榛仁免疫活性肽分离纯化及结构鉴定

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

以榛仁分离蛋白水解肽经超滤获得的分子质量小于3 k Da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-GluAsp-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。

Hazelnut protein isolate hydrolysate was ultrafiltrated to obtain peptides with molecular weight less than3 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 to100.0 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.