| 摘要: |
| 于1995年3—7月采用本实验室培养的绿色巴夫藻，经沸水提取及去除蛋白、小分子物等杂质后，通过Sephadex G-50柱层析进行分离、纯化实验。结果表明，分离纯化所得到的两种多糖—4P1和4P2，通过醋酸纤维素薄膜电泳及Sephadex G-100凝胶过滤检验纯度后，证实其均为单一含硒多糖，且都带负电。两种硒多糖含硒量都较低，分别为77mg/g和220mg/g。 |
| 关键词: 硒多糖 分离 纯化 |
| DOI：10.11693/hyhz200006009009 |
| 分类号: |
| 基金项目:国家“八五”科技攻关资助项目，85-08-07-04号 |
附件 |
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| EXTRACTION, ISOLATION, AND PURIFICATION OF SELENIUM POLYSACCHARIDE IN PAVLOVA VIRDIS |
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ZHANG Xin- yu1, WANG Lei1, LI Guang- you2, ZHU Yan-xiong3
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1.Institute of Oceanology,The Chinese Academy of Sciences,Qingdao,266071;2.First Institute of Oceanology , State Oceanic Administration, Qingdao, 266003;3.The Centre of Science and Technology Information , Qingdao, 266003
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| Abstract: |
| A kind of marine microalgae, Pavlova virdis, which we entitled 4P, was used to extract selenium-polysaccharide in laboratory. It was extracted three times in boiling water. After centrifugal operation ( 3000 round per minute, 20 minutes) , the supernatant was collected and further isolated with Savag method (6–10 times) and dialysis method ( in running water for 24 hours) in order to wipe out proteins and other little impurity. Then through purifying it with Sephadex G-50 gel filtration (diameter 20mm; length 920mm) we obtained two kinds of pure selenium-polysaccharides, which were entitled 4P1 and 4P2. They were checked with Sephadex G-100 gel filtration and cellulose acetate film electrophoresis. The results showed that they were homogeneous and negatively charged. The electrophoresis condition was barbitone buffer (pH= 8.6, Ion intensity= 0.09) , constant 120 Volt for 20 minutes, 0.5% toluidine blue dying for 5–10 minutes, and 0.5% HAc solution decolouring several times.
Through this experiment, we measured the selenium (atom-fluorescence method) and sugar (Vitriol- Phenolmethod) content after each procedure. The ratio of gaining selenium from 4P marine microalga was only 8.5%. It contained 77mg selenium per gram 4P1, and 220mg selenium per gram 4P2. The selenium content in selenium-polysaccharide is relatively small.
In conclusion, Pavlova virdis contained two kinds of pure selenium-polysaccharide, 4P1 and 4P2, it also showed that Pavlova virdis has very good ability to collect selenium during its growth. |
| Key words: Selenium-polysaccharide, Isolation, Purification |
