Nutr Res Pract.  2026 Apr;20(2):201-219. 10.4162/nrp.2026.20.2.201.

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

• Lee S
• Suh HJ
• Jung EJ
• Choi HS

• KMID: 2580127
• DOI: http://doi.org/10.4162/nrp.2026.20.2.201

Abstract

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) β, C/EBPα, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein (SREBP) 1c, while upregulating Krüppel-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.

Keyword

AVPYP; adipogenesis; lipid accumulation; skim milk hydrolysate; 3T3-L1 cells