食品科学 ›› 2020, Vol. 41 ›› Issue (21): 113-120.doi: 10.7506/spkx1002-6630-20191024-255

• 营养卫生 • 上一篇    下一篇

体外模拟胃肠消化及碱性蛋白酶处理后蛋清肽抗氧化活性差异及肽序列解析

马思彤,刘静波,张婷,王莹,孙惠炎,魏以恒,王寒颖,雷洪辉,刘博群   

  1. (1.吉林省营养与功能食品重点实验室,吉林 长春 130062;2.吉林大学食品科学与工程学院,吉林 长春 130062;3.吉林工程技术师范学院食品工程学院,吉林 长春 130052)
  • 发布日期:2020-11-23
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFD0400300);吉林省省校共建计划专项(SF2017-6-4);吉林省科技厅技术攻关项目(20190301015NY);吉林大学创新团队项目(2017TD-29);中央高校基本科研业务费专项资金项目

Differences in Antioxidant Activity and Sequence Analysis of Egg White Peptides Derived from Simulated Gastrointestinal Digestion and Alkaline Protease Treatment

MA Sitong, LIU Jingbo, ZHANG Ting, WANG Ying, SUN Huiyan, WEI Yiheng, WANG Hanying, LEI Honghui, LIU Boqun   

  1. 1. Jilin Provincial Key Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, China; 2. College of Food Science and Engineering, Jilin University, Changchun 130062, China; 3. College of Food Engineering, Jilin Engineering Normal University, Changchun 130052, China
  • Published:2020-11-23

摘要: 为明确体内消化和体外加工处理后蛋白肽抗氧化活性差异,验证蛋白肽体外加工的必要性,本实验以蛋清蛋白为原料,利用体外化学手段对比了体外模拟胃肠消化及碱性蛋白酶处理后蛋清肽抗氧化活性变化,通过构建双氧水损伤HepG2细胞模型,验证高活性组分(分子质量小于1 kDa的碱性蛋白酶酶解物)的细胞抗氧化能力。结果表明,碱性蛋白酶酶解物的抗氧化活性明显高于体外模拟胃肠消化产物及未经处理的蛋清蛋白,酶解和消化产物的抗氧化活性与其分子质量呈负相关并表现出明显的量效关系。其中分子质量小于1 kDa的碱性蛋白酶酶解物能够有效抑制H2O2诱导的HepG2细胞内活性氧的产生,进而阻止细胞的氧化应激损伤和凋亡。利用超高效液相色谱-串联质谱,从中解析出3 条四肽(FYCP、YCPI、YLPR),均具有良好的抗氧化活性。本研究在一定程度上为蛋品深加工的方法和意义提供了实验依据。

关键词: 体外加工;模拟胃肠消化;蛋清蛋白;肽;抗氧化活性

Abstract: The aim of the research was to clarify the difference in the antioxidant activity of peptides derived from the in vivo digestion and in vitro processing of proteins, and further to verify the necessity of the in vitro processing of proteins for preparing antioxidant peptides. The antioxidant activity of egg white peptides derived from simulated gastrointestinal digestion and alkaline protease treatment were compared by chemical methods in vitro. Potent bioactive peptides with a molecular mass less than 1 kDa were isolated and sequenced from the alkaline protease hydrolysate and their protective effect on oxidative damage induced by H2O2 in human hepatoma cells (HepG2) was evaluated. The results showed that the antioxidant activity of the alkaline protease hydrolysate was significantly higher than that of the simulated gastrointestinal digestive products and untreated egg white protein. The antioxidant activity of both the enzymatic hydrolysate and the digestive products was negatively related to their molecular masses and significantly dose-dependent. The peptides with a molecular mass less than 1 kDa from the alkaline protease hydrolysate could effectively inhibit the production of reactive oxygen species induced by H2O2 in HepG2 cells, thereby preventing oxidative stress injury and apoptosis in cells. Three tetra-peptides, FYCP, YCPI and YLPR, were identified by ultra performance liquid chromatography/tandem mass spectrometry, all of which had good antioxidant activity. To a certain extent, this study provides an experimental basis for egg deep processing.

Key words: in vitro processing; simulated gastrointestinal digestion; egg white protein; peptide; antioxidant activity

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