Abstract: 【Objective】The production of short peptide from zein was investigated to greatly promote the application of zein.【Method】The conditions such as extraction time, temperature, ethanol concentration and ratio of liquid to solid were optimized by orthogonal test. Zein was hydrolyzed by protease K and alkaline protease in this study.【Result and conclusion】The optimal conditions of extracting zein were as follows: the ratio of solid to liquid was 1∶4; the extracting time was 3 hours; the extracting temperature was 60 ℃ and the concentration of ethanol was 75%. Results showed that the zein could be hydrolyzed more efficiently by combining two enzymes than by only using one of the two enzymes. Zein hydrolyzed by protease K and alkaline protease in turn is the best enzymatic hydrolysis method.