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 microbial production of substances that have the ability to inhibit the growth of other microorganisms is possibly the most common defense strategy developed in nature. Microorganisms produce a variable collection of microbial defense systems, which include antibiotics, metabolic by-products, lytic agents, bacteriocins and others. The aim of the present study was to isolate and identify Enterococcus spp. and its most prevalent species from food samples and determine its antibacterial activity against Staphylococcus aureus isolates. A total of 50 food samples from different sources (dairy products (20 samples) and vegetables and fish (15 samples each)) were collected from different local markets in Baghdad and
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spectra and J>hysical methods,selected metals,which were Cu11
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The Escherichia coli isolated from UTIs, were identified with biochemical tests and vitek test.The ethanolic extracts of Zingiberofficinale(Z.officinale) were tested against E.coli by using the good agar diffusion test, the alcoholic extracts from (25 -100) mg/ml showed antimicrobial activity against tested microorganism. The diameter of inhibition zone increase at high concentrations and rang from (18-20mm) , and these results compared with antibiotics sensitivity discs were used by discs diffusion method against E.coliisolates, they were resisted to all antibiotics used in this study. It could be concluded that alcoholic extract of Z.officinalehad good antimicrobial effects, and may be able to use for treatment UTIs caused by E.c
... Show MoreOut of 150 different specimens, 67 S. aureus isolate were isolated. However, 16sRNA gene was located only in 60 isolates. Moreover, mecA gene was located in 48 isolates; thereby MRSA covered 80% of all S. aureus isolates. Of considerable interest, pvl gene was detected in only six isolates (10%). Hence, the present work emphasizes the notion suggested that pvl is not an indicative of CA-MRSA.
98 samples were collected from various clinical sources included (Burns, wounds, urines, sputums, blood) From the city of Baghdad, After performing the biochemical and microscopic examination, 52 isolates were obtained for Pseudomonas aeruginosa, 17 (32.7%) isolates from burn infection, 12 (23%) isolates from Wound infection 11 (21.2%) isolates from urine infection, 7 (13.5%) isolates of sputum and 5 (9.6%) isolates from blood. Bacteria susceptibility to form biofilm has been detectedby microtiter plate method, The results showed that 80% of the bacterial isolates were produced the biofilm with different proportions, alg D gene (alginate production) has been detected by polymerase chain reaction (PCR) Which plays an essential role in the fo
... Show MoreStaphylococcus aureus, which includes the methicillin-resistant S. aureus (MRSA), is a significant human pathogen producing different toxins and results in many different infection types, which include bacteremia, soft-tissue infections, as well as staphylococcal food poisoning. S. aureus is an important food-borne pathogen of humans due to ingestion of food containing enterotoxigenic strains. Detecting S. aureus femA and mecA genes was evaluated with the use of a Loop-mediated Isothermal Amplification Method (LAMP). The accuracy of this approach was similar to that attained using the approach of the conventional polymerase chain (PCR). Those two methods characterized 43 isolates of MRSA which
... Show MoreThe study included isolate and diagnose fungus Fusarium solani of the local soil and purified and development in the PDB medium and the filtrate extracted using a solvent (Ethyl acetate) to obtain the fungal secondary metabolites extract. This extract has shown bioactivity against both reference isolates (E.coli (ATCC25922) and S.aureus(NCTC6571)) and pathogenic isolates S.pyogenes, K. pneumonia and S.typhimurium using agar disk diffusion technique , The diameters of the inhibition zones of fungal secondary metabolites24.0 mm against E.coli and 31.5 mm against S.aureus,and 34.0 mm against K.pneumoniae and 18.0 mm against S.pyogenes and 33.5mm against S.typhimurium. The test revealed the minimum inhibitory concentration (MIC) of the fungal
... Show MoreThis study aimed to evaluate the anticancer activity and cell division arresting by dandelion methanolic extract on breast cancer cell line MCF-7 cancer cell line. For achieving this goal, cytotoxicity assay (MTT assay), multipara system assay: High Content Screening (HCS) which include (viable cell count VCC; membrane permeability MP; cellular mitochondrial permeability CMP; nuclear intensity NI and cytochrome C releasing ), reactive oxygen species detection and cell cycle phases division were tested. The results of this study showed the ability of the plant to reduce cancer cell viability in a dose-dependant manner within IC50 (141.0) in comparison to IC50 of (334.4) on the
... Show MoreThe cytotoxicity of different concentrations of purified methionine γ- lyase from Pseudomonas putida on cancer cell lines (RD, AMN3 and AMGM) at 96 hr was studied. The bacterial enzyme with concentration 1000µg/ml was revealed highly cytotoxicity against cancer cell lines in comparison with other concentrations whereas slight cytotoxicity was observed on normal cell (REF).