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海洋真菌多糖YCP的酶法降解及其产物分析

Enzymatic degradation of marine polysaccharide YCP and analysis of YCP-derived fragments

  • 摘要: 目的: 研究海洋真菌多糖YCP经酶法降解后理化性质及免疫活性的改变,探讨其结构和活性之间的关系。 方法: 采用人唾液α-淀粉酶降解海洋真菌多糖YCP,经凝胶色谱分离纯化;通过HPLC、红外光谱以及薄层色谱等对其理化性质进行分析;通过小鼠腹腔巨噬细胞NO诱生实验,中性红吞噬实验以及与巨噬细胞表面YCP相关受体竞争性结合实验分析其免疫活性的改变。 结果: 海洋真菌多糖YCP经人唾液α-淀粉酶降解后得到相对分子质量约为6.6×103的单一组分,该组分的单糖组成以及红外光谱特征与YCP相同,对巨噬细胞分泌NO以及吞噬中性红的能力没有显著的促进作用,对YCP与巨噬细胞表面相关受体的结合几乎不产生抑制作用。 结论: 海洋真菌多糖YCP经人唾液α-淀粉酶部分降解后免疫活性显著降低,说明YCP的完整分子可能对维持YCP的免疫活性具有重要作用。

     

    Abstract: Abstract Aim: To explore the relationship between structure and immunological activity of marine polysaccharide YCP,the physicochemical property and immunological activity of the YCP-derived fragment were studied. Methods: YCP was hydrolyzed by α-amylase from human saliva.The hydrolysate was purified to obtain an polysaccharide fragment by gel filtration chromatography.The physicochemical properties of this YCP-derived fragment was characterized by HPLC,FT-IR and TLC.In addition,changes of phagocytic activity,production of reactive nitrogen and macrophage binding were investigated. Results: The relative molecular weight of YCP-derived fragment was approximately 6.6×103.The monosaccharide composition and FT-IR of the YCP-derived fragment were identical to YCP.No significant effect of the YCP-derived fragment on NO production and murine macrophage phagocyte were observed.And this fragment was not able to compete the binding between YCP and macrophages. Conclusion: The remarkable decrease of immunological activity of YCP-derived fragments degraded by α-amylase of human saliva suggests that the complete structure and high molecule weight of YCP are essential for its immuno-modulatory activity.

     

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