Sonochemical effects on fabrication, characterization and antioxidant activities of β-lactoglobulin-chlorogenic acid conjugates

The β-lactoglobulin-chlorogenic acid (LG-CA) conjugate was explored to be formed through ultrasonication, redox-pair method and their combination, the ultrasonication used a probe ultrasonic machine with a 6 mm probe at 270 W, and the frequency was 20-25 kHz. The formation of the conjugate was confirmed by SDS-PAGE with a larger molecular weight. Besides, Fourier infrared spectroscopy (FTIR) and Circular dichroism (CD) indicated changes in the secondary structure of the LG-CA conjugate. The α-helix and β-sheet contents of LG decreased and the unordered content increased significantly after the formation of covalent complexes. In addition, both the ultrasonic treatment and its combination with redox-pair method could significantly improve the antioxidant properties of LG. The former increased to 23.16 μmol Trolox/g sample, the latter 82-106 μmol Trolox/g sample. Therefore, ultrasonication could be used both individually and in combination with the redox-pair method to produce LG-CA conjugates with stronger antioxidant activities.