(II) and Co(II) ions were successfully synthesized. The malonic acid dihydrazide was converted into the Schiff base ligand by combining it with 1-hydroxy-2-naphthaldehyde, and the last step required reacting it with the appropriate metal(II) chloride to produce pure target complexes. The generated complexes were thoroughly characterized using FTIR, 1H-NMR, 13C-NMR, GC-mass, and UV-Vis spectroscopies. In order to photo-stabilize polystyrene (PS) and reduce the photodegradation of its polymeric chains, these chemicals have been used in this work. The efficiency of the generated complexes as photo-stabilizers was evaluated using a variety of techniques, including FTIR, weight loss, visc
... Show MoreBidentate Schiff base ligand 3-(3,4-Dihydroxy-phenyl)-2-[(4-dimethylamino-benzylidene)-amino]-2-methyl-propionic acid was prepared and characterized by spectroscopic techniques studies and elemental analysis. The Cd(II), Ni(II), Cu(II), Co(II), Cr(III),and Fe(III) of mixed-ligand complexes were structural explicate through Moler conductance , [FT-IR, UV-Vis & AAS], chloride contents, , and magnetic susceptibility measurements. Octahedral geometries have been suggested for all complexes. The Schiff base and its complexes were tested against various bacterial species, two of {gram(G+) and gram(G-)} were shown weak to good activity against all bacteria.
First record of Myxobolus insignis Eiras, Malta, Varella, Pavanelli, 2005 (Myxozoa: Myxobolidae) in Iraq from gills of the common carp Cyprinus carpio
New series of imidazole[1,2-a]pyridine-sulfonamides was designed and synthesized from 2-aminopyridine, which was reacted with p-bromo phenacyl bromide in the present of MgO to produce the corresponding imidazole[1,2-a]pyridine, which was then reacted with chlorosulfonic acid to produce 2-(4-bromophenyl)imidazole[1,2-a]pyridine-3-sulfonyl chloride [2]. Following that, treatment of (2) with different amines using the grand method to generate imidazole [1,2-a] pyridine sulfonamides. All the synthesized compounds have been characterized by FTIR, 1HNMR and 13CNMR and C.H.N analysis. The DFT, POM analysis and molecular docking were carried out on for all final compounds to investigate drug like attributes, and the results revealed showed that the
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Corrosion of Zn-Ni alloy coatings on stainless steel 316 SS in a chloride-sulfate bath with the addition of either triethanolamine or sucrose was examined. A constant cathode potential was used to deposit zinc-nickel alloys, while cyclic voltammetry and potentio-dynamic polarization were used to measure corrosion. In addition, scanning electron microscopy was utilized to analyse Zn-Ni alloy coating surface layers formed with¬out and with additives. The outcomes discovered that the corrosion resistance of Zn-Ni alloy coat¬ings in 3.5 % NaCl solution was highly influenced by adding triethanolamine or sucrose. Decreasing the Zn:Ni molar ratio led to an increase in corrosion resistance. All Zn-Ni alloy coatings were superior to pure Z
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