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.
Medicinal plants are a source for a wide variety of natural active compounds and are used for the treatment of diseases throughout the world. Conocarpus erectus L. widely planted all over Iraq and has different secondary metabolites, which has been used in treatment of anemia, cancer, fever and diarrhea. The present study aims to estimate the antibacterial activity of Conocarpus erectus leaves extracts on some microorganisms collected from patients with burn infection. The study began with the collection of Conocarpus erectus leaves in June 2018 from the trees in university of Baghdad. Maceration method was used to prepare aqueous extract, while Soxhelt apparatus was used to prepare methanolic extract. The results of phytochemical test show
... Show MoreAntibacterial activity of CNSs against Staphylococcus aureus and Escherichia coli was estimated. Higher inhibition zone of 18 mm and 20 mm were observed against S. aureus and E.coli, respectively, at a concentration of 2 mg/ml of carbon nanosphere after 24 hrs of incubation at 37 ºC. In vitro cytotoxicity experiment was performed on two parasite strains of Leishmania donovani and Leishmania tropica by using MTT assay. L. donovani revealed more sensitiv to the CNSs than L. tropica. An intermediate level of cytotoxicity of 51.31 % was observed when 2.4 mg/ml of CNSs was incubated with L. donovani, while weak cytotoxicity of 37.20 % was shown when the
... Show MoreLipase enzyme has attracted a lot of attention in recent years because of its diverse biotechnological applications. The present study was conducted to screen germinated seeds of four crops, namely sunflower (Helianthus annuus), flaxor linseed (Linum usitatissimum ), peanut (Arachis hypogaea ) and castor bean (Ricinus communis), for the activity of their lipases. to the study also included the extraction and purification of lipase from the seeds of the most promising crop using different solvents. The results indicated that the maximum enzymatic activity (0.669 U/ml) was obtained when 0.1 M Tris-HCl buffer extract was used after 3 days of seed germination of all the tested species, as compared to the other test solvents
... Show MoreIn this research , phthallic anhydride ring is opened with 4-methyl aniline and acetone as a solvent to results the compound [I] that reacted with dimethyl sulphate and anhydrous sodium carbonate formation to phathalate ester [II], while the acid hydrazide compound [III], was obtained from mixed the compound [II]with hydrazine hydrate, Synthesis four type of shiff bases[IV]a-d was synthesized from the reaction of acid hydrazide [III] with aromatic aldehyde or ketone , when reacted Shiff bases with phthalic anhydride or naphthalicanhydride,I get eight derivatives of oxazepine [V]a-d , [VI]a-d. The bacterial activity of the new compounds studied by four species of bacteria: Esherichia Coli, Enterobactecloacae (Gram negative) and staphylococcu
... Show MoreObjective: Synthesis, Characterization of formazan derivatives and studies the antioxidant activity of prepared compounds and molecular docking. Methods: In this study, formazan compounds (III–XIV) were produced by combining Schiff base compounds (I), (II) with diazonium salts resulting from reactions of different aromatic amines with sodium nitrate in the presence of Con.HCl at 0–5°C. When isonicotinic acid hydrazide reacts with (N,N-dimethylbenzaldehyde, 4-hydroxy-3-methoxybenzaldehyde) in the presence glacial acetic acid as a solvent Schiff base compounds are created. Results: The prepared compounds were identified by FT-IR, 1H NMR, 13C NMR, then the antioxidant activity of the derivatives and molecular docking were studied. D
... Show MoreThe objective of this study was to investigate the prophylactic roles of human enteric derived Lactobacillus plantarum L1 (Ll) and Lactobacillus paracasei L2 (L2), on EHEC O157:H7 infection in rodent models (In vivo). The Lactobacillus suspensions (L1 and L2) were individually and orally administered to experimental rats at a daily two consecutives of 100 μl (108 CFU/ ml/rat) for up to two weeks. Thereafter, on the 8th day of experiment rats were orally challenged with one dose infection of EHEC (105 CFU/ml/rat). Animals mortality and illness symptoms have been monitored. There was no fatal EHEC infection in rats that had been pre‑colonized with the Lactobacillus strains, while most of EHEC infected rats were died (90%). The
... Show MoreCeliac disease (CD) is the most common genetically - based disease in correlation with food intolerance. The aim of this study is to measure the activity of ALT enzyme and purify enzyme from sera women with celiac disease. Alanine aminotransferase (ALT) activity has been assayed in (30) women serum samples with celiac disease, age range between (20-40) year and (30) serum of healthy women as control group, age range between (22-38) year. In the present study, the mean value of ALT activity was significantly higher in patients with celiac disease than healthy group (p<0.01). The ALT enzyme was partial purified from sera women with celiac disease by dialysis, gel filtration using Sephadex G- 50 and ion exchange chromatography using DEAE- cell
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