食品科学 ›› 2019, Vol. 40 ›› Issue (2): 52-58.doi: 10.7506/spkx1002-6630-20171116-208

• 食品化学 • 上一篇    下一篇

雨生红球藻多糖的分离纯化和免疫活性组分鉴定

刘涵,张苗,刘晓娟*,曹庸   

  1. (华南农业大学食品学院,广东省功能食品活性物重点实验室,广东省天然活性物工程技术研究中心,广东?广州 510642)
  • 出版日期:2019-01-25 发布日期:2019-01-22
  • 基金资助:
    广州市科技计划项目(201707010035);广东省自然科学基金项目(2017A030313150); 国家自然科学基金青年科学基金项目(31401481)

Isolation, Purification and Identification of Immunologically Active Polysaccharides from Haematococcus pluvialis

LIU Han, ZHANG Miao, LIU Xiaojuan*, CAO Yong   

  1. (Guangdong Provincial Key Laboratory of Nutraceuticals and Functional Foods, Research Center of Natural Active Compound Engineering Technology of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou 510642, China)
  • Online:2019-01-25 Published:2019-01-22

摘要: 以虾青素提取后的雨生红球藻渣为原料,在体外免疫细胞模型活性跟踪下,对超声辅助提取的雨生红球藻粗多糖(Haematococcus pluvialis polysaccharide,HPP)采用DEAE-52纤维素阴离子交换柱层析和Sephacryl S400葡聚糖凝胶柱层析进行分离纯化,使用高效凝胶渗透色谱法测定各组分多糖分子质量。结果表明:HPP经DEAE-52柱层析分离得到HPP-c1、HPP-c2、HPP-c3、HPP-c4和HPP-c5五个多糖组分,其中HPP-c3免疫活性较好且含量最高;HPP-c3经Sephacryl?S400柱层析分离得到HPP-c3-s1、HPP-c3-s2和HPP-c3-s3,其中HPP-c3-s1在0~250?μg/mL质量浓度范围内极显著刺激小鼠脾细胞的增殖(P<0.01),并呈现剂量效应,最高刺激指数达1.34,高于HPP-c3-s2和HPP-c3-s3。经纯度鉴定,HPP-c3-s1为均一多糖,分子质量为23?413?kDa。研究结果可为雨生红球藻的高值化综合利用提供有利指导。

关键词: 雨生红球藻, 多糖, 分离纯化, 免疫活性

Abstract: An in vitro immune cell model was used for bioactivity-guided fractionation of the crude polysaccharides ultrasonically extracted from Haematococcus pluvialis residue left after the extraction of astaxanthin by sequential DEAE-52 anion exchange and Sephacryl S400 column chromatographies. The molecular masses of the isolated polysaccharides were determined by high performance gel permeation chromatography (HPGPC). The results showed that five polysaccharides, named HPP-c1, HPP-c2, HPP-c3, HPP-c4 and HPP-c5, were obtained by DEAE-52 column chromatography. Of these, HPP-c3 had the highest immunological activity. Further, HPP-c3-s1, HPP-c3-s2, and HPP-c3-s3 were isolated from HPP-c3 by Sephacryl S400 column chromatography. HPP-c3-s1 could significantly stimulate the proliferation of spleen cells (P < 0.01) in the concentration range of 0–250 μg/mL in a dose-dependent fashion. HPP-c3-s1 had the highest stimulation index (SI) of 1.34 among the HPP-c3 fractions. The purity of HPP-c3-s1 was found to be homologous with a molecular mass of 23 413 kDa. These results provide a valuable guidance for the high-value comprehensive utilization of H. pluvialis.

Key words: Haematococcus pluvialis, polysaccharide, isolation and purification, immunological activity

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