菜籽饼超微粉酶解制备抗氧化活性肽
TS201.1; 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)...
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| Published in | 农业工程学报 Vol. 40; no. 4; pp. 284 - 291 |
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| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
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
01.02.2024
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1002-6819 |
| DOI | 10.11975/j.issn.1002-6819.202311100 |
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| Abstract | TS201.1; 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. |
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| AbstractList | TS201.1; 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. |
| Abstract_FL | 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 using alkaline protease and flavourzyme.Both hydrolytic products showed better ABTS free radical scavenging and total antioxidant at 1 mg/mL,but the lower DPPH free radical scavenging.Moreover,the hydrolytic products of flavourzyme were mainly composed of small peptides with a molecular weight of less than 1 kDa.The alkaline protease hydrolysis was dominated by peptides from 5 to 10 kDa fractions.The antioxidant activities of peptides were related to their molecular weight distribution.Specifically,the low molecular weight fraction of alkaline protease hydrolysis(＜1 kDa)showed better ABTS free radical scavenging at(89.73±0.33)%and total antioxidant activity at(2 669.5±2.82)μmol/g at 1 mg/mL.Furthermore,the active peptide fraction(J3)was achieved using Sephadex G-15 gel filtration chromatography.The peptide content of the J3 fraction was about 32.5%.Five peptides were selected,namely RF,LPF,PAGPF,VF,and LATPF.All five peptides showed a content of over 50%hydrophobic amino acids with phenylalanine as the N-terminal.Specially,RAGPF,VF,and LATPF were newly identified.In summary,the rapeseed meal can be used as a source of prepared antioxidant peptides,which would broaden their applicability.This finding can provide a theoretical reference to further develop the rapeseed/canola meal protein resources through direct enzymatic hydrolysis of rapeseed meal ultra-fine powder. |
| Author | 李媛媛 梁亮 裴海生 翟晓娜 胡龙彪 张志民 胡雪芳 王萍 班世栋 |
| AuthorAffiliation | 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 |
| AuthorAffiliation_xml | – name: 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 |
| Author_FL | ZHANG Zhimin ZHAI Xiaona LI Yuanyuan BAN Shidong HU Xuefang HU Longbiao WANG Ping LIANG Liang PEI Haisheng |
| Author_FL_xml | – sequence: 1 fullname: ZHAI Xiaona – sequence: 2 fullname: HU Longbiao – sequence: 3 fullname: LI Yuanyuan – sequence: 4 fullname: LIANG Liang – sequence: 5 fullname: HU Xuefang – sequence: 6 fullname: ZHANG Zhimin – sequence: 7 fullname: BAN Shidong – sequence: 8 fullname: WANG Ping – sequence: 9 fullname: PEI Haisheng |
| Author_xml | – sequence: 1 fullname: 翟晓娜 – sequence: 2 fullname: 胡龙彪 – sequence: 3 fullname: 李媛媛 – sequence: 4 fullname: 梁亮 – sequence: 5 fullname: 胡雪芳 – sequence: 6 fullname: 张志民 – sequence: 7 fullname: 班世栋 – sequence: 8 fullname: 王萍 – sequence: 9 fullname: 裴海生 |
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| Keywords | 膜分离 films 菜籽饼 flavourzyme 氨基酸序列 风味蛋白酶 enzymes 抗氧化活性 antioxidant activity 碱性蛋白酶 amino acid sequence 酶 rapeseed meal alkaline protease 膜 membrane separation |
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| Publisher | 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 |
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| Title | 菜籽饼超微粉酶解制备抗氧化活性肽 |
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