This research was aimed to the purification and characterization of cytosine deaminase as a medically important enzyme from locally isolated Escherichia coli; then studying its cytotoxic anticancer effects against colon cancer cell line. Cytosine deaminase was subjected to three purification steps including precipitation with 90% ammonium sulfate saturation, ion exchange chromatography on DEAE-cellulose column, and gel filtration chromatography throughout Sephadex G-200 column. Specific activity of the purified enzyme was increased up to 9 U/mg with 12.85 folds of purification and 30.85% enzyme recovery. Characterization study of purified enzyme revealed that the molecular weight of cytosine deaminase produced by E. coli was about 48 KDa, the highest enzyme activity at pH 8.5, and is most stable at pH 7.5 - 9, the enzyme also showed a full activity at a range of temperatures between 45-60 0C. Enzyme activity was strongly inhibited in the presence of mercuric chloride and copper sulphate, when added individually at a constant concentration. However, calcium chloride, manganese chloride and ferric chloride caused a little increase in enzyme activity while sodium azide had no effect on enzyme activity. Upon cytotoxic effect study through micro-cultured tetrazolium assay (MTT) against Caco-2 cell line. Purified cytosine deaminase was found to inhibit the growth of Caco-2 cancer cell line with an IC50 of 242.5 ?g/ml in a comparison to an IC50 of 1864 ?g/ml for crude enzyme. Besides, the enzyme didn’t show significant effect on WRL normal cell line.
Four hundred and fifty urine samples were collected from patients suffering from urinary tract infection from the General Azadi hospital in Kurkuk province ,during the period of october 2007 till march 2008 . Results of bacteriological culture revealed that (168) out of (450) studied samples (37.3%) gave positive culture using blood agar and macConkey agar ; different species of bacterial isolates were detected using morphological and biochemical tests ,from these isolates the highest percentage of the isolates were from Escherichia coli when it was (100) isolates out of (190) isolate (52.63%) . one hundred isolates were distributed between (77) from females and (23) from
... Show MoreA total of 165 clinical sample included Urine, Swab wounds and Burns were collected from Baghdad Governorate. Results showed that rate all isolates of E. coli was 50(30.3%) and rate of urine infection was 46(92%) and rate of swab wounds infection 4(8%). Where was diagnostic based on streaked on MacConkey agar, then single colony was transferred to Eosin Methylene Blue (EMB). Identification some of the biochemical test included: Catalase test, Oxidase test, Indole test, Methyl red, Vogues - Proskauer test and Citrate Utilization test. Then confirmed by the Vitek - 2 Compact System. The ability of E.coli isolate to biofilm formation to be studied it is considered one of the most important factors of virulence and has role in causing injury an
... Show MoreThis study was conducted to investigate activity and some kinetic
and thermodynamic parameters of adenosine deaminnse (ADA)in
serum in patient affected with P-thalassaemia and compared with that of healthy individuals .Serum AD levels were found to be significantly higher in patient with P-thalassaemia (98±9.15 IU/L) than in healthy individuals (22± 2.04 IUIL)the study was carried in optimum pH value 6.5 and 37c by which the enzyme possess highest activity .The study was concentrated to comprehensive determina
... Show MoreThis study was conducted with the aim to extract and purify a polyphenolic compound “ Resveratrol†from the skin of black grapes Vitis vinifera cultivated in Iraq. The purified resveratrol is obtained after ethanolic extraction with 80% v/v solution for fresh grape skin, followed by acid hydrolysis with 10% HCl solution then the aglycon moiety was taken with organic solvent
( chloroform). Using silica gel G60 packed glass column chromatography with mobile phase benzene: methanol: acetic acid 20:4:1 a
... Show MoreThe detection for Single Escherichia Coli Bacteria has attracted great interest and in biology and physics applications. A nanostructured porous silicon (PS) is designed for rapid capture and detection of Escherichia coli bacteria inside the micropore. PS has attracted more attention due to its unique properties. Several works are concerning the properties of nanostructured porous silicon. In this study PS is fabricated by an electrochemical anodization process. The surface morphology of PS films has been studied by scanning electron microscope (SEM) and atomic force microscope (AFM). The structure of porous silicon was studied by energy-dispersive X-ray spectroscopy (EDX). Details of experimental methods and results are given and discussed
... Show MoreIn this paper to isolate and study the properties of the cyclooxygenase-2 (EC: 1.14.99.1) enzyme in the blood of a patient suffering from rheumatoid arthritis and study the effect of natural products of the Soapwort on the activity of purified enzyme. The study involves taking 30 ml of blood from an adult woman 40 years old, who suffers from rheumatoid arthritis disease for 13 years. Serum is separated and subjected to a series of purification processes including: precipitation by ammonium sulfate, filtration by centrifugation radiator, dialysis in presence of ammonium bicarbonate, separation using the technology of ion exchange, lipholization and then estimating approximate molecular weight of the enzyme using gel filtration techni
... Show MoreBackground: The antimicrobial resistance is one of the most serious and expanding health problems world -wide in the last decades. The esbl escherichia coli. (extended – spectrum beta-lactamase e.coli) represents an important aspect of it .Objectives: To get an overview on the esbl e.coli prevalence profile in general. Also to assess the antibiotic sensitivity of esbl e. coli trying to specify the most effective antibiotics in combating this micro-organism.Methods: this study tries to focus on this problem in Iraq which through a prospective study approach by taking 35 clinical samples from various sources (urine, blood, abscess, eye ,vagina ,stool and others),and after confirming the presence of e.coli, the presence of esbl e.coli and
... Show MoreSome Factors determining the virulence of Escherichia coli ( E. coli ) isolates were studied ,of 25 isolates , 17(group A) uropathogenic E. coli ,6 (group B) infected gastrointestinal tract , 2 (group C) infected wound , beside these group we use the standard strain E. coli HB101 as control group. The twenty five isolates were tested for adherence capability to human buccal cavity epithelial cells by in vitro experiment . The results showed that all isolates have different adhesion capability with mean ranging from (14.35±11.39) to (33.80 ± 22.68) bacteria / epithelial cell It was noticed that isolates EU9, ES6, EW17 displayed high adhesive capability with mean value (33.80 ± 22.68), (32.60 ± 21.19), (29.90±22.50) bacteria /epithelial
... Show More
Leuconostoc bacteria was isolated from local pickled cabbage (Brassica oleracea capitata) and identified as Leuconostoc mesenteroides by morphology,biochemical and physiological. The local isolated L. mesenteroides bacteria under the optimal conditions of dextran production showed that, the highly production of dextran was 7.7g achieved by using a modified natural media comprised of 100ml whey, 10g refined sugar, 0.5g heated yeast extract, 0.01g CaCl2, 0.001g MgSO4, 0.001g MnCl2 and 0.001g NaCl at pH 6 and 25̊C for 24 hr of fermentation and by using 1ᵡ106 cell/ml as initial inoculums volume. Some applications in food technology (Ice cream, Loaf, Ketchup and Beef preservation) have been performed with processed dextran. The result
As major nosocomial pathogens,
In this study, 20