Free radical scavenging and antiproliferative activities of peptide from monkey’s head mushroom (Hericium erinaceus)

Abstract

Free radicals have been identified as a major trigger of various syndromes in living systems. Oxidative stress occurs when free radicals are excessive and antioxidants are insufficient, resulting in the necessary consumption of additional antioxidants. Peptide derived from monkey’s head mushroom seeds was hydrolysed using various concentrations (1, 2.5, and 5%) of proteases (Alcalase, Favourzyme, and Neutrase), and the 1% Alcalase hydrolysate exhibited the highest radical scavenging activities (DPPH, ABTS, and NO assay) compared to other hydrolysates. After ultrafiltration, Mw < 0.65 kDa showed the strongest activity. Then, Mw < 0.65 kDa was purified using gel filtration chromatography and separated into two fractions (F1, and F2), of which fraction F1 exhibited the highest activity and this was further purified by reversed phase high performance liquid chromatography. Four fractions (F11, F12, F13, and F14) from RP-HPLC were isolated. Thus, four antioxidant peptides were identified by quadrupole time-of-flight (Q-TOF) mass spectrometer. The cytotoxicity activity of the fraction F1 was determined by MTT assay in five cell lines. The results showed that F1 exhibited greater inhibition against the proliferation of Chago-K1 cell lines. Moreover, the fraction F1 has the protective activity of the hydroxy radical-induced DNA damage as shown in pBR322, pKS and pUC19. The apoptosis of F1 was measured by an FITC Annexin V Apoptosis Detection Kit with PI using flow cytometry and caspase 3, 8 and 9 activities were determined in Chago-K1 cells for 24, 48, and 72 hours. The findings indicate that monkey’s head mushroom is a source of antioxidant peptides. Consequently, this suggests the importance of monkey’s head mushroom as a source of antioxidant peptides.