Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol.12, Issue 1, 2010; Page No.(115- 120)

PRODUCTION AND PURIFICATION OF BACTERIOCIN FROM LACTOBACILLUS BREVIS

V. MUTHUMANI AND V. GUNASUNDARI

Abstract

This study was to determine the production of Bacteriocin from Lactobacillus brevis. The different colonies were collected from cow milk and gram staining was performed for identification of gram positive lactic acid bacteria. The test organisms were inoculated into lactobacillus agar medium for pure culture. Lactobacillus brevis was selected test organism for the production of bacteriocin, confirmed by using gram staining and biochemical tests, then the Lactobacillus brevis cells were taken in different time points such as 12 hrs, 24 hrs, 36 hrs, 48 hrs and 72 hrs to study the time kinetics of the strain of interest. The best growing culture were centrifuged at 12000 rpm for 10 minutes at 4 oC. After 36 hrs, the Lactobacillus brevis culture were subjected into Heat shock treatment at 55 oC, Cold shock treatment at 4 oC and pH variation of the medium maintained at pH 9 for bacteriocin production. The produced bacteriocins were allowed to antimicrobial assay.Then bacteriocin was partially purified by using Column chromatography technique. Again SDS PAGE was performed to determine the molecular weight of the bacteriocin.

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