Characterization of the chemical and antimicrobial properties of piscicolin 126, a bacteriocin produced by Carnobacterium piscicola JG126

Abstract

A novel peptide bacteriocin produced by the lactic acid bacterium Carnobacterium piscicola JG126 isolated from spoiled ham was purified and characterized. This bacteriocin, designated piscicolin 126, inhibited the growth of several gram-positive bacteria, especially the food-borne pathogen Listeria monocytogenes, but had no effect on the growth of a number of yeasts and gram-negative bacteria. Bactericidal activity was not destroyed by exposure to elevated temperatures at low pH values; however, bactericidal activity was lost at high pH values, especially when high pH values were combined with an elevated temperature. Piscicolin 126 activity was not affected by catalase, lipase, or lysozyme but was destroyed by exposure to a range of proteolytic enzymes. Piscicolin 126 was purified to homogeneity and was found to be a peptide having a molecular weight of 4,416.6 ؎ 1.9. A sequence analysis revealed that this compound is a cystibiotic (class IIa) bacteriocin containing 44 amino acid residues and one intrapeptide disulfide ring. Piscicolin 126 has regions of homology with some other bacteriocins obtained from lactic acid bacteria and is most closely related to sakacin P and pediocin PA-1 (levels of identity, 75 and 55%, respectively). Addition of piscicolin 126 to a devilled ham paste test food system inhibited the growth of L. monocytogenes for at least 14 days. Piscicolin 126 was more effective than two commercially available bacteriocin preparations tested in the same system. MATERIALS AND METHODS Materials. Chemicals and other products were obtained from the following suppliers. Growth media and agar were obtained from Oxoid, Basingstoke, United Kingdom. Catalase was obtained from Boehringer, Mannheim, Germany. ␣-Chymotrypsin, ␤-chymotrypsin, lipase, lysozyme, pepsin, trypsin, protease type I, protease type XIV, protease type XXIII, and ␣-cyano-4-hydroxycinnamic acid were obtained from Sigma Chemical Co., St. Louis, Mo. Nisin (as Nisaplin) was obtained from Aplin and Barrett, Trowbridge, United Kingdom. ALTA 2431 was obtained from Quest International, Sydney, New South Wales, Australia. Devilled ham paste was obtained from Master Foods, Sydney, New South Wales, Australia. Plasmid pGEM-T was obtained from Promega Corp., Madison, Wis. CM-Sepharose Fast Flow was obtained from Pharmacia Biotech, Uppsala, Sweden. 2-Mercaptoethanol was obtained from BDH,

Isolation of aCarnobacterium maltaromaticum-like bacterium from systemically infected lake whitefish (Coregonus clupeaformis)

“…Whether the observed serine to leucine amino acid substitution has any functional implications on the protein itself and whether this protein is actually being produced by lake whitefish isolates are currently unknown. The piscicolin-126 protein, a class II bacteriocin, is bactericidal to some Gram-positive bacteria, such as Listeria monocytogenes (Jack et al, 1996). Thus, in addition to the potential for disease causation in lake whitefish, the C. maltaromaticum-like isolates recovered in this study may also be inhibitive to other closely related bacterial pathogens under some conditions, although further investigation is required.…”

Anti-bacterial activity of Lactobacillus plantarum strain SK1 against Listeria monocytogenes is due to lactic acid production

“…However, the anti-microbial activity of the peptide nisin [produced by certain strains of Lactococcus lactis ssp. lactis (Daba et al 1991)] and the bacteriocin produced by C. piscicola JG126, have been shown to be influenced by pH (Benkerroum and Sandine 1988;Jack et al 1996). Therefore, the possibility still existed that Lact.…”

“…These include Leuconostoc gelidum (Harding and Shaw 1990), Leuconostoc spp. (Choi et al 1999), Carnobacterium piscicola (Jack et al 1996), Staphylococcus xylosus (Villani et al 1994), Lactobacillus sakei (Katla et al 2002), Enterococcus mundtii (Kawamoto et al 2002) and Enterococcus faecium (Ennahar et al 1998). All the above organisms were found to produce bacteriocin-like substances that inhibited L. monocytogenes.…”

Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

“…Then, colonies were picked at random, purified by streaking and re-streaking on fresh media, and examined for inhibition against the cultures of L. garvieae and S. iniae using the cross-streak method described by Robertson, O'Dowd, Burrells, Williams & Austin (2000). Antibacterial activity was also examined using the double-layer method described by Jack, Wan, Gordon, Harmark, Davidson, Hillier, Wettenhall, Hickey & Coventry (1996). For this, the bacterial isolates were grown in 10 mL of tryptone soya broth (TSB; Oxoid) for 24 h at 20°C.…”