Anti-inflammatory Properties of Bioactive Peptide Derived from Gastropod Influenced by Enzymatic Hydrolysis.

Abstract

The visceral mass of the gastropod, Harpa ventricosa was hydrolysed using trypsin, alcalase and pepsin for 12h to produce protein hydrolysates. Subsequently, the active hydrolysate was observed in the 3rd hour of tryptic hydrolysate (29.17±0.62 and 34.85±0.55%) using human red blood cell (HRBC) membrane stabilization and albumin denaturation (AD) assays. The active hydrolysate was fractionated by membrane filtration unit, where <10-kDa fraction revealed better anti-inflammatory activity with IC50 value 6.27±0.05 and 5.38±0.02mg/ml for HRBC and AD assays, respectively. Additionally, the active fraction contains essential and non-essential (aspartic acid, arginine, glutamic acid and leucine) amino acids and, sequentially, the active fraction was further purified using consecutive chromatography, in which fraction C-II exhibited strong anti-inflammatory activity (HRBC 56.02±0.52 and AD 50.71±1.10% assays). The non-toxic, low molecular weight (690.2Da) hexapeptide (Ala-Lys-Gly-Thr-Trp-Lys) suppressed the nitric oxide (NO) and pro-cytokine production in a dose-dependent manner on THP-1 cell lines.