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.
Many diseases can produce cardiac overload, of these disease hypertension, valve disease congenital anomaly in addition to many other disease. One of the most common diseases causing left ventricle overload is hypertension. A long term hypertension can cause myocardium hypertrophy leading to changes in the cardiac contractility and reduced efficiency. The investigations were carried out using conventional echocardiography techniques in addition to the tissue Doppler imaging (TDI) from which many noninvasive measurements can be readily obtained. The study has involved the effect of hypertension on the myocardium stiffness index through the measurement of early diastolic filling (E) and the early velocity of lateral mitral annulus (E
... Show MoreBacteria could produce bacterial nanocellulose through a procedure steps: polymerization and crystallization, that occur in the cytoplasm of the bacteria, the residues of glucose polymerize to (β-1,4) lineal glucan chains that produced from bacterial cell extracellularly, these lineal glucan are converted to microfbrils, after that these microfbrils collected together to shape very pure three dimensional pored net. It could be obtained a pure cellulose that created by some M.O, from the one of the active producer organism like Acetic acid bacteria (AAB), that it is a gram -ve, motile and live in aerobic condition. The bacterial nanocellulose (BNC) have great consideration in many fields because of its flexible properties, features
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This investigation deals with the use of orange peel (OP) waste as adsorbent for removal of nitrate (NO3) from simulated wastewater. Orange peel prepared in two conditions dried at 60C° (OPD) and burning at 500 °C (OPB). The effect of pH: 2-10, contact time: 30- 180 min, sorbent weight: 0.5- 3.0 g were considered. The optimal pH value for NO3 adsorption was found to be 2.0 for both adsorbents. The equilibrium data were analyzed using Langmuir and Freundlich isotherm models. Freundlich model was found to fit the equilibrium data very well with high-correlation coefficient (R2). The adsorption kinetics was found to follow pseudo-second-order rate kinetic model, with a good correlation (R2
... Show MoreThe main objectives of this research is to extract essential oil from: orange ( citrus sinensis), lemon( citrus limon) and mandarin( citrus reticulata) peels by two methods: steam distillation (SD) and microwave assisted steam distillation (MASD), study the effect of extraction conditions (weight of the sample, extraction time, and microwave power, citrus peel type) on oil yield and compare the results of the two methods, the resulting essential oil was analyzed by Gas Chromatography (GC).
Essential oils are highly concentrated substances used for their flavor and therapeutic or odoriferous properties, in a wide selection of products such as foods, medicines and cosmetics. Extracti
... Show MoreThe effect of crude colicin extracted from E. coli isolated from
urinary tract infection patients on phagocytosis in vitro was studied . Results showed that the effect of crude colicin on phagocytic cells and their activities were concentration dependent. Low concentration (50
Mg/ml) have no significant effect (p>O.OS) on shape, size, migration and engulfment activity of phagocytic cells, while (50, 100 Mg/ml)
enhanced killing activity and increase superoxide ( 02 ) production as indicated by (NBT) test. But high concentration (500 mg/ml) of crude colicin caused inhibition of phagocytic cel
... Show MoreSome mechanical and thermal properties of mullite samples prepared by mixing different phases of alumina and silica powders have been studied according to ASTM methods the cold crushing strength of the sintcred bodies.With different porosity, at room temperature was in the range(18-54)Mpa
Pharmaceutical-instigated pollution is a major concern, especially in relation to aquatic environments and drugs such as meropenem antibiotics. Adsorbents, such as multi-walled carbon nanotubes, offer potential as means of removing polluting meropenem antibiotics and other similar compounds from water. In order to evaluate the effectiveness of multi-walled carbon nanotubes in this capacity, various experimental parameters, including contact time, initial concentration, pH, temperature and the dose of adsorbent have been investigated. The Langmuir and the Freundlich isotherm models have been used. The data obtained using a modified Langmuir model have been consistent with the experimental ones; the best pH value has been obtained to have the
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In this study, chalcones were synthesis by condensing 2-acetylpyridine with aromatic aldehyde derivatives in dilute ethanolic potassium hydroxide solution at room temperature according to Claisen-Schmidt condensation. After that, new heterocyclic derivatives such as Oxazine, Thiazine and Pyrazol were synthesis by reaction between chalcones with urea, thiourea and hydrazine hydrate respectively scheme 1. All these compounds wrer characterization by FTIR, 1H-NMR spectroscopy and elemental analysis.