Plasmid-associated Bacteriocin Production by Leuconostoc sp. LAB145-3A Isolated from Kimchi

Leuconostoc sp. LAB145-3A isolated from kimchi produced a bacteriocin which was active against food pathogens, such as Listeria monocytogenes, Enterococcus faecalis, and E. faecium. Bacteriocin production occurred during the early exponential phase of growth and was stable upto the late stationary phase of growth. Optimum conditions for bacteriocin production were $37^{\circ}C$ with an initial pH of 7.0. The bacteriocin of LAB145-3A was sensitive to proteases, but stable for solvents, pH change and heat treatment. It was stable even at autoclaving temperature for 15 min. The bacteriocin exhibited a bactericidal mode of action against Lactobacillus curvatus LAB170-12. The bacteriocin produced by Leuconostoc sp. LAB145-3A was purified by CM-cellulose cation exchange column chromatography and Sephadex G-50 gel filtration. The purification resulted in an approximate 10,000-fold increase in the specific activity. Approximately 4% of the initial activity was recovered. Purified bacteriocin exhibited a single band on the SDS-PAGE with an apparent molecular weight of 4,400 daltons. This bacteriocin was named leucocin K. Leuconostoc sp. LAB145-3A had two residential plasmids with molecular sizes of 23 kb and 48 kb. A comparison of plasmid profiles between LAB145-3A and its mutants revealed that the 23 kb plasmid (pCA23) was responsible for bacteriocin production and immunity to the bacteriocin in Leuconostoc sp. LAB145-3A.