Heterologous expression of lacticin 3147 in Enterococcus faecalis: comparison of biological activity with cytolysin.
Lacticin 3147 is a broad-spectrum, two-component, lanthionine-containing bacteriocin produced by Lactococcus lactis DPC3147 which has widespread food and biomedical applications as a natural antimicrobial. Other two-component lantibiotics described to date include cytolysin and staphylococcin C55. Interestingly, cytolysin, produced by Enterococcus faecalis, has an associated haemolytic activity. The objective of this study was to compare the biological activity of lacticin 3147 with cytolysin. The lacticin 3147-encoding determinants were heterologously expressed in Ent. faecalis FA2-2, a plasmid-free strain, to generate Ent. faecalis pOM02, thereby facilitating a direct comparison with Ent. faecalis FA2-2.pAD1, a cytolysin producer. Both heterologously expressed lacticin 3147 and cytolysin exhibited a broad spectrum of activity against bacterial targets. Furthermore, enterococci expressing active lacticin 3147 did not exhibit a haemolytic activity against equine blood cells. The results thus indicate that the lacticin 3147 biosynthetic machinery can be heterologously expressed in an enterococcal background resulting in the production of the bacteriocin with no detectable haemolytic activity.
Ryan
M P
MP
Dairy Products Research Centre, Teagasc, Moorepark, Fermoy, Co. Cork, Ireland.
McAuliffe
O
O
Ross
R P
RP
Hill
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000187:drug effects*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D001430:Bacteriocins; D003603:Cytotoxins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D018274:Electroporation; D013293:Enterococcus faecalis / Q000235:genetics* / Q000378:metabolism; D020440:Gene Duplication; D005798:Genes, Bacterial; D006461:Hemolysis / Q000187:drug effects*; D006736:Horses; D013294:Lactococcus lactis / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D016415:Sequence Alignment; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D001430:Bacteriocins; D003603:Cytotoxins; D011994:Recombinant Proteins; C120351:cytolysin, Enterococcus faecalis; C082508:lacticin 481