食品科学 ›› 2018, Vol. 39 ›› Issue (12): 191-198.doi: 10.7506/spkx1002-6630-201812030

• 成分分析 • 上一篇    下一篇

HPGFC双柱串联检测右旋糖酐蔗糖酶催化产物及右旋糖酐聚合过程

黄双霞1,侯殿志1,2,陈华磊1,于玥1,陈山1,3,*   

  1. (1.广西大学轻工与食品工程学院,广西?南宁 530004;2.中国农业大学食品科学与营养工程学院,北京 100083;3.广西蔗糖产业协同创新中心,广西?南宁 530004)
  • 出版日期:2018-06-25 发布日期:2018-06-15
  • 基金资助:
    国家自然科学基金地区科学基金项目(21768001)

Determination of Dextransucrase-Catalyzed Reaction Products of Sucrose by High Performance Gel Filtration Chromatography with Tandem Two Columns and Analysis of the Dextran Polymerization Process

HUANG Shuangxia1, HOU Dianzhi1,2, CHEN Hualei1, YU Yue1, CHEN Shan1,3,*   

  1. (1. College of Light Industry and Food Engineering, Guangxi University, Nanning 530004, China; 2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; 3. Collaborative Innovation Center for Guangxi Sugar Industry, Nanning 530004, China)
  • Online:2018-06-25 Published:2018-06-15

摘要: 建立一种能够精密检测产物右旋糖酐分子质量变化及定量跟踪检测蔗糖、葡萄糖及果糖浓度的方法。通过对比高效凝胶过滤色谱单柱和双柱串联对右旋糖酐标准品及催化产物的检测效果,探寻得到右旋糖酐分子质量检测的有效方法。同时,通过构建右旋糖酐蔗糖酶耦合两级膜探索得到右旋糖酐的合成机理是葡萄糖基在右旋糖酐蔗糖酶活性位点上,不断聚合,直至达到一定的分子质量(mw>106 Da)才脱落,而不是一个从小到大、慢慢聚合、不断脱落的过程。

关键词: 高效凝胶过滤色谱, 双柱串联, 右旋糖酐分子质量, 聚合过程

Abstract: A precise method for the determination of changes in the molecular mass of dextran and the quantification of sucrose, fructose and glucose concentration during the dextransucrase-catalyzed reaction of sucrose was presented in this paper. High performance gel filtration chromatography (HPGFC) with one column and with two tandem columns were compared for their efficiencies in the determination of dextran standard and the catalytic reaction products in order to develop a robust method to measure the molecular mass of dextran. By dextransucrase combined with two-stage membrane treatment, we investigated the mechanism underlying dextran synthesis. It was found that the synthesis process was continuous aggregation of glucosyl group at the active site of dextransucrase until reaching a specific molecular mass (> 106 Da) before detachment rather than stepwise aggregation and then detachment.

Key words: high performance gel filtration chromatography (HPGFC), two tandem columns, molecular mass of dextran, aggregation process

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