罗非鱼肌浆蛋白源抗氧化肽的制备、分离纯化及其体外抗氧化活性

doi:10.7506/spkx1002-6630-20200116-193

食品科学 ›› 2021, Vol. 42 ›› Issue (3): 63-70.doi: 10.7506/spkx1002-6630-20200116-193

罗非鱼肌浆蛋白源抗氧化肽的制备、分离纯化及其体外抗氧化活性

胡晓，周雅，陈星星，李来好，杨贤庆，陈胜军，吴燕燕，杨少玲

（1.中国水产科学研究院南海水产研究所，农业农村部水产品加工重点研究室，广东广州 510300；2.上海海洋大学食品学院，上海 201306）

发布日期:2021-02-25
基金资助:
现代农业产业技术体系建设专项（CARS-46）；茂名市引进创新创业团队项目（200201095834980）；广东省基础与应用基础研究基金项目（2019A1515011588）；广东省自然科学基金项目（2017A030313164）；广东省科技创新战略专项资金（纵向协同管理方向）计划项目（2018S0044）；中国水产科学研究院基本科研业务费资助项目（2020TD69）

Preparation, Purification and in Vitro Evaluation of Antioxidant Peptides from Tilapia (Oreochromis niloticus Sarcoplasmic Protein

HU Xiao, ZHOU Ya, CHEN Xingxing, LI Laihao, YANG Xianqing, CHEN Shengjun, WU Yanyan, YANG Shaoling

(1. Key Laboratory of Aquatic Product Processing, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China; 2. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China)

Published:2021-02-25

摘要/Abstract

摘要： 采用木瓜蛋白酶对罗非鱼肌肉蛋白组分（肌浆蛋白、肌原纤维蛋白、基质蛋白）进行酶解，研究罗非鱼不同蛋白组分酶解产物的抗氧化活性，并通过超滤、凝胶过滤色谱、反相高效液相色谱对高活性肌浆蛋白组分抗氧化肽进行分离纯化，同时对纯化后的抗氧化肽氨基酸组成予以分析。结果表明：罗非鱼肌浆蛋白酶解物（tilapia sarcoplasmic protein hydrolysates，TSPH）抗氧化活性高于肌质与肌原纤维蛋白酶解物，当肌浆蛋白酶解4 h时，TSPH（6 mg/mL）对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基、羟自由基清除率及其还原力均达到最高，分别为84.61%、50.01%和0.629；经超滤分离后，其分子质量小于5 kDa的组分具有较高的抗氧化活性；该组分经凝胶色谱分离后，富集得到E1～E4共4 个组分，其中E3组分（0.8～0.3 kDa）的抗氧化活性最高， 3 mg/mL E3组分对DPPH自由基清除率达83.61%，还原力为0.953；E3组分再经反相高效液相色谱分离纯化后，富集得到F1～F9共9 个组分，其中F4组分的抗氧化活性最高，其DPPH自由基的半清除质量浓度为0.78 mg/mL，且经氨基酸组成分析发现色氨酸（Trp）、甘氨酸（Gly）、组氨酸（His）为其含量较高的氨基酸，相对含量分别高达31.20%、27.25%和8.97%。

关键词: 罗非鱼；肌浆蛋白；抗氧化肽；制备；分离纯化

Abstract: Papain was selected to hydrolyze three tilapia muscle proteins (sarcoplasmic, myofibrillar and stroma proteins) and the antioxidant activities of the resulting hydrolysates were investigated. The antioxidant peptides derived from tilapia sarcoplasmic protein were separated and purified by using ultrafiltration, gel filtration chromatography and reversed-phase high performance liquid chromatography (RP-HPLC), and their amino acid composition was determined. The results showed that the antioxidant activity of tilapia sarcoplasmic protein hydrolysates (TSPH) was higher than that of myofibrillar and stroma protein hydrolysates. The TSPH obtained by after 4 h of hydrolysis (6 mg/mL) exhibited the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate (84.61%), hydroxyl radical scavenging rate (50.01%) and reducing power (0.629) of the various hydrolysates tested. It was then separated by ultrafiltration, and the fraction with molecular masses of less than 5 kDa was found to possess high antioxidant activity. This fraction was further separated into four sub-fractions (E1–E4) by gel filtration chromatography. It was found that sub-fraction E3 (0.8–0.3 kDa) possessed the highest antioxidant activity, which was determined to be 83.61% and 0.953 in DPPH radical scavenging capacity and reducing power at a dose of 3 mg/mL. Finally, E3 was separated into nine fractions (F1–F9) by RP-HPLC, and F4 exhibited the highest DPPH radical scavenging activity with a half-maximal inhibitory concentration (IC50) of 0.78 mg/mL. Its amino acid composition was dominated by Trp, Gly and His, which accounted for 31.20%, 27.25% and 8.97% of the total amino acids, respectively.

Key words: tilapia; sarcoplasmic protein; antioxidant peptides; preparation; purification

中图分类号:

TS254.1

胡晓，周雅，陈星星，李来好，杨贤庆，陈胜军，吴燕燕，杨少玲. 罗非鱼肌浆蛋白源抗氧化肽的制备、分离纯化及其体外抗氧化活性[J]. 食品科学, 2021, 42(3): 63-70.

HU Xiao, ZHOU Ya, CHEN Xingxing, LI Laihao, YANG Xianqing, CHEN Shengjun, WU Yanyan, YANG Shaoling. Preparation, Purification and in Vitro Evaluation of Antioxidant Peptides from Tilapia (Oreochromis niloticus Sarcoplasmic Protein[J]. FOOD SCIENCE, 2021, 42(3): 63-70.

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罗非鱼肌浆蛋白源抗氧化肽的制备、分离纯化及其体外抗氧化活性

Preparation, Purification and in Vitro Evaluation of Antioxidant Peptides from Tilapia (Oreochromis niloticus Sarcoplasmic Protein

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