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.
Abstract This research investigates how activated carbon (AC) was synthesized from potato peel waste (PPW). Different ACs were synthesized under the atmosphere's conditions during carbonation via two activation methods: first, chemical activation, and second, carbon dioxide-physical activation. The influence of the drying period on the preparation of the precursor and the methods of activation were investigated. The specific surface area and pore volume of the activated carbon were estimated using the Brunauer–Emmett–Teller method. The AC produced using physical activation had a surface area as high as 1210 m2/g with a pore volume of 0.37 cm3/g, whereas the chemical activation had a surface area of 1210 m2/g with a pore volume of 0.34 c
... Show More
Schiff bases were prepared prepared Baaan NMR to some elements of which have contributed to the results of different methods in diagnosis prove structural formulas of compounds prepared
In this study, the adsorption of Zn (NO3)2 is carried out by using surfaces of malvaparviflora. The validity of the adsorption is evaluated by using atomic absorption Spectrophotometry through determination the amount of adsorbed Zn (NO3)2. Various parameters such as PH, adsorbent weight and contact time are studied in terms of their effect on the reaction progress. Furthermore, Lagergren’s equation is used to determine adsorption kinetics. It is observed that high removal of Zn (NO3)2 is obtained at PH=2. High removal of Zn (NO3)2 is at the time equivalent of 60 min and reaches equilibrium,where 0.25gm is the best weight of adsorbant . For kinetics the reaction onto malvaparviflora follows pseudo first order Lagergren’s equation.
(1)
In the present study waste aluminium cans were recycled and converted to produce alumina catalyst. These cans contain more than 98% aluminum oxide in their structure and were successfully synthesized to produce nano sized gamma alumina under mild conditions. A comprehensive study was carried out in order to examine the effect of several important parameters on maximum yield of alumina that can be produced. These parameters were reactants mole ratios (1.5, 1.5, 2, 3, 4 and 5), sodium hydroxide concentrations (10, 20, 30, 40, 50 and 55%) and weights of aluminum cans (2, 4, 6, 8 and 10 g). The compositions of alumina solution were determined by Atomic absorption spectroscopy (AAS); and maximum yield of alumina solution was 96.3% obtain
... Show MoreAdsorption experiments were carried out using two different low-cost sorbent materials, date seeds and olive seeds. These sorbents used as a single phase (not as mixture) to remove cadmium ions from simulated wastewater by adsorption process. The equilibrium time was found at 2 hr. The experiments include different parameters such sorbent type and weight and contact time. It was found that both of olive seed and date seed have approximately the same adsorption capacity (qm) with 15.644 mg/g and 15.2112 mg/g, respectively. Equilibrium isotherms and kinetic studies have been carried out. Langmuir isotherm model better fits the experimental data compared with the Freundlich isotherm for olive seed, while Freundlich isotherm fits for date se
... Show MoreThe aim of the research is to identify the methods of achieving mental health from an Islamic perspective by using an analytical approach. The methods that were explained: strengthen the spiritual side to control the motives and emotion s overcome the whims of the soul, fear of God and treat mental illness from an Islamic point of view by recognizing self, assurance, foresight, learning and acquiring new trends. Conclusion: we can achieve mental health by optimism, not despair, the compatibility of the Muslim with himself and with others, Consistency, emotional balance, and patience in difficult situations. Islam has attributes that make an individual feels psychological security. Thus, all these elements achiev
... Show MoreReaction of,2- [( 4- amio phenyl ) diazenyl] 1,3,4- thiadiazole -5- thiol (S1) with p- chlorobenzeldehyde,3,4 – dimethoxy benzaldehyde and pyrrol-2- carbonxaldehyde gave -5- [{4-(4-chlorobenzylidene amino) phenyl} diezenyl]-1,3,4- thiadiazole-2- thiol (S2),5-[{ 4-[(3,4- dimethoxybenzyldene )amino phenyl ] diazenyl)-1,3,4- thiadiazole-2-thiol,(S3) and -5- [4-(1,H – pyrrol -2- yl- methylene)amino phenyl] diazenyl)-1,3,4- thiadiazole-2- thiol (S4) respectively as schiff's bases compounds. On the same route-2-[(4-amino-1- naphthyl ) diazenyl] -1,3,4- thiadiazole -5- thiol (S5) reacts with –p- chloro benzaldehyde and –m- nitrobenzaldehyde to give the follwing schiff's bases -5-[{ 4-(4- chloro benzylidene ) amino -1- naphthyl} diazenyl]
... Show More
(1)
In this study, the circulating fluidized bed was used to remove the Tetracycline from wastewater utilizing a pistachio shell coated with ZnO nanoparticles. Several parameters including, Tetracycline solution flowrate, initial static bed height, Tetracycline initial concentration and airflow rate were systematically examined to show their effect on the breakthrough curve and the required time to reach the adsorption capacity and thus draw the fully saturated curve of the adsorbent. Results showed that using ZnO nanoparticles will increase the adsorbent surface area and pores and as a result the adsorption increased, also the required time for adsorbent saturation increased and thus the removal efficiency may be achieved at mi
... Show More
(21)
Animal fats are a good, promising and ethical alternative source for biodiesel production, but they need more complex treatments than vegetable oils. Iraqi butchery plants waste fats (sheep fat) which are suggested as feedstock to produce biodiesel. This type of fat contains a large quantity of free fatty acids (FFAs) (acid number 49.13 mg KOH/g of fat). The direct transesterification of such fats produce high amount of soap instead of desired biodiesel, so a pre-treatment step (to reduce FFAs) is necessary before transesterification. This step was done by esterification of the free fatty acids in the fat by adding ethanol and using 1% acid catalyst (H2SO4) for 30 minutes. The results showed that the acid number of sheep fat after pre-tr
... Show More