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化学学报
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化学学报 ›› 2009, Vol. 67 ›› Issue (9): 951-956. 上一篇    下一篇

研究论文

荧光辅助糖电泳用于果胶水解产物中寡糖的分析研究

孙 静a 李健军a,b 黄琳娟a 王仲孚*,a

  

  1. (a西北大学生命科学学院 西部资源生物与现代生物技术省部共建教育部重点实验室
    西北大学糖生物学与糖工程研究中心 西安 710069)
    (b西安交通大学理学院化学系 西安 710049)

  • 投稿日期:2008-06-27 修回日期:2008-11-08 发布日期:2009-05-14
  • 通讯作者: 王仲孚

Studies on Pectic Oligosaccharides Analysis by Fluorophore-assisted Carbohydrate Electrophoresis

Sun, Jing a Li, Jianjun a,b Huang, Linjuan a Wang, Zhongfu *,a   

  1. (a Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, ＆ Glycobiology and Glycotechnology Research Center, Northwest University, Xi’an 710069)
    (b Department of Chemistry, College of Science, Xi’an Jiaotong University, Xi’an 710049)
  • Received:2008-06-27 Revised:2008-11-08 Published:2009-05-14
  • Contact: Wang, Zhongfu

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2345

被引次数

32 | 5

荧光辅助糖电泳(FACE)是首先用荧光衍生化试剂对糖类分子的还原端进行衍生化标记, 然后在一定浓度的聚丙烯酰胺凝胶上进行分离的分析方法, 可同时分析中性糖与酸性糖. 将该方法用于果胶寡糖的分离分析, 对影响FACE的诸多因素如荧光试剂的种类、用量, 荧光衍生化时间、温度, 分离胶浓度及盐、酸等进行了考察, 对果胶寡糖的FACE条件进行了优化, 结果显示: 每1.2 mg无酸且不含盐的果胶寡糖中, 加入0.2 mol/L的ANTS溶液3.75 μL, 1.0 mol/L的NaBH3CN溶液5 μL, 40 ℃衍生化反应16 h后, 在浓度为38%的分离胶上电泳分离, 取得良好的分离效果. 在该实验条件下, 果胶多糖酸水解后得到聚合度为2～16的果胶寡糖混合物, 与质谱分析结果基本一致. 该方法快速、简捷、灵敏、分辨率高, 费用低, 为果胶多糖可控性降解的监测和果胶寡糖的分离分析提供了技术手段.

关键词: 果胶, 寡糖衍生物, 荧光辅助糖电泳, 8-氨基萘-1,3,6-三磺酸, 2-氨基吖啶酮

Fluorophore-assisted carbohydrate electrophoresis (FACE) was used to analyse pectic oligosaccharides. The influence of different factors on the separation of oligosaccharides by FACE was investigated. The condition including the type and the content of fluorescent reagent, labelling time and temperature, salt, acid and the concentration of resolving gel was optimized as follows: 1.2 mg of salty-free and acid-free pectic oligosaccharides were labeled by 3.75 μL of 0.2 mol/L ANTS solution and 5 μL of 1.0 mol/L NaBH3CN solution. The derivative reaction was accomplished at 40 ℃ after 16 h, and the concentration of resolving gel was 38%. In this condition, a pectic oligosaccharide mixture was obtained with the range of DP 2～16. The FACE result was accordant with that of MS analysis. So the FACE procedure described is useful for the rapid monitoring of pectic polysaccharide depolymerization and acdic oligosaccharide analysis.

Key words: pectin, oligosaccharide derivative, fluorophore-assisted carbohydrate electrophoresis, 8-aminonaphthalene-1,3,6-trisulfonate, 2-aminoacridone