Skim milk hydrolysate (SMH-AP) containing AVPYP and GLPQE ameliorates adipogenic lipid accumulation in 3T3-L1 cells

BACKGROUND/OBJECTIVES: This study evaluated the anti-adipogenic effects of skim milk hydrolysate with alcalase and prozyme (SMH-AP) in 3T3-L1 cells and identified its bioactive peptides.MATERIALS/METHODS: After adipogenic differentiation, 3T3-L1 adipocytes were treated with SMH-AP.RESULTS: SMH-AP, produced via sequential hydrolysis with Alcalase and Prozyme, significantly inhibited lipid accumulation and modulated adipogenic and lipogenic biomarkers.It downregulated CCAAT/enhancer-binding protein (C/EBP) , C/EBP, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein (SREBP) 1c, while upregulating Krppel-like factor 2. SMH-AP suppressed the cholesterogenic markers (SREBP2 and 3-hydroxy-3-methylglutaryl-coenzyme A reductase) and activated AMP-activated protein kinase alpha, which phosphorylated acetyl-CoA carboxylase and activated hormone-sensitive lipase.Molecular analysis confirmed extensive proteolysis into small, cell-permeable peptides (~330 Da).Liquid chromatography-mass spectrometry/mass spectrometry identified Ala-Val-Pro-Tyr-Pro and Gly-Leu-Pro-Gln-Glu as major bioactive peptides.CONCLUSION: These findings suggest that SMH-AP may serve as a promising functional food ingredient for preventing obesity.