Potential of enterocin from Enterococcus durans MF5 in controlling Listeria species

This research paper presents the characterization of an enterocin-producing Enterococcus durans MF5 isolate and the determination of the in vitro antilisterial activity of enterocin produced by this isolate, named Ent-MF5. PCR-based screening for bacteriocin biosynthetic genes revealed that E. durans MF5 harbors multiple enterocin-encoding genes (ent A, B, P and X), classified as class II bacteriocins and enterocin-P of Enterococcus faecium (sharing up to 99% similarity at the genetic level). E. durans MF5 is sensitive to eight clinically important antibiotics and does not possess cytolysin activator -cylA, gelatinase -gelE and hyaluronidase -hylA virulence genes. The antilisterial activity of Ent-MF5 was abolished by trypsin, α-chymotrypsin, protease and proteinase-K. Ent-MF5 showed thermal and pH stability. In addition, the activity of Ent-MF5 was unaffected in the presence of various surfactants (1% SDS, Triton X-100, Tween 20, and Tween 80). Ent-MF5 exhibited antimicrobial activity against Listeria monocytogenes, Listeria innocua, Listeria ivanovii and Listeria seeligeri at concentrations as low as 0.13 μg/ml. Ent-MF5 had a bactericidal effect against L. monocytogenes with a significant reduction in surviving cells at concentrations equal to or greater than 0.13 μg/ml. A 75-100% reduction in L. monocytogenes growth and bactericidal effect determined by CFU counts was observed following treatment with Ent-MF5 at 4.47 μg/ml at time points starting at 2 and 4 h, respectively. Ent-MF5 action is associated with Listeria cell membrane damage, as observed by flow cytometry and fluorescence microscopy. Thus, the effective antilisterial activity and stability of Ent-MF5 presents promising perspectives for application as biopreservatives in the food industry.