Controlled hydrolysis and EGCG conjugation enhance the ADH/ALDH activation activity of chia seed meal protein...

This study examines the effects of hydrolysis duration (20-100 min using flavourzyme) and EGCG conjugation on the structure and bioactivity of chia seed meal protein hydrolysates (CSPH) through multi-spectroscopic techniques and physicochemical property evaluation. Subsequently, the activation effects of EGCG-conjugated peptides on alcohol metabolism-related enzymes were further analyzed through the integration of peptidomics, bioinformatics, and computational chemistry. It was found that with the extension of hydrolysis time, the thermal stability of CSPH diminishes, its rigid structure becomes more flexible, and its crystallinity decreases (by up to 27.19 %). Meanwhile, the activation effects on alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity were significantly enhanced (P < 0.05). CSPH hydrolyzed for 60 min demonstrated the highest binding affinity for EGCG, primarily driven by hydrophobic interactions and hydrogen bonds. The CSPH-EGCG conjugate exhibited enhanced physicochemical properties and significantly elevated activation of ADH and ALDH, with ADH activation rising from 22.66 % to 95.56 % and ALDH activation from 9.45 % to 30.93 %, compared to unmodified CSPH. Seven active peptides were identified from PE-60 by peptidomics and bioinformatics. Computer docking optimized selected three optimal peptides (IPW, FPIH, and IYP). Two-dimensional and three-dimensional interaction analyses showed that these peptides bind to EGCG, ADH, and ALDH via hydrogen bonds, hydrophobic interactions, and salt bridges. These findings highlight the potential of controlled hydrolysis with flavourzyme and EGCG incorporation to enhance CSPH's properties and bioactivities and offer insights into the practical applications of CSPH and its EGCG complexes in food processing and therapeutic systems.