酶法水解荞麦清蛋白制备抗氧化活性肽的研究  

Preparation of Antioxidant Peptides from Buckwheat Albumin by Enzymatic Hydrolysis

作　　者：付媛[1,2] 张美莉[1] 侯文娟[1] 

机构地区：[1]内蒙古农业大学食品科学与工程学院,内蒙古呼和浩特010018 [2]内蒙古农业大学理学院,内蒙古呼和浩特010018

出　　处：《食品科学》2009年第15期142-147,共6页Food Science

基　　金：国家自然科学基金项目(30760151)

摘　　要：以水解度和对O·清除率为指标,采用正交试验对碱性蛋白酶酶解荞麦清蛋白工艺进行探索,实验结果表明,最佳工艺为:温度50℃、pH10.0、底物浓度20g/L、酶与底物比为10%。在此条件下水解,水解度可达17.57%,对O·清除率达37.06%。对酶解产物进行Sephadex G-25柱层析分离,得到组分Ⅰ和组分Ⅱ。其分子量范围分别为20.89～204.2kD,219～691D。组分Ⅰ和组分Ⅱ对O·、·OH、DPPH·三种自由基的清除能力要强于清蛋白酶解产物,且组分Ⅱ的清除能力最强。组分Ⅱ对三种自由基清除能力表现为O·>·OH>DPPH·,其IC50分别为:0.072、0.19、0.32mg/ml。We investigated the enzymatic hydrolysis of buckwheat albumin to produce antioxidant peptides. In order to obtain the highest degree of hydrolysis （DH） and O2^-· scavenging activity of buckwheat albumin hydrolysates, an optimal enzyme was screened from alkaline protease, trypsin and papain, and the parameters of hydrolysis catalyzed by screened enzyme were optimized by single-factor method and orthogonal array design. Subsequendy, the buckwheat albumin hydrolysates were separated by Sephadex G-25 column chromatography, and fractions obtained were analyzed for their molecular weight and O2^-·,· OH and DPPH · scavenging activities. Higher DH and O2^-· scavenging activity of buckwheat albumin hydrolysate were achieved using alkaline protease in comparison with other two enzymes. The optimal parameters for improved hydrolysis of buckwheat albumin by alkaline protease were determined as follows： temperature 50 ℃, pH 10, substrate concentration 20 g/L, enzyme concentration 10% and hydrolysis time 2 h, and the DH reached 17.57% and the O2^-· scavenging ratio was 37.06% under these conditions. Two fractions, namely fractions Ⅰ and Ⅱ, were obtained from the hydrolysates, whose molecular weights were 20.89 - 204.2 kD and 219 - 691 D, respectively. The O2^-·, · OH and DPPH · scavenging activities of fractions Ⅰ and Ⅱ were all higher than those of the hydrolysates and higher activities of scavenging the three free radicals were found in fraction Ⅱ when compared with fraction Ⅰ The radical scavenging efficiency of fraction Ⅱ decreased in the following order of O2^-· 〉 · OH 〉 DPPH · and the IC50s were 0.072, 0.19 and 0.32 mg/ml, respectively.

关 键 词：荞麦 清蛋白 活性肽 蛋白酶 抗氧化 自由基 

分 类 号：TS201.25[轻工技术与工程—食品科学]