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.
The genus Larra Fabricius, 1793 (Hymenoptera: Crabronidae) is recorded for the first time from Vietnam. Three species and two subspecies belonging to this genus as follows: L. amplipennis (F. Smith, 1873); L. carbonaria (F. Smith, 1858); L. fenchihuensis Tsuneki, 1967; L. polita polita (F. Smith, 1858) and L. polita luzonensis Rohwer, 1919 are presented. Keys to both sexes of the three species and two subspecies reported here are provided.
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In this paper flotation method experiments were performed to investigate the removal of lead and zinc. Various parameters such as pH, air flow rate, collector concentrations, collector type and initial metal concentrations were tested in a bubble column of 6 cm inside diameter. High recoveries of the two metals have been obtained by applying the foam flotation process, and at relatively short time 45 minutes . The results show that the best removal of lead about 95% was achieved at pH value of 8 and the best removal of zinc about 93% was achieved
at pH value of 10 by using 100 mg/l of Sodium dodecylsulfate (SDS) as a collector and 1% ethanol as a frother. The results show that the removal efficiency increased with increasing initial m
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Mutans streptococci (MS) are a group of oral bacteria considered as the main cariogenic organisms. MS consists of several species of genus Streptococcus which are sharing similar phenotypes and genotypes. The aim of this study is to determine the genetic diversity of the core species of clinical strains of Streptococcus mutans, Streptococcus sobrinus and Streptococcus downei by using repitative extragenic palindromic (REP) primer. The DNA of the clinical strains of S. mutans (n=10), S. sobrinus (n=05) and S. downei (n=04) have been employed in the present study, which have been previously isolated from caries active subjects. The DNA of the clinical and reference strains was
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Lactiplantibacillus plantarum, one of lactic acid bacteria (LAB), is found in various foods, including dairy products, meat, and vegetables, and most of these bacteria offer beneficial effects to humans and animals as potential probiotics with broad-spectrum antimicrobial activities. The aim of this study was evaluating the antibacterial efficacy of L. plantarum against some foodborne bacteria isolated from dairy products. This research involved 34 dairy products, including local and imported milk, cheese, and yogurt sold locally in Baghdad province, Iraq, during May 2022. For the isolation of L. plantarum, a special medium called MRS (de Man Rogosa and Sharpe) was applied. Colonie
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Due to the deliberate disposal of industrial waste, a great amount of petroleum hydrocarbons pollute the soil and aquatic environments. Bioremediation that depends on the microorganisms in the removal of pollutants is more efficient and cost-effective technology. In this study, five rhizobacteria were isolated from Phragmites australis roots and exposed to real wastewater from Al-Daura refinery with 70 mg/L total petroleum hydrocarbons (TPH) concentration. The five selected rhizobacteria were examined in a biodegradation test for seven days to remove TPH. The results showed that 80% TPH degradation as the maximum value by Sphingomonas Paucimobilis as identified with Vitek® 2 Compact (France).
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The Catharanthus roseus plant was extracted and converted to nanoparticles in this work. The Soxhlet method was used to extract alkaloid compounds from the Catharanthus roseus plant and converted them to the nanoscale. Chitosan polymer was used as a linking material and converted to Chitosan nanoparticles (CSNPs). The extracted alkaloids were linked with Chitosan nanoparticles by maleic anhydride to get the final product (CSNPs-Linker-alkaloids). The pure Chitosan, Chitosan nanoparticles, and CSNPs-Linker-alkaloids were characterized by X-ray diffractometer, and Fourier Transform Infrared spectroscopy. X-ray results show that all samples have an orthorhombic structure with crystallite size in nanodimensions. FTIR spectra prove that
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