This study was to find an effective process to extract bioactive peptides from mushroom foot and deter- mine their effects on activation of alcohol metabolic enzymes in vitro. The optimum extraction assisted by ultra-high-pressure processing of mushroom foot peptides was obtained with a pressure of 400 MPa and a processing time of 10 min. After ultrafiltration, peptides with molecular weight of 0–3 kDa had the highest activity to activate alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) by 70.79%and 71.35%, respectively. Following dextran gel chromatography, two peaks (p-I and p-II) appeared and the activation activities on ADH and ALDH of p-I were 72.00% and 73.43%, both higher than p-II. Nine pep- tides were found in p-I as determined by LC–MS/MS, and two of them (IPLH and IPIVLL) were synthe- sized. IPLH activated ADH and ALDH by 42.7% and 29.2% respectively, which were higher than IPIVLL.