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毛木耳子实体中活性多糖APPⅡA的分离纯化与结构初探

Isolation,purification and structural characterization of bioactive polysaccharide APPⅡA from Auricularia polytricha

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【作者】 清源余梦瑶罗霞江南郑林用葛绍荣

【Author】 QING Yuan1 YU Meng-Yao2, 3 LUO Xia2, 3 JIANG Nan2, 3 ZHENG Lin-Yong2, 4 GE Shao-Rong2 1Department of Food Science, Xichang College, Xichang 615013, China 2Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, School of Life Science, Sichuan University, Chengdu 610064, China 3Cytological and Molecular Lab of Sichuan Institute of Chinese Materia Medica, Chengdu 610064, China 4Sichuan Academy of Agricultural Science, Chengdu 610064, China

【机构】 西昌学院食品科学系四川大学生命科学院生物资源与生态环境教育部重点实验室四川省中医药研究院中药细胞与分子生物学实验室四川省农业科学院

【摘要】 毛木耳Auricularia polytricha子实体500g经热水浸提、乙醇沉淀、Sevage法脱蛋白后,得到粗多糖3.94g,得率为0.79%。经小鼠S180载体实验证实,50mg/kg该粗多糖对肿瘤的抑制率达47.89%,呈极显著差异。利用离子交换柱对其进行分离,共得到三个部分,分别命名为APPⅠ、APPⅡ、APPⅢ。其中,APPⅡ对小鼠S180的抑制率达到56.36%,明显优于其他两部分。通过SephacrylS300凝胶过滤层析柱后,APPⅡ被进一步分离为A和B两个部分,APPⅡA的抑瘤率为53.61%,远高于APPⅡB,呈极显著差异。通过紫外光谱、红外光谱等方法对该单一多糖组分进行结构分析,表明APPⅡA为一相对分子质量约为110,000Da、可能同时具有α和β构象的杂多糖,主要有木糖和甘露糖组成,且纯度较高,含有硫酸基。

【Abstract】 The water-soluble crude polysaccharide (APP) was isolated from fruit bodies of Auricularia polytricha by hot water extraction, ethanol precipitation, and deproteination with Sevage method. It was found that APP could markedly inhibit growth of Sarcoma 180 in mice. APPⅠ, Ⅱ and Ⅲ was separated from APP by ion-exchange on DEAE-Cellulose-52, and APPⅡ couldsignificantly exhibit the anti-tumor effect. APPⅡ was further purified by gel-filtration chromatography on Sephacryl S-300 columns and separated into APPⅡA and APPⅡB. It was confirmed that APPⅡA could significantly inhibit tumor growth. APPⅡA was characterized by thin-layer chromatography (TLC), agarose gel electrophoresis (AGE), high performance liquid chromatography (HPLC), and UV, IR and 1H-NMR spectroscopy. The results demonstrated that APPⅡA was homogeneous and its relative molecular weight was approximately 110,000Da. Monosaccharide analysis showed that APPⅡA was a heteropolysaccharide which was composed of xylose and mannose. The glycosidic linkages of APPⅡA was estimated to contains both α- and β- pyranoside form.

【关键词】 粗多糖抑瘤作用木糖甘露糖
【Key words】 crude polysaccharideanti-tumor effectxylosemannose
  • 【分类号】S646.6
  • 【被引频次】18
  • 【下载频次】255
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