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.
This research was conducted to measure the safety of heat stable enterotoxin a (STa) produced by enterotoxigenic Escherichia coli, through studying its toxic effect on mice since it showed a promising effect in reducing the proliferation of colorectal cancer cells. The cytogenetic effect was determined after giving five different doses (100, 200, 400, 800 and 1600)μg/Kg in comparison with negative (phosphate buffer saline / PBS) and positive (mitomycin C/ MMC, at doses of 2 and 5μg/Kg) controls on mouse bone marrow cells by employing the following parameters: mitotic index, chromosomal aberrations and micronucleus, also, the serum level of liver functional enzymes (GOT, GPT, ALP) was recorded. In addition, lethal dose 50 (LD 50) with cert
... 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 MoreNanotechnology is a continually expanding field for its uses and applications in multiple areas i.e. medicine, science, and engineering. Biosynthesis is straightforward, less-toxicity, and cost-effective technology. TiO2 NPs biosynthesis has attained consideration in recent decades. In this study, probiotic bacteria were isolated from cow’s raw milk samples, and then were identified by using the Vitek2 system; as Leuconostoc spp. included Leuconostoc mesenteroides subsp. mesenteroides (Leu.1), Leuconostoc mesenteroides subsp. cremoris (Leu.4), and Leuconostoc pseudomesenteroides (Leu.14). All Leuconostoc spp. isolates showed an ability for TiO2 NPs bio-production, after being incubated at anaerobic conditions (30 o C/ 24 h) in DeM
... Show MoreMedicinal 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 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
... 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 chr
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