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

The visceral mass of the gastropod, Harpa ventricosa was hydrolysed using trypsin, alcalase and pepsin for 12 h 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 ± 0.02 mg/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 (690.2 Da) 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.