食品科学 ›› 2018, Vol. 39 ›› Issue (3): 200-205.doi: 10.7506/spkx1002-6630-201803031

• 营养卫生 • 上一篇    下一篇

榛仁免疫活性肽分离纯化及结构鉴定

王 鹏,王明爽,刘春雷,方 丽,王 辑,刘秀奇,闵伟红*   

  1. 吉林农业大学食品科学与工程学院,小麦和玉米深加工国家工程实验室,吉林 长春 130118
  • 出版日期:2018-02-15 发布日期:2018-01-30
  • 基金资助:
    国家高技术研究发展计划(863计划)项目(2013AA102206)

Isolation, Purification and Structural Identification of Immunoactive Peptides Derived from Hazelnut (Corylus heterophylla Fisch.) Protein

WANG Peng, WANG Mingshuang, LIU Chunlei, FANG Li, WANG Ji, LIU Xiuqi, MIN Weihong*   

  1. National Engineering Laboratory on Wheat and Corn Further Processing, College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China
  • Online:2018-02-15 Published:2018-01-30

摘要: 以榛仁分离蛋白水解肽经超滤获得的分子质量小于3 kDa的组分为研究对象,采用凝胶色谱及反相高效 液相色谱对其进行分离纯化,并对所得组分进行结构鉴定和验证。结果显示:经Sephadex G-15分离得到的组分B1 能极显著降低小鼠RAW264.7巨噬细胞肿瘤坏死因子和白介素-6水平(P<0.01);经质谱解析筛选出的肽段Pro-Glu- Asp-Glu-Phe-Arg(PEDEFR)对细胞无毒性作用,高浓度PEDEFR(>50 μmol/L)能提高小鼠RAW264.7巨噬细胞吞噬能 力;当浓度达到100.0 μmol/L时,促进脾淋巴细胞增殖率达到44.21%,在伴刀豆蛋白A共同作用下,增殖率达到53.22%。 PEDEFR对小鼠RAW264.7巨噬细胞具有较好的免疫调节能力,本研究为榛仁免疫活性肽的开发提供了理论依据。

关键词: 榛仁水解肽, 免疫活性, 分离纯化, 结构鉴定

Abstract: Hazelnut protein isolate hydrolysate was ultrafiltrated to obtain peptides with molecular weight less than 3 kDa. Further purification and structural identification were carried out using gel filtration chromatography and reversedphase high-performance liquid chromatography (RP-HPLC). The results showed that fraction B1, separated on a column of Sephadex G-15, could significantly decrease the production of TNF-α and IL-6 (P < 0.01) in RAW264.7 cells. The peptide Pro-Glu-Asp-Glu-Phe-Arg (PEDEFR) as identified by mass spectrometry (MS) had no cytotoxicity on cells. PEDEFR at a high concentration (> 50 μmol/L) could enhance the phagocytosis of RAW264.7 cells. When the concentration was up to 100.0 μmol/L, the proliferation rate of spleen lymphocytes was 44.21% and reached up to 53.22% in the presence of concanavalin A. PEDEFR had an immunomodulatory effect on RAW264.7 cells. This study can provide an experimental basis for the development and utilization of immunoactive peptides derived from hazelnut protein.

Key words: hazelnut peptides, immunoactivity, isolation and purifiscation, structural identification

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