title: The antimicrobial mechanism of action of epsilon-poly-l-lysine.
abstract: Epsilon-poly-l-lysine (ε-PL) is a natural antimicrobial cationic peptide which is generally regarded as safe (GRAS) as a food preservative. Although its antimicrobial activity is well documented, its mechanism of action is only vaguely described. The aim of this study was to clarify ε-PL's mechanism of action using Escherichia coli and Listeria innocua as model organisms. We examined ε-PL's effect on cell morphology and membrane integrity and used an array of E. coli deletion mutants to study how specific outer membrane components affected the action of ε-PL. We furthermore studied its interaction with lipid bilayers using membrane models. In vitro cell studies indicated that divalent cations and the heptose I and II phosphate groups in the lipopolysaccharide layer of E. coli are critical for ε-PL's binding efficiency. ε-PL removed the lipopolysaccharide layer and affected cell morphology of E. coli, while L. innocua underwent minor morphological changes. Propidium iodide staining showed that ε-PL permeabilized the cytoplasmic membrane in both species, indicating the membrane as the site of attack. We compared the interaction with neutral or negatively charged membrane systems and showed that the interaction with ε-PL relied on negative charges on the membrane. Suspended membrane vesicles were disrupted by ε-PL, and a detergent-like disruption of E. coli membrane was confirmed by atomic force microscopy imaging of supported lipid bilayers. We hypothesize that ε-PL destabilizes membranes in a carpet-like mechanism by interacting with negatively charged phospholipid head groups, which displace divalent cations and enforce a negative curvature folding on membranes that leads to formation of vesicles/micelles.
authors.affiliation: Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark DuPont, Brabrand, Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Hyldgaard
authors.affiliation: DuPont, Brabrand, Denmark.
authors.forename: Tina
authors.initials: T
authors.lastname: Mygind
authors.affiliation: Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Brian S
authors.initials: BS
authors.lastname: Vad
authors.affiliation: Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Marcel
authors.initials: M
authors.lastname: Stenvang
authors.affiliation: Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Otzen
authors.affiliation: Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark Department of Bioscience, Aarhus University, Aarhus, Denmark rikke.meyer@inano.au.dk.
authors.forename: Rikke L
authors.identifier: http://orcid.org/0000-0002-6485-5134
authors.initials: RL
authors.lastname: Meyer
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology*; D001692:Biological Transport; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D008087:Listeria / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D011107:Polylysine / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D011107:Polylysine