文章摘要
一种新型乌贼墨糖胺聚糖的分离纯化、理化性质及其抗氧化活性研究
Separation, purification and analyses on physicochemistry properties, antioxidative activity of a novel glycosaminoglycans from Sepia esculenta
投稿时间:2014-10-09  修订日期:2015-01-14
DOI:
中文关键词: 乌贼墨  糖胺聚糖  单糖组成  理化性质  抗氧化活性
英文关键词: Sepia esculenta ink  glycosaminoglycans  monosaccharide composition  physicochemical property  antioxidative activity
基金项目:海洋药物教育部重点实验室基金
作者单位E-mail
牛庆凤 浙江海洋学院 342457383@qq.com 
王斌 浙江海洋学院 食品与医药学院  
李涛 浙江海洋学院 食品与医药学院  
张申微 浙江海洋学院 食品与医药学院  
陈荫 浙江海洋学院 食品与医药学院 mojojo1984@163.com 
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中文摘要:
      目的 从金乌贼墨(Sepia esculenta ink)中提取其活性成分乌贼墨多糖。并首次对其理化性质及体外抗氧化活性进行研究。方法 依次用两步酶解法提取乌贼墨粗多糖,用Q Sepharose 4 Fast Flow 阴离子交换层析柱对其进行分离纯化,获得主要组分。通过高效液相(HPLC)、紫外光谱(UV)、红外光谱(IR)、离子色谱法对乌贼墨多糖进行理化性质研究以及初步的结构判定,采用ABTS法对乌贼墨进行总的体外抗氧化活性研究。结果 两步酶解后乌贼墨粗多糖的得率为3%,分离纯化后得到0.05、0.25、0.5 mol/L NaCl三个洗脱组分,分别命名为SE-1、SE-2、SE-3,其中以SE-2为最主要组分,故主要对SE-2进行分析。经过离子交换柱层析,SE-2得率为50%。SE-2主要含有半乳糖胺(GalN)、岩藻糖(Fuc)、葡萄糖胺(GlcN)、乙酰氨基半乳糖(GalNAc)、葡萄糖醛酸(GlcUA)以及甘露糖(Man)等单糖,紫外扫描无蛋白质的特征吸收峰;该多糖为均一性多糖,分子量8.5 kD;SE-2的总糖含量、蛋白质含量、硫酸根含量、氨基己糖含量分别为54.2%、3.92%、7.74%、20.5%;IR光谱进一步证明该糖为SE-2糖胺聚糖,构象为D-吡啶环结构,并有硫酸根的存在;SE-2对ABTS自由基的清除能力较强,其半数清除质量浓度为0.21mg/mL。结论 金乌贼墨多糖SE-2是一种硫酸酯化的新型糖胺聚糖,具有较强的体外抗氧化性活性。
英文摘要:
      Objective To extract , isolate and purify polysaccharides from Sepia esculenta ink and analyze physicochemical properties and antioxidant activity of these polysaccharides. Methods The main component was fractionated by a Q Sepharose Fast Flow column(3.5 × 30 cm) from crude polysaccharide. HPLC, HPGPC, UV spectrophotometer, IR spectrometry, ion chromatography were used to determinate the physicochemical and stuctural properties; ABTS was used to analyse the total antioxidative activity of main polysaccharide. Results After two-step enzyme solution, the yield of the crude polysaccharide was 3%. Three components which were eluted with a step-wise gradient of 0.05, 0.25 and 0.5 mol/L NaCl were named SE-1, SE-2, SE-3 while SE-2 was the main component. Through ion exchange column chromatography,the yield of SE-2 relative to the pure polysaccharide was 50%. The fraction of SE-2 was found to be composed of galactosamine, fucose, glucosamine, N-acetylgalactosamine, glucuronic acid andmannose; From HPGPC,it was proved to be homogeneous and had a molecular weight of 8.5 kD; The concents of total sugar, protein, sulfate and aminohexose of SE-2 was 54.2%、3.92%、7.74% and 20.5% respectively; In the IR spectrogram,it was shown the typical absorption of sugar residues, glycosaminoglycans and sulfate ester;The ability of scanvenging the ABTS free radicals of SE-2 is very strong. The half maximal inhibitory concentrations(IC50) against ABTS free radicals was 0.21 mg/mL respectively. Conclusion The research shows SE-2 which was isolated from Sepia esculenta ink proved to be a novel sulfated glycosaminoglycans and had excellent antioxidative activity on ABTS free radicals.
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