This essay compared the enzymatic hydrolysis effect of glutamine peptide prepared from wheat gluten with Alcalase 2. 4 L FG in aqueous system and ethanol system respectively. According to the hydrolysates' protein content,degree of hydrolysis,peptides content and amide nitrogen content,it turned out that the best system for enzymatic hydrolysis was ethanol system of single concentration. Moreover,using the amide nitrogen content as the main index,along with the changes in the degree of hydrolysis,we did the single-factor and orthogonal experiments to determine the optimum conditions for enzymatic hydrolysis. The results showed that the degree of hydrolysis was 14. 26% and amide nitrogen content was 3. 07 mmol/g under the conditions as follows: ethanol concentration was 40 %,temperature was 47. 5 ℃,substrate concentration was 14 %,the amount of enzyme was 7 000 U/g,and the reaction time was 3. 5 h.