CN108033997B - 具有抗氧化功能的多肽fkgpaca及其用途 - Google Patents
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Abstract
本发明公开了一种具有抗氧化功能的多肽FKGPACA，该多肽FKGPACA的氨基酸序列为：Phe‑Lys‑Gly‑Pro‑Ala‑Gys‑Ala。本发明还同时提供了上述具有抗氧化功能的多肽FKGPACA的用途：作为抗氧化剂。该多肽(抗氧化肽)具有较强的抗氧化作用，且具有安全无毒副作用、易于吸收等优点，可以作为抗氧化剂应用于食品添加剂，也可以作为功能成分，用于功能食品、保健品以及护肤品中。
Description
技术领域
本发明属于生物技术领域，特别涉及一个具有抗氧化活性的七肽及其应用。
背景技术
人体的生理代谢会产生一些含氧自由基，这些含氧自由基如不能及时清除，就会与人体的DNA、蛋白质等生物大分子物质发生反应，生成氧化物或过氧化物，影响人的正常生理代谢，进而产生癌症、冠心病、糖尿病等疾病。另外食品储运过程中，食品尤其是油脂类食品易发生氧化反应，造成食品的风味、营养成分的改变，甚至产生一些有害物质，降低食品品质和营养价值，对人体产生一定危害。
抗氧化剂能够清除自由基，减少自由基造成的氧化损伤，对人体健康，以及保持食品品质具有重要作用。目前，市场普遍使用的抗氧化剂如叔丁基对羟基茴香醚(BHA)、二叔丁基对甲酚(BHT)、叔丁基对二苯酚(TBHQ)，虽然能有效抑制氧化,但对人体健康也有一定的潜在安全影响，目前各国也都制定了使用限量。因此发现安全、健康的抗氧化剂越来越受到研究人员关注。
抗氧化肽指具有抗氧化活性的多肽，大多由3～20个氨基酸组成，抗氧化肽不仅安全、无毒副作用，且具有易吸收，易溶解、低黏度等良好的营养和加工特性，在食品、保健品及药品中具有良好的应用前景。目前已有很多研究者从动物或者植物组织中制备得到抗氧化肽。近几年研究也发现蚕蛹蛋白酶解肽具有一定的抗氧化活性，但具体的活性肽组分还不清楚，因此进一步从蚕蛹蛋白酶解液中分离、鉴定新的具有抗氧化生物活性的多肽，对于拓展新的功能食品、药品及食品添加剂领域具有一定的促进作用。
发明内容
本发明要解决的技术问题是提供一种新的具有良好抗氧化活性的肽序列及其用途。
为了解决上述技术问题，本发明一种具有抗氧化功能的多肽FKGPACA，该多肽(七肽)FKGPACA的氨基酸序列为：Phe-Lys-Gly-Pro-Ala-Gys-Ala。
该多肽FKGPACA从蚕蛹蛋白酶解液中鉴定而得；本发明的多肽FKGPACA可通过蚕蛹蛋白酶解液分离制备，也可由多肽合成公司通过化学合成制备。
本发明还提供了上述具有抗氧化功能的多肽FKGPACA的用途：作为抗氧化剂。
该抗氧化剂以此抗氧化肽(FKGPACA)为主要成分。
本发明提供的抗氧化肽具有较强的抗氧化作用，且具有安全无毒副作用、易于吸收等优点，可以作为抗氧化剂应用于食品添加剂，也可以作为功能成分，用于功能食品、保健品以及护肤品中。
本发明的多肽(七肽)FKGPACA，其用法和用量为：口服，用量约为1～2g/人.天。
本发明所报道的七肽序列是针对总抗氧化能力(ABTS法)和抑制羟自由基能力作为活性强弱指标，从而表现出具有抗氧化功能的特性。
本发明中所涉及的抗氧化活性的检测方法：
1)、总抗氧化能力测定：本实验使用南京建成生物工程研究所生产的总抗氧化能力测定试剂盒(ABTS法，产品序号A015-2)进行总抗氧化能力测定，即在96孔板上加入170μLABTS工作液，20μL应用液，再加入10μL样品溶液(浓度为1mg/mL)，混匀，室温反应6min，测定414nm的光密度值，以蒸馏水代替样品做空白。同时以不同浓度(浓度为0.15、0.3、0.6、0.9、1.2)的Trolox溶液做标准对照溶液，以相当于mmoL/LTrolox溶液来定义抗氧化能力，抗氧化能力以mmoL/L TE表示。
2)、羟自由清除能力测定：本实验使用南京建成生物工程研究所生产的羟自由基测试盒(产品序号A018)测定，具体操作步骤如下：
混匀，37℃反应1分钟，从加完试剂三开始到1分钟结束，立即加入显色剂终止反应(一次做一只管子)，室温放置20分钟后，550nm下测定吸光度。
