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.
Pectin is available in many plants and in this study, the peels of tomatoes and beet were used to be an economical source of pectin production instead of dumping it with waste or using it as animal feed. The pectin extracted from the peels using different solutions, namely citric acid (2 M), oxalic acid (2%) and hydrochloric acid (0.5 M) the outcome of the extraction methods, 7. 1%, 6% and 11% respectively for tomatoes peels, while the pectin of beet peels were 8%, 6.5%, and 8.3%, and the highest percentage obtained in the manner of hydrochloric acid adopted in the manufacture of yogurt.Yogurt was manufactured with four treatments, in the first treatment standard pectin was added and the second treatment in addition to the pectin extracted
... Show MoreThis research was carried out to study the effect of plants on the wetted area for two soil types in Iraq and predict an equation to determine the wetted radius and depth for two different soil types cultivated with different types of plants, the wetting patterns for the soils were predicted at every thirty minute for a total irrigation time equal to 3 hr. Five defferent discharges of emitter and five initial volumetric soil moisture contents were used ranged between field capacity and wilting point were utilized to simulate the wetting patterns. The simulation of the water flow from a single point emitter was completed by utilized HYDRUS-2D/3D software, version 2.05. Two methods were used in developing equations to predict the domains o
... Show MoreObjective: The current study aimed identifying the impact of rehabilitative exercises combined with ultrasonic waves on reducing pain in people with carpal tunnel compression and determining how these activities affect range of motion of the upper limb for those suffering from carpal tunnel compression. Research methodology: With pre- and post-tests, the researchers employed the experimental method in the form of two equal groups, the experimental and the control. The scientific community and sample are among the priorities that fall on the researcher, so The scientific community is determined by those suffering from carpal tunnel compression, numbering (14) patients. (12) Patients were approved and two were excluded from the resear
... Show MoreThe removal of fluoride ions from aqueous solution onto algal biomass as biosorbent in batch and continuous fluidized bed systems was studied. Batch system was used to study the effects of process parameters such as, pH (2-3.5), influent fluoride ions concentration (10- 50 mg/l), algal biomass dose (0–1.5 g/ 200 ml solution), to determine the best operating conditions. These conditions were pH=2.5, influent fluoride ions concentration= 10 mg/l, and algal biomass dose=3.5 mg/l. While, in continuous fluidized bed system, different operating conditions were used; flow rate (0.667- 0.800 l/min), bed depth (8-15 cm) corresponded to bed weight of (80- 150 g). The results show that the breakthrough time increases with the inc
... Show MoreSynthesis of a new class of Schiff-base ligand with a tetrazole moiety to form polymeric metal complexes with CoII, NiII, ZnII, and CdII ions has been demonstrated. The ligand was synthesised by a multi-steps by treating 5-amino-2-chlorobenzonitrile and cyclohexane -1,3-dione, the 5,5'-(((1E,3E)-cyclohexane-1,3-diylidene)bis(azanylylidene))bis(2-chlorobenzonitrile) was obtained. The precursor (M) was prepared from the reaction 5,5'-(((1E,3E)-cyclohexane-1,3-diylidene)bis(azanylylidene))bis(2-chlorobenzonitrile) with NaN3 to obtained (1E,3E)-N1,N3-bis(4-chloro-3-(1H-tetrazol-5-yl)phenyl)cyclohexane-1,3-diimine (N). By reacting the precursor (M) with CS2
... Show MoreIn this work, microbubble dispersed air flotation technique was applied for cadmium ions removal from wastewater aqueous solution. Experiments parameters such as pH (3, 4, 5, and 6), initial Cd(II) ions concentration (40, 80, and 120 mg/l) contact time( 2, 5, 10 , 15, and 20min), and surfactant (10, 20and 40mg/l) were studied in order to optimize the best conditions .The experimental results indicate that microbubbles were quite effective in removing cadmium ions and the anionic surfactant SDS was found to be more efficient than cationic CTAB in flotation process. 92.3% maximum removal efficiency achieved through 15min at pH 5, SDS surfactant concentration 20mg/l, flow rate250 cm3/min and at 40mg/l Cd(II) ions initial co
... Show MoreCo+2, Ni+2, Cu+2 as well Zn+2 compounds mixed ligand from 8-hydroxyquinoline(8-HQ) also tributylphosphine (PBu3) have been attended at aquatic ethyl alcohol for (1:2:2) (M:8-HQ:PBu3). Produced complexes have been identified by utilizing atomic absorption flame, FT-IR as well UV-Vis spectrum manners also magnetic susceptibility as well as conductivity methods. At addendum antibacterial efficiency from the ligands as well complexes oboist three species about bacteria have been as well examined. Ligands and their complexes show good bacterial efficiencies. Of the gained datum the octahedral geometry was proposed into whole prepared complexes
The synthesized ligand [4-chloro-5-(N-(5,5-dimethyl-3-oxocyclohex-1-en-1-yl)sulfamoyl)-2-((furan-2-ylmethyl)amino)benzoic acid] (H2L1) was identified utilizing Fourier transform infrared spectroscopy (FT-IR), 1 H, 13 C – NMR, (C.H.N), Mass spectra, UVVis methods based on spectroscopy. To detect mixed ligand complexes, analytical and spectroscopic approaches such as micro-analysis, conductance, UV-Visible, magnetic susceptibility, and FT-IR spectra were utilized. Its mixed ligand complexes [M(L1)(Q)Cl2] [ where M= Co(II), Ni(II) , and Cd(II)] and complexes [Pd(L1)(Q)] and [Pt(L1)(Q)Cl2]; [H2L1] =β-enaminone ligand =L1 and Q= 8-Hydroxyquinoline = L2]. The results showed that the complexes were synthesised utilizing the molar ratio M: L1
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