本文旨在应用响应曲面实验设计优化酶联合挤压豌豆蛋白酶解条件，以期得到最优的抗氧化肽和准确的酶解预测模型，并分离得到具有高自由基清除率的豌豆蛋白抗氧化肽。以含水量为30%的豌豆蛋为原料，以O2-.清除率为评价指标，利用响应曲面设计优化中性蛋白酶挤出物的酶解条件，得出酶解最佳条件为：加酶量12.23%、温度40.23 ℃、pH 6.99、底物浓度6.92%。在此条件下，O2-.清除率为75.93%。采用透析和高效液相相结合的方法，对酶解产物进行分离及分子量的确定。分离得到小于1 ku、1~3.5 ku、3.5~7 ku和大于7 ku的酶解液。经测定发现分子量小于1 ku的酶解液抗氧化活性最强，达78.31%，其分子量集中在200~800 u左右，为2~8肽。经验证，所建模型预测准确可靠，酶解产物抗氧化性高，可为相关生产和工艺产品开发提供参考价值。透析可显著提高肽的自由基清除率。

The purpose of this paper was to optimize the enzymolysis conditions of pea protein with enzyme combined extrusion by response surface methodology, so as to obtain the optimal antioxidant peptide with high free radicals scavenging capacity and an accurate enzymolysis prediction model. The enzymolysis conditions of neutral protease were optimized with the scavenging ratio of O2-.as evaluation index and with water content of 30% pea protein as raw by response surface methodology. The optimal enzymolysis conditions were as follows: enzyme amount 12.23% (E/S), temperature 40.23℃, pH 6.99, and substrate concentration 6.92% (g/mL). Under these conditions, the O2-.scavenging rate was 75.93%. The separation and determination of molecular weight of enzymolysis products were studied by the combination of dialysis and HPLC. The enzymolysis liquids with molar weights <1 ku, 1~3.5 ku, 3.5~7 ku, and<7 ku were separated. The results showed that the antioxidant activity of enzymolysis liquid less than 1 ku was the strongest, reaching 78.31%. The molecular weight was concentrated in about 200 ~ 800 u as 2 ~ 8 peptide. It is confirmed that the prediction model is accurate and reliable, and hydrolysates has high antioxidant activity, which can provides the reference value for the development of related production and process products. Dialysis can significantly improve the free radical scavenging rate.

山东省重点研究开发计划（2017YYSP033）；山东省自然科学基金联合专项（ZR2016BL03）