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.
ABSTRACT The role of specific amino acids namely cysteine, methionine, threonine and asparagine in the protection provided by vamin solution against B-lactam inhibition to E. coli was evaluated in vitro. In minimal medium, Cells were treated with 32 ug/ml of penicillin G, carbencillin, hostacillin, cloxacillin and cephalotin in the presence of specific amino acid supplementations. Deletion of specific amino acids from the media abolished the protection provided by vamin. Threonine was essential for the protection of cells against all tested antibiotics, while cysteine was essential for protection against carbencillin and cephalotin Deletion of methionine or asparagine abolished the protec- tion against carbencillin and to a less extent ce
... Show MoreAim: To evaluation the effect of Lactobacillus acidophilus on Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 with detection of some virulence factors. Methods: Two hundred and fifty specimens (stool) from children under five years for both sexes were collected from some hospitals. All isolates were diagnosed according to morphological characteristics, biochemical tests. Monoplex pattern of PCR was used also for detection different genes in (7) Escherichia coli )O157:H7 (isolates; include 16SrRNA, eae, lifA, Stx1,Stx2 that encoded for ribosomal RNA, intimin, lymphocyte inhibitory factor, shiga toxins. Three types of probiotics strains were obtained, Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus acidophilus (A
... Show MoreColorectal cancer (CRC) is the most common disease and cause of death globally. The aim of the study is investigation and detection of some bacterial interfering with CRC occurrence and progression. The study conducted between September 2022 till February 2023, a total of 50 specimens were collected from confirmed CRC patients. In addition, 50 stool specimens were collected from Healthy volunteers, considers as control group. Isolation and identification of bacteria in all collected specimens were done by using cultural and differential media (blood agar, macconkey agar and Pfizer agar), as well as the VITEK- 2 compact system. The bacterial species, in the specimens of control were ( Escherichia coli 50 (86.20%), Klebsiella Pneumoni
... Show MoreSummary The aim of this study is the evaluation the resistance of S. marcescence obtained from soil and water to metals chlorides (Zn+2, Hg+2, Fe+2, Al+3, and Pb+2). Four isolates, identified as Serratia marcescence and S. marcescena (S4) were selected for this study according to their resistance to five heavy metals. The ability of S. marcescena (S4) to grow in different concentrations of metals chloride (200-1200 µg/ml) was tested, the highest concentration that S. marcescence (S4) tolerate was 1000 µg/ml for Zn+2, Hg+2, Fe+2, AL+3, pb+2 and 300 µg/ml for Hg+2 through 24 hrs incubation at 37 Co. The effects of temperature and pH on bacteria growth during 72 hrs were also studied. S. marcescence (S4) was affected by ZnCl2, PbCl2, FeC12
... Show MoreAlkaloids are regarded as important nitrogen-containing chemical compounds that serve as a rich source for discovering and developing new drugs where most plant-origin alkaloids have antiproliferation effects on different kinds of cancers. Alkaloids’ continence of Calotropis procera leaves are detected by two biochemical alkaloid reagents. Also GC-MS analysis for leaf alkaloid extract was done that showed the existence of one type of alkaloid compound at retention time12.8min detected as colchicine (C22H25N06( by comparing it with colchicine standard reference (Sigma Aldrich) with M.wt 399g/mol and percentage area 7.1%. Furthermore, identification, separation, and purification
... Show MoreIn this work, we synthesized thirteen compounds of 1-(2-furoyl)thiourea derivatives 1-13 by conversion of 2-furoyl chloride to 2-furoyl isothiocyanate by reacting it with potassium thiocyanate in dry acetone in a quite short reflux time then, in the same pot, different of (primary and secondary amines) were added individually to achieve thiourea derivatives. The products were characterized spectroscopically using (FT-IR, 1H NMR and 13C NMR) techniques. Some of them were evaluated as antioxidant agents using DPPH radical scavenging method, and all were examined theoretically as enzyme inhibitors against Bacillus pasteurii urease (pdb id: 4ubp) and by studying molecular docking using Autodock (4.2.6) software.
The research includes the synthesis and identification of the mixed ligands complexes of M 2 Ions in general composition ,[M(Leu) 2 (SMX)] Where L leucine (C 6 H 13 NO 2 )symbolized (LeuH) as a primary ligand and Sulfamethoxazole C 10 H 11 N 3 O 3 S) symbolized (SMX)) as a secondary ligand . The ligands and the metal chlorides were brought in to reaction at room temperature in(v/v) ethanol /water as solvent containing NaOH. The reaction required the following [(metal: 2(Na Leu --): (SMX )] molar ratios with M(II) ions, Were M ( Mn ( II),Co (II),Ni(II),Cu( II),Zn (II),Cd(II)and Hg( The UV Vis and magnetic moment data revealed an octahedral geometry around M(II), The conductivity data show a non electrolytic nature of the complexes . The
... Show MoreA first step in this research was to synthesize Schiff's bases(1-3)using an Amoxcilline intensification reaction with different aromatic aldehydes in absolute ethanol. In benzene and refluxing conditions,Schiff's bases were cyclized with succinic and Phthalic anhydride to give a new sequence of 1,3-oxazepine derivatives(4-6) and (7-9),respectively.The last step,cyclization reactions with sodium azide in THF solvent resulted in the formation of [10 and 11], which are supposed to be biologically significant.FT.IR, 1H-NMR and 13C-NMR (for compound 4,7,9, and 11),as well as melting points reported, were used to characterize these prepared compounds ,Bacillus (G+), Staphylococcus (G+), and E.Coli (G-)were screened against these compounds. . To i
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