RESUMEN

The aim of this study was to identify dipeptidyl peptidase IV (DPP-IV) inhibitory peptides from salmon skin collagen hydrolysate, and to evaluate the possible inhibition mechanism of DPP-IV and peptide. Salmon skin collagen was hydrolyzed by pepsin, trypsin, papain, or Alcalase 2.4 L, separately. Trypsin hydrolysate (10 mg/mL) showed the highest inhibitory activity of 66.12 ± 0.68%. The hydrolysate was separated into three fractions by ultrafiltration, and the inhibitory IC50 of M1 (molecular weight <3 kDa) was 1.54 ± 0.06 mg/mL. M1 was separated by gel chromatography and RP-HPLC; A10 was the highest inhibitory fraction in the 12 fractions, i.e., IC50 was 0.79 ± 0.13 mg/mL. A novel peptide LDKVFR with the IC50 value of 0.1 ± 0.03 mg/mL (128.71 µM) was identified from A10. Molecular docking revealed that six hydrogen bonds and eight hydrophobic interactions between LDKVFR and DPP-IV were contributed to DPP-IV inhibition.

Asunto(s)

Inhibidores de la Dipeptidil-Peptidasa IV , Salmo salar , Animales , Dipeptidil Peptidasa 4 , Inhibidores de la Dipeptidil-Peptidasa IV/farmacología , Simulación del Acoplamiento Molecular , Péptidos

RESUMEN

Extracted salmon skin collagen was hydrolysed with the free or immobilized extracellular protease of Vibrio sp. SQS2-3. The hydrolysate exhibited anti-freezing activity (>3 kDa) and antioxidant activity (<3000 Da) after ultrafiltration. The antioxidant peptide was further purified by size-exclusion chromatography and found to scavenge DPPH (73.29 ±â€¯1.03%), OH (72.73 ±â€¯3.34%,), and intracellular ROS in HUVECs; protect DNA against oxidation-induced damage; and have an ORAC of 2.78 ±â€¯0.28 mmol TE/g. The antioxidant peptide fraction was identified using mass spectrometry, and nineteen salmon collagen-sourced peptides were obtained. Of these, the peptide Pro-Met-Arg-Gly-Gly-Gly-Gly-Tyr-His-Tyr is a novel sequence and was the major component; this peptide was shown to have antioxidant activity via the ORAC assay (2.51 ±â€¯0.14 mmol TE/g). These results suggested that the protease from Vibrio sp. SQS2-3 is suitable for preparation of anti-freezing peptides and antioxidant peptides in a single step and represents a comprehensive use of fish skin collagen.

Asunto(s)

Proteínas Anticongelantes/farmacología , Antioxidantes/farmacología , Colágeno/química , Proteínas de Peces/química , Salmón , Animales , Proteínas Anticongelantes/química , Antioxidantes/química , Cromatografía en Gel , Proteínas de Peces/aislamiento & purificación , Proteínas de Peces/farmacología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Hidrólisis , Péptido Hidrolasas/química , Péptido Hidrolasas/metabolismo , Hidrolisados de Proteína/química , Especies Reactivas de Oxígeno/metabolismo , Piel/química , Ultrafiltración , Vibrio/enzimología