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.
Some physical properties enthalpy (?H), entropy (?s), free energy (?G),capacities(?cp?) and Pka values) for valine in dimethyl foramideover the temperature range 293.15-318.15K, were determined by direct conductance measurements. The acid dissociation at six temperature was examined at solvent composition x2) involving 0.141 of dimethyl foramide . As results, calculated values have been used to determine the dissociation constant and the associated thermodynamic function for the valine in the solvent mixture over temperatures in the range 293.15-318.15 k. The Pka1, and Pka2 were increased with increasing temperature.
In this work Aquatic plant (Nile rose) was used to study adsorption of industrial dye (safranin-O from aqueous solution within several operation conditions. The dried leaves of Nile rose plant were used as adsorbents safranin-O from aqueous solution after different activations such as wet and dry enhancements. The data show increasing in dye solution removal percentage for both activation methods of the adsorbent and also dye removal percentage that was obtained by using adsorbent without any treatment with the progress contact time. The dye removal percentages at equilibrium time 40 minutes were 88.7% at non-activation, 92.3% at thermal activation, and 98.3% at acidic activation. The samples adsorbents before and after adsorption which wer
... Show More
(10)
(4)
Language as a means of communication has long been the concern of many conversation analysts in their studies such as: Sacks et al. (1974), Schegloff et al. (1977), Duncan (1972), Grice (1975) and Burton (1980). Burton has attempted analyzing the first ten transitions of the play “The Dumb Waiter” for mere a presentation of her approach. This paper aims at analyzing the conversational structure of forum on the subject of literary fiction and genre fiction by applying Burton’s model (1980) of analysis to answer the question to what extent this model is applicable in analyzing the presented text. The findings of the investigation have proved the applicability of the structure of conversation formulated by Burton (1980) in her model wit
... Show MoreThe development of economic and environmentally friendly extractants to recover cobalt metal is required due to the increasing demand for this metal. In this study, solvent extraction of Co(II) from aqueous solution using a mixture of N,N0-carbonyl difatty amides (CDFAs) synthesised from palm oil as the extractant was carried out. The effects of various parameters such as acid, contact time, extractant concentration, metal ion concentration and stripping agent and the separation of Co(II) from other metal ions such as Fe(II), Ni(II), Zn(III) and Cd(II) were investigated. It was found that the extraction of Co(II) into the organic phase involved the formation of 1:1 complexes. Co(II) was successfully separated from commonly associated metal
... Show More
(27)
(14)
In this study, the acid-alkaline transesterification of refined coconut seed oil (RCOSO) to fatty acid methyl ester was followed by the production of a trimethylolpropane-based thermosensitive biolubricant using potassium hydroxide, and its physicochemical characteristics were evaluated. The American Standard Test for Materials (ASTM) was employed to ascertain the biolubricant's pour point and index of viscosity, which were found to be -4 oC and 283.75, respectively. The opposite connection between lubricant viscosity and temperature was shown by the measured viscosities at varied transesterification to be transformed into biodiesel. Following this, a biolubricant was created by further transesterifiedtemperature. The ester gr
... Show More
(1)
One of the most important problems facing the world today is the energy problem. The solution was in finding renewable energy sources such as solar energy. The solar energy applications in Iraq is facing many problems . One of the most important problems is the accumulation of dust on the solar panels surface which causes decreasing its performance sharply. In the present work, a new technique was presented by using two-axis solar tracking system to reduce the accumulated dust on the solar panel surface and compared it with the fixed solar panels which installed at tilt angles 30° and 45°. The results indicated that the maximum losses of the output power due to accumulation of dust on the fixed solar panels is about 31.4% and 23.1% res
... Show MorePoly urea formaldehyde –Bentonite (PUF-Bentonite) composite was tested as new adsorbent
for removal of mefenamic acid (MA) from simulated wastewater in batch adsorption
procedure. Developed a method for preparing poly urea formaldehyde gel in basic media by
using condensation polymerization. Adsorption experiments were carried out as a function of
water pH, temperature, contact time, adsorbent dose and initial MA concentration .Effect of
sharing surface with other analgesic pharmaceuticals at different pH also studied. The
adsorption of MA was found to be strongly dependent to pH. The Freundlich isotherm model
showed a good fit to the equilibrium adsorption data. From Dubinin–Radushkevich model the
mean free
Liquid-liquid membrane extraction technique, pertraction, using three types of solvents (methyl isobutyl ketone, n-butyl acetate, and n-amyl acetate) was used for recovery of penicillin V from simulated fermentation broth under various operating conditions of pH value (4-6) for feed and (6-8) for receiver phase, time (0-40 min), and agitation speed (300-500 rpm) in a batch laboratory unit system. The optimum conditions for extraction were at pH of 4 for feed, and 8 for receiver phase, rotation speed of 500 rpm, time of 40 min, and solvent of MIBK as membrane, where more than 98% of penicillin was extracted.
Leaching process applied for the extraction of bio active compounds from dried roots of (Elecampane) Inula helenium. Ethanol, hexane and distillated water were used as solvents. Roots were soaked with ethanol (5% w/v) with various concentration of ethanol (30 to 98%) at one day to know effect concentration of the solvent with concentration of bio active compound in Inula helenium. The same procedure was done using hexane as solvent. Also distilled water was used as solvent for extraction 5%(w/v) where plant material was soaked in water at different temperatures (25, 40, 65, 80, and 90) C. In all solvents undertaken, the effect of time duration on active ingredient (Thymol, Isoalatolactone, Alatolactone, 10-isobutyryl-oxy 8-9-epoxy thymol is
... Show More
(3)