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 IC50value 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.
Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur, Chennai, Tamilnadu, 603 203, India.
Ila
I
Joshi
Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur, Chennai, Tamilnadu, 603 203, India.
Sekar
S
Sudhakar
Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur, Chennai, Tamilnadu, 603 203, India. nazeer.ra@ktr.srmuniv.ac.in.
Rasool Abdul
RA
Nazeer
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005591:Chemical Fractionation; D049851:Gastropoda / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D053583:Interleukin-1beta / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008970:Molecular Weight; D009569:Nitric Oxide / Q000096:biosynthesis; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology*; D011492:Protein Hydrolysates / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D021241:Spectrometry, Mass, Electrospray Ionization; D014357:Trypsin / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000596:Amino Acids; D000893:Anti-Inflammatory Agents; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D010455:Peptides; D011492:Protein Hydrolysates; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D014357:Trypsin
Anti-inflammatory; ESI-MS/MS; Gastropod; Hydrolysate; Peptide