Abstract
We carried out the purification of C-phycocyanin and allophycocyanin from Spirulina platensis taking advantage of the adsorption properties of the expanded beds. Initially, phycobiliproteins were released from the microalga cells by osmotic shock. Next, phycocyanins were recovered by applying the centrifuged cell suspension directly to the anion exchanger Streamline-DEAE using expanded bed columns, equilibrated with 50 mM sodium phosphate buffer, pH 7.0. After adsorption, washing was carried out in the expanded-bed mode. Having removed unbound proteins and cellular debris, the bed was allowed to sediment and phycocyanins rich solution was eluted with a downward flow of 500 mM sodium phosphate buffer, pH 7.0. Finally, we utilized conventional gel filtration and ion exchange chromatography methods for separation and purification of C-phycocyanin and allophycocyanin. The purification steps were monitored using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the purity of recovered phycocyanins was confirmed by absorption and emission spectroscopy. The main advantage of this new method is the high yield achieved in the steps of product extraction and adsorption by expanded bed adsorption, so reducing both processing times and costs.
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References
Glazer AN (1988) Methods Enzymol 167:291–303
Glazer AN (1999). Phycobiliproteins. In: Cohen Z (ed) Chemicals from Microalgae, Taylor and Francis Ltd, UK, pp. 262–280
Kronick MN, Grossman AR (1983) Clin Chem 29:1582–1586
Glazer AN, Stryer L (1984) Trends Biochem Sci 9:423–427
Glazer AN (1994) J Appl Phycol 6:105–112
González R, Rodríguez S, Romay C, Ancheta O, González A, Armesto J, Remirez D, Merino N (1999) Pharmacol Res 39:55–59
Dainippon Ink and Chemicals Inc. (1979) Japanese Patent 95:770
Dainippon Ink and Chemicals Inc, (1987) Japanese Patent 06:691
Arad S, Yaron A (1992) Trends Food Sci Techol 3:92–97
Swanson RV, Glazer AN (1990) J Mol Biol 214:787–796
Brecj K, Ficner R, Huber R, Steinbacher S (1995) J Mol Biol 249:424–440
Shapiro HM, Glazer AN, Christenson L, Willians JM, Strom TB (1983) Cytometry 4:276–279
Parks DR, Hardy RR, Herzenberg LA (1984) Cytometry 5:159–169
Loken MR, Keij J, Kelley KA (1987) Cytometry 8:96–100
Ficner R, Lobeck K, Schmidt G, Huber R (1992) Mol Biol 228:935–950
Campanella L, Crescentini G, Avino P, Angiello L (2000) Ann Chim 90:153–161
Bermejo R, Talavera EM, Alvarez-Pez JM, Orte JC (1997) J Chromatogr A 778:441–450
Bermejo R, Talavera EM, Alvarez-Pez JM (2001) J Chromatogr A 917:135–145
Tchernov AA, Minkova KM, Houbavenska NB, Kovacheva N (1999) J Biotechnol 69:69–73
Saxena AM (1988) J Mol Biol 200: 579–591
Chase HA (1994) Trends Biotechnol 12:296–303
Chang YK, Chase HA (1996) Biotechnol Bioeng 49:512–526
Wang G (2002) Chromatographia 56:509–513
Bermejo R, Acién FG, Ibáñez MJ, Fernández JM, Molina E, Alvarez-Pez JM (2003) J Chromatogr B 790:317–325
Siegelman H, Kycia JH (1978) Algal biliproteins. In: Hellebust JA, Craigie JS (eds) Handbook of Phycological Methods: Physiological Biochemistry Methods, Cambridge University Press, London, pp 71–79
Laemmli UK (1970) Nature 277:680–686
Güzeltunc E, Ulgen KÖ (2001) J Chromatogr A 914:67–76
Chang YK, McCreath GM, Draeger NM, Chase H (1993) Trans Inst Chem Eg. 71:299–303
Oi VT, Glazer AN, Stryer L (1982) J Cell Biol 93:981–986
MacColl R, Csatorday K, Berns DS, Traeger E (1980) Biochemistry 19:2817–2820
Mimuro M, Rümbeli R, Füglistaller P, Zuber H (1986) Biochim Biophys Acta 851:447–456
Huang C, Berns DS, MacColl R (1987) Biochemistry 26:243–245
Jung TM, Dailey MO (1989) J Immunol Methods 121:9–18
Szantolai B, Gombos Z, Csizmadia V, Csatorday K, Lutz M (1989) Biochem 28:6467–6472
Bermejo R (1997) Caracterización espectroscópica y estructural de biliproteínas procedentes de Spirulina platensis. Aplicaciones como marcadores fluorescentes, Ph. D. Thesis, University of Granada, Granada (Spain)
Bermejo R, Talavera EM, Del Valle C, Alvarez-Pez JM (2000) Coll Surf B Biointerf 18:51–59
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Bermejo, R., Felipe, M.A., Talavera, E.M. et al. Expanded Bed Adsorption Chromatography for Recovery of Phycocyanins from the Microalga Spirulina Platensis . Chroma 63, 59–66 (2006). https://doi.org/10.1365/s10337-005-0702-9
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DOI: https://doi.org/10.1365/s10337-005-0702-9