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.
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Cupressus sempervirens L., Cupressaceae, that is known as evergreen cypress, Mediterranean cypress and in Arabic called “al -Sarw. It is an evergreen, medium sized, longevity, and wide distributed over all the world. The plant represents an important member of conifer plants which characterized with aromatic leaves and cones. Cupressus sempervirens have been ethnobotanical uses as an antiseptic, relief of cough, astringent, antispasmodic, wound healing and anti-inflammatory. Aims of this work are phytochemical analysis, isolation and structural identification of Quercitroside (quercitrin) and essential oil in Iraqi C. sempervirens. Isolation of quercitrin was
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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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Ten blue swimming crabs Portunus segnis (Forskål, 1775) were collected from the north west of the Arabian Gulf off the Iraqi marine waters from October to November 2017 at 29ᵒ 37′ N to 48ᵒ 47′ E.
The barnacle Octolasmis angulata (Aurivillius, 1894) was found on the gills of the present species of crab, the mean incidence of infestation was 30%, while the mean Intensity of infestation was 12.3. The barnacle have a long and slim shaped calcareous plate with the presence of carina and the absence of tergum, in addition to the elongated shape of carina and scutum. The current study represents the first record of the barnacle O. angulata in the Ar
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The current study aimed to isolate and diagnose Candida spp yeasts that cause candidiasis with a PCR device from patients reviewed for some hospitals in Baghdad city and by 190 samples, the study recorded 123 isolates and the total percentage of infection was 64.7% .Samples were taken from different clinical cases of the vagina, blood and mouth and the Candida spp were (70.37%, 41.26%, 86.95%) respectively. Five types of yeasts were isolated and diagnosed, namely C. albicans, C. tropicalis, C. parapsilosis, C. krusei and C.glabarta. They were confirmed by PCR device and the most notable were yeast C. albicans, where 91 isolates were found, 73.98%, while the lowest infection was recorded. C.glabartawith 3 isolates, at 2.43%, significant diff
... Show MoreThe aim of the work is synthesis and characterization of bidentate ligand [dipotassium sodium7-((E)-2-(2-((Z)-1-carboxylatoethylideneamino)thiazol-4-yl)-2 (carboxylatemethoxyimino) acet amido)-8-oxo-3-vinyl-5- thia-1-azabicyclo[4.2.0] oct-2- ene-2- carboxylate] [Nak2L], from the reaction of cefixime with sodium pyruvet to produce the ligand [Nak2L], the reaction was carried out in methanol as a solvent under reflux. The prepared ligand [Nak2L] which was characterized by FT-IR, UV-Vis spectroscopy, 1H, 13C-NMR spectra, Mass spectra, (C.H.N) and melting point. The mixed ligand complexes were prepared from ligand [Nak2L] was used as a primary ligand while 8-hydroxy quinoline [Q] was used as a secondary ligand with metal ion M(?).Where M(?) =
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Fatty Acid Methyl Ester (FAME) produced from biomass offers several advantages such as renewability and sustainability. The typical production process of FAME is accompanied by various impurities such as alcohol, soap, glycerol, and the spent catalyst. Therefore, the most challenging part of the FAME production is the purification process. In this work, a novel application of bulk liquid membrane (BLM) developed from conventional solvent extraction methods was investigated for the removal of glycerol from FAME. The extraction and stripping processes are combined into a single system, allowing for simultaneous solvent recovery whereby low-cost quaternary ammonium salt-glycerol-based deep eutectic solvent (DES) is used as the membrane phase.
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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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This study focuses on the biodegradation of oxymatrine insecticide by some soil fungi isolated from four agriculture stations. The results showed that the highest degradation rate 94.66% was recorded by Ulocladium sp. at 10 days and A. niger recorded the lowest degradation rate 45.86%, while at 20 days Ulocladium sp. also showed the highest degradation rate 94.98% and the lowest degradation rate reached to 82.49% with A.niger. The mix (Exerohilum sp.+Ulocladium sp.) recorded the highest degradation rate of oxymatrine insecticide 90.22%, 88.51%, 85.34% at 4, 8 and 12 ppm.The use of mixed isolates enhanced the biodegradation process. There is no study of oxymatrine biodegradation
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Thirty one samples of gum swabs were collected from patients with tooth caries (5-30 years old) from the College of Science (Biology department )- University of Baghdad- Iraq for the period from October 2018 to December 2018. , The samples were transported, after inoculation in a transport media (nutrient broth), to the laboratory of the College of Science and then cultured on mannitol salt agar and blood agar). The isolates belonging to Staphylococcus spp. were identified by biochemical tests and Vitek 2 compact system, while the more antibiotic resistant isolates were identified by using Polymerase Chain Reaction(ï´¾PCR) and sequencing of 16SrRNA . The results showed sharp UV absorption peaks at 330 - 340nm and AFM at 5
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