灵芝菌丝体多糖的分离纯化及其单糖组成分析与分子量测定
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(1.江南大学食品学院,江苏 无锡 214022;2.江南大学食品科学与技术国家重点实验室,江苏 无锡 214022)

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王海燕(1990-),女,江南大学在读硕士研究生。E-mail: yichenwhy@163.com

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国家自然科学基金(编号:31171688)


Separation, monosaccharide composition and molecular weight analysis of polysaccharide from G. lucidum mycelium
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(1. Food Academy of Jiangnan University, Wuxi, Jiangsu 214022, China; 2. State Key Laboratory of Food Science and Technology in Jiangnan University, Wuxi, Jiangsu 214022, China)

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    摘要:

    前期杂交优化后赤芝菌种经液体深层发酵后,提取灵芝菌丝体多糖,并过DEAE-Sepharose Fast Flow柱分离纯化,利用高效体积排阻色谱(HPSEC)检测多糖级分的纯度,采用完全酸水解PMP柱前衍生化RP—HPLC测定多糖级分的单糖组成,多角度光散射仪联用装置(SEC—MALLS)测定其绝对重均分子量(Mw),并且根据分子旋转半径与分子摩尔数的关系曲线斜率初步推断其空间构象。结果显示:分离纯化得到3个多糖级分GLMP1、GLMP2和GLMP3,HPSEC检测其峰面积百分比分别为93.58%,97.64%,99.19%,单糖组成分析结果表明GLMP1、GLMP2和GLMP3均含有甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖和岩藻糖,但单糖摩尔比各异。SEC—MALLS测试GLMP1、GLMP2和GLMP3的Mw分别为4.526×105,4.603×104,3.760×103 g/mol,3个多糖级分构象可能均为高度紧缩且具有分支结构的聚合物。

    Abstract:

    The polysaccharide was extracted from G. lucidum mycelium by submerged fermentation after previous hybird optimization. The polysaccharide of G. lucidum mycelium was separated by DEAE Sepharose fast flow column. Then we used high volume exclusion chromatography (HPSEC) to explore purity of three polysaccharide fractions and complete acid hydrolysis and PMP pre column derivatization RP—HPLC to measure the monosaccharide composition of them. At last, multi angle light scattering instrument combined with device (SEC—MALLS) was used to measure the Mw of polysaccharide fractions, while the slope of curves between the molecular rotation radius and molecular molar of moles inferred the space conformation of them. HPSEC results showed that the purified of GLMP1, GLMP2 and GLMP3 were 93.58%, 97.64% and 99.19%. Monosaccharide composition analysis showed that mannose, rhamnose, galacturonic acid, glucose, galactose, xylose, arabinose and fucose were contained in GLMP1, GLMP2 and GLMP3, however they had different molar ratios of monosaccharide. SEC—MALLS tested the absolute weight-average molecular weights (Mw) of GLMP1, GLMP2 and GLMP3 respectively were 4.526×105, 4.603×104 and 3.760×103 g/mol and it suggested that the conformations of the three polysaccharide fractions maybe were polymers which were high tightening and having branched structures.

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王海燕,戴军,陈尚卫.灵芝菌丝体多糖的分离纯化及其单糖组成分析与分子量测定[J].食品与机械,2015,31(5):201-205.
WANGHaiyan, DAIJun, CHENShangwei. Separation, monosaccharide composition and molecular weight analysis of polysaccharide from G. lucidum mycelium[J]. Food & Machinery,2015,31(5):201-205.

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  • 收稿日期:2015-07-30
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  • 在线发布日期: 2023-03-17
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