Alcalase Enzymolysis of Red Bean (adzuki) Ferritin Achieves Nanoencapsulation of Food Nutrients in a Mild Condition

Classical methods to fabricate ferritin-nutrients shell–core nanoparticles usually apply extremely acid/alkaline pH transition, which may cause the activity loss of nutrients or the formation of insoluble aggregates. In this work, we prepared an extension peptide (EP) deleted red bean (adzuki) ferritin (apoRBFΔEP) by Alcalase 3.0T enzymolysis. Such enzymolysis could delete the EP domain and remain the typical shell-like structure of the ferritin. Meanwhile, the α-helix content of apoRBFΔEP was decreased by 5.5%, and the transition temperature (Tm) was decreased by 4.1 °C. Interestingly, the apoRBFΔEP can be disassembled into subunits under a benign condition at pH 4.0 and is assembled to form an intact cage protein when the pH was increased to 6.7. By using this novel route, the epigallocatechin gallate (EGCG) molecules were successfully encapsulated into the apoRBFΔEP cage with an encapsulation ratio of 11.6% (w/w), which was comparable with that by the traditional pH 2.0 transition. The newly prepared EGCG-loaded apoRBFΔEP exhibited a similarly protective effect on the EGCG upon simulated gastrointestinal tract and thermal treatment as compared with the control. In addition, the EGCG-loaded apoRBFΔEP could significantly relieve the ferritin association induced by pH transition, which was superior to traditional method. The thinking of this work will be especially suitable for encapsulating pH-sensitive molecules based on ferritin in a benign condition.