鸡腿蘑胞外多糖的形貌结构及分子量动态变化与抗氧化的相关性研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0471

摘要/Abstract

摘要：

对鸡腿蘑多糖的结构进行检测,并在此基础上探讨结构与活性的关系,对深度发掘鸡腿蘑多糖的功效具有重要意义。制备发酵时间为72 h、96 h和120 h的鸡腿蘑胞外粗多糖,采用PMP柱前衍生化-HPLC法分析其单糖组成,结果表明发酵72 h、96 h和120 h胞外多糖均由D-甘露糖、L-鼠李糖、D-葡萄糖、D-半乳糖、D-阿拉伯糖组成,但各单糖的相对摩尔比不同。采用扫描电子显微镜和原子力显微镜对不同发酵时间的多糖形貌结构进行分析,结果显示发酵120 h多糖的板状结构更加规则,多糖分子的聚集作用更强。采用高效凝胶过滤色谱法对不同发酵时间的鸡腿蘑胞外多糖的分子量分布情况进行分析,结果显示发酵120 h的多糖分子量更大,分布范围更广。体外抗氧化试验结果显示,在相同浓度下,发酵120 h多糖的抗氧化活性较发酵72 h和96 h的多糖强。研究结果表明鸡腿蘑胞外多糖的抗氧化活性可能与其形貌结构和分子量分布有关。

关键词: 鸡腿蘑胞外多糖, 单糖分析, 形貌结构, 分子量, 抗氧化

Abstract:

Detecting the structure of Coprinus comatus polysaccharides and exploring the relationship between structure and activity are of great significance for exploring its efficacy in depth. Three Coprinus comatus extracellular crude polysaccharides with fermentation times of 72 h,96 h and 120 h were prepared,and their monosaccharide composition were determined by the method of PMP pre-column derivatization combined with HPLC. The results showed that the extracellular polysaccharides fermented for 72 h,96 h and 120 h were all composed of D-mannose,L-rhamnose,D-glucose,D-galactose,and D-arabinose,but the relative molar ratio of each monosaccharide was different. Scanning electron microscopy and atomic force microscopy were used to analyze the morphology structure of polysaccharides,and the results showed that the plate-like structure of polysaccharides fermented 120 h was more regular and the molecules aggregation was stronger. High performance gel filtration chromatography was employed to analyze the molecular weight distribution,and the results demonstrated that the molecular weight of polysaccharides fermented for 120 h was larger and the distribution range was wider. In vitro antioxidant test results suggested that the polysaccharide fermented for 120 h displayed stronger antioxidant activity than that of polysaccharides fermented for 72 h and 96 h at the same concentration. The results indicate that the antioxidant activity of C. comatus extracellular polysaccharide may be related to its morphological structure and molecular weight distribution.

Key words: Coprinus comatus extracellular polysaccharides, monosaccharide analysis, morphological structure, molecular weight, antioxidant

王珊珊, 孙敏, 王永霞, 李惟栋, 韩春超. 鸡腿蘑胞外多糖的形貌结构及分子量动态变化与抗氧化的相关性研究[J]. 生物技术通报, 2021, 37(2): 129-137.

WANG Shan-shan, SUN Min, WANG Yong-xia, LI Wei-dong, HAN Chun-chao. Correlation Detween the Dynamic Changes of Morphological Structure and Molecular Weight of Coprinus comatus Extracellular Polysaccharides and Their Antioxidant Activity[J]. Biotechnology Bulletin, 2021, 37(2): 129-137.

图/表 7

图1 混合单糖与鸡腿蘑胞外多糖的HPLC色谱图 S1：混合单糖;S2、S3和S4：分别为发酵72 h,96 h和120 h鸡腿蘑胞外多糖;1：D-甘露糖;2：L-鼠李糖;3：D-葡萄糖;4：D-半乳糖;5：D-阿拉伯糖

表1 五种单糖的线性范围和回归方程

单糖	回归方程	相关系数
D-甘露糖D-mannose	y=182277x-127844	R²=0.999 8
L-鼠李糖L-Rhamnose	y=155433x-101943	R²=0.999 2
D-葡萄糖D-glucose	y=98305x-22473	R²=0.999 7
D-半乳糖D-galactose	y=47785x+33756	R²=0.999 2
D-阿拉伯糖Arabinose	y=78214x-112323	R²=0.999 6

表2 鸡腿蘑胞外多糖的单糖组成及摩尔比

发酵时间/h	单糖组成	摩尔比
72	甘露糖-鼠李糖-葡萄糖-半乳糖-阿拉伯糖	0.019∶0.052∶0.137∶0.703∶0.183
96	甘露糖-鼠李糖-葡萄糖-半乳糖-阿拉伯糖	0.013∶0.036∶0.246∶1.010∶0.166
120	甘露糖-鼠李糖-葡萄糖-半乳糖-阿拉伯糖	0.028∶0.051∶0.183∶0.646∶0.156

图2 鸡腿蘑胞外多糖的SEM图像 A、B和C：分别为发酵72 h、96 h和120 h鸡腿蘑胞外多糖

图3 鸡腿蘑胞外多糖的AFM图像 A、B 和 C ：分别为发酵 72 h、96 h 和 120 h 鸡腿蘑胞外多糖的平面和三维图像

图4 鸡腿蘑胞外多糖的分子量分布 A、B和C：分别为发酵72 h、96 h和120 h鸡腿蘑胞外多糖

图5 鸡腿蘑胞外多糖的抗氧化活性 A：总抗氧化能力;B：DPPH自由基清除能力

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