Abstract:To enhance the utilization of wheat resources， a two-step enzymatic hydrolysis method was employed to process wheat gluten. The effects of four proteases on the protein yield from wheat gluten were investigated. Protease， which resulted in higher protein yields， was selected for single-factor and orthogonal tests，which was conducted to optimze the two-step enzymatic hydrolysis sequence and duration. The results indicated that the optimal conditions of the first step was neutral protease hydrolysis at pH7.0， enzyme addition of 8 000 U/g， substrate concentration of 10%， hydrolysis temperature of 55 ℃， and hydrolysis time of 1.5 h. The second step was alkaline protease hydrolysis at pH8.0， enzyme addition of 8 000 U/g， hydrolysis temperature of 60 ℃， and hydrolysis time of 1.5 h. Under the optimized conditions， the protein yield exceeded 90%.