Bacteriocin-producing lactobacilli in Spanish-style fermented sausages: Characterization of bacteriocins

Three bacteriocins were studied. Sakacin K, a bacteriocin from Lactobacillus sake CTC494 was purl fled to homogeneity by a four-step system involving ammonium sulphate precipitation binding to a cation exchanger, hydrophobic interaction and reverse phase chromatography in FPLC (fast performance liquid chromatography) system. The peptide sequence was determined by Edman degradation. The first 30 amino acid residues from the N-terminus of sakacin K were identical to those of curvacin A from Lactobacillus curvatus LTH1174 and sakacin A from Lactobacillus sakei Lb706 The structural gene of sakacin K in Lb. sake CTC494 was located on a 60 Kbp plasmid (as has been previously reported) by curvacin A in Lb. curvatus LTH1174 and sakacin A in Lb. sakei Lb706. Plantaricin D, a bacteriocin-like compound isolated from Lb. plantarum CTC305 was purified and sequenced using the same procedure as described for sakacin K. The first 15 amino acid residues of plantaricin 5) were identical to those obtained from the bacteriocin inducer peptide (termed plantaricin A) of the bacteriocinogenic system in Lb. plantarum CII. The structural gene of the plantaricin D peptide was located on a similar EcoRI chromosomal fragment when compared to plantaricin A in Lb, plantarum Cli! These two isolated bacteriocin-like peptides (sakacin K and plantaricin DJ were purified from two new different strains obtained from fermented meat, confirming the ecological importance of these substances. Lactobacillus sakei CTC372, the third bacteriocinogenic strain selected in this study from fermented sausages, produced a bacteriocin named sakacin T The bacteriocin is not found free in the supernatant of the cells, it is active against Listeria monocytogenes and Staphylococcus aureus. The genetic determinant of this bacteriocin was localized in a 84.4 Kbp plasmid by conjugative transfer and curing assays. (C) 2000 Academic Press.