n this study, we prepared enzymatic extracts using trypsin and pepsin from Hermetia illucens (THI and PHI) and evaluated their effects on oxidative stress in ultraviolet B (UVB)-stimulated keratinocytes. PHI showed a higher extraction yield than THI, but the protein content of THI was higher. When the antioxidant activity was assessed in vitro, both indicated an effect of 25% or more compared to the positive control (L-ascorbic acid). In addition, both THI and PHI decreased the generation of intracellular reactive oxygen species (ROS) in UVB-stimulated keratinocytes, without cytotoxicity at all concentrations used. We explored the efficacy of THI. Results demonstrated that THI effectively reduced apoptosis and sub-G1 DNA content. In addition, it was found that UVB-stimulated keratinocytes protect skin keratinocytes from UVB by regulating the apoptotic pathway, MAPKs, and NF-kB signaling pathways. In conclusion, these results show that trypsin hydrolysate of H. illucens protects cells from oxidative stress in UVB-irradiated human keratinocytes.