计算公式：
具体实施方式
下面结合具体实施例对本发明进行进一步描述，但本发明的保护范围并不仅限于此：
实施例1
七肽FKGPACA在1mg/mL的浓度下的总抗氧化活性：
检测方法：将通过化学合成法获得的此七肽FKGPACA，进行活性检测(检测方法同上)。此时七肽FKGPACA浓度为1.0mg/mL。
结果：七肽FKGPACA在1.0mg/mL时的总抗氧化活性相当于0.83mmoL/L TE，抑制羟自由基能力为12.3U/mL。
通过实施例1的活性数据，说明此七肽FKGPACA具有较好的抗氧化活性，此七肽结构具有抗氧化活性未见报道，属于新的具有抗氧化活性功能肽。
对比例1-1、设定如表1所述的七肽，按照上述实验方法进行总抗氧化能力测定和羟自由清除能力测定(浓度为1.0mg/mL)；所得结果与本发明实施例1所得结果的对比，如表1所述。
表1
| 七肽 | 总抗氧化活性(mmoL/L TE) | 抑制羟自由基能力(U/mL) |
| FKGPACA(实施例1) | 0.83 | 12.3 |
| GGFGFGG | 0.012 | 1.6 |
| FKGCGCA | 0.16 | 3.1 |
| CKGPTCA | 0.11 | 2.1 |
| AGGPACA | 0.09 | 1.8 |
对比例1-2、设定如表2所述的多肽，按照上述实验方法进行总抗氧化能力测定和羟自由清除能力测定(浓度为1.0mg/mL)；所得结果与本发明实施例1所得结果的对比如表2所述。
表2
| 多肽 | 总抗氧化活性(mmoL/L TE) | 抑制羟自由基能力(U/mL) |
| FKGPACA(实施例1) | 0.83 | 12.3 |
| GPP | 0.36 | 6.8 |
| FK | 0.43 | 9.1 |
| KF | 0.49 | 9.5 |
实施例2
抗氧化肽的应用
1、按40％的抗氧化肽，28％的乳糖，30％结晶纤维素，2％的水充分混合后，通过压片机进行压片，制造成具有抗氧化作用的片剂。
2、抗氧化功能营养饮料：将饮料各成分按以下比例进行混合：果汁10％，柠檬酸0.3％，白砂糖10％，抗氧化肽为1％，水78.7％，制成具有抗氧化功能的果汁饮料。
最后，还需要注意的是，以上列举的仅是本发明的若干个具体实施例。显然，本发明不限于以上实施例，还可以有许多变形，比如不同蛋白质来源降解分离所得的FKGPACA结构及其衍生化结构。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形，均应认为是本发明的保护范围。
序列表
<110> 浙江省农业科学院
<120> 具有抗氧化功能的多肽 FKGPACA及其用途
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Phe Lys Gly Pro Ala Cys Ala
1 5
Claims (2)
1.具有抗氧化功能的多肽FKGPACA，其特征是：该多肽FKGPACA的氨基酸序列为：Phe-Lys-Gly-Pro-Ala-Gys-Ala。
2.如权利要求1所述的具有抗氧化功能的多肽FKGPACA的用途，其特征是：作为抗氧化剂。
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| CN109055468B (zh) * | 2018-08-21 | 2021-05-07 | 浙江大学 | 桑蚕蛹抗癌活性肽bpp-1及其制备方法与应用 |
| CN109182428B (zh) * | 2018-08-21 | 2021-05-07 | 浙江大学 | 桑蚕蛹抗癌活性肽bpp-2及其制备方法与应用 |
| CN110713516A (zh) * | 2019-11-14 | 2020-01-21 | 浙江省农业科学院 | 具有抗氧化功能的多肽vvc及其应用 |
| CN113527422B (zh) * | 2021-07-10 | 2024-02-09 | 浙江省农业科学院 | 具有抗氧化功能的多肽flcqwp及其用途 |
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