摘要:
从发酵的富锗金针菇(Flammulina velutipes)菌丝体分离纯化得到多糖Flammulina velutipes polysaccharide 1（FVP1），并对其进行结构初步鉴定和体外活性分析。利用凝胶渗透色谱(GPC)测定相对分子质量；气相色谱法(GC)检测单糖组成；运用高碘酸氧化-Smith降解法、甲基化方法对多糖结构初步解析。体外分析FVP1对小鼠巨噬细胞的活化作用。结果显示，FVP1是一种高度分支的多糖，相对分子质量约为140 kD，单糖组成包括甘露糖、葡萄糖、半乳糖、盐藻糖及鼠李糖，其物质的量比为2∶4∶5∶1∶1；FVP1的主链结构包括1,6-葡萄糖、1,6-半乳糖、1,3-半乳糖，并在1,3,6-甘露糖少量的1,3,4-鼠李糖产生支链结构。结果发现，FVP1能与小鼠腹腔巨噬细胞靶向性结合，并可显著促进其分泌NO。FVP1在体外展示了良好的免疫调节活性，具有潜在的生物学功能。
Abstract:
A new fraction ofFlammulina velutipes polysaccharide 1 (FVP1) was separated and purified from Flammulina velutipes mycelium with several processes.Its structural features and bioactivities in vitro were also detected.The molecular mass of FVP1 was determined by HPGPC；the monosaccharide composition was identified according to the result of gas chromatography (GC)；the glycosidic bonds between monosaccharides were analyzed by periodic acid oxidation-Smith degradation and methylation analysis.Fluorescamine was bound with FVP1 and fluorescent activated cell sorting (FACS) was used to analyze the binding of FVP1-FLA to peritoneal macrophages.The molecular mass is about 140 kD.FVP1 consists of mannose (Man),glucose (Glc),galactose(Gal),fucose(Fuc) and rhomnose (Rha) in a molar ratio of 2:4:5:1:1.It was found that FVP1 is a complicated polysaccharide with a main composition of 1,4-Glc,1,3-Gal,1,3-Fuc,1,6-Gal,1,3,6-Man,1,6-Glc,1,3,4-Rha by methylation analysis and gas chromatography-mass spectrometry (GC-MS).Finally,bioactivity analysis showed that FVP1 acted as an immunomodulator,which could bind to peritoneal macrophages and strongly stimulate them to produce NOin vitro.FVP1 was a highly branched polysaccharide with great potential as an immunomodulator.
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