摘要 | 采用4℃反复浸提、离心、硫酸铵沉淀、DEAE-Sepharose Fast F low离子交换柱层析,从紫球藻(Porphyridium cruentum Naegeli)冻干粉中分离纯化藻红蛋白,分离纯度达到4.85,总收率51.9%;经羟基磷灰石柱层析纯化,藻红蛋白纯度达到5.10,总收率34.0%,聚丙烯酰胺凝胶电泳显示1条带。所分离纯化的藻红蛋白含有3个亚基(α、β、γ),在可见光区545 nm和560 nm处有2个吸收峰,在498 nm处有1个吸收肩峰。实验结果说明所分离纯化的藻红蛋白纯度符合要求。 |
Abstract | The freeze dried powder of Porphyridium cruentum Naegeli was extracted with distillated water at 4℃ , centrifugilized, precipitated by(NH4)2SO4, chromatographed on a DEAE-Sepharose Fast Flow ion exchange column. As a result, the B-phycoerythrin was got with a purity of 4. 85 and a yield of 51. 9%, and with a purity of 5. 10 and a yield of34. 0% when it was further purified by hydroxyl column chromatography. When the purified B-phycoerythrin was electrophoresed with polyacrylamide gel electrophoresis, only one bandwas observed. The purified B-phycoerythrin had three subunits (α,β,γ) and possessed two peaks at 545 nm and 560 nm, and a shoulder at 498 nm. The results showed that phycoerythrin from P. cruentum could be separated and purified quickly on the DEAE-Sepharose Fast Flow ion exchange column and hydroxyl column. |
关键词 | 紫球藻; 藻红蛋白; 分离纯化; 光谱学特性; |
Key words | Porphyridium cruentum Naegeli; phycoerythrin; isolation and purification; spectrum characteristics |
作者 | 黄 键1,王 娟1,陈必链1 ,王明兹1,梁世中2 |
所在单位 | 1.福建师范大学生命科学学院,福建福州350007; 2.华南理工大学生物科学与工程学院,广东广州510640 |
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基金项目 | 福建省发展和改革委员会资助课题(闽计投资[2003]203); |