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.
Recently, important efforts have been made in an attempt to search for the cheapest and ecofriendly alternatives adsorbents. In the present work, waste molasses from Iraqi date palm (Zahdi) had been used as a provenance to produce charcoal for the removal of methylene blue (MB) dye from water. The optimum prepared charcoal was obtained at 150 C, by increasing temperature to 175 C, the charcoal had almost converted to ash. The obtained charcoal have been inspected for properties using scanning electron microscope (SEM), atomic force microscope (AFM), porosity and surface area. Adsorption data were optimized to Langmuir and Freundlich and adsorption parameters have been evaluated. The thermodynamic parameters like a change
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This work was conducted to study the extraction of eucalyptus oil from natural plants (Eucalyptus camaldulensis leaves) using water distillation method by Clevenger apparatus. The effects of main operating parameters were studied: time to reach equilibrium, temperature (70 to100°C), solvent to solid ratio (4:1 to 8:1 (v/w)), agitation speed (0 to 900 rpm), and particle size (0.5 to 2.5 cm) of the fresh leaves, to find the best processing conditions for achieving maximum oil yield. The results showed that the agitation speed of 900 rpm, temperature 100° C, with solvent to solid ratio 5:1 (v/w) of particle size 0.5 cm for 160 minute give the highest percentage of oil (46.25 wt.%). The extracted oil was examined by HPLC.
A total of 437 individuals of Turbellarin Platyhelminthes were sorted from benthic samples collected monthly for a period of seven months( April to November 2013 ) from AL-Dalmage lake, a part of middle section for main outfall drain south of Baghdad. They were identified as Gyratrix hermaphroditus, Stenostomum leucops ,Stenostomum unicolar and Stenostomum bryophilum ,The relative abundance of worms decreased during hot season which (May to September) ,where they start rising again. The species were studied alive , the identification criteria were illustrated by photos. G. hermaphroditus was the most abundant species among the four species.
Siderophores are low molecular weight organic compounds produced by microorganisms growing under low iron concentration.In this study we describe the detection, production and extraction of siderophores secreted by Acinetobacter baumannii (Multiple-drug resistant ) pathogens. One hundered twenty Gram –negative non lactose fermenter bacilli isolates have been collected from three hospitals at Baghdad city over three months. Primary identification of these isolates is performed by standard diagnostic methods (biochemical tests and API 20 NE); 19 clinical isolates of A. baumannii are cultured on CHROMagar (highly selective medium for detection of MDR Acinetobacter) as well as diagnoses is documented by using Vitek 2 system. Isolates are exa
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This work was conducted to study the recovery of catalyst and desirable components from tar formed in phenol production unit and more particularly relates to such a method whereby better recovery of copper salts, phenol, benzoic acid and benzoate salts from tar by aqueous acid solution was accomplished.
The effect of solvent type, solvent concentration (5, 10, 15, 20, 25 and 30 wt%), agitation speed (100, 200, 300 and 400 rpm), agitation time (5, 10, 15, 20 and 25 min), temperature (90, 100, 110, 120, 130 and 140 oC) , phase ratio (1/1, 2/1, 3/1, 4/1 and 5/1) and number of extraction (1, 2, 3, 4, and 5) were examined in order to increase the catalyst and desirable components extraction.
Four types of solvent were used; hydrochloric
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Hazardous materials, heavy metals, and organic toxins released into the environment have caused considerable harm to microbes, plants, animals, and humans. Wastewater is one of the most contaminated ecosystems due to heavy metals emitted mostly by human activity. Bioremediation of wastewater is an ecologically acceptable and cost-effective method of removing heavy metals from sewage; the general purpose of this study is to analyse the dependability of anaerobic sludge biomass in removing sulfur compounds and heavy metals from waste water. The anaerobic sludge biomass evaluated in this work was taken from a wastewater treatment plant (WWTP) in Al-Rustumiya, Baghdad, and grown in the mineral medium for anaerobic growth. In serum bottl
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A mixture of algae biomass (Chrysophyta, Cyanophyta, and Chlorophyte) has been investigated for its possible adsorption removal of cationic dyes (methylene blue, MB). Effect of pH (1-8), biosorbent dosage (0.2-2 g/100ml), agitated speed (100-300), particle size (1304-89μm), temperature (20-40˚C), initial dye concentration (20-300 mg/L), and sorption–desorption were investigated to assess the algal-dye sorption mechanism. Different pre-treatments, alkali, protonation, and CaCl2 have been experienced in order to enhance the adsorption capacity as well as the stability of the algal biomass. Equilibrium isotherm data were analyzed using Langmuir, Freundlich, and Temkin models. The maximum dye-sorption capacity was 26.65 mg/g at pH= 5, 25
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