Preparation and Purification of Dipeptidyl Peptidase-Ⅳ Inhibitory Peptides from Pearl Mussel Muscle

Bioactive peptides with an inhibitory effect on dipeptidyl peptidase-Ⅳ (DPP-Ⅳ) were prepared from pearl mussel muscle.

Two-step enzymatic hydrolysis was adopted and enzymatic hydrolysis conditions were optimized. Fractions with in vitro DPP-Ⅳ inhibitory activity were concentrated by ultrafiltration, and purified by Superdex^TM peptide 10/300 GL gel filtration column chromatography and reverse phase-high performance liquid chromatography (RP-HPLC). DPP-Ⅳ inhibitory peptides were purified from the enzymatic hydrolysate and their amino acid sequences were identified by liquid chromatography-tandem mass spectrometry.

Both neutral protease and alkaline protease could effectively hydrolyze pearl mussel muscle. After concentration using a 3 kDa cut-off ultrafiltration membrane, the DPP-Ⅳ inhibition rate of the enzymatic hydrolysate reached 67.4%. After separation by gel filtration column and purification by RP-HPLC, 6 active peptides were obtained, whose sequences were FNAPAM, FIPNY, IYNPPTPF, LAMPYP, FFVVMP and LAGMP, respectively. Furthermore, the interactions between these peptides and DPP-Ⅳ were analyzed by molecular docking.

Our present study provides a theoretical reference for the effective utilization of fish processing byproducts as raw materials for functional food production.