一种龙须菜中纯化藻红蛋白的新方法

A New Purification Method of Phycoerythrin from Gracilariopsis lemaneiformis

藻红蛋白(Phycoerythrin,PE)是龙须菜主要的捕光色素蛋白之一,呈红色,具有高荧光量子产率,被广泛应用于生物医学、食品和化妆品行业。本文交替运用疏水层析和分子筛层析分离纯化龙须菜中的PE。先使用低温组织捣碎法对龙须菜细胞进行破壁作为粗提液,硫酸铵分级沉淀处理后经磷酸盐缓冲液重悬,经疏水层析和分子筛层析,最终获得纯度(A_(565)/A_(280))达到3.8的PE,同时表征了最终纯化产物的可见光吸收光谱、荧光发射光谱及Gauss解叠运算分析,证明最终纯化得到的PE保持了天然的光学活性和蛋白构象。此纯化工艺为今后从龙须菜中大规模提取PE,开发相关产品提供了良好的借鉴。

Phycoerythrin(PE)is one of the major light-harvesting pigment proteins in Gracilariopsis lemaneiformis with bright red color and high fluorescence quantum yield.PE can be widely used in biomedical,food and cosmetic industries.In this paper,hydrophobic chromatography and gel filtration chromatography were used to purify PE from G.lemaneiformis.Algae cells were broken by low temperature tissue crushing,after fractional precipitation with ammonium sulfate,it was resuspended in phosphate buffer and subjected to hydrophobic chromatography and gel filtration chromatography.Finally,the purity(A_(565)/A_(280)) reached 3.8 after treatment of hydrophobic chromatography column and gel filtration chromatography,which the supernatant containing PE was used as loading sample.The analysis of visible light absorption spectrum,fluorescence emission spectrum and Gauss dissolution analysis demonstrated that the corrected functions in energy transfer and natural optical activities of purified PE.This new purification protocol provided a good strategic on large-scale extraction of PE from G.lemaneiformis.After fractional precipitation with ammonium sulfate,it was resuspended in phosphate buffer and subjected to hydrophobic chromatography and gel filtration chromatography.