The Degree of Hydrolysis and Peptide Profile Affect the Anti-Fatigue Activities of Whey Protein Hydrolysates in Promoting Energy Metabolism in Exercise Mice

The purpose of this study was to investigate the effects of characteristics of whey protein hydrolysates (WPHs) on energy metabolism in exercise mice. Results showed that high-degree of hydrolysis (DH) hydrolysates (22%, H-Alc and H-AXH) showed better anti-fatigue effects than low-DH hydrolysates (10%, L-Alc and L-AXH) in enhancing energy substances and reducing metabolic byproducts. It might be related to the higher content of components less than 3 kDa in H-Alc and H-AXH (92.35 and 81.05%, respectively) and higher intensities of small peptides containing two to nine residues. Moreover, Western blot results revealed that WPHs maintained the energy balance in exercise mice by regulating the AMP-activated protein kinase (AMPK) and mTOR signaling pathways. Notably, H-Alc had higher intensities of peptides containing two to five residues than H-AXH and these peptides were rich in essential amino acids, which might explain why H-Alc exhibited better effects in decreasing protein metabolites. Meanwhile, H-AXH contained more free amino acids, especially Leu, which might contribute to its ability to promote glucose consumption in muscle. Furthermore, 40 peptides with two to nine residues and high intensities (>5 × 105) were screened from H-Alc and H-AXH and predicted by bioinformatics tools. Among them, LLL, LLF, GTW, AGTW, and ALPM showed high bioavailability, cell permeability, and potential bioactivity.