Abstract: This study aimed to screen and identify bioactive peptides from Tabanus, characterize their structural and functional features, and provide a theoretical basis for the development of novel natural peptide-based therapeutics. Initially, bioactive peptides were extracted using phosphate buffered saline (PBS), followed by protein precipitation with cold ethanol and purification via solid-phase extraction (SPE). Peptide identification and sequence analysis were performed based on LC-MS/MS data using PEAKS Studio software. Peptide activity potential was then predicted using Peptide Ranker, and a comprehensive bioinformatics analysis was conducted to evaluate key functional characteristics, including active sites, solubility, stability, toxicity, and intestinal absorption capacity. A total of 10 378 peptide sequences were identified, with lengths predominantly ranging from 6 to 26 amino acids. Most peptides had a molecular weight below 1 500 Da, and 97.49% were below 2 500 Da. Based on Peptide Ranker score (≥ 0.80), 95 candidate bioactive peptides were selected. Among them, 59 peptides were identified as having neuroregulatory potential via the BIOPEP-UWM database, due to the presence of key functional residues. Bioinformatics prediction further revealed that among these 59 peptides, 29 exhibited good water solubility, 22 were predicted to be stable, 43 had favorable intestinal absorption potential, and none were toxic, highlighting their suitability for further functional studies and therapeutic development. Molecular docking analysis demonstrated that FPPRDF, the highest-ranking peptide according to Peptide Ranker, formed stable interactions with dipeptidyl peptidase Ⅲ (DPP Ⅲ), suggesting its potential neuroregulatory and antitumor activities. Overall, this study established a systematic and efficient workflow for the screening and functional prediction of bioactive peptides from natural sources. The proposed strategy provides robust technical support for subsequent experimental validation, mechanistic studies, and the development of novel peptide-based drugs.