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.
Extraction of copper (Cu) from aqueous solution utilizing Liquid Membrane technology (LM) is more effective than precipitation method that forms sludge and must be disposed of in landfills. In this work, we have formulated a liquid surfactant membrane (LSM) that uses kerosene oil as the main diluent of LSM to remove copper ions from the aqueous waste solution through di- (2-ethylhexyl) phosphoric acid - D2EHPA- as a carrier. This technique displays several advantages including one-stage extraction and stripping process, simple operation, low energy requirement, and. In this study, the LSM process was used to transport Cu (II) ions from the feed phase to the stripping phase, which was prepared, using H2SO4. For LSM p
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Calendula officinalis L. (Asteraceae) known as marigold is known to have several pharmacological activities and used for the treatment of several diseases as measles, jaundice, constipation and several inflammations. Marigold flowers contain several chemical constituents mainly flavonoids, triterpenoids and essential oil. In this study marigold flowers cultivated in Iraq had been investigated for its flavonoids content. The study revealed the presence of quercetin and kaempferol glycosides and the absence of myricetin glycosides. The flowers were extracted with ethanol 70% fractionated with different solvent and the flavonoids were isolated by preparative HPLC. The isolated flavonoids were identified by measuring melting points, UV, IR,
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Wohlfahrtia longicorpuris sp. nov., from Iraq described, illustrated and distinguished from related species. The adults were reared from larvae collected from ulcer of a human face. Wohlfahrtia Brauer and Bergenstam is one of most important genus,which contains 19 species (Pape, 1998), some of these produce myiasis in mammals (Verves,1985).Taxonomic revision of this genus has been carried out by the following authors: Rohdendrof (1956), Zumpt (1965) and Pape (1996).
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Feasibility of biosorbent of England bamboo plant origin was tested for removal of priority metal ions such as Cu and Zn from aqueous solutions in single metal state. Batch single metal state experiments were performed to determine the effect of dosage (0.5, 1 and 1.5 g), pH (3, 4, 4.5, 5 and 6), mixing speed (90, 111, 131, 156 and 170 rpm), temperature (20, 25, 30 and 35 °C) and metal ion concentration (10, 50, 70, 90 and 100 mg/L) on the ability of dried biomass to remove metal from solutions which were investigated. Dried powder of bamboo removed (for single metal state) about 74 % Cu and 69% Zn and maximum uptake of Cu and Zn was 7.39 mg/g and 6.96 mg/g respectively, from 100 mg/L of synthetic metal solution in 120 min. of contact t
... Show MoreThe objective of this study was to investigate the release profile of different fat and water soluble bases using diazepam as a model drug , and then to develop a satisfactory formula with a rapid release of diazepam from suppository bases .The study was conducted using theobroma oil ,glycerol-gelatin and glycerol-PEG1540 bases using conventional mold method for preparation .while the later base was utilized to incorporate diazepam ( buffered solution ) in a hollow type suppositories. The results indicated that all types of bases can be utilized to formulate diazepam as rectal suppositories with acceptable disintegration time ( 12, 10, 6, and 6min.), respectively . While 100% of the released drug had been shown differen
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ABSTRACT: Pathogenic bacteria responsible for the causation of many common diseases have been identified on currency notes. The present investigation was carried out on one hundred currency notes of all the denominations (50, 100, 250, 500 and 1000RY), obtained from different occupational mainly bus drivers, hawker street, vegetable vendor, restaurants and butchers and fish seller groups in Taiz city,Yemen. Identification and characterization revealed active participation of the following species of organisms in the ascending order of percentage as E. coli(50.28 %),Staphylococci aureus(14.04 %), Klebsiellaspp(4.39 %),proteus(4.39 %), salmonella(1.25 %), shigella(0.72 %), Coagulase negative staphylococcus(0.60 %), pseudomonas(0.50 %),
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