Grafted calcium pectinate-whey protein isolate covalent immobilizers: Optimization, kinetics, thermodynamics, and application

Abstract

Whey protein isolate (WPI) was incorporated within calcium pectinate (CPT) beads in order to boost their anionic qualities and meliorate their glutaraldehyde (GA)-polyethyleneimine (PEI) grafting process. The Box-Behnken Design (BBD) verified that WPI inclusion significantly raised the GA-PEI-CPT-WPI beads immobilized β-D-galactosidase (iβ-GLD) activity. The BBD also revealed the optimal settings for WPI concentration, PEI pH, PEI concentration, and GA concentration, which were 2.91 %, 10.8, 3.5 %, and 2.24 %, respectively. The GA-PEI-CPT-WPI beads grafting process was scrutinized via FTIR, EDX, and SEM. The optimal GA-PEI-CPT-WPI immobilizers provided fine β-GLD immobilization efficiencies, which reached up to 65.28 %. The free and GA-PEI-CPT-WPI iβ-GLDs pH and temperature profiles were scrutinized. It was also unveiled that the thermal stability of the iβ-GLD surpassed that of its free compeer as it provided lesser kd and ΔS values and larger t1/2, D-values, Ed, ΔH, and ΔG values. Furthermore, the iβ-GLD provided 92.00±3.39 % activity after 42 storage days, which denoted its fine storage stability. The iβ-GLD short duration (15 min) operational stability was also inspected, and 82.70±0.78 % activity was provided during the fifteenth degradation run. Moreover, the iβ-GLD long duration (24 h) operational stability was inspected while degrading the lactose of buffered lactose solution (BLS) and cheese whey (CW). It was unveiled that 81.86±0.96 % and 73.58±2.24 % of the initial glucose were detected during the sixth degradation runs, respectively.