杏鲍菇多糖组分的分离纯化及结构鉴定

Isolation, purification and structural identification of Pleurotus eryngii polysaccharides

中文关键词:  杏鲍菇多糖  岩藻糖  分离纯化  结构鉴定

英文关键词:Pleurotus eryngii polysaccharides  fucose  separation and purification  structural identification

中文摘要:

      目的 对含有岩藻糖(fucose, Fuc)的杏鲍菇多糖进行分离纯化, 并分析其结构。方法 通过水提分级醇沉得到60%乙醇醇沉杏鲍菇多糖(Pleurotus eryngii polysaccharides, PEP)。杏鲍菇多糖经离子交换层析和凝胶层析分离纯化, 采用凝胶渗透色谱法(gel permeation chromatography, GPC)测定PEP各组分的相对分子质量; 高效液相色谱法(high performance liquid chromatography, HPLC)检测岩藻糖; 傅里叶变换红外光谱(fourier transform infrared spectrum, FT-IR)、X-射线衍射(x-ray diffractometer, XRD)分析、原子力显微镜(atomic force microscope, AFM)观察等方法对PEP各组分进行结构表征。结果 PEP经分离纯化得到PEP-1、PEP-2和PEP-3共3种多糖组分, 分子量依次为1.585×106、4.266×104和1.995×103 Da。岩藻糖存在于PEP-1组分中; FT-IR显示PEP-1和PEP-2存在C-O-C键拉伸和吡喃环构型; PEP-2存在β型糖苷键, PEP-3为β-葡聚糖。XRD结果显示3个组分多糖结晶指数分别为38.89%、47.22%和20.00%。AFM观察结果显示, PEP-1整体呈现流星状结构, 多糖粒子聚集; PEP-2为链状螺旋结构且以小球状体聚集; PEP-3呈现小螺旋棒状结构。结论 PEP分离纯化得到3个组分多糖, 岩藻糖存在PEP-1组分中。

英文摘要:

      Objective To isolate and purify Pleurotus eryngii polysaccharides containing fucose, and analyze its structure. Methods The 60% alcohol precipitated Pleurotus eryngii polysaccharides (PEP) was obtained by water extraction, grading and alcohol precipitation. Pleurotus eryngii polysaccharides were separated and purified by ionexchange and gel column chromatography, the molecular weight of PEP components was determined by gel permeation chromatography (GPC); fucose was detected by high performance liquid chromatography (HPLC); the structures of PEP were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and atomic force microscope (AFM). Results Three polysaccharides PEP-1, PEP-2 and PEP-3 were isolated and purified from PEP, with molecular weight of 1.585×106, 4.266×104, 1.995×103 Da. Fucose existed in PEP-1 component; FT-IR results showed that PEP-1 and PEP-2 had C-O-C bond stretching and pyran ring configuration; PEP-2 presented β-type glycosidic bond, PEP-3 was β-dextran. XRD results showed that the crystallization indexes of the 3 components were 38.89%, 47.22% and 20.00%, respectively. AFM observation indicated that PEP-1 showed meteor like structure and polysaccharide particles gathered; PEP-2 was a chain spiral structure and aggregates in small globules; PEP-3 presented a small spiral rod structure. Conclusion PEP is separated and purified to obtain 3 component polysaccharides; fucose exists in the PEP-1 component.