ISOLATION AND CHARACTERIZATION OF BACTERIOCIN FROM BACILLUS CEREUS MTCC 1307
Publication Date : 19/12/2016
Bacteriocins are antibacterial proteins produced by bacteria that inhibit the growth of other bacteria. In the present study a bacteriocin produced from Bacillus cereus MTCC1307 (Microbial Type Culture Collection) was isolated and characterized. The bacteriocin was extracted from culture supernatants of 40% saturation with Ammonium Sulfate and further concentrated by Chloroform/Methanol mixture in 2:1 ratio. The purified product resolved a single band on SDS-PAGE with a molecular weight of 23 kDa. The bacteriocin was characterized by testing the bactericidal activity at different temperatures, pH, organic solvents, enzymes and storage. It was found that bactericidal activity was stable up to 60oc for 10 minutes but lost its activity at 100oc. The action of bacteriocin was found to be stable at pH 3.5, 4.5 and 7.0 but the activity was completely lost at pH 9.0. Bacteriocin was insensitive to organic solvents such as chloroform and n-propanol, where as sensitive to proteolytic enzymes such as trypsin and proteinase K. They were found to be stable for a period of 30 days at -20oc and +4oc, but on storage at room temperature resulted in loss of activity after 15 days. The antibacterial activity of the isolated bacteriocin was determined against typed E.coli isolates of poultry origin by agar spot assay. This study has shown the way of exploring the possibilities of using these bacteriocins as antimicrobial agents against the infections caused by E.coli in young animals and birds by incorporating them in feed or water.
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