短蛸章鱼多糖的提取分离和基本性质分析

Extraction,separation and physicochemical properties analyses of Octopus ocellatus polysaccharides

目的从短蛸(Octopus ocellatus)的胴体和腕部提取多糖,对短蛸不同部位提取的粗多糖的基本理化性质进行分析和比较,并对粗多糖进行分离和纯化。方法依次采用冷水,木瓜蛋白酶和胰蛋白酶联合酶解的方法提取多糖,采用Sevage法和等电点法去除粗多糖中的蛋白。对短蛸不同部位提取粗多糖的总糖、蛋白、糖醛酸、硫酸根含量和重均相对分子质量、单糖组成等性质进行分析和比较,并采用Sephacryl S-300凝胶柱层析和QSepharose4 Fast Flow离子交换层析方法对粗多糖进行分离和纯化。结果从短蛸胴体中提取的粗多糖中总糖和糖醛酸的含量明显高于腕部粗多糖,且蛋白含量较腕部多糖低;水提多糖中总糖和糖醛酸的含量明显低于酶提多糖,且蛋白含量高于酶提多糖;短蛸胴体粗多糖的单糖组成相对简单,主要含有Glc和少量Man、GlcNAc、GlcUA,而腕部粗多糖的单糖组成较为复杂,除主要含有Man、GlcNAc、GlcUA和GlcN外,还含有少量Gal和Fuc等;各多糖的重均相对分子质量在127.9～266.4kD之间;水提多糖和酶提多糖在SephacrylS-300凝胶柱层析上的洗脱曲线也明显不同。结论采用不同方法、从不同部位提取的多糖基本理化性质无论是总糖、糖醛酸和蛋白的含量,还是单糖组成和在凝胶柱层析上的洗脱曲线都存在明显的差异。

Objective To compare the physico-chemical properties of polysaccharides which extracted and separated from the naked body and wrist of Octopus ocellatus,respectively,and purify these polysaccharides by gel permeation chromatography and ion exchange chromatography.Methods Polysaccharides from different parts of Octopus ocellatus were extracted in order by distilled water,enzymolysis with combination of papain and trypsin digestion,and then the proteins existed in polysaccharides were removed by Sevage and isoelectric precipitation methods.The contents of total sugars,protein,uronic acid,sulfate,molecular weight and monosaccharide composition of Octopus ocellatus polysaccharides were determined,and these polysaccharides were purified further by Sephacryl S-300 and Q Sepharose 4 Fast Flow chromatography.Results The contents of total sugar and uronic acid of polysaccharides from the naked body were higher than that from wrist,and the contents of protein were lower than that from wrist.The contents of total sugar and uronic acid of polysaccharides extracted by water were lower than that extracted by enzymolysis,and the contents of protein were high.Monosaccharide compositions of polysaccharides extracted from naked body were simple,the major component was Glc with a little of Man,GlcNAc and GlcUA,while monosaccharide compositions of polysaccharides extracted from wrist were complicated,including Man,GlcNAc,GlcUA,GlcN as well as small amounts of Gal,Fuc and Fru.The average molecular weights of these polysaccharides were ranged from 127.9 to 266.4 kD.Elution curves of polysaccharides extracted by distilled water and enzymolysis on Sephacryl S-300 chromatography were quite different also.Conclusion The physico-chemical properties of polysaccharides,not only the contents of total sugar,protein,uronic acid,but also the monosaccharide compositions and elution curves are different obviously,which extracted by different methods and from different parts of Octopus ocellatus.