Development of a novel strategy for controlled release of lysozyme from whey protein isolate based active food packaging films

The purpose of this study is to develop a novel controlled release system based on pH-responsive polyacrylic acid (PAA)/lysozyme (LYS) complexes incorporated within a hydrophilic whey protein isolate (WPI) film matrix for active food packaging applications. Complex formation is simple under benign conditions that are suitable for preserving antimicrobial activity of the lysozyme. In addition, the pH-dependent charge density of complexes allowed a uniform distribution in the matrix. The properties of the complexes such as size, surface charge and hydrophilicity were varied by changing PAA/LYS ratio (0.1 and 0.3 w/w) and PAA molecular weight (2 kDa and 450 kDa). The effects of complex properties as well as mode of lysozyme incorporation into the films (100%-free, 50%-free+50%-PAA/LYS complex and 100%-PAA/LYS complex) on the LYS release rate, activity and antimicrobial efficacy of the films were investigated. The results have shown that similar to 100% LYS loading into the complexes is possible regardless of PAA molecular weight or PAA/LYS ratio. Incorporating lysozyme into the film in complexed form extended its release time from less than 24 h up to 500 h and reduced its diffusivity from similar to 10(-9) to similar to 10(-13) cm(2)/s. The films including 50%-free-LYS+50%-PAA/LYS complex showed a 5.7 log reduction in bacterial population within 72 h whereas 100%-free-LYS containing film could not suppress Listeria innocua growth after 24 h. Overall, the results suggest that complexation of lysozyme with weak polyelectrolytes can be used as an effective strategy to achieve a long-lasting antimicrobial effect and that films prepared with such complexes have great potential as food packaging materials. (C) 2016 Elsevier Ltd. All rights reserved.