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从可食用红海藻蜈蚣藻中一步纯化R-藻红蛋白。

One-step purification of R-phycoerythrin from the red edible seaweed Grateloupia turuturu.

作者信息

Munier Mathilde, Morançais Michèle, Dumay Justine, Jaouen Pascal, Fleurence Joël

机构信息

FR CNRS 3473 IUML, Mer Molécule Santé (MMS), EA 2160, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.

FR CNRS 3473 IUML, UMR-CNRS 6144 (GEPEA), Université de Nantes, CRTT 37 boulevard de l'Université, BP 406, 44602 Saint-Nazaire Cedex, France.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Jun 15;992:23-9. doi: 10.1016/j.jchromb.2015.04.012. Epub 2015 Apr 17.

Abstract

A one-step chromatographic method for the purification of R-phycoerythrin (R-PE) of Grateloupia turuturu Yamada is described. Native R-PE was obtained with a purity index of 2.89 and a recovery yield of 27% using DEAE-Sepharose Fast Flow chromatography with a three-step increase in ionic strength. The analysis by SDS electrophoresis showed a broad band between 18 and 21kDa in size corresponding to subunits α and β and a low intensity band of 29kDa corresponding to the γ subunit. Two forms of R-PE were identified by gel filtration chromatography: a native form with a molecular weight of 260±5kDa and a dissociated form with a molecular weight of 60±2kDa. The native form presented the characteristic absorption spectrum of R-PE with three absorbance maxima at 498, 540 and 565nm, whereas the dissociated form presented only the 498 and 540nm peaks. Moreover, the two forms displayed two different fluorescence maxima.

摘要

描述了一种用于纯化海膜(Grateloupia turuturu Yamada)中R-藻红蛋白(R-PE)的一步色谱法。使用DEAE-琼脂糖快速流动色谱法,通过三步增加离子强度,获得了纯度指数为2.89且回收率为27%的天然R-PE。SDS电泳分析显示,在18至21kDa之间有一条宽带,对应于α和β亚基,还有一条强度较低的29kDa带,对应于γ亚基。通过凝胶过滤色谱法鉴定出两种形式的R-PE:一种分子量为260±5kDa的天然形式和一种分子量为60±2kDa的解离形式。天然形式呈现出R-PE的特征吸收光谱,在498、540和565nm处有三个吸光度最大值,而解离形式仅呈现498和540nm的峰。此外,这两种形式表现出两个不同的荧光最大值。

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