脱脂羊脑蛋白水解多肽的分离纯化及抗氧化活性  

Purification and Antioxidant Activity of Peptides Derived from Defatted Goat Brain Protein

作　　者：常飞[1] 杨雪果[1] 肖士成 蒋鹏飞[1] 黄慧娜[1] 段旭昌[1] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100 [2]陕西杨陵华兴羊产业科技发展有限公司,陕西杨凌712100

出　　处：《食品科学》2016年第1期33-39,共7页Food Science

基　　金："十二五"国家科技支撑计划项目(2011BAD28B05-3)

摘　　要：为制备羊脑蛋白抗氧化肽,本实验对脱脂羊脑蛋白含量及氨基酸组成进行了分析;采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)对枯草芽孢杆菌中性蛋白酶不同酶解时间的酶解液分子质量进行了分析;采用交联葡聚糖凝胶Sephadex G-25和Sephadex G-15对羊脑酶解产物进行了逐级分离纯化,以羟自由基(·OH)和亚硝酸根离子清除能力为指标对分离组分进行抗氧活性评价,并对纯化后的组分抗氧化活性进行了测定。结果表明,脱脂羊脑粉中蛋白含量为60.55%,在测定的17种氨基酸中,谷氨酸和天冬氨酸这两种酸性氨基酸含量最高,且含有7种必需氨基酸;羊脑蛋白经酶解后,分子质量集中在10 k D以下;经Sephadex G-25纯化后,得到了6个组分,其中组分F4的抗氧化活性最强,组分F4经Sephadex G-15纯化后,得到3个组分,其中组分F4-2的抗氧化活性最强,组分F4-2对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、·OH、超氧阴离子自由基(O_2^-·)、亚硝酸根离子的半数抑制率IC_(50)分别为1.64、2.47、7.98、5.14 mg/m L。This research was designed to prepare antioxidant peptides derived from goat brain protein. The protein content and amino acids composition of defatted goat brain were determined. The molecular mass distribution of protein hydrolysates by neutral protease from Bacillus subtilis at different times was analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis（SDS-PAGE）, and goat brain protein hydrolysates were separated and purified by Sephadex G-25 gel filtration chromatography and Sephadex G-15 gel filtration chromatography. The antioxidant activity of the resulting fractions was evaluated by DPPH, hydroxyl radical, superoxide anion radical, and nitrite radical scavenging assays. The results showed that the protein content of defatted goat brain was 60.55% and contained seven essential amino acids among which glutamic acid and aspartic acid were the most predominant. The molecular mass of the hydrolysates was below 10 k D. After purification by Sephadex G-25 gel filtration chromatography, the six fractions of the hydrolysates were obtained and F4 fraction showed the strongest antioxidant activity. F4 was further purified into three fractions by Sephadex G-15 gel filtration chromatography, including F4-2 showing the strongest antioxidant activity. The IC_（50） values of F4-2 for scavenging DPPH, hydroxyl radical, superoxide anion radical, and nitrite radicals were 1.64, 2.47, 7.98 and 5.14 mg/m L respectively.

关 键 词：羊脑蛋白 多肽 分离纯化 抗氧化活性 

分 类 号：TS209[轻工技术与工程—食品科学]