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.
Out of 120 isolates from different clinical cases, only 75 were found and confirmed that they belong to the Pseudomonas aeruginosa bacteria. The result revealed that the LasB virulent gene was present in 63 isolates with 63% percentage. The gel electrophoresis showed that the molecular weight of LasB gene was 300 bp. DNA sequences of LasB gene was done, and the results showed the presence of some gene mutations like substitution, addition and deletion with 97% identity with the Refseq gene. From the other side, the results of identities of translated nucleotides sequence with the original sequence of amino acids revealed that there are no effects of gene mutations on translation of the product protein.
Abstract A total of 207 specimens were collected from different sources including patients, health care staff and hospital environment in Ibb city, Yemen. The study used the bacteriocin produced from active producer strains in typing of Staphylococcus aureus. Depending on the morphological, cultural and biochemical characteristics, 54 (26.09%) isolates of Staphylococcus aureus were identified. An antibiotic sensitivity test was done for the bacterial isolates, and the results showed that there were multiple resistant antibiotics. The Staphylococcin production of these isolates has been detected by using wells assay. Fifty one isolates were Staphylococcin producer. Four isolates (staph19, staph25, staph28 and staph43) were chosen as go
... Show MoreThe genus Bacillus bacterium isolated from soil and tests their ability to produce glutamic acid, the bacteria are diagnosed by traditional methods and VITEK-2 system. And we studied the optimal conditions for production and try to improve the productivity of the isolation by manipulating the components of the production medium and of the circumstances environmental of production medium. There is 43 isolates of the 70 bacterial isolates belonging to the genus Bacillus depending on phenotypic characteristics of the colonies and microscopic characteristics and found that 13 isolated revert to species B.subtilis depending on physiological and biochemical characteristics. As well as on microscopic and biochemical tests, 13 isolates were subj
... Show MoreThe objective was to study the effect of prepared ginkgo biloba extracts against Candida albicans isolated from healthy persons. Conducting susceptibility test, biofilm formation test, phytochemical screening test, and antioxidant activity test. One hundred oral swabs sample were obtained from healthy persons with oral lesion attending dentistry teaching hospital in dentistry college, their age ranged from 1-30 years of both sexex. The studied samples collected through 8 months (April - December / 2018). This study included two different types of ginkgo bilola extracts were prepared as aqueous and ethanolic extracts. Many tests were used, which included isolation and identification of C.albicans, conduct susceptibility test, biofilm form
... Show MoreOne of the most important problems confronts hospitals is the strains emergence of Enterococcus spp. with multiple resistance to antibiotics, which propel researchers to modify or produce new antibiotics or combination between two antibiotics so that to be more effective against Enterococcus . This study was aimed to susceptibility some of local Enterococcus spp. Isolates with of 21 antibiotic using disc diffusion method. The results showed absolute resistant 100% toward (Cephalexin , Gentamycin , Amikacin ,Erythromycin and Nalidixic acid), while showed a high sensitivity toward (Vancomycin and Impenem ) at percentage of 92.3% for each . Also highl
... Show MoreThe study was conducted to isolate and identify endophytic fungi from leaves and twigs of lebbeck (Albizia lebbeck) and study their antagonistic activity against some plant fungal pathogens. Results showed isolation of 75 isolate endophytic fungi from 240 segments of leaves and twigs representing colonization frequency of 47.43%. Isolated fungi included different species of Aspergillus which prevail over other species (6 species), and three different isolates of Penicillium, one more species of Paecilomyces, Drechslera, Fusarium, Curvularia, Nigrospora and hyaline sterile fungus. Results showed variation in colonization frequencies within all species isolated from the two plant parts. Dual culture methods for
... Show MoreIn this paper we define and study new concepts of fibrewise topological spaces over B namely, fibrewise Lindelöf and locally Lindelöf topological spaces, which are generalizations of will-known concepts: Lindelöf topological space (1) "A topological space X is called a Lindelöf space if for every open cover of X has a countable subcover" and locally Lindelöf topological space (1) "A topological space X is called a locally Lindelöf space if for every point x in X, there exist a nbd U of x such that the closure of U in X is Lindelöf space". Either the new concepts are: "A fibrewise topological space X over B is called a fibrewise Lindelöf if the projection function p : X→B is Lindelöf" and "The fibrewise topological space X over B
... Show MoreProteinases (E.C.3.4.21) family are widely distributed in the nature; it was present in animals tissues , plants and microbial cell . Protease was purified from Zahdi seed (Phoenix dactylifera L.) by several steps included ammonium sulphite ppt (75%) saturation and dialyzed against the 80mM sodium phosphate buffer at pH 7.5 . The enzyme specific activity was 407.62 unit/mg protein. The obtained extract was purified by DEAE-Cellulose column followed by gel filtration through Sephacyl S-200 column .The enzyme specific activity ,yield and purification fold were 1873.49 unit/mg protein, 22.99 and 58.42% respectively. The results of protease characterization showed that the molecular weight was 25118 daltons as determined by gel f
... Show MoreIntroduction and Aim: Klebsiella pneumoniae is a Gram-negative bacterium responsible for a wide range of infections, including respiratory tract infections (RTIs). This research was aimed to study the antibacterial and anti-biofilm effect of AgNPs produced by Gram positive and negative bacteria on RTIs associated with K. pneumoniae. Materials and Methods: The biofilm formation of K. pneumoniae was determined by tube method qualitatively from select bacterial species characterized by UV-Visible spectroscopy. The antibacterial susceptibility of the bacteria AgNPs was tested for their antibacterial and antibiofilm activity on a clinical isolate of K. pneumoniae. Results: K. pneumoniae isolated from RTIs were strong biofilm prod
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