Abstract: In this work, a bioactive peptide with inhibitory effect on dipeptidyl peptidase-IV (DPP-IV) was prepared from corn protein meal. The protease with the best inhibitory effect on DPP-IV was selected by enzymatic preparation technique, and the optimal enzymatic hydrolysis conditions were determined by single factor and response surface optimization test. The molecular weight distribution, ultrafiltration classification and amino acid composition of the obtained inhibitory peptides were further analyzed. The results showed that the neutral protease had the best effect on the preparation of DPP-IV inhibitory peptide from corn protein source. The optimal enzymolysis conditions were substrate concentration of 10% (w/v), enzyme dosage of 640 U/g, enzymolysis time of 157 min, enzymolysis temperature of 45 ℃ and enzymolysis pH of 7.0. The inhibitory concentration (IC₅₀) value of DPP-IV for the inhibitory peptide obtained under this condition was 0.53±0.06 mg/mL. After ultrafiltration, the molecular weight of components less than 1 kDa accounted for 58.9% of the original inhibitory peptide, and the inhibitory activity against DPP-IV was higher than that of the original inhibitory peptide, which was related to its amino acid composition. The results of enzymatic reaction kinetics showed that the inhibition type of DPP-IV was competitive and reversible. The results of this study provide a theoretical basis for the development and utilization of DPP-IV inhibitory peptide from corn protein source, and provide a new idea for the high-value utilization of corn protein.