祝 贺,张玉倩,郎银芝,等.3种鱼鳃中糖胺聚糖的分离纯化及细微结构比较分析[J].中国海洋药物,2014,33(5):1-8. |
3种鱼鳃中糖胺聚糖的分离纯化及细微结构比较分析 |
Isolation, purification and fine structure comparison analysis of glycosaminoglycans from three fish gills |
投稿时间:2014-03-04 修订日期:2014-03-27 |
DOI: |
中文关键词: 鱼鳃 糖胺聚糖 杂合硫酸软骨素 二糖组成 核磁共振氢谱 |
English Keywords:Fish gill Glycosaminoglycan Hybrid chondroitin sulfate Disaccharide composition Protonnuclear magnetic resonance |
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中文摘要: |
目的 从狭鳕 (Alaska pollack)、褐牙鲆 (Bastard halibut) 和真鲷 (Red seabream) 三种鱼鳃中提取分离糖胺聚糖并对其细微结构进行比较分析。方法 利用酶法降解并以阴离子交换色谱法分离纯化糖胺聚糖,通过单糖组成和醋酸纤维素薄膜电泳对其种类进行鉴别,采用高效液相色谱法对其相对分子量和二糖组成进行比较分析,最后利用核磁共振波谱法 (1H-NMR) 对其结构进行表征。结果 从3种鱼鳃组织中提取分离得到54种多糖,其中18种是电荷和分子量均一的糖胺聚糖,它们集中在0.8 mol?L-1 NaCl洗脱部分 (P4) 和1.0 mol?L-1 NaCl洗脱部分 (P5),占多糖总量的17.4~58.4%;P4主要为硫酸软骨素C (CSC),P5主要为硫酸软骨素A (CSA) 和杂合硫酸软骨素 (HyCS),尤其,与牛气管来源CSA比较,HyCS的4S/6S比和电荷密度均较大。结论 狭鳕、褐牙鲆和真鲷鱼鳃组织中的糖胺聚糖主要为CSA、CSC和HyCS。 |
English Summary: |
Objective To extract and purify glycosaminoglycans (GAGs) from three fish gills of Red seabream, Bastard halibut and Alaska pollack and characterize their fine structures. Methods The fish gills were firstly digested with enzymes, and the GAGs were isolated and purified by anion exchange chromatography. The sorts of GAGs were identified by monosaccharide composition analysis and cellulose acetate membrane electrophoresis, and the disaccharide composition and relative molecular mass were determined and compared by high performance liquid chromatography. At last, protonnuclear magnetic resonance spectroscopy was applied to characterize their structures. Results Fifty-four polysaccharides were acquired, of which eighteen molecular size and charge homogeneous GAGs were eluted at 0.8 mol?L-1 NaCl (P4) and 1.0 mol?L-1 NaCl (P5) on Q-Sepharose Fast Flow column from three fish gill tissues, and the yield account for 17.4-58.4% of total polysaccharide, respectively. Fraction P4 was mainly identified as chondroitin sulfate C (CSC), and P5 was mainly identified as CSA and hybrid chondroitin sulfate (HyCS). Especially, HyCS had a bigger ratio of 4-sulfate disaccharide to 6-sulfate disaccharide (4S/6S) and higher charge density than CSA purified from bovine trachea. Conclusion GAGs contained in fish gill tissues of Alaska pollack, Bastard halibut and Red seabream were mainly CSA, CSC and HyCS . |
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