研究Alcalase水解蛋白酶和Flavourzyme风味蛋白酶分步水解脱酚棉籽蛋白的最佳条件。以水解度为评价指标，采用Plackett-Burman(PB)设计和Box-Behnken响应面分析法对影响双酶分步水解棉籽蛋白的9个工艺条件进行筛选优化。双酶分步水解脱酚棉籽蛋白的最优操作条件为：先用Alcalase水解蛋白酶在底物浓度(w/v) 9.66%，温度60.5℃、pH 8.0、酶用量25000 U/g的条件下水解150 min，灭酶后再用Flavourzyme风味蛋白酶在温度50℃、pH 6.0、酶用量26576 U/g的条件下水解120 min。在此条件下水解液的水解度可达到42.73%。试验结果可为棉籽蛋白多肽的开发利用提供依据。

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.