Large-scale isolation and purification of C-phycocyanin from the cyanobacteria Anabaena marina using expanded bed adsorption chromatography

• Published in: Journal of chemical technology and biotechnology (1986) Vol. 85; no. 6; pp. 783 - 792
• Main Authors: Ramos, Amparo, Acién, F.Gabriel, Fernández-Sevilla, José M, González, Cynthia V, Bermejo, Ruperto
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.06.2010; Wiley
• Subjects: Adsorption; Anabaena marina; anion-exchange chromatography; Applied sciences; C-phycocyanin; Chemical engineering; Chromatography; Columns (process); Cost engineering; Exact sciences and technology; expanded bed adsorption chromatography; Extraction; Ion exchange; Macromolecules; Marinas; phycobiliproteins; preparative protein purification; Purification; Viscosity; Anabaena marina; Purification; Scale effect; Biomass; Adsorption chromatography; preparative protein purification; Ion exchange chromatography; Extrapolation; expanded bed adsorption chromatography; Adsorption; C-phycocyanin; Ion exchange; anion-exchange chromatography; phycobiliproteins
• ISSN: 0268-2575; 1097-4660; 1097-4660
• DOI: 10.1002/jctb.2361

BACKGROUND: Phycobiliproteins are water soluble proteins useful as fluorescent markers of cells and macromolecules, and as natural colorants, and are anticarcinogenic. Although phycobiliproteins have many applications, their use is limited by the high cost of the purified macromolecules, mainly related with the cost of extraction and purification. In this study a fast and scalable method for preparative extraction and purification of C-phycocyanin (C-PC) from Anabaena marina is developed.RESULTS: The method developed consists in the extraction of phycobiliproteins using repeated single contact strategy, separation being performed by expanded bed adsorption (EBA) chromatography using Streamline-DEAE. Optimal conditions for EBA were obtained at small scale, using a 15 mm internal diameter column, these being a sample load of 0.9 mg C-PC mL⁻¹ adsorbent, an expanded bed volume twice the settled bed volume and a sample viscosity of 1.109 mP. The process was then scaled up 36 times, the success of the scale-up process being verified. Finally, to obtain pure C-PC conventional ion-exchange chromatography was utilized.CONCLUSION: Small diameter columns was shown to be useful to simulate the behavior of larger diameter columns for use in scaled up systems. Expanded bed adsorption was demonstrated to be a scalable technology allowing large quantities of C-PC to be obtained, maintaining high protein recovery while reducing both processing cost and time. The proposed methodology allows recovery of more than 62% of the C-PC contained in the biomass in the form of pure C-PC concentrates. Copyright