Glycation mechanism of lactoferrin–chitosan oligosaccharide conjugates with improved antioxidant activity revealed by high-resolution mass spectroscopy

Abstract

Glycosylation has a great effect on the antioxidant ability of proteins, which is due to the structural conformational change of peptides in the protein. In this study, a chitosan oligosaccharide (COS) was selected as the saccharide for glycation with lactoferrin (LF) by a wet-heat method, and a new stripe at a higher molecular zone in the gel of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a change in the bond revealed by Fourier transform infrared (FT-IR) and fluorescence spectroscopy analyses were used to confirm that the chitosan oligosaccharide was successfully covalently bound to lactoferrin. The ABTS and oxygen radical absorbance capacity (ORAC) assays indicated that lactoferrin exhibited a stronger antioxidant activity after glycation with the chitosan oligosaccharide. High-resolution mass spectroscopy further illuminated the relationship of enhanced antioxidant capacity and structural conformational change of peptides in lactoferrin at a molecular level.