菜籽饼超微粉酶解制备抗氧化活性肽  

Preparing antioxidant peptides from ultra-fine powder of rapeseed meal

作　　者：翟晓娜 胡龙彪 李媛媛 梁亮 胡雪芳 张志民 班世栋 王萍 裴海生 ZHAI Xiaona;HU Longbiao;LI Yuanyuan;LIANG Liang;HU Xuefang;ZHANG Zhimin;BAN Shidong;WANG Ping;PEI Haisheng(Key Laboratory of Agro-Products Primary Processing,Ministry of Agriculture and Rural Affairs,Academy of Agricultural Planning and Engineering,MARA,Beijing 100121,China)

机构地区：[1]农业农村部规划设计研究院农业农村部农产品产地初加工重点实验室,北京100121

出　　处：《农业工程学报》2024年第4期284-291,共8页Transactions of the Chinese Society of Agricultural Engineering

基　　金：农规英才计划(QNYC-2021-10)新型功能原料绿色制备关键技术研究及应用。

摘　　要：为探究菜籽饼超微粉直接酶解制备抗氧化肽技术,该研究以菜籽饼超微粉为原料,利用碱性蛋白酶、风味蛋白酶对其进行酶解,通过测定2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)自由基清除能力、1,1-二苯基-2-苦基肼自由基清除能力和总抗氧化能力表征酶解产物体抗氧化活性,使用膜分离、Sephadex G-15凝胶层析对酶解产物进行分离纯化,利用高效液相色谱-串联质谱联用技术分析鉴定抗氧化活性组分的氨基酸序列。结果表明,风味蛋白酶酶解产物的肽含量较高,并以分子量小于1 kDa肽段为主,碱性蛋白酶解产物以5~10 kDa肽段为主,但同等浓度下(1 mg/mL)的碱性蛋白酶酶解产物抗氧化活性较高;膜分离后,同种酶解产物中低分子量(<1 kDa)组分具有更好的ABTS自由基清除能力和总抗氧化活性;通过凝胶层析分离纯化得到的高抗氧化活性组分J3经氨基酸序列鉴定,结合PeptideRanker生物活性预测筛选得到5个肽段,肽段的N-端均为苯丙氨酸且肽段中疏水性氨基酸的含量占比达50%以上。该结果可为深入开发菜籽饼蛋白资源提供理论参考。Plant-derived peptides have been the high-quality resources of plant protein,because of their natural availability,unique nutritional and physiological functions,particularly without any toxic or side effects.The high protein content can also be balanced in the composition of amino acids.Among them,rapeseed/canola meal is one of the by-products in the extraction of rapeseed oil,while commonly used as animal feed,resulting in low economic benefit.Rapeseed protein hydrolysates have been proven to process antioxidant activity.High-value use of rapeseed protein source is very necessary.Enzymatic hydrolysis can be preferred to produce the bioactive peptides,due to the high specificity,mild conditions,and no by-products.However,the preparation of rapeseed active peptides usually requires the rapeseed protein prior to enzymatic hydrolysis,which is relatively complicated and consumed.Our previous study found that the rapeseed meals with superfine grinding effectively increased their protein dissolution rate.In this study,more attempts were made to directly prepare the active peptides by enzymatic approaches.The raw material was taken as the rapeseed meal powder that was treated with superfine grinding.A comparison was made on the molecular weight distribution and antioxidant activity of the products after rapeseed meal enzymatic hydrolysis by different enzymes.Furthermore,the high-purity antioxidant peptides were purified from the enzymatically hydrolyzed products by membrane separation and Sephadex G-15 column chromatography.The antioxidant activity was evaluated by DPPH(2,2-Diphenyl-1-picrylhydrazyl)radical scavenging capacity,ABTS(2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)radical scavenging capacity,and total antioxidant activity(T-AOC).Finally,the amino acid sequence of the peptide was determined by LC-MS/MS.The results showed that the proteolytic products of rapeseed/canola meal with the higher peptide content were obtained by direct proteolytic hydrolysis of the rapeseed/canola mean ultrafine powder u

关 键 词：酶 膜 菜籽饼 风味蛋白酶 碱性蛋白酶 膜分离 氨基酸序列 抗氧化活性 

分 类 号：TS201.1[轻工技术与工程—食品科学]