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.
This study aimed to study the effect of Ziziphus spina christi Aqueous cold and Alcoholic leaves and fruits extracts on the growth and activities of the following types of bacteria :( Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pyogenes ). The results appeared outweigh the alcoholic extract of leaves and fruits of Sidr that prepared by saxholate extractor by addition of ethanol 95% significant superiority as compared with aqueous extract that prepared by using distilled water as was its influence inhibitor to the growth and effectiveness of bacteria , about the treatment of in-vivo to cause injury to these types of bacteria diagnosed laboratory mice and treated with alcoholic extract of the leaves o
... Show MoreEnzyme activity were studied in the sera of children with leukemia than healthy children, where 31 cases were studied, including 21 cases of patients with acute lymphatic leukemia
Colibactin is a genotoxin produced by Enterobacteriaceae via a polyketide synthase (pks) island cluster. There is less knowledge regarding the distribution of colibactin genes in E. coli isolates in Iraq and its correlation with biofilm and antibiotic susceptibility. Therefore, this study aimed to investigate the frequency of some colibactin genes (CIbA and CIbQ) in uropathogenic E. coli in Iraq and evaluate the correlation with biofilm and antimicrobial resistance. Between October 2023 and January 2024, 70 E. coli isolates were isolated from 120 females diagnosed with UTIs. Isolates were identified first by biochemical methods and confirmed molecularly by amplification of 16S rRNA gene with specific primers. PCR was employed to detect the
... Show MoreSpecific microorganisms can produce bacterial nanocellulose (BNC), with acetic acid bacteria (AAB) being the most active producer. The family Acetobacteraceae includes the obligate aerobic, motile acetic acid bacteria. The BNC has attracted a lot of interest across a wide range of industries, including pharmaceuticals, due to its flexible characteristics, properties, and advantages. The present study was conducted to purify and characterize BNC produced from AAB isolated from apple vinegar. Bacterial nanocellulose was synthesized using a natural date palm liquid medium at pH 6 at 30°C for 8–10 days. The bacterial cellulose produced was then purified using a technique involving 0.1 M sodium hydroxide. To ascertain the surface mor
... Show MoreCoumarin is a natural substance isolated from different plants. It belonges to a group of benzobyrones which consists of a benzene ring joined to a pyrone nucleus. In the present research, a new series of coumarin derivatives were formed. Compound (1) (7-hydroxy-4-methyl Coumarin) was converted into 4-methylquinolin-2(H) derivative (2) by reaction with acetamide, and then reaction of (2) with thiosemicarbazide in ethanol leads to the synthesize of hydrazincarbothioamide derivative (3).The reaction of (3) with ethylchloroacetate in presence of sodium acetate leads to closure ring to get [(1-(5-oxo-2-thioxoimidazolidin-1-ylimino) ethyl)]quinolin-2(1H)-one (4). Mannich bases were prepared through the reaction of (4) with primary
... Show MoreA new set of metal complexes by the general formula [M(C)2(H2O)2]Cl2 has been prepared through the interaction of the new Ligand [N1, N4-bis(4-chlorophenyl)succinamide] (C) derived from succinyl chloride with 4-Chloroaniline with the transition metal ions Mn(II), Co(II), Ni(II), Hg(II), Cu(II) and Cd(II). Compounds diagnosed by TGA, 1 H, 13CNMR and Mass spectra (for (C)), Fourier-transform infrared and Electronic spectrum, Magnetic measurement, molar conduct, (%M, %C, %H, %N). These measurements indicate that (C) is associated with the metal ion in a bi-dentate fashion by nitrogen atoms (the amide group) and the octahedral composition of these complexes is suggested. The anti-bacterial action of the compounds towards three types of bacteria
... Show MoreThe members of the family of Eentrobacteriaceae harbour a gene cluster called polyketide synthase (pks) island. This cluster is responsible for the synthesis of the genotoxin colibactin that might have an important role in the induction of double-strand DNA breaks, leading to promote human colorectal cancer (CRC). Eleven out of the eighty eight isolates (12.5%) were pks+, distributed as 7 (8%) isolates of E. coli, 2 (2.25%) of K. pneumoniae and 2 (2.25%) of E. aerogenes. The cytotoxic effects of selected pks+ isolates (E. coli and E. aerogenes) on HeLa cells were represented by decreasing cell numbers and enlarged cell nuclei in comparison to the untreated cells. Cytological changes were observed when the infected HeLa cells culture
... Show MoreThe members of the family of Eentrobacteriaceae harbour a gene cluster called polyketide synthase (pks) island. This cluster is responsible for the synthesis of the genotoxin colibactin that might have an important role in the induction of double-strand DNA breaks, leading to promote human colorectal cancer (CRC). Eleven out of the eighty eight isolates (12.5%) were pks+, distributed as 7 (8%) isolates of E. coli, 2 (2.25%) of K. pneumoniae and 2 (2.25%) of E. aerogenes. The cytotoxic effects of selected pks+ isolates (E. coli and E. aerogenes) on HeLa cells were represented by decreasing cell numbers and enlarged cell nuclei in comparison to the untreated cells. Cyt
... Show MoreThis study aimed for isolation and identification of Candida glabrata and identifying some virulence factors. The distribution of patients with candidemia thrush showed that the age group 50-65 years old recorded the highest incidence of candidiasis in female and male with leukemia by 50% and 37.9 % respectively compared to the lowest incidence of candidiasis in the age group under 17 years old in female and male 8.8% and 13.5%, respectively. While the age group between 5-8 years was high, reaching 18 cases of oral candidiasis and 42% of children with leukemia compared with the age group, which was the least, that reached 9 cases, 21%. The highest incidence of C. glabrata was 59 isolates of females and males with leukemia, while C. kefyer w
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