Abstract:

To optimize the stepwise double-enzyme hydrolysis condition of cottonseed protein by Alacalase protease and Flavourzyme protease. Plackett-Burman (PB) design and Box-Behnken Design were applied to screening and optimizing 9 factors for the hydrolysis of cottonseed protein by Alacalase protease and Flavourzyme protease, in which the degree of hydrolysis (DH) was regarded as an index. It was shown that the optimum hydrolysis conditions were as follows: substrate concentration 9.66%(w/v), firstly, Alacalase hydrolyzed for 150 min under the conditions of 60.5℃, pH 8.0 and enzyme/substrate 25000 U/g, then Flavorzyme hydrolyzed for 120 min under the conditions of 50℃, pH 6.0 and enzyme/substrate 26576 U/g. Under this condition, the DH of enzyme-hydrolyzed product was 42.73%. The experimental results provide the basic data for the utilization of cottonseed protein.