紫芝菌丝体多糖的结构鉴定及免疫活性研究  

Characterization of the Structure and Immunomodulatory Activities of Polysaccharides from Ganoderma Lucidum Mycelium

作　　者：张婷[1] 吴晖[1] 赖富饶[1] 张晓元[1,2] 闵甜[1] 董洲[1] 李慧娴[1] 

机构地区：[1]华南理工大学食品科学与工程学院,广东广州510640 [2]韶关市华工高新技术产业研究院,广东韶关512027

出　　处：《现代食品科技》2017年第4期52-60,228,共10页Modern Food Science and Technology

基　　金：广州省科技计划项目(2016A040402020);广东省科技计划项目(2016B010121014)

摘　　要：本采用热水浸提紫芝菌丝体得到粗多糖后经DEAE-sepharose Fast Flow和Sephacryl S-200 HR分离纯化出一种新型的灵芝多糖LZ1。通过采用凝胶渗透色谱法、电镜、紫外、红外光谱、气相色谱、核磁共振、高碘酸氧化和Smith降解方法等方法对LZ1进行结构鉴定和解析,结果表明LZ1是不含核酸、蛋白质的均一多糖,其分子量为7498 u;其是由鼠李糖、岩藻糖、甘露糖、葡萄糖、半乳糖和阿拉伯糖以0.94:0.50:1.68:26.91:4.80:17.12的摩尔比组成的酸性杂多糖,其糖苷键为1→4、1→2或1→6、1→3;刚果红实验表明LZ1没有三螺旋结构;通过小鼠巨噬细胞RAW264.7体外免疫模型,研究LZ1对TNF-α、IL-6、NO三种细胞因子的分泌情况的影响发现LZ1对这三种细胞因子的分泌有明显的促进作用,且呈现浓度相关性,表现出明显的免疫活性;通过AAPH诱导的红细胞氧化性溶血模型对LZ1的抗氧化活性进行评价,发现LZ1有很好的抑制AAPH诱导的红细胞氧化溶血的特性。A new polysaccharide LZ1 was isolated and purified using DEAE-Sepharose Fast Flow and Sephacryl S-200 HR from crude polysaccharide extract obtained from Ganoderma lucidum mycelium via hot-water extraction. The structure of LZ1 was identified and elucidated by gel permeation chromatography（GPC）, scanning electron microscopy（SEM）, ultraviolet spectroscopy, Fourier transform infrared（FT-IR） spectroscopy, gas chromatography（GC）, nuclear magnetic resonance（NMR） spectroscopy, periodate oxidation, and Smith degradation. The results showed that LZ1 was a homogeneous polysaccharide and did not contain impurities such as proteins and nucleic acids. The molecular weight（Mw） was determined to be 7498 u by GPC. LZ1 was an acidic polysaccharide and consisted of rhamnose, fucose, mannose, glucose, galactose, and arabinose at a molar ratio of 0.94：0.50：1.68：26.91：4.80：17.12. The main glycosidic linkage types were 1→4 and 1→2 or 1→6 and 1→3. Congo red test suggested that there were no triple helical structures in LZ1. The effect of LZ1 on the expression of tumor necrosis factor-α（TNF-α）, interleukin 6（IL-6）, and nitric oxide（NO） was studied using murine macrophage RAW264.7 cells as an in vitro immunomodulatory model. The results showed that LZ1 significantly promoted the secretion of TNF-α, IL-6, and NO in a concentration-dependent manner, and exhibited excellent immunomodulatory activity. Evaluation of the antioxidant activity of LZ1 by a 2,2’-azobis（2-amidinopropane） dihydrochloride（AAPH）-induced oxidative hemolysis model in erythrocytes suggested that LZ1 could effectively inhibit the AAPH-induced oxidative hemolysis of erythrocytes.

关 键 词：菌丝体 多糖 结构 免疫调节 抗溶血 

分 类 号：TS201.2[轻工技术与工程—食品科学]