Isolation and identification of antioxidative peptides from frog (Hylarana guentheri) protein hydrolysate by consecutive chromatography and electrospray ionization mass spectrometry

Appl Biochem Biotechnol. 2014 Jul;173(5):1169-82. doi: 10.1007/s12010-014-0793-6. Epub 2014 Apr 29.

Abstract

Frog (Hylarana guentheri) proteins were hydrolyzed by papain and Flavourzyme to obtain antioxidative peptides. The antioxidant activities of the frog protein hydrolysates (FPHs) were measured, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (IC50=9.94±0.13 mg/mL), reducing power (0.39±0.01 at 5.0 mg/mL), and oxygen radical absorbance capacity (ORAC) value (789.15±75.10 μmol Trolox equivalents/g). The hydrolysates were purified by ultrafiltration, ion exchange chromatography, gel filtration chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). Through analysis of ESI-MS/MS, two dipeptides were identified as Leu/Ile-Lys (259.1607 Da) and Phe-Lys (293.1446 Da), respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amphibian Proteins / metabolism*
  • Animals
  • Chromatography*
  • Free Radical Scavengers / analysis*
  • Free Radical Scavengers / chemistry
  • Free Radical Scavengers / isolation & purification*
  • Free Radical Scavengers / pharmacology
  • Hydrolysis
  • Peptide Fragments / analysis*
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification*
  • Peptide Fragments / pharmacology
  • Ranidae
  • Spectrometry, Mass, Electrospray Ionization*
  • Ultrafiltration

Substances

  • Amphibian Proteins
  • Free Radical Scavengers
  • Peptide Fragments