梯次酶解法制备核桃抗氧化肽工艺优化

Optimization of preparation process of Juglans regia L. antioxidant peptide by echelon enzymatic hydrolysis

中文关键词:  脱脂核桃粕粉  核桃酶解工艺  响应面分析  正交试验

英文关键词:defatted Juglans regia L. meal powder  Juglans regia L. enzymatic hydrolysis process  response surface analysis  orthogonal experiment

中文摘要:

      目的 优化梯次酶解法制备核桃抗氧化肽的工艺条件。方法 以脱脂核桃粉为原料, 依次采用碱性蛋白酶、中性蛋白酶和酸性蛋白酶梯次酶解方式制备核桃抗氧化肽, 以抗氧化活性和水解度为指标, 通过单因素结合正交试验及响应面优化酶解工艺。结果 中性蛋白酶的最佳工艺为酶解温度51 ℃, pH为6.87, 底物浓度5%, 酶添加量28000 U, 酶解时间3 h, 酸性蛋白酶酶解的最佳工艺为酶解温度50.0 ℃, pH为5, 底物浓度5.0%, 酶添加量30000 U, 酶解时间3 h, 水解度为43.74%, 2,2’-联氮基双(3-乙基苯并噻唑啉-6-磺酸)二铵盐[diammonium 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonate), ABTS]阳离子自由基清除率92.14%, 1,1-二苯基-2-苦基肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)清除率为87.55%。结论 本研究优化了蛋白梯次酶解工艺, 可作为核桃抗氧化肽制备的一种可行策略。

英文摘要:

      Objective To optimize the process conditions for the preparation of Juglans regia L. antioxidant peptides by echelon enzymatic hydrolysis. Methods Juglans regia L. antioxidant peptides were prepared from defatted Juglans regia L. powder by alkaline protease, neutral protease and acid protease in a consecutive manner. The enzymatic hydrolysis process was optimized by single factor combined with orthogonal experiment and response surface method with antioxidant activity and hydrolysis degree as indexes. Results The optimum process of neutral protease was enzymatic hydrolysis temperature 51 ℃, pH 6.87, substrate concentration 5%, enzyme addition 28000 U, enzymatic hydrolysis time 3 h. The optimum process of acid protease was enzymatic hydrolysis temperature 50.0 ℃, pH 5, substrate concentration 5.0%, enzyme addition 30000 U, enzymatic hydrolysis time 3 h. The degree of hydrolysis was 43.74%, the cation radical scavenging rate of 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) was 92.14%, and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging rate was 87.55%. Conclusion This study optimizes the protein echelon enzymatic hydrolysis process, which can be used as a feasible strategy for the preparation of Juglans regia L. antioxidant peptides.