食品科学

• 基础研究 • 上一篇    下一篇

DEAE琼脂糖凝胶纯化龙胆多糖及其分子特性

肖 健,曹荣安*,贾 建,梁茜茜,李泽伟,魏恭禄,王 巍   

  1. 黑龙江八一农垦大学食品学院,黑龙江 大庆 163319
  • 出版日期:2016-08-15 发布日期:2016-08-30

Purification and Molecular Characterization of Polysaccharides from Gentianae Radix et Rhizoma by DEAE Sepharose Fast Flow Chromatography

XIAO Jian, CAO Rongan*, JIA Jian, LIANG Xixi, LI Zewei, WEI Gonglu, WANG Wei   

  1. College of Food Science, Heilongjiang Bayi Agricultural University, Daqing 163319, China
  • Online:2016-08-15 Published:2016-08-30

摘要:

以龙胆为原料提取龙胆多糖,纯化后对其分子特性进行研究。通过正交试验确定了龙胆多糖的最佳提取参数:浸提时间2.5 h、浸提温度90 ℃、液料比20∶1(V/m),得率为12.80%。利用DEAE琼脂糖凝胶柱对龙胆多糖粗品进行纯化,得到F1和F2两个分离组分,得率分别为14.1%和63.4%。化学组成分析表明龙胆多糖粗品、F1和F2是由不同含量的总糖、蛋白质、硫酸根和糖醛酸组成的,单糖组成主要包括阿拉伯糖和半乳糖,同时含有少量的葡萄糖、鼠李糖、甘露糖和木糖。采用高效尺寸排阻色谱-紫外检测器-多角度激光光散射仪-示差折光检测器联机系统对多糖的分子质量及分布进行分析,结果表明龙胆粗品、F1和F2分子质量(Mw)为67.2×103~788.4×103 u,回转半径(Rg)为90.2~321.1 nm。研究确定了龙胆多糖的提取参数,并通过离子交换柱进行纯化,分析了龙胆多糖的分子特性,为下一步的生物活性研究奠定了基础。

关键词: 龙胆, 多糖, 分离纯化, 分子特性

Abstract:

The molecular characterization of the polysaccharides purified from Gentianae Radix et Rhizoma was investigated
in this study. By using orthogonal array design, the optimal extraction parameters that provided the maximum yield of
polysaccharides of 12.80% were determined as follows: extraction time 2.5 h, temperature 90 ℃, and ratio of liquid to solid
20:1 (V/m). The crude polysaccharide was purified by DEAE sepharose fast flow chromatography to obtain two fractions,
F1 and F2, with a yield of 14.1% and 63.4%, respectively. All the crude polysaccharide and purified fractions were composed
of neutral sugar, protein, sulfate and uronic acid with various ratios. Arabinose and galactose were the major monosaccharide
units along with a small portion of glucose, rhamnose, mannose and xylose. The molecular weights (Mw) and radii of gyration
(Rg) of all three samples ranged from 67.2 × 103 to 788.4 × 103 u and from 90.2 to 321.1 nm, respectively, as determined by
HPSEC-UV-MALLS-RI system. The results obtained in this study can provide the basis for further bioactivity studies.

Key words: Gentianae Radix et Rhizoma, polysaccharides, separation and purification, molecular characterization

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