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PURIFICATION AND CHARACTERIZATION OF BACTERIOCIN FROM WEISSELLA PARAMESENTEROIDES DFR‐8, AN ISOLATE FROM CUCUMBER ( CUCUMIS SATIVUS )
Author(s) -
PAL AJAY,
RAMANA K.V.
Publication year - 2010
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2010.00340.x
Subject(s) - bacteriocin , listeria monocytogenes , antimicrobial , cucumis , microbiology and biotechnology , bacteria , biology , food science , chemistry , botany , genetics
Bacteriocin from Weissella paramesenteroides DFR‐8 isolated from cucumber ( Cucumis sativus ) was purified by using only two steps, viz., pH‐mediated cell adsorption–desorption method and gel permeation chromatography. A single peak observed in the purity check by analytical Reverse Phase‐High Performance Liquid Chromatography (Waters 600 analytical HPLC system, Milford, MA) and a single band (molecular weight ∼ 3.74 kDa) shown on SDS‐PAGE analysis strongly indicated the homogeneity of the bacteriocin preparation. Treatment with proteolytic enzymes abolished the antimicrobial activity indicating the proteinaceous nature of bacteriocin. The purified bacteriocin exhibited a broad inhibitory spectrum against foodborne pathogens and spoilage microorganisms, including gram‐negative bacteria such as Salmonella typhimurium, Vibrio parahaemolyticus, Aeromonas hydrophila and Listeria monocytogenes. Response surface methodology was employed to study the interactive effect of temperature and pH on bacteriocin activity, and a regression equation was developed. The bacteriocin retained full activity after storage at − 20C for 90 days, while partial and complete activity loss was observed when stored at 4 and 37C, respectively.PRACTICAL APPLICATION In recent years, bacteriocins of lactic acid bacteria have gained much attention as food biopreservatives because of their origin from generally regarded as safe organisms. In spite of various bacteriocins studied worldwide, studies on bacteriocins of Weissella paramesenteroides remain rare. The present work involves the purification of bacteriocin up to absolute homogeneity from W. paramesenteroides , an isolate first time reported from cucumber ( Cucumis sativus ). The purified bacteriocin (molecular weight ∼3.74 kDa) was found to inhibit a large number of foodborne pathogens, including Listeria monocytogenes , which is resistant to commercially available bacteriocin, i.e., nisin. The application of central composite rotatable design enabled us to design a regression equation from which the residual activity of bacteriocin can be predicted at any given conditions of temperature and pH within the experimental domain. The broad inhibitory spectrum and thermostability of bacteriocin suggest its potential application in food preservation.