胰蛋白酶制备鹰嘴豆抗氧化肽的条件优化  

Optimal Preparation of Antioxidant Peptides by Enzymatic Hysrolysis of Chickpea Protein Isolate with Trypsin

作　　者：施文卫 王伟[2] 胡冰[1] 周莉[1] 曾晓雄[1] 孙怡[1] 

机构地区：[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

出　　处：《食品科学》2016年第15期185-191,共7页Food Science

基　　金：江苏高校优势学科建设工程资助项目

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℃、p H 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 k D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。Response surface methodology was employed to optimize the preparation conditions of antioxidant peptidesby enzymatic hydrolysis of chickpea protein isolate （CPI）, prepared by alkaline extraction and acid precipitation fromchickpea seeds, with trypsin. The response variable was the percentage scavenging of superoxide anion radical. The resultsshowed that all samples collected throughout the hydrolysis process had significant antioxidant activity and the optimalconditions were determined as substrate concentration 2%, enzyme/substrate （[E]/[S]） ratio 1 800 U/g, reaction time35 min, temperature 32 ℃ and pH 7.0. Under these conditions, the percentage scavenging of superoxide anion radicalby chickpea antioxidant peptides was 67.59%. Chickpea peptide fractions with molecular weight cut-off of 1 kD fromchickpea protein hydrolysates by ultrafiltration were analyzed by liquid chromatography-mass spectroscopy （LC-MS）.A pentapeptide with molecular weight of 668.40 in chickpea peptide fractions was achieved, whose amino acidsequence was determined to be RQLPR.

关 键 词：鹰嘴豆分离蛋白 胰蛋白酶 抗氧化 响应面法 

分 类 号：TS201.2[轻工技术与工程—食品科学]