Aislamiento, purificación y caracterización parcial de la C-ficocianina-620 obtenida a partir de la Spirulina platensis

Abstract

Spirulina platensis has a big quantity of phycobiliproteins, such as phycocyanin and allophycocyanin. They are components of a macromolecular complex called phycobilisome. The phycobiliproteins are colored and fluorescent pigment-proteins which constitute the light-harvesting antenna complexes in cyanobacteria (blue-green algae). These chromoproteins have covalently attached linear tetrapyrroles, bilins, structurally linked to the biliverdin. For their fluorescent properties, phycobiliproteins are used in industry, biotechnology and medicine. The extraction method of phycobiliproteins from biomass of Spirulina platensis was by freeze-unfreeze at -20°C. The C-phycocyanin was purified by anionic exchange chromatography using gel DEAE Sepharose Fast Flow and then
its purity was verified by electrophoresis (8 to 25 % polyacrylamide) and high pressure liquid chromatography. For the partial characterization of the C-phycocyanin hexamer, its alpha and beta subunits, were separated by gel filtration. Their purity was proved by electrophoresis (8 to 25 % polyacrylamide). Later, the molecular weight of the hexamer and its subunits, was estimated. The pure C-phycocyanin with 229 kDa and the alpha and beta subunits, also pure, with 17 .92 and 19 .85 kDa were obtained respectively.