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.
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2013 - 2012
... Show MoreThe first aim of the present study was performed to assay the activity of arginase in sera of women with uterine fibroid.. This study consisted of(50) women with uterine fibroid as patient's group and (30) healthy women as control group. The age ranged between (30-55) years for the two groups. The results showed that highly significant increas (P< 0.0001) in the arginase activity in sera of women with uterine fibroid (7.99± 0.23) I.U/L is found when compared with healthy group (0.52±0.02) I.U/L. The second aim was performed to isolate arginase from sera of women with uterine fibroids. The purification is done by addition of ammonium sulfate, dialysis, gel filtration chromatography by using sephadex G-50 and ion exchange chromatography by
... Show MoreForty one isolates of genus Proteus were collected from 140 clinical specimens such as urine, stool, wound, burn, and ear swabs from patients of both sex. These isolates were identified to three Proteus spp. P. mirabilis, P. vulgaris and P. penneri .The ability of these bacteria to produce L-asparaginase II by using semi quantitative and quantitative methods was determined. P. vulgaris Pv.U.92 was distinguished for high level of L-asparaginase II production with specific activity 1.97 U/mg. Optimum conditions for enzyme production were determined; D medium with 0.3% of L-asparagine at pH 7.5 with temperature degree 35°C for incubation. Ultrasonication was used to destroy the P. vulgaris Pv.U.92 cells then ASNase II was extracted and pu
... Show MoreThe first aim of the present study was performed to assay the activity of arginase in sera of women with uterine fibroid.. This study consisted of(50) women with uterine fibroid as patient's group and (30) healthy women as control group. The age ranged between (30-55) years for the two groups. The results showed that highly significant increas (P< 0.0001) in the arginase activity in sera of women with uterine fibroid (7.99± 0.23) I.U/L is found when compared with healthy group (0.52±0.02) I.U/L. The second aim was performed to isolate arginase from sera of women with uterine fibroids. The purification is done by addition of ammonium sulfate, dialysis, gel filtration chromatography by using sephadex G-50 and ion exch
... Show MoreThe presence and prevalence of V. cholerae were investigated in forty five water samples collected from different locations of Tiger River/ Baghdad city. Twenty one isolates were isolated by adopting a simple isolation techniques. The final identification revealed that only three isolates were confirmed as V. cholerae. They were named 1J, 1R and Dial 131 which are all serogrouped as non-O1. Toxin Coregulated Pili (TCP) and heat labile enterotoxin (LT) were determined in only the environmental isolate 1J while non of the isolates produced heat stabile toxin (ST). The purification scheme was improved, few steps were adopted to include back extraction of ammonium sulfate, saturation between 80-20%, desalting through Sephadex G25, and gel filt
... Show MoreThe objective of this study was to investigate the release profile of different fat and water soluble bases using diazepam as a model drug , and then to develop a satisfactory formula with a rapid release of diazepam from suppository bases .The study was conducted using theobroma oil ,glycerol-gelatin and glycerol-PEG1540 bases using conventional mold method for preparation .while the later base was utilized to incorporate diazepam ( buffered solution ) in a hollow type suppositories. The results indicated that all types of bases can be utilized to formulate diazepam as rectal suppositories with acceptable disintegration time ( 12, 10, 6, and 6min.), respectively . While 100% of the released drug had been shown differen
... Show MoreFifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity. The DNA of isolate M3 was extracted and lipase gene was amplified through PCR tec
... Show MoreThis work was conducted to study the extraction of pelletierine sulphate from Punica granatum L. roots by liquid membrane techniques. Pelletierine sulphate is used widely in medicine. The general behavior of extraction process indicates that pelletierine conversion increased with increasing the number of stages and the discs rotation speed but high rotation speed was not favored because of the increased risk of droplet formation during the operation. The pH of feed and acceptor solution was also important. The results exhibit that the highest pelletierine conversion was obtained when using two stages,(10 rpm) discs speed of stainless steel discs,(pH= 9.5) of feed solution and (pH= 2) of acceptor solution in n-decane. Assuming the existence
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