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.
ABSTRACT
This study was conducted to determine the effect of various levels of hump fat (HF) used in manufacturing of camel, beef and chicken sausage to understand the effect of (HF) on physicochemical composition sausage, Different levels of hump fat (5, 7, and 10 %) were used, physicochemical compositions like (moisture, protein, fat, Ash, water holding capacity, shrinkage, cooking loss and pH) were determined. Results of the study revealed that moisture content showed high significant differences (P≤0.01)among treatments groups, Camel sausage and beef sausage tended to have highest values while chicken sausage reported the lowest value. The study showed no significant difference (P≤0.05) among the
... Show MoreSince its first description as a cytotoxic agent, Olea europaea leaves extract gained significant popularity against human breast cancer, ethyl acetate extract of Olea europaea leaves obtained by acid hydrolysis method was evaluated in vitro as cytotoxic agent against new human breast cancer (AMJ13) cell line, using the MTT assay. One main pentacyclic triterpenoid; oleanolic acid, was isolated from leaves of Olea europaea by well-known two different methods, but not used for this compound before, the acidic hydrolysis method and basic acidic method. The presence of oleanolic acid was proved in both methods with qualitative and quantitative d
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This study aims to suggest an alternative to the use of quality agricultural soil in the brick industry (Iraq). The Late Miocene claystone bed in the Injana Formation in central Iraq was targeted through the study of 18 exposed sections that were sampled by using the trench sampling method. The claystones are characterized by quartz (36.4%) followed by calcite (32.8%), quartz (36.4%) feldspar (2.6%), gypsum (1.3%) and dolomite (0.7%), kaolinite (10.5%), illite (7.7%), chlorite (6.7%), palygorskite (6.0%) and montmorillonite (0.7%). New thermal mineral phases were formed at 950°C, including diopside (62.9%), quartz (18.4%), wollastonite (8.28%), akermanite (7.6%), Anorthite (6.25%), Nosean (4.9%), gehlenite (3.75%) and Lazurite (3.1
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The problem of research is to identify the uses of Iraqi women for Facebook and the motives of these uses and the innovations that have been achieved from them, in view of the increasing role-played by social networking sites in the lives of individuals in general and women in particular.The research seeks to identify the following: Identify the extent to which Iraqi women use Facebook and the usage motives and innovations achieved by the interviewees When they using Facebook. This study belongs to the descriptive studies that are interested in monitoring the characteristics of a particular phenomenon to identify its characteristics and characteristics.This study adopted the survey methodology to test the hypotheses of the study and how
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The Catharanthus roseus plant was extracted and converted to nanoparticles in this work. The Soxhlet method was used to extract alkaloid compounds from the Catharanthus roseus plant and converted them to the nanoscale. Chitosan polymer was used as a linking material and converted to Chitosan nanoparticles (CSNPs). The extracted alkaloids were linked with Chitosan nanoparticles by maleic anhydride to get the final product (CSNPs-Linker-alkaloids). The pure Chitosan, Chitosan nanoparticles, and CSNPs-Linker-alkaloids were characterized by X-ray diffractometer, and Fourier Transform Infrared spectroscopy. X-ray results show that all samples have an orthorhombic structure with crystallite size in nanodimensions. FTIR spectra prove that
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The electrospun nanofibers membranes have gained considerable interest in water filtration applications. In this work, the fabrication and characterization of the electrospun polyacrylonitrile-based nonwoven nanofibers membrane are reported. Then, the membrane's performance and antifouling properties were evaluated in removing emulsified oil using a cross flow filtration system. The membranes were fabricated with different polyacrylonitrile (PAN) concentrations (8, 11, and 14 wt. %) in N, N-Dimethylformamide (DMF) solvent resulted in various average fiber sizes, porosity, contact angle, permeability, oil rejection, and antifouling properties. Analyses of surface morphology of the fabricated membranes before and after oil removal revealed
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This work was conducted to study the extraction of pelletierine sulphate from Punica granatum L. roots by liquid membrane techniques. Pelletierine sulphate is used widely in medicine. The general behavior of extraction process indicates that pelletierine conversion increased with increasing the number of stages and the discs rotation speed but high rotation speed was not favored because of the increased risk of droplet formation during the operation. The pH of feed and acceptor solution was also important. The results exhibit that the highest pelletierine conversion was obtained when using two stages,(10 rpm) discs speed of stainless steel discs,(pH= 9.5) of feed solution and (pH= 2) of acceptor solution in n-decane. Assuming the existence
... Show MoreIn this work, the extraction of glycyrrhizin from Licorice using bulk liquid membrane technique was developed and optimized. The effect of various parameters such as pH of stripping and donor solutions, temperature, stirring speed and kinetic parameters were investigated. Moreover, to study the impact of the polarity of membrane solvent, two types of extraction solvents were used as a membrane solvent: n-Hexane was used as a non-polar solvent and 1-Hexanol was as a polar solvent. The optimum extraction condition was found (95.53%) using 1-Hexanol, rotating speed was 400 rpm, and pH of the acceptor and donor solutions were 8 and 5.5, respectively. The reaction kinetics constants ( and ) for the transport of glycyrrhizin from the donor pha
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