Effect of enzymatic cross-linking of naringenin-loaded β-casein micelles on their release properties and fate in in vitro digestion

The microbial transglutaminase (mTG) was used to improve the stability of the naringenin-loaded β-casein micelles (CNMs). The formation of cross-linked CNMs was confirmed by SDS-PAGE electrophoresis, showing a decrease in monomeric β-CN levels with increasing crosslinking time. Dynamic light scattering (DLS) showed that after crosslinking the particle size distribution did not change upon dilution, suggesting occurrence of intra-crosslinking. Fluorescence spectroscopy and circular dichroism (CD) showed that crosslinking induced only minor changes in the structure. Finally, release of naringenin in buffer at pH 7.4 demonstrated a slower release from the cross-linked micelles compared to the untreated micelles. In addition, the cross-linked micelles exhibited a partial resistance to pepsin enzyme. We conclude that crosslinking with mTG is a suitable method to modulate naringenin release kinetics from β-CN micelles and improves the potential of these micelles as delivery systems targeted to the small intestine.