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.
The best optimum temperature for the isolate was 30○C while the pH for the maximum mineral removal was 6. The best primary mineral removal was 100mg/L, while the maximum removal for all minerals was obtained after 8 hrs, and the maximum removal efficiency was obtained after 24 hrs. The results have proved that the best aeration for maximum removal was obtained at rotation speed of 150 rpm/ minute. Inoculums of 5ml/ 100ml which contained 106 cell/ ml showed maximum removal for the isolate.
Background: Toxin-producing Shiga Escherichia coli has been identified as a new foodborne pathogen that poses a significant health risk to humans. Shiga toxin-producing Escherichia coli can be found in raw cow milk and its derivatives. A small number of Escherichia coli strains that produce shiga toxin are pathogenic. Aim of study: The study aimed to see if there were any virulence genes in 50 milk samples that were typical of Entero-haemorrhagic E. coli and evaluate the Myrtus communis effects on these bacteria. Materials and Method: Milk samples were used to isolate E. coli bacteria (n= 27), biochemically analyzed, and genetically screened for virulence genes using a multiplex (PCR). The hydro-alcoholic extraction of Myrtus communis leave
... Show MoreABSTRACT Fifty extremely halophilic bacteria were isolated from local high salient soils named Al-Massab Al-Aam in south of iraq and were identified by using numerical taxonomy. Fourty strains were belong to the genus Halobacterium which included Hb. halobium (10%). Hb. salinarium (12.5%), Hb.cutirubrum (17.5%), Hb-saccharovorum (12.5%), Hb. valismortis (10%) and Hb. volcanii (37.5%). Growth curves were determined. Generation time (hr) in complex media and logarithmic phase were measured and found to be 10.37±0.59 for Hb. salinarium. 6.49 ± 0.24 for Hb.cutirubrum. 6.70±0.48 for Hb-valismonis, and 11.24 ± 0.96 for Hb. volcanii
A total of nine swab samples were collected from inflamed teeth and gingiva of human’soral cavity from a dentist clinic in Baghdad. All specimens were cultured in Mitis Salivarius agar medium and the isolated bacterial pure colonies werethen identified by using VITEK2. Three samples were diagnosed and identified as Staphylococcus lentus. One of the three isolates which showed a distinctive heavy growth on the media was selected for further analysis in this study. Paper disk diffusion method was used to detect the antibacterial activityof three of mouthwash solutions (Zak, Colgate and Listerine). The results showed that “Colgate†was the most active solution with antibacterial activity compared with the other two s
... Show MoreMicrobial lipases today occupy a place of prominence among biocatalysts owing to their ability to catalyze awide variety of reactions in aqueous and non- aqueous media, A.baumannii were isolated from different clinical specimens from hospitalized patients from Baghdad hospitals and were detected by biochemical tests and API20E system. The percentage of isolation was (16.6%), A. baumannii is an increasingly multidrug – resistant (MDR), it showed high level of resistant to Ceftriaxon, Colistin, Piperacillin, Co-trimoxazol, Tertracycline, Carbenicillin, Amoxicillin, Penicillin G, Gentamicin and Ceftazidim , wherease the isolates were highly sensitive to Imipenem, Ciprofloxacin, Meropenem, Amikacin, and Cefotaxime.
... Show MoreNew derivatives of the anti-inflammatory, leprostatic drug dapsone 4 are synthesized, characterized and biologically screened by the treating the drug dapsone with chloroacetyl chloride in the presence of base. Both amino groups are acylated to give compound 6. The symmetrical acylated product then treated with Phenol, N-Acetyl-p-aminophenol, p-Chlorophenol, m-Chlorophenol, o-Hydroxybezoic acid and m-Hydroxybezoic acid to give compounds 8(a-f). The antimicrobial activity was tested for the synthesized compounds; activates were good compared to the parent drug. All the new compounds have scanned for their biological activities toward gram ‒ve and gram +ve (M. tuberculosis, S. pneumoniae, E. coli and P. mirabilis) bacteria, the synthesized
... Show MoreThis study aimed to study the inhibition activity of purified bacteriocin produced from the local isolation Lactococcuslactis ssp. lactis against pathogenic bacteria species isolated from clinical samples in some hospitals Baghdad city. Screening of L. lactis ssp. Lactis and isolated from the intestines fish and raw milk was performed in well diffusion method. The results showed that L. lactis ssp. lactis (Lc4) was the most efficient isolate in producing the bacteriocin as well observed inhibitory activity the increased that companied with the concentration, the concentration of the twice filtrate was better in obtaining higher inhibition diameters compared to the one-fold concentration. The concentrate
... Show More- coli K12 and B. subtilis 168 were investigated for their cadmium and mercury tolerance abilities. They were developed by UV mutagenesis technique to increase their tolerances either to cadmium or mercury, and their names then were designated depend on the name and concentration of metals. E. coli K12 Cd3R exhibited bioremediation amount of 6.5 mg Cd/g dry biomass cell. At the same time, its wild-type (E. coli K12 Cd3) was able to remove 5.2 mg Cd/g dry biomass cell in treatment of 17 mg Cd /L within 72 hours of incubation at 37 °C (pH=7) in vitro assays. The results show that E.coli K12 Hg 20 was able to remove 0.050 µg Hg/g dry biomass cell