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 novel heterocyclic organozinc compounds were prepared from the reaction of diazonum salt cytosine zinc chloride with thymol and vanilin as coupler components. The prepared compounds were characterized by elemental analysis and UV-Vis, FTIR and 1HMNR spectroscopic techniques. The biological activity was also studied for all prepared compounds.
Agricultural chemicals on a large scale of use throughout the world, and there are many studies about these chemicals and its disadvantages, but most of them were limited to its impact on mammals such as rodents in determine. This study was designed to determine the impact of these chemicals on DNA damage for E. coli bacteria from Iraq. The DNA is similar in terms of structure and function in all living organisms with a different in number and sequence of the nitrogenous bases among living organisms. This study showed that snails and slugs killer material Metaldehyde are strongly bind with the DNA extracted from bacteria, and herbicides Glyph
... Show MoreSome of the characters of the Staphylolysin A and D enzymes purified from Pseudomonas aeruginosa P16 and P5 respectively were studied, the molecular weights of Staphylolysin A and D were 20.417 kilo dalton and 23.988 kilo Dalton respectively by SDS- polyacryl amide gel electrophoresis. The optimum pH for staphylolysin A activity was found to be 8 which gives higher activity reaches 150 unit/ml, and for enzyme stability was 7.5-8.5 in which the enzyme nearly retained its full activity, while it was 9.5 for staphylolysin D that gives higher activity of 16 unit/ml,and 8.5-9.5 for enzyme stability in which the enzyme nearly retained its full activity, Maximum activity of two enzymes was obtained at 40C in which the specific activity for st
... Show MoreThe aim of this study is to evaluating the antibacterial activity of Laurus nobilis leaves extract on E. coli isolates. Maceration and Soxhlet apparatus were used to prepare aqueous and methanolic extracts; total phenolic content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were conducted to determine the active compounds in the extracts. The results showed that both Laurus nobilis methanolic and aqueous extracts have a noticeable effect on scavenging free radicals. Free radical scavenging activity. The total phenolic contents were 28.60 ±0.12 and 16.58 ±0.11mg/g in 50 mg/ml, in methanolic and aqueous extracts respectively. The antibacterial activity of Laurus nobilis leaves extracts showed that the methanolic extract was more effective than
... Show MoreThe present study aims to detect CTX-M-type ESBL from Escherichia coli clinical isolates and to analyze their antibotic susceptibility patterns. One hundred of E. coli isolates were collected from different clinical samples from a tertiary hospital. ESBL positivity was determined by the disk diffusion method. PCR used for amplification of CTX-M-type ESBL produced by E. coli. Out of 100 E. coli isolates, twenty-four isolates (24%) were ESBL-producers. E. coli isolated from pus was the most frequent clinical specimen that produced ESBL (41.66%) followed by urine (34.21%), respiratory (22.23%), and blood (19.05%). After PCR amplification of these 24 isolates, 10 (41.66%) isolates were found to possess CTX-M genes. The CTX-M type ESBL
... Show MoreSeven [35%] and five [25%] Serratia marcescens isolates were obtained out of 20 samples of lettuce and 20 samples of spinach, respectively, taken from different locations in a farm in Baghdad city. The isolate that produced chitinase in higher level was chosen to purify chitinase through several stages of purification including: ammonium sulfate precipitation, DEAE- sephadex ion exchange chromatograpgy and sephadex G-200 gel filtration with 89.5- fold purification and 30% recovery. The purified chitinase was characterized and the molecular weight of enzyme was 59000 daltons by using gel filtration chromatography. The optimum pH and temperature of the purified chitinase were 6.0 and 50°
... Show MoreThe effect of crude colicin extracted from E. coli isolated from
urinary tract infection patients on phagocytosis in vitro was studied . Results showed that the effect of crude colicin on phagocytic cells and their activities were concentration dependent. Low concentration (50
Mg/ml) have no significant effect (p>O.OS) on shape, size, migration and engulfment activity of phagocytic cells, while (50, 100 Mg/ml)
enhanced killing activity and increase superoxide ( 02 ) production as indicated by (NBT) test. But high concentration (500 mg/ml) of crude colicin caused inhibition of phagocytic cel
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