摘要
通过对灰树花(Grifola frondosa)深层发酵的胞外多糖(EXGFP)进行分离纯化,研究其性质和免疫活性.灰树花胞外粗多糖经醇沉、酶–sevag脱蛋白和Sephadex G-100色谱柱分离得到纯化组分EXGFP-A.通过Sepharose 4B凝胶柱法和HPLC法鉴定EXGFP-A的纯度,结果表明EXGFP-A是均一多糖组分,纯度为92.68%,.理化性质实验结果表明,EXGFP-A是一种非淀粉类、不含糖醛酸及多酚类物质,含有α–D–葡萄糖苷键和吡喃糖环的中性多糖.MTT实验表明,当EXGFP-A质量浓度为80,μg/m L、作用48,h时,小鼠巨噬细胞RAW264.7细胞增殖指数达到最大值,为137.5%,.吞噬活性实验结果证实EXGFP-A能够提高RAW264.7细胞对中性红的吞噬能力,扫描电子显微镜结果发现RAW264.7细胞经过EXGFP-A处理后,细胞表现出了明显的活化特征.
An extracellular polysaccharide（EXGFP）was obtained by submerged culture of Grifola frondosa,and its propertyand immunomodulatory activity were investigated.The final purified fraction,EXGFP-A,was obtained by using alcoholfor precipitation,enzymes-sevag for deproteinization and Sephadex G-100,gel column for further isolation and purification.The purity of EXGFP-A was tested with Sepharose 4B gel column and HPLC.The results show that EXGFP-A is ahomogeneous polysaccharide,and its purity is about 92.68%,.Tests of its physicochemical properties indicate that EXGFP-Ais a non-starch neutral polysaccharide,excludes uronic acid and polyphenols,and contains α-D-glucoside bond and pyranosering.The result of MTT assay shows that the cell proliferation index reached a maximum of 137.5%, when EXGFP-A was ata concentration of 80,μg/mL and the treatment time was 48,h.The neutral red phagocytosis experiment indicates that thephagocytic activity of RAW264.7,cells was significantly enhanced by EXGFP-A.According to scanning electronmicroscope,the activation of RAW264.7,cells could also be promoted by adding EXGFP-A.
出处
《天津科技大学学报》
CAS
北大核心
2016年第4期25-29,共5页
Journal of Tianjin University of Science & Technology
基金
国家科技支撑计划资助项目(2012BAD33B04)
天津市应用基础与前沿技术研究计划资助项目(13JCZDJC29800)
关键词
灰树花
胞外多糖
分离纯化
小鼠巨噬细胞RAW264.7
免疫活性
Grifola frondosa
extracellular polysaccharide
purification
mouse macrophage RAW264.7
immunomodulatoryactivity
