Earthworms, regarded as a valuable resource, are rich in proteins (up to 70%) and have huge potential to produce high-value products. This study selected five proteases to enzymatically hydrolyze earthworms, and key factors including the enzyme dosage, hydrolysis temperature and time were optimized. The extracted earthworm-derived peptides with different molecular weights were purified and separated using membrane separation method, and their antioxidant properties were evaluated through DPPH and ABTS assay. The results showed that alcalase protease, trypsin and papain could achieve good hydrolysis degrees with a peptide yield of up to 22.6%. The majority of the earthworm-derived peptides had molecular weights less than 3 kDa (97.67%), with over half having molecular weights smaller than 1 kDa (55%). The overall amount of amino acids was found to be 676.60 mg/g, with a contribution of 24.6% from amino acids known for their strong antioxidant properties. Following the identification of peptide sequences, a total of 548 peptides were recognized, with short peptides containing 3-7 amino acids representing 64% of the total. This work aims to contribute towards the future utilization of earthworm bioactive peptides in food, cosmetic and pharmaceutical industries.