Ultrasound and enzyme assisted preparation of novel lactoferrin-cereal phenolic acid conjugates: structural, physicochemical and functional properties

Abstract

The effects of covalent binding of protocatechuic acid (PA) and gallic acid (GA) to lactoferrin (LF) on the structure, functional, and antioxidant properties of the protein conjugate were investigated. These protein-phenolic conjugates were produced by laccase cross-linking and ultrasound-assisted free radical grafting, which were characterized using turbidity, particle size, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses. Structural changes in conjugates were monitored by endogenous fluorescence spectroscopy, fourier transform infrared spectroscopy (FTIR), and circular dichroism (CD). The antioxidant capacities and pH stability were determined using DPPH, ABTS, FRAP, and potentiometric analysis. The enzymatic cross-linking and free radical grafting yielded LF-PA/GA conjugates with altered hydrodynamic diameter and zeta-potential. Spectroscopic and chromatographic analyses revealed that binding to PA/GA altered the molecular structure of LF, with a decrease in LF isoelectric point post binding to PA/GA, without affecting antioxidant activities. In conclusion, LF-PA/GA conjugates present potential applications in the food industry.

Keywords: Antioxidant; Conjugation; Gallic acid; Laccase; Lactoferrin; Protocatechuic acid.