Formation of Lysozyme-Caseinate Heteroprotein Complexes for Encapsulation of Lysozyme by Spray-Drying: Effect of Mass Ratio and Temperature

The formation of heteroprotein complexes obtained by the interactions between sodium caseinate (CAS) and lysozyme (LYS) at pH 7 was investigated by using turbidimetric analysis, particle size distribution, and zeta potential at different CAS/LYS ratios. Moreover, isothermal titration calorimetry (ITC) was used to determine the type and magnitude of the energies involved in the CAS/LYS complexation process and evaluated the thermodynamic behavior of their complexation. Results revealed that the structure of CAS/LYS complexes drastically changed when CAS/LYS ratio increased to 1.0 and the structuring stages were characterized by exothermic signals and were controlled by favorable enthalpy changes due to electrostatic interactions between both proteins. In addition, the interaction between two proteins was temperature-dependent and mainly entropy-driven, which was verified by molecular dynamics (MD) simulations, and the hydrophobic interactions and hydrogen bonding were shown to play an important role in CAS/LYS interactions. Furthermore, CAS/LYS complexes showed minimum LYS enzymatic activity at CAS/LYS ratio 1.0. Though spray-drying of CAS/LYS complexes with ratio 1.0, the LYS activity in reconstituted solution was recovered >80 % of initial activity after calcium chloride addition. The present study provides useful information about CAS/LYS complexation and binding processes, which could facilitate their application in antimicrobial edible food packaging.