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.
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Prodigiosin is a ‘natural red pigment produced by Serratia marcescens which exhibits immunosuppressive and anticancer properties in addition to antimicrobial activities. This work presents an attempt to maximize the production of prodigiosin by two different strategies: one factor at time (OFAT) and statistical optimization. The result of OFAT revealed that sucrose and peptone were the best carbon and nitrogen sources for pigment production with concentration of prodigiosin of about 135 mg/ L. This value was increased to 331.6mg/ L with an optimized ratio of C/N (60:40) and reached 356.8 with pH 6 and 2% inoculum size at end of classical optimization. Statistical experimental design based on Response surface methodology was co
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?Natural Occurrence of aflatoxin Bi and B2 was investigated monthly in paddy rice stored for duration of six months. Aflatoxin B| was detected at the beginning of storage. The concentration of aflatoxin Bi in paddy and polished rice was found to be (0.3 and ()) ppb respectively while aflatoxin B2 W'as not detected in both paddy and polished rice afler six months of storage, concentration of aflatoxin Bi ‘ ’ ' reached 3 ppb in paddy rice and 0.9 ppb in polished rice, while the study proved that the concentration of aflatoxin in paddy rice was higher than polished rice as a result of processing which reduce the toxin
Abstract
This study aimed to survey fungi associated with the product Indomie and Chips being the trades Iargely by a very important segment of society who are the children, beside consumed by adults, but less so, as the survey results to accompany some fungui samples sterile showed proportions presence included various fungi like. Aspergillus flavus, Aspergillus niger, Penicillium Spp., Fusarium graminearum, F.moniliforme, Alternaria alternate and Rhizopus Spp., and other fungi sterile are not diagnosed. The results showed large dominion fungi A. niger by presence sterile samples of both producers, followed by infection in Fusarium Spp., Penicillium Spp., and A. alternata by infection percentage 55, 20 and 17% respectively for the pr
Gestational diabetes mellitus (GDM) is a complication of gestation that is characterized by impaired glucose tolerance with first recognition during gestation. It develops when ?- cell of pancreas fail to compensate the diminished insulin sensitivity during gestation. This study aims to investigate the relationship between mother adiponectin level and ?- cell dysfunction with development gestational diabetes mellitus (GDM) and other parameters in the last trimester of pregnancy. This study includes (80) subjects ( pregnant women) in the third trimester of pregnancy, (40) healthy pregnant individuals as control group aged between (17 - 42) years and (40) gestational diabetes mellitus patients with aged between (20 - 42) years. The f
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In this research, carbon nanotubes (CNTs) is prepared through the Hummers method with a slight change in some of the work steps, thus, a new method has been created for preparing carbon nanotubes which is similar to the original Hummers method that is used to prepare graphene oxide. Then, the suspension carbon nanotubes is transferred to a simple electrode position platform consisting of two electrodes and the cell body for the coating and reduction of the carbon nanotubes on ITO glass which represents the cathode electrode while platinum represents the anode electrode. The deposited layer of carbon nanotubes is examined through the scanning electron microscope technique (SEM), and the images throughout the research show the
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The angiotensin converting enzyme (ACE) I\D gene polymorphism influences the blood ACE enzyme activity. Renoprotective effect of ACE inhibitors (ACEIs) varies among patients due to genetic variation, particularly in Renin-Angiotensin-Aldosterone System genes. This study investigates the genetic variations of ACE I\D and AGT1RA1166C gene polymorphisms in the antiproteinuric effect of ACEI therapy in type 2 diabetes mellitus (T2DM) patients. This is a cross-sectional study that included 76 T2DM patients who are ACEI users, divided into two groups: T2DM without diabetic kidney disease (DKD) included 31 patients, and T2DM with DKD included 45 patients. Urine samples were taken for measurement of urine albumin and creatinine, then calcul
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Previous studies indicated that supplementation with antioxidants has a protective effects against oxidative stress–induced damage in type 2 diabetes. In this study we evaluated the antioxidant effects of melatonin on the oxidative stress parameters and microalbuminuria in type 2 DM patients. 30 patients with type 2 DM were treated with 3mg/day melatonin for 90 days. Erythrocytes and plasma MDA and glutathione, fasting plasma glucose, %HbAIC, microalbuminuria, total plasma protein and lipid profile were measured each 30 days and compared with those obtained from 20 healthy controls.
A decrease in MDA levels associated with the elevation in GSH levels were observed, compared with the pre–treatment levels.
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The current work was designed to investigate serum angiopoietin like protein-8 and hyaluronic acid among Iraqi hemodialysis patients with and without type 2 diabetes mellitus, and to find relationship between them, as well as if these patients are at risk of kidney fibrosis. Subjects & Methods: in this study, serum samples were obtained from (60) Iraqis patients with end stage renal diseases (ESRD)on hemodialysis (HD) (30 patients with T2DM (G2) and 30 patients withoutT2DM (G3)) in addition to (30) healthy individuals as a control group (G1), their ages ranged from (35-65) years. The patients attended the Al-Yarmouk Teaching Hospital, Baghdad. Results: the results in this study showed a highly a significant elevation inserum angiopoietin li
... Show MoreBackground: Rheumatoid arthritis is a chronic destructive inflammatory disease associated with destruction of joint connective tissues and bones, affecting 0.5%–1% of the population worldwide reporting higher prevalence of periodontitis among rheumatoid arthritis patients. The purpose of this study is to estimate level of salivary C-reactive protein in relation to the occurrence and severity of the periodontal disease and other oral parameters among group of patients with rheumatoid arthritis Material and methods: Fifty women patients with rheumatoid arthritis; twenty five on Methotrexate treatment and twenty five on combination treatment of Methotrexate and Etanercept selected as study groups with an age range (30-40) years old and
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