Structural characterization, HepG2 cell cytoprotective ability, and antioxidant mechanism of novel antioxidant peptides identified from black soldier fly larvae (Hermetia illucens L.).

ABSTRACT

This study isolated a novel antioxidant peptide from black soldier fly larvae (BSFL) using enzymatic hydrolysis. Firstly, the BSFL enzymatic hydrolysate was fractionated through ultrafiltration, with the <3 kDa fraction exhibiting the strongest DPPH and ABTS radical scavenging activity. Subsequently, this fraction was further fractionated through gel filtration chromatography and RP-HPLC. Totally, 153 peptides were identified through LC-MS/MS analysis, from which a novel peptide EDEGTYKCVLS (Pep6) was screened according to activity prediction and verification. Pep6 exhibited high radical scavenging capacity and cytoprotective effect on HepG2 cells against H2O2 damage, meanwhile significantly increasing the intracellular antioxidant enzymes activity. Molecular docking analysis indicated that Pep6 competitively bound to Keap1, thereby inhibiting the formation of Keap1-Nrf2 complex, ultimately protecting cells from oxidative stress damage. In this study, a novel antioxidant peptide Pep6 was identified from BSFL, and its antioxidant mechanism was elucidated, providing a theoretical basis for its use as a natural antioxidant.