Activity retention after nisin entrapment in a polyethylene oxide brush layer

The cationic, amphiphilic peptide nisin is an effective inhibitor of gram-positive bacteria whose mode of action does not encourage pathogenic resistance, and its proper incorporation into food packaging could enhance food stability, safety, and quality in a number of circumstances. Sufficiently small peptides have been shown to integrate into otherwise nonfouling polyethylene oxide (PEO) brush layers in accordance with their amphiphilicity and ordered structure, including nisin, and we have recently shown that nisin entrapment within a PEO layer does not compromise the nonfouling character of that layer. In this work we test the hypothesis that surface-bound, pendant PEO chains will inhibit displacement of entrapped nisin by competing proteins and, in this way, prolong retention of nisin activity at the interface. For this purpose, the antimicrobial activity of nisinloaded, PEO-coated surfaces was evaluated against the gram-positive indicator strain, Pediococcus pentosaceous. The retained antimicrobial activity of nisin layers was evaluated on uncoated and PEO-coated surfaces after incubation in the presence of bovine serum albumin for contact periods up to 1 week. Nisin-loaded, uncoated and PEO-coated samples were withdrawn at selected times and were incubated on plates inoculated with P. pentosaceous to quantify nisin activity by determination of kill zone radii. Our results indicate that nisin activity is retained at a higher level for a longer period of time after entrapment within PEO than after direct adsorption in the absence of PEO, owing to inhibition of nisin exchange with dissolved protein afforded by the pendant PEO chains.