[关键词]
[摘要]
目的分离纯化转基因西洋参冠瘿组织中多糖组分,为从该转基因材料中寻找重要生物活性前体多糖药物提供科学依据。方法用水醇提取法,以DEAE Sepharose FF和Sephacryl S-100柱色谱进行多糖的分离纯化,采用高效液相色谱测定单糖组成,色谱和光谱学方法鉴定其纯度和化学结构。结果分离得到4个精多糖(PPQ50-1、PPQ50-2、PPQ70-1和PPQ70-2),凝胶色谱法(Sephadex G-200)和旋光法证实了它们为化学均一性多糖;凝胶色谱法测得4种精多糖的相对分子质量分别为4.7×104,2.9×105,7.5×104和1.3×105;对得量较多的PPQ50-2和PPQ70-2进行了结构分析,证明二者均为葡聚糖;并利用NaIO4氧化反应和核磁共振谱等方法对PPQ50-2的化学结构进行了初步的糖基键合分析。结论首次从转基因西洋参冠瘿组织中分离得到4种化学均一性多糖成分,其中PPQ50-2和PPQ70-2均为葡聚糖。
[Key word]
[Abstract]
Objective To investigate the polysaccharides constituents from the transgenic crown gall cultures of Panax quinquefolium. Methods The crude polysaccharides PPQ50 and PPQ70 were extracted by water-ethanol method from the crown gall of P. quinquefolium. Four polysaccharides, named PPQ50-1, PPQ50-2, PPQ70-1, and PQ70-2, were isolated and purified by DEAE Sepharose and Sephacry S-100 column chromatography. Results The homogeneity of four polysaccharides was examined by gel chromatography and polarimetry. Their molecular weights were determined by gel filtration chromatography. They were 4.7×104, 2.9×105, 7.5×104, and 1.3×105, respectively. The sugar composition was analyzed by HPLC. The polysaccharides are mainly composed of neutral sugar particularly of D-glucose. The structure elucidation of PPQ50-2 was also supported by 1H-NMR and 13C-NMR spectroscopy. Conclusion All of the four polysaccharides are first isolated and purified from the transgenic crown gall cultures of P. quinquefolium.The result shows the PPQ50-2 and PPQ70-2 are both glucans.
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[基金项目]
教育部科技计划重点项目(104180);广东省自然科学基金项目(31891);广东省中医药局科研项目(103041)