摘要
目的从大海马中提取和纯化糖蛋白,并对其结构进行初步鉴定,为大海马糖蛋白抗氧化特性研究提供依据。方法采用Sevage法从大海马中提取粗糖蛋白,并用DEAE-52阴离子交换柱和葡聚糖G-100进行粗糖蛋白纯化,采用苯酚硫酸法和考马斯亮蓝法测定纯化后的组分中糖和蛋白质含量,并对纯化后的组分进行单糖成分、氨基酸组成、糖肽键和初级结构的鉴定及清除自由基的能力的测定。结果从大海马粗糖蛋白中纯化得到2个海马糖蛋白组分HG-11和HG-21,其糖和蛋白质含量分别为56.80%和30.55%、39.48%和51.75%,HG-11和HG-21中单糖主要是甘露糖、核糖、葡萄糖和半乳糖,糖和蛋白质组分是通过O-糖苷键连接的,HG-11检测出15种氨基酸,HG-21检测出16种氨基酸,通过红外光谱两者都发现了糖和蛋白的特征吸收峰,并且多糖含有α型吡喃糖,蛋白质的二级结构具有α螺旋结构。结论海马糖蛋白对自由基的清除能力是由多糖和蛋白质共同决定的,与葡萄糖、半乳糖、疏水性氨基酸、芳香族氨基酸、糖和蛋白质之间的链接方式及空间结构都有着密切的关系。
Objective The glycoprotein was extracted and purified from the hippocampus,and its structure was preliminarily identified,which provided the basis for the study of the antioxidant properties of hippocampal glycoprotein.Methods Novel glycoprotein were isolated from the yellow seahorse＇s by Sevage reagent crude extracts,and purified by DEAE-52 anion exchange column and Sephadex G-100 gel column.The content of sugar and protein in the purified fraction was determined by phenol sulfuric acid method and Coomassie brilliant blue method.The chemical analyses（including monosaccharide composition,amino acid composition,glycan peptide bond etc.）of HG-11 and HG-21 were identified and crude glycoprotein scavenging free radical activities were determined.Results After purification,HG-11 and HG-21 were obtained from the novel glycoprotein,HG-11 contained 56.80% sugar moiety and 30.55% protein,and HG-21 contained 39.48% sugar moiety and 51.75% protein.The composition of monosaccharides consists of mannose,ribose,glucose and galactose.The existences of O-glycosidic linkage in HG-11 and HG-21 were demonstrated with aβ-elimination reaction.Fifteen and 16 amino acids were identified in HG-11 and HG-21,respectively.The FT-IR spectrum of HG-11 and HG-21 showed typical characteristics of polysaccharide and protein,and polysaccharides containingα-type pyranose,secondary structure of proteins havingα-helix structure.Conclusion The glycoprotein scavenging free radical activities were affected by polysaccharides and proteins,which were closely related with glucose,galactose,hydrophobic amino acids,aromatic amino acid,glycan peptide bond and spatial structure.The free radical scavenging activities and structure-function relationship of glycoprotein are important in the free radicals medicine.
出处
《中国海洋药物》
CAS
CSCD
2017年第6期39-46,共8页
Chinese Journal of Marine Drugs
基金
国家自然科学基金项目(41276123)
浙江省新苗计划项目(2016R405090)资助
关键词
大海马
糖蛋白
化学分析
抗氧化特性
Hippocampus kuda Bleeker
glycoprotein
chemical analyses
antioxidant properties
