Abstract
This paper describes simple procedures for the purification of large amounts of phycocyanin and allophycocyanin from the cyanobacterium Microcystis aeruginosa. A homogeneous natural bloom of this organism provided hundreds of kilograms of cells. Large samples of cells were broken by freezing and thawing. Repeated extraction of the broken cells with distilled water released phycocyanin first, then allophycocyanin, and provides supporting evidence for the current models of phycobilisome structure. The very low ionic strength of the aqueous extracts allowed allophycocyanin release in a particulate form so that this protein could be easily concentrated by centrifugation. Other proteins in the extract were enriched and concentrated by large scale membrane filtration. The biliproteins were purified to homogeneity by chromatography on DEAE cellulose. Purity was established by HPLC and by N-terminal amino acid sequence analysis. The proteins were examined for stability at various pHs and exposures to visible light.
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Abbreviations
- Aλ:
-
absorbance at wavelength in nanometers
- DEAE cellulose:
-
diethylamino ethyl cellulose
- HPLC:
-
high pressure liquid chromatography
- UV:
-
ultraviolet
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Padgett, M.P., Krogmann, D.W. Large scale preparation of pure phycobiliproteins. Photosynth Res 11, 225–235 (1987). https://doi.org/10.1007/BF00055062
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DOI: https://doi.org/10.1007/BF00055062