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.
The present study examines the extraction of lead (Pb), cadmium (Cd) and nickel (Ni) from a contaminated soil by washing process. Ethylenediaminetetraacetic acid disodium salt (Na2EDTA) and hydrochloric acid (HCl) solution were used as extractants. Soil washing is one of the most suitable in-situ/ ex-situ remediation method in removing heavy metals. Soil was artificially contaminated with 500 mg/kg (Pb , Cd and Ni ). A set of batch experiments were carried out at different conditions of extractant concentration , contact time, pH and agitation speed. The results showed that the maximum removal efficiencies of (Cd, Pb and Ni ) were (97, 88 and 24 )&nbs
... Show MoreThe removal of Anit-Inflammatory drugs, namely; Acetaminophen (ACTP), from wastewater by bulk liquid membrane (BLM) process using Aliquat 336 (QCl) as a carrier was investigated. The effects of several parameters on the extraction efficiency were studied in this research, such as the initial feed phase concentration (10-50) ppm of ACTP, stripping phase (NaCl) concentration (0.3,0.5,0.7 M), temperature (30-50oC), the volume ratio of feed phase to membrane phase (200-400ml/80ml), agitation speed of the feed phase (75-125 rpm), membrane stirring speed (0, 100, 150 rpm), carrier concentration (1, 5, 9 wt%), the pH of feed (2, 4, 6, 8, 10), and solvent type (CCl4 and n-Heptane). The study shows that high ext
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This research has been prepared to isolate and diagnose one of the most important vegetable oils from the plant medical clove is the famous with Alaeugenol oil and used in many pharmaceuticals were the isolation process using a technique ultrasonic extraction and distillation technology simple
Solanum americanum is a new annual shrubby plant seen recently in fields and gardens of Baghdad city. A new species is described and illustrated, inhabit wet or semi dry places and have consequently a mesophytic habit. A detailed morphological study of the stems, leaves, Inflorescence, flower, male and female reproductive organs and fruits has been done, revealed several interesting taxonomic characteristics, which have not previously been studied in Iraq. Also, anatomical studies reveals constant taxonomical characteristics such as the presence of anthocayanine in outer row of epidermis, distinct chlorenchyma in whole cortex, the wide pith of stems, and presence of distinct mesophyll that differentiated into palisade layer and spongy laye
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Refractories are mineral and chemical-, based, materials with excellent heat resistance, making them ideal for use in the construction of ovens, furnace walls industries. According to this our research is concerned to study the effect of addition of (4% CaO) and (5% graphite) on the silica brick properties. Different amounts of CaO and Graphite were included in the white sand (raw ingredients) of silica bricks as a binder to prepare the composition then the composition were sintered using Different sintering temperatures ranging from (1000–1400)𝛐C under static air. Density, thermal conductivity, porosity, and water absorption Compression there was power tested after sintering. XRD analysis was used to identify raw materials’
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This work included synthesis of several new polymers of polyacryloyl chloride in two steps . The first step the included the reaction of N-( sub. or un sub. benzoyl and sub. or un sub. acetyl ) amidyl sub. 2,6- diamino -4-methyl-1,3,5-triazine (1-5) by condensation of many substituted acid chlorides with 2,6- diamino -4-methyl-1,3,5-triazine . While the second step included the reaction of polyacryloyl chloride with the produced compounds (1-5) in step (1) in the presence amount triethyl amine (Et3N) to obtain new polyimides (6-10). The prepared compounds were characterized by UV. , FT-IR, and some of them by 1H-NMR and 13C- NMR spectroscopy.
In the present study, a low cost adsorbent is developed from the naturally available sawdust
which is biodegradable. The removal capacity of chromium(VI) from the synthetically prepared
industrial effluent of electroplating and tannery industrial is obtained.
Two modes of operation are used, batch mode and fixed bed mode. In batch experiment the
effect of Sawdust dose (4- 24g/L) with constant initial chromium(VI) concentration of 50 mg/L and
constant particle size less than1.8 mm were studied.
Batch kinetics experiments showed that the adsorption rate of chromium(VI) ion by Sawdust
was rapid and reached equilibrium within 120 min. The three models (Freundlich, Langmuir and
Freundlich-Langmuir) were fitted to exper
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Introduction. Intraoperative rupture (IOR) of an aneurysm is a frightful complication that causes significant morbidity and mortality worldwide. IOR can be attributed to various parameters, including hypertension, increased intracranial pressure (ICP), fragility of the vessels, and inadequate anaesthesia. IOR due to insufficient anaesthesia is scarcely reported in the literature. Here, we describe a re-ruptured anterior communicating artery (ACoA) after incomplete clipping of the neck during craniotomy closure due to unintended early wake-up from anaesthesia with a discussion about the management. Case description. A 38-year-old male suddenly developed a severe headache, a brief loss of consciousness, and vomiting. Computed tomogr
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This study was aimed to evaluate atotal phenolic content, antibacterial activity, and antioxidant activity of M. communis callus extracts were evaluated. Callus induction in general Murashige and Skoog (MS) media is completed by the Benzil adenine's unique knowledge of callus formation. A well diffusion experiment was used to examine antibacterial interest in Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. The DPPH radical scavenging activity test was used to measure antioxidant activity. FTIR and HPLC have been used to pinpoint the presence of polyphenol compounds in calluses. The total phenol content of plant leaves extract (0.1, 0.5, and 1) mg/ml was 42.12, 94.08, and 189 mg of Gallic ac
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In the present work advanced oxidation process, photo-Fenton (UV/H2O2/Fe+2) system, for the treatment of wastewater contaminated with oil was investigated. The reaction was influenced by the input concentration of hydrogen peroxide H2O2, the initial amount of the iron catalyst Fe+2, pH, temperature and the concentration of oil in the wastewater. The removal efficiency for the system UV/ H2O2/Fe+2 at the optimal conditions and dosage (H2O2 = 400mg/L, Fe+2 = 40mg/L, pH=3, temperature =30o C) for 1000mg/L load was found to be 72%.
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