Development of pH-controlled triggering mechanisms for controlled release of lysozyme

Abstract

In this study, zein and whey protein (WP) based films and their blends and composites have been prepared to obtain pH-controlled triggering mechanisms for controlled release of lysozyme. The total amount of lysozyme released from standard zein films was not considerably affected from changes in pH between 7.3 and 5.3 since this hydrophobic biopolymer lacked charged ionisable groups to bind lysozyme ionically. In contrast, incorporation of lentil protein isolate (LPI) into zein created a composite structure and caused binding of positively charged lysozyme (pI>9.0) on negatively charged groups of LPI (pI: 4.5-6) within the film matrix in release medium with pH between 5.3 and 7.3. The amount of bound lysozyme in zein-LPI composites increased linearly as LPI concentration increased between 1,5 and 4,5 mg/cm2 at pH 7.3. The release of bound enzyme could be triggered by reducing pH of release medium from 7.3 to 4.3, down below pI of LPI. On the other hand, films of WP (pI≈5.2) bind considerable amount of lysozyme due to their inherent net negative charges close to neutrality. The release of bind lysozyme could be trigged as pH of release medium reduced from 6.0 to 3.0, down below the pI of WP. The preparation of WP-oleic acid blend and WP-bees wax composites increased the film porosity and amounts of released lysozyme from films at pH 4.5 and 5.0, by 2-4 and 1.2-1.5 folds, respectively. The zein and WP based films containing 0.7 to 1.4 mg/cm2 lysozyme showed good antimicrobial activity against Listeria innocua. This work showed the potential of creating pH-controlled release systems during antimicrobial packaging of food.