高等学校化学学报 ›› 2014, Vol. 35 ›› Issue (12): 2547.doi: 10.7503/cjcu20140681

• 有机化学 • 上一篇    下一篇

基于电喷雾串联质谱的蓝藻寡糖脱果糖分析及系列α-1,2-葡聚寡糖的制备

张洪涛1, 朱莉2, 詹晓北1()   

  1. 1. 江南大学生物工程学院糖化学与生物技术教育部重点实验室, 无锡 214122
    2. 江苏瑞光生物科技有限公司, 无锡 214125
  • 收稿日期:2014-07-24 出版日期:2014-12-10 发布日期:2014-11-29
  • 作者简介:联系人简介: 詹晓北, 男, 博士, 教授, 博士生导师, 主要从事多糖的发酵法生产研究. E-mail:xbzhan@yahoo.com
  • 基金资助:
    国家自然科学基金(批准号: 31201384, 3117164)、 国家博士后科学基金(批准号: 2012M20996)、 江苏省博士后科研资助计划项目(批准号: 1301011B)、 国家建设高水平大学公派研究生项目(批准号: 2008679005)、 高等学校学科创新引智计划(批准号: 111-2-06)和中央高校基本科研业务费专项资金(批准号: JUSRP1007)资助

Assignment of Removing Fructose in Reducing Terminal of Nostoc Oligosaccharides Based on ESI-CID-MS/MS and Preparation of Full Series of α-1,2-Gluco-oligosaccharide

ZHANG Hongtao1, ZHU Li2, ZHAN Xiaobei1,*()   

  1. 1. Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Biotechnology,Jiangnan University, Wuxi 214122, China
    2. Jiangsu Rayguang Biotech Co. Ltd., Wuxi 214125, China
  • Received:2014-07-24 Online:2014-12-10 Published:2014-11-29
  • Contact: ZHAN Xiaobei E-mail:xbzhan@yahoo.com
  • Supported by:
    † Supported by the National Natural Science Foundation of China(Nos.31201384, 3117164), the Postdoctoral Science Foundation of China(No.2012M20996), the Jiangsu Planned Projects for Postdoctoral Research Funds, China(No.1301011B), the China Scholarship Council Program(No.2008679005), the “111” Project(No.111-2-06) and the Fundamental Research Funds for the Central Universities of China(No.JUSRP1007)

摘要:

为获得系列α-1,2-葡聚寡糖, 首先以蓝藻寡糖六糖、 八糖、 九糖和十糖为原料, 在0.5 mol/L的三氟乙酸(TFA)中于95 ℃酸解9 min以脱去还原端果糖, 经低压凝胶色谱分离纯化, 用电喷雾离子化-碰撞诱导解离-串联质谱(ESI-CID-MS/MS)和基质辅助激光解吸电离质谱(MALDI-MS)鉴定和序列表征, 获得了除去末端果糖的α-1,2-五、 七、 八和九糖; 然后在0.5 mol/L的TFA中于95 ℃对混合蓝藻寡糖六糖和八糖酸水解45 min, 用Bio-Gel P2凝胶柱对混合物进行分离和纯化, 并通过ESI-MS和MALDI-MS对获得的每个寡糖组份进行表征, 获得了聚合度为2, 3, 4和6的α-1,2-葡聚寡糖. 本研究为利用糖生物芯片技术进行α-1,2-葡聚寡糖的功能筛选及分析其与靶标蛋白之间相互作用的特异性提供了葡聚寡糖物质基础.

关键词: 蓝藻寡糖, 脱果糖, α-1,2-葡聚寡糖, 电喷雾串联质谱

Abstract:

Carbohydrate microarray has become a powerful tool to explore the structure-function relationship between protein-carbohydrate, but it depends on enough reducing terminal oligosaccharides with aldehyde group. Nostoc oligosaccharides are a kind of oligo(1→2)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranoside. To acquire a full series of α-1,2-gluco-oligosaccharides from oligo(1→2)-α-D-glucopyranosyl-(1→2)-β-D- fructofuranoside, the terminal fructose of four Nostoc oligosaccharides were removed using acid hydrolysis methods under the condition of 0.5 mol/L trifluoroacetic acid(TFA) treated at 95 ℃ for 9 min, respectively. Following hydrolyzed products were separated and purified using high performance liquid chromatography(HPLC), and then electrospray ionization-collision-induce dissociation tandem mass spectrometry(ESI-CID-MS/MS) and matrix assisted laser desorption ionization(MALDI)-MS were used to identify the removing of terminal fructose and sequenced, α-1,2-Glc5, Glc7, Glc8 and Glc9 were acquired. The mixture of Nostoc-Octa and Nostoc-Hexa treated with 0.5 mol/L TFA at 95 ℃ for 45 min were used to acquire low degree of polymerization(DP) of α-1,2-gluco-oligosaccharides, which were separated with P2 column, ESI-MS and MALDI-MS were used to analysis these oligosaccharides, α-1,2-gluco-oligosaccharides with the degree of polymerization 2, 3, 4 and 6 were obtained. Finally, full series of α-1-2-gluco-oligosaccharides from 2 mer to 9 mer were acquired successfully.

Key words: Nostoc oligosaccharide, Fructose removing, α-1,2-Gluco-oligosaccharide, Electrospray ionization-collision-induce dissociation tandem mass spectrometry(ESI-CID-MS/MS)

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