紫球藻藻红蛋白的分离纯化及光谱学特性

Isolation and purification of phycoerythrin from Porphyridium cruentum and its spectrum characteristics

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(PorphyridiumcruentumNaegeli)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。

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 of 34.0% when it was further purified by hydroxyl column chromatography. When the purified B-phycoerythrin was electrophoresed with polyacrylamide gel electrophoresis, only one band was 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 hvdroxvl column.