灰树花水溶性β-葡聚糖的制备和结构研究-生药学专业论文.docxVIP

灰树花水溶性B一葡聚糖的制备和结构研究摘 灰树花水溶性B一葡聚糖的制备和结构研究 摘 要 灰树花(Gr帕lafrondisa)是一种名贵食用与药用真菌，具有抗肿瘤等多 种生物活性。目前，已经能用发酵工程技术对其进行大规模发酵，获得大量 菌丝体。 本文以灰树花发酵菌丝体为原料，经过乙醇脱脂，用热水提取获得GFMl 水溶性杂多糖组分；用冷碱提取分离得到水溶性0一葡聚糖组分GFM3；用热 碱提取分离得到水溶性0一葡聚糖组分GFM4。经进一步分离纯化，从冷碱提 取组分GFM3中得到GFM3P1与GFM3P2组分； 从热碱提取组分GFM4中 得到了GFM4P2组分。 经各种化学与波谱分析表明，GFM3P1与GFM4P2 均属于B一葡聚糖，而GFM3P2则是一种B一葡聚糖肽，且GFM3P2中糖链 与肽链之间的连接方式主要为O一连接。 经过对该肽链氨基酸的组成分析表 明， 该肽链含有16种氨基酸，其中酸性氨基酸含量丰富。经刚果红结合实 验表明，GFM3P1、GFM3P2的超螺旋结构容易被0．2mol／L NaOH破坏， 而 GFM4P2则相反，其超螺旋结构不能被2m01]LNaOH水溶液破坏。结构研究 表明，GFM4P2为13—1，6主链及B一1，3支链的新型B一葡聚糖，分枝度为 36．4％，该结构属于首次报导。 本文进一步利用结构确定的纯品多糖GFM3P1与GFM4P2，初步评价了 它们抗肿瘤活性。结果表明，GFM3P1的抑瘤率是65％，GFM4P2的抑瘤率 是73％，均高于临庆应用的香菇多糖的抑制率(35％)：且具有提高小鼠免疫 的功能，提示该类多糖具有开发为抗肿瘤药物的前景。 关键词：灰树花菌丝体：B一葡聚糖：纯化； 结构；抗肿瘤活性 PreparatioN PreparatioN And Character i zation of Water Soluble 13一GIucans from Gr ifola Frandosa MyceI iUilll Abstract Gr咖lafrondosa is all edible／medical fungus with a renowned fame and it has anticancer etc．activities．It has been liquid cultured on a large—scale to acquire mycelium at present． This thesis involves in the preparation and characterization of the water-soluble polysaccharides from Grifolafrondosa．The mycelium was extracted with 85％ethanol and then extracted sequentially with hot water,cold and hot sodium hydroxide．One water soluble heteropolysaccharide(GFM 1)was acquired using hot water extraction，and two water soluble p—glucan(OFM3，GFM4)were acquired using cold and hot alkali extraction，respectively． GFM3 was further fractioned into GFM3P1 and GFM3P2．GFM3P1 is a 13-glucan and GFM3P2 is a B—glycopeptides．T11e structure of GFM3P1 and GFM3P2 WaS similar to that of GFM4P2．The linkage of glucan to peptide is O·linkage in GFM3P2 and the peptide contained 16 kinds of amino acid，and acid amino acid is very abandant．The super helix snucture ofGFM3P1 and GFM3P2 is easily destroyed by 0．2mol／1NaOH，but that of GFM4P2 is hardly destroyed by 2mol／L NaOH in the Congo red binding assay．The structllre of GFM4P2 is proved to be beta-l，6一glucan with bet