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.
Due to wind wave actions, ships impacts, high-speed vehicles and others resources of loading, structures such as high buildings rise bridge and electric transmission towers undergo significant coupled moment loads. In this study, the effect of increasing the value of coupled moment and increasing the rigidity of raft footing on the horizontal deflection by using 3-D finite element using ABAQUS program. The results showed that the increasing the coupled moment value leads to an increase in lateral deflection and increase in the rotational angle (α◦). The rotational angle increases from (0.014, 0.15 to 0.19) at coupled moment (120 kN.m), (0.29, 0.31 and 0.49) at coupled moment (240 kN.m) and (0.57, 0.63 and 1.03) at cou
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In this work, we studied the effect of power variation on inductively coupled plasma parameters using numerical simulation. Different values were used for input power (750 W-1500 W), gas temperature 300K, gas pressure (0.02torr), 5 tourns of the copper coil and the plasma was produced at radio frequency (RF) 13.56 MHZ on the coil above the quartz chamber. For the previous purpose, a computer simulation in two dimensions axisymmetric, based on finite element method, was implemented for argon plasma. Based on the results we were able to obtain plasma with a higher density, which was represented by obtaining the plasma parameters (electron density, electric potential, total power, number density of argon ions, el
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This study was conducted to evaluate the efficacy of Saccharomyces cerevesiae as a growth promoting agent in tomato. Soaking the seeds in yeast suspension at 5 g/L for 12h increased germination percentage, root length, root fresh and dry weight, plant height, foliage fresh and dry weight, attained 88.5% ; 8.1 cm ; 84.3 mg ; 7.03 mg ; 10.75 cm ; 839 mg and 37.75 mg compared with 80% ; 5.33 cm ; 39 mg ; 4.8 mg ; 7.35 cm ; 608 mg and 25.5 mg in seedlings grown from non treated seeds respectively. Similar results were obtained with seedling from seeds soaked in S. cerevesiae filtrate for 12 hrs. with values of 77.5% ; 6.875 cm ; 91.5 mg ; 7.5 mg ; 9.5 cm ; 777 mg and 40.35 mg compared to 66% ; 5.8 cm ; 57.7 mg ; 5.03 mg ; 5.9 cm ; 493 mg
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Abstract. The main technique for removing bacteria from water for various applications is chemical disinfection. However, this method has many disadvantages such as producing disinfectant by-products (DBPs), biofilm formation and either rendering the water unpotable (at high residual disinfection) or leaving a potential for lethal diseases such as Cholera (if the residual disinfection is too low). Recently, a process was developed for continuous removal of bacteria from water using the principle of froth flotation through compressed air only without any chemicals (Hassan, 2015). This work examines the extent to which chemical free froth flotation can purify drinking water. The experiments were carried out using two flotation columns
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Nowadays, there is increased interest in the biosynthesis of microbial melanin related to their numerous biological functions and applications in many fields, especially in medical fields, including immune-modulating, antimicrobial antibiotic, antiviral antivenin, anticancer, antitumor activity, and anti-biofilm activity. Pyomelanin is a hydrophobic macromolecule that is typically dark brown or black in color, formed by the oxidative polymerization of phenolic or indolic compounds. Pyomelanin is reported to be safe for consumption, thus providing a crucial strategy for biocontrol of biofilm. Furthermore, natural pyomelanin is known as a potent antioxidant, photoprotective, and free radical scavenging. Objective: This study focuses on the
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The removal of commercial orange G dye from its aqueous solution by adsorption on tobacco leaves (TL) was studied in respect to different factor that affected the adsorption process. These factors including the tobacco leaves does, period of orange G adsorption, pH, and initial orange G dye concentration .Different types of isotherm models were used to describe the orange G dye adsorption onto the tobacco leaves. The experimental results were compared using Langmuir, and frundlich adsorption isotherm, the constants for these two isotherm models was determined. The results fitted frundlich model with value of correlation coefficient equal to (0.981). The capacity of adsorption for the orange G dye was carried out using various kinetic models
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In this part of programme , different bacterial isolates mainly Salmonella spp, Shigella spp and Escherichia coli were used for antagonism with Saccharomyces boulardii under different conditions . S.boulardii was grown under aerobic conditions and antagonized with young overnight nutrient broth cultures of test bacterial isolates and other kept in refrigerator for a week after full growth . Young cultures were more susceptible to antagonistic effect of yeast compared to old cultures and on isolates grown on solid medium for 24 hr. S.boulardii grown under aerobic and microaerobic conditions and antagonized with overnight broth cultures of test bacterial isolates , The results revealed that aerobic cultures of yeast had more inhibito
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Halobacterium saccharovorum was isolated from local highsalinity souls named Al-Massab Al-Aam. A growth curve was determined. The average generation time during logarith- mic phase was 17.80±0.62 hr. Bacteriorhodopsin was 1808 lated from the purple membrane, its concentration was 4.8 mg/ml and H.W was 26000. The pattern of other membrane Bpoteins was studied and compared with those of other Boletes. Several unique proteins were isolated and their molecular weights were determined.