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Efficient separation and purification of allophycocyanin from Spirulina (Arthrospira) platensis

Su, Hai-Nan, Xie, Bin-Bin, Chen, Xiu-Lan, Wang, Jin-Xia, Zhang, Xi-Ying, Zhou, Bai-Cheng, Zhang, Yu-Zhong
Journal of applied phycology 2010 v.22 no.1 pp. 65-70
Arthrospira, Rhodophyta, Spirulina, absorbance, ammonium sulfate, anion exchange chromatography, fractionation, pH, phosphates, phycobiliprotein, polyacrylamide gel electrophoresis, sodium, sodium dodecyl sulfate
Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.