食品科学 ›› 2020, Vol. 41 ›› Issue (16): 37-44.doi: 10.7506/spkx1002-6630-20190822-231

• 食品化学 • 上一篇    

末水坛紫菜蛋白源抗氧化肽的制备、分离纯化与体外抗氧化活性

胡晓,于娇,陈胜军,杨贤庆,李来好,吴燕燕   

  1. (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实验室,广东 广州 510300;2.上海海洋大学食品学院,上海 201306)
  • 发布日期:2020-08-19
  • 基金资助:
    现代农业产业技术体系建设专项(CARS-50);广东省现代农业产业技术体系创新团队建设专项(2019KJ151); 农业部水产品加工重点实验室开放基金项目(NYJG201706); 广东省海洋与渔业发展专项资金(技术推广)项目(2017A0001); 国家水产品质量安全风险评估专项(GJFP201800904)

Preparation and Purification of Antioxidant Peptide from Porphyra haitanensis Protein and Its Antioxidant Activities in Vitro

HU Xiao, YU Jiao, CHEN Shengjun, YANG Xianqing, LI Laihao, WU Yanyan   

  1. (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:2020-08-19

摘要: 通过单因素试验、Box-Behnken设计响应面分析法优化酸性蛋白酶水解末水坛紫菜蛋白工艺,并测定不同水解时间的酶解产物体外抗氧化活性与分子质量分布。结果表明:当水解时间4 h、底物质量浓度2 g/100 mL、加酶量4 240 U/g、pH 3.7、温度54 ℃的条件下,水解度可高达11.40%。此外,通过调节水解时间以控制水解度,探讨了水解度与酶解产物抗氧化活性的关系,发现当水解时间为4 h、水解度为11.40%时,酶解产物对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除活性最强,其清除DPPH自由基的半抑制率质量浓度为0.68 mg/mL,分子质量小于1 kDa的小分子肽占总量的73.46%。同时,采用凝胶色谱层析对末水坛紫菜酶解产物进行了分离纯化,得到末水坛紫菜抗氧化肽(Porphyra haitanensis antioxidant peptide,PHAP),其质量浓度为1 mg/mL时对DPPH自由基的清除率为73.32%,对羟自由基的清除率为30.15%。最后通过建立H2O2诱导人肝癌HepG2细胞的氧化应激模型,证实了PHAP对HepG2细胞的氧化损伤具有修复作用,可增强超氧化物歧化酶的活性,降低脂肪氧化产物丙二醛的含量。

关键词: 坛紫菜;酶解;制备;分离纯化;抗氧化肽

Abstract: In this study, single factor experiment and response surface methodology (RSM) based on Box-Behnken design (BBD) were used together to optimize the hydrolysis process of Porphyra haitanensis protein by acid protease. The antioxidant activity and molecular mass distribution of the hydrolysates at different hydrolysis times were measured. The results showed that under the optimized hydrolysis conditions (substrate concentration of 2 g/100 mL, enzyme/substrate of 4 240 U/g, pH value of 3.7, temperature of 54 ℃, and hydrolysis time of 4 h), the highest degree of hydrolysis (DH) of 11.40% was obtained. The relationship between DH and antioxidant activity of the hydrolysates was investigated. It was found that the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the 4 h hydrolysate (DH = 11.40%) was the strongest with a half maximum inhibition concentration (IC50) of 0.68 mg/mL, and that small molecular mass peptides (< 1 kDa) accounted for 73.46% of the total peptides in the hydrolysate. This hydrolysate was fractionated by gel filtration chromatography, yielding an antioxidant peptide (named PHAP). PHAP at 1 mg/mL concentration scavenged DPPH and hydroxyl radicals by 73.32% and 30.15%, respectively. It was confirmed that PHAP could repair H2O2-induced oxidative damage in HepG2 cells by enhancing the activity of superoxide dismutase and decreasing the content of malondialdehyde.

Key words: Porphyra haitanensis; enzymatic hydrolysis; preparation; purification; antioxidant peptide

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