食品科学 ›› 2019, Vol. 40 ›› Issue (7): 40-46.doi: 10.7506/spkx1002-6630-20180319-241

• 基础研究 • 上一篇    下一篇

辣木籽抗氧化肽的制备及其对氧化损伤红细胞的保护作用

林恋竹,朱启源,赵谋明   

  1. 华南理工大学食品科学与工程学院,广东 广州 510640
  • 出版日期:2019-04-15 发布日期:2019-05-05
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2017YFD0400200)

Preparation of Antioxidant Peptide from Moringa oleifera Seeds and Its Protective Effects on Oxidatively Damaged Erythrocytes

LIN Lianzhu, ZHU Qiyuan, ZHAO Mouming   

  1. School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China
  • Online:2019-04-15 Published:2019-05-05

摘要: 采用酶法制备辣木籽水解物,以抗氧化活性为跟踪指标,通过乙醇分级沉淀和大孔树脂柱层析法定向制备辣木籽抗氧化肽,构建模拟胃、肠道消化模型,评价辣木籽抗氧化肽胃、肠道消化物的氧自由基吸收能力(oxygen radical absorbance capacity,ORAC),利用超高效液相色谱-四极杆-高分辨飞行时间串联质谱技术解析消化物中多肽组成与结构,建立2,2’-偶氮二异丁脒盐酸盐(2,2’-azobis(2-methylpropionamidine) dihydrochloride,AAPH)诱导大鼠红细胞氧化损伤模型,研究经胃、肠道消化后的辣木籽抗氧化肽对氧化损伤红细胞溶血的抑制作用。结果表明:采用20%(体积分数,下同)、40%乙醇分级沉淀辣木籽水解物去除杂质,取60%乙醇分级沉淀组分进行柱层析分离纯化,经水、20%乙醇、40%乙醇洗脱除杂,采用60%乙醇洗脱,定向制备得到富含抗氧化氨基酸、疏水性氨基酸的辣木籽抗氧化肽(蛋白质量分数92.84%,1,1-二苯基-2-三硝基苯肼自由基清除能力、ORAC分别为39.95、1454.57 μmol/g);辣木籽抗氧化肽经模拟胃、肠道消化后,抗氧化活性增强,释放出更多短肽段,包括11条二肽、5 条三肽;辣木籽抗氧化肽消化物、Gln-Met、Leu-Phe通过清除胞内活性氧,下调丙二醛含量与超氧化物歧化酶活力,拮抗AAPH所致氧化应激,有效抑制氧化损伤红细胞发生溶血。

关键词: 辣木籽, 抗氧化肽, 生物可接受度, 结构鉴定, 氧化损伤红细胞

Abstract: An antioxidant peptide was purified from enzymatic hydrolysate of Moringa oleifera seed proteins by antioxidant activity-guided separation using fractional precipitation with 20%, 40% and 60% ethanol and macroporous resin column chromatography. Simulated gastrointestinal digest of the antioxidant peptide was evaluated for oxygen radical absorbance capacity (ORAC). The peptide profile and structure of the digest was identified by ultra performance liquid chromatographyquadrupole time of flight-tandem mass spectrometry (UPLC-QTOF-MS/MS). The anti-hemolysis activity and protective effect of the digest against 2,2’-azobis(2-methylpropionamidine) dihydrochloride (AAPH)-induced oxidative damage in erythrocytes were studied. The 60% ethanol precipitate was harvested and purified using column chromatography by sequential elution with pure water, 20%, 40% and 60% ethanol, yielding an antioxidant peptide rich in antioxidant and hydrophobic amino acids (which contained 92.84% protein and exhibited 1,1-diphenyl-2-picrylhydrazyl scavenging capacity and ORAC value of 39.95 and 1 454.57 μmol/g, respectively). The antioxidant activity of the peptide was enhanced after gastrointestinal digestion, and short-chain peptides including 11 dipeptides and 5 tripeptides were released. The resulting digest, Gln-Met and Leu-Phe could effectively inhibit oxidative stress-induced hemolysis through scavenging intracellular reactive oxygen species (ROS), down-regulating malondialdehyde (MDA) level and superoxide dismutase (SOD) activity, and inhibiting AAPH-induced oxidative stress.

Key words: Moringa oleifera seed, antioxidant peptides, bioaccessibility, structure identification, oxidatively damaged erythrocytes

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