胰蛋白酶制备鹰嘴豆抗氧化肽的条件优化.pdfVIP

※生物工程 食品科学 2016, Vol.37, No.15 185 胰蛋白酶制备鹰嘴豆抗氧化肽的条件优化 1 2 1 1 1 1,* 施文卫 ，王 伟，胡 冰，周 莉，曾晓雄，孙 怡 （1.南京农业大学食品科技学院，江苏 南京 210095 ；2.新疆农业大学食品科学与药学学院，新疆 乌鲁木齐 830052 ） 摘  要：以碱溶酸沉法制备的鹰嘴豆分离蛋白为酶解底物，以超氧阴离子自由基清除率为响应值，采用响应面法优 化胰蛋白酶制备鹰嘴豆抗氧化活性肽的条件。结果表明：在整个酶解过程中，所制备的鹰嘴豆抗氧化肽具有显著的 抗氧化活性，胰蛋白酶制备鹰嘴豆抗氧化肽的最佳酶解条件为：底物质量分数2%、加酶量（[E]/[S] ）1 800 U/g、温 度32 ℃、pH 7.0 、时间35 min 。此条件下所制备的鹰嘴豆抗氧化肽的超氧阴离子自由基清除率为67.59%。鹰嘴豆分 离蛋白水解液经超滤膜系统纯化获得了截留相对分子质量1 kD以下的水解液，并通过液相色谱-质谱联用分析，发 现了一种相对分子质量为668.40、序列为RQLPR 的亲水性五肽。 关键词：鹰嘴豆分离蛋白；胰蛋白酶；抗氧化；响应面法 Optimal Preparation of Antioxidant Peptides by Enzymatic Hysrolysis of Chickpea Protein Isolate with Trypsin 1 2 1 1 1 1,* SHI Wenwei , WANG Wei , HU Bing , ZHOU Li , ZENG Xiaoxiong , SUN Yi (1. College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; 2. College of Food Science and Pharmacy, Xinjiang Agricultural University, Ürümqi 830052, China) Abstract: Response surface methodology was employed to optimize the preparation conditions of antioxidant peptides by enzymatic hydrolysis of chickpea protein isolate (CPI), prepared by alkaline extraction and acid precipitation from chickpea seeds, with trypsin. The response variable was the percentage scavenging of superoxide anion radical. The results showed that all samples collected throughout the hydrolysis process had significant antioxidant activity and the optimal conditions were determined as substrate concentration 2%, enzyme/substrate ([E]/[S]) ratio 1 800 U/g, reaction time 35 min, temperature 32 ℃ and pH 7.0. Under these conditions, the percentage scavenging of sup