Enhancing zein functionality through sequential limited Alcalase hydrolysis and transglutaminase treatment:...

This study investigated the effects of sequential enzymatic hydrolysis using Alcalase, followed by transglutaminase conjugation on the secondary and tertiary structures, hydrophobicity, free amine content, protein-protein interactions, and functional properties of zein. Fourier-transform infrared spectroscopy showed that the most significant secondary structural changes, characterized by a decrease in α-helix content and an increase in β-turns, occurred at a higher degree of hydrolysis. At a 2 % degree of hydrolysis, it revealed notable emulsifying activity (65.96 m²/g), while at 5 % hydrolysis, it achieved the highest solubility (75.06 %). Additionally, the zein hydrolysate with a 7 % hydrolysis degree, treated with transglutaminase, demonstrated improved H₀ values (2992.33), enhanced foam capacity (65.95 %), and increased solubilized protein content in a dithiothreitol extractant (31.35 %). Meanwhile, native zein treated with transglutaminase showed the highest water holding capacity (4.47 g/g). Overall, the combined enzymatic approach modified zein structure and properties, suggesting potential for improving functionality in plant-based food applications.