This experiment used two kinds enzymes （papain and pancreatin） to hydrolyze corn gluten meal for preparing corn anti-alcoholism peptides， the single factor experiment was aimed to explore the influence of different liquid-to-material， the quantity of enzyme， bi-enzyme papain / pancreatin and reaction time during the enzymolysis on the protein recovery ratio and the activation ratio of ADH of corn anti-alcoholism peptides. The optimum technology conditions were as follows： the liquid-to-material ratio was 10 ∶ 1， the quantity of enzyme was 2%， the bi-enzyme papain/ pancreatin was 1 ∶ 1， reaction time was 8 h. Under such condition， the protein recovery ratio was 69.89% and the ADH activation ratio was 65.72%. Then corn anti-alcoholism peptides were separated and enriched by ethanol with various concentrations， and alcohol-sinking component with 80% ethanol had the highest ADH activation ratio（75.08%）. Sephadex G-15 was used for the second step separation and enrichment of the ethanol precipitation component of 80% ethanol， the four peaks were eluted， the overall recovery ratio was 84.52%. the percentage points of 4 classes were as followed： 24.36%， 47.79%， 3.25%， 9.12%， respectively. From the anti-alcohol experiments of 4 classes， the ADH activation ratio of the G4 fraction was as high as 90.12%， so corn peptide with facilitating alcohol metabolism activity was enriched efficiently. Ultra-performance liquid chromatography tandem mass-spectrometry （UPLC-ESI-Q-TOF-MS/MS） was applied for structural identification of the G4 fraction separated by Sephadex G-15. Six peptides with length ranging from 7 to 9 amino acids were found by searching against the corn data from Uniport database， including GQAIILP， GGTALPA， QAKGILP， YQQPIIGGA， VAAGGPA， VAVGGVP， 3 of them contained leucine， all of them contain alanine， These two amino acids played an important role in facilitating alcohol metabolism activity. In addition， Proline also had a positive effect on facilitating alcohol metabolism. In conclusion， it was suggested that the corn hydrolysates could be used in the potential functional ingredients with anti-alcoholism effect. This will greatly help to provide theoretical and technical guidance preparation of corn peptides with excellent anti-alcoholism properties.