Production and characterization of enterocin 900, a bacteriocin produced by Enterococcus faecium BFE 900 from black olives

Enterococcus faecium BFE 900 isolated from black olives produced a bacteriocin termed enterocin 900, which was antagonistic towards Lactobacillus sake, Clostridium butyricum, enterococci as well as Listeria spp. including Listeria monocytogenes. Enterocin 900 was inactivated by pepsin, cu-chymotrypsin, proteinase K and trypsin but not by catalase, alpha-amylase, or other non-proteolytic enzymes tested. The bacteriocin was heat stable, retaining activity after heating at 121 degrees C for 15 min. Enterocin 900 was active at pH values ranging from 2.0-10.0, with highest activity at pH 6.0. Bacteriocin production occurred in the late logarithmic growth phase when culture density was ca. log 8.0 CFU ml(-1). Enterocin 900 was produced in media with initial pH ranging from 6.0-10.0, but not in media with a pH lower than 6.0. Medium composition, especially the concentrations of peptone and yeast extract influenced bacteriocin production, with no bacteriocin being produced in the absence of either of these compounds. No plasmids could be isolated from Enterococcus faecium BFE 900, indicating that the gene for bacteriocin activity is located on the chromosome.