豌豆蛋白O^2-清除肽酶解优化及分离制备工艺研究

Study on Enzymatic Hydrolysis and Separation and Preparation of Pea Protein O^2- Scavenging Peptide

摘要: 本研究以含水量为30%的豌豆蛋白为主要原材料，选用中性蛋白酶对其进行酶解，利用响应曲面实验设计优化酶解条件及在此条件下的O²⁻清除率。采用透析对酶解产物进行分离，高效液相进行分子量的确定，以期得到具有高抗氧化活性的豌豆蛋白肽。经实验可得酶解最佳条件为：加酶量10.6% (E/S)、温度40.3℃、pH 7.02、底物浓度6.80% (g/ml)，此条件下，O²⁻清除率为78.1%。分子量小于1 ku的酶解液抗氧化活性最强，为81.3%。其分子量集中在200~800 u左右，为2~8肽。实验得到的模型准确可靠，酶解产物抗氧化性高，可用于指导实际生产中豌豆蛋白酶解条件的参数设定及分离方式的选定。透析能显著提高抗氧化肽对自由基的清除能力。

Abstract: The enzymatic conditions of pea protein with water content of 30% by neutral protease were optimized to the clearance ratio for O²⁻ by response surface methodology. The separation and molecular weight determination of the hydrolysates were determined by dialysis and high performance liquid phase, in order to get high antioxidant pea protein peptide. The optimum conditions were as follows: enzyme amount 10.6% (E/S), temperature 40.3˚C, pH 7.02 and the substrate concentration 6.80% (g/ml). Under this condition, the best O²⁻ clearance rate was 78.1%. The antioxidant activity of the hydrolysates with molecular weight less than 1 ku was 81.3%. And its molecular weight concentrated in about 200~800 u, was 2~8 peptides. The model obtained from the experiment is accurate and reliable, hydrolysates with high antioxidant activity, and it can be used to guide the parameter setting and separation selecting of pea protein in actual production. Dialysis could significantly improve clearance ability to free radicals of antioxidant peptides.