红藻中r -藻红蛋白的提取与纯化

Huu Phuoc Trang Nguyen, M. Morançais, J. Fleurence, T. Tran, J. Dumay
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引用次数: 7

摘要

r -藻红蛋白(R-PE)是三种被广泛用作荧光探针、食品和化妆品生产中的天然着色剂的藻胆蛋白之一,由红藻制备而成。星状乳香被认为是卡拉胶和蛋白质的丰富来源。本研究采用20 mM磷酸盐缓冲液(pH 7.1)提取红藻Mastocarpus stellatus中的R-PE,并采用DEAE sepharose Fast Flow色谱法进行离子强度三步提高纯化。NaCl洗脱步骤将粗提物分离成AE-150、AE-200和AE-1000三个馏分。采用分光光度法、凝胶过滤法等不同的监测方法,考察了R-PE回收率的质量和数量。AE-200馏分A565/A280的纯度指数(PI)为1.91,是粗提物的14倍,是硫酸铵饱和度85%沉淀法的8倍左右。
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Extracting and Purifying Pigment R-phycoerythrin from the Red alga Mastocarpus Stellatus
R-phycoerythrin (R-PE) is one of the three phycobiliproteins which are extensively employed as fluorescent probes, natural colorant for the production of food and cosmetics and it is prepared from red macroalgae. Mastocarpus stellatus is known as a rich source of carrageenan and protein. In this study, the R-PE in the red seaweed Mastocarpus stellatus was extracted in 20 mM phosphate buffer (pH 7.1) and purified by DEAE sepharose Fast Flow chromatography with a three-step increase in ionic strength. The NaCl elution step allowed the crude extracts to be separated into three fractions: AE-150, AE-200 and AE-1000. The quality and quantity of R-PE recovery were indicated different methods to monitor each fraction obtained, such as spectrophotometric indicators, gel filtration. The A565/A280 ratio (Purity Index, PI) obtained after the purification step was 1.91 in the fraction AE-200 and this value was higher 14 times than the PI of the crude extract and about 8 times than the sample precipitation by ammonium sulfate saturation of 85%.
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