Isolation, purification and characteristics of R-phycoerythrin from a marine macroalga Heterosiphonia japonica |
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Authors: | Sun Li Wang Shumei Gong Xueqin Zhao Mingri Fu Xuejun Wang Lu |
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Institution: | aCollege of Chemistry Engineering and Biology Sciences, Yantai University, No. 30 Qingquan Road, Yantai 264005, PR China;bCollege of Photo-electronic Information Science and Technology, Yantai University, Yantai 264005, PR China |
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Abstract: | R-phycoerythrin is one of the three phycobiliproteins which are extensively employed as fluorescent probes, and it is prepared from red macroalgae. Phycobiliproteins in the marine red macroalga Heterosiphonia japonica were extracted in 50 mM phosphate buffer (pH 7.0) and precipitated by salting-out. The R-phycoerythrin was isolated by gel filtration with Sepharose CL-4B and Sephadex G-200. Then it was purified by ion exchange chromatography on DEAE Sepharose Fast Flow which was developed by linear ionic strength gradients. The purified R-phycoerythrin gave a ratio of A565 to A280 of 4.89. It showed a single band and a pI of 4.8 on the examination by polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing. The polypeptide analysis of the purified R-phycoerythrin by SDS–PAGE demonstrated that it contains four chromophore-carrying subunits and no colorless polypeptide and has two hexameric aggregates. The preparative procedures of the R-phycoerythrin purification established based on the experiments exhibit advantages and can offer a reference for R-phycoerythrin preparation from other marine red macroalga. |
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Keywords: | Phycobiliprotein R-phycoerythrin Heterosiphonia japonica Chromatography Gel electrophoresis Isoelectric focusing |
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