Single-step chromatography for simultaneous purification of C-phycocyanin and allophycocyanin with high purity and recovery from Spirulina (Arthrospira) platensis
- 832 Downloads
The cyanobacterium Spirulina (Arthrospira) platensis is a good source of phycobiliprotein purification. C-phycocyanin (C-PC) is the major phycobiliprotein, while allophycocyanin (APC) is less abundant in S. platensis. Previously reported methods for C-PC purification are only able to offer either high purity or high efficiency. This paper describes one-step anion exchange chromatography method with continuous pH gradient elution for simultaneous purification of C-PC and APC with high purity and high recovery. Crude C-PC and APC were extracted and concentrated by ammonium sulfate fractionation at saturation of 25% and 60%, then purified on a DEAE-Sepharose Fast Flow chromatography column with continuous pH gradient elution from pH 5.0 to 3.6. After this single-step chromatography, C-PC and APC with high purity and recovery were simultaneously obtained. The purity ratios of C-PC and APC reached 5.59 (A620/A280) and 5.19 (A650/A280), respectively. Their purity was further demonstrated by electrophoresis and fluorescence emission spectroscopy. Moreover, the total recovery yield of pure C-PC and APC were 67.04% and 80.0%, representing 111.83 and 29.28 mg·g−1 lyophilized weight, respectively. The obtained C-PC and APC remained stable over a pH range of 4–9. This purification method for high purity and recovery of C-PC and APC proved to be fairly efficient compared with previously reported methods.
KeywordsC-phycocyanin Allophycocyanin Anion exchange chromatography pH gradient Purification
The work was supported by the Hi-Tech Research and Development program of China (2008AA09Z404), the National Natural Science Foundation of China (40676078), and the Scientific and Technological Development Program of Shandong Province (2006GG2205009).
- Glazer AN (1981) Photosynthetic accessory proteins with bilin prosthetic groups. In: Hatch MD, Boardman NK (eds) The biochemistry of plants, vol. 8, Photosynthesis. Academic, New York, pp 51–96Google Scholar
- Glazer AN, Stryer L (1984) Phycofluor probes. Trends Biochem Sci 9:423–427Google Scholar
- Haugland RP (1996) Handbook of fluorescent probes and research chemicals, 6th edn. Molecular Probes, EugeneGoogle Scholar
- Market Corporation (2005). http://www.marketbio.com (consult date: October 2005)
- Qureshi MA, Garlich JD, Kidd MT (1996) Dietary Spirulina platensis enhances humoral and cell-mediated immune functions in chickens. Immunopharmacol Immunotoxicol 18:465–476Google Scholar
- Romay C, Gonzalez R (2000) Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals. J Pharm Pharmacol 52:367–368Google Scholar
- Siegelman HW, Kycia JH (1978) Algal biliproteins. In: Hellebust JA, Craigie JS (eds) Handbook of phycological methods. Physiological and biochemical methods. Cambridge University Press, Cambridge, pp 71–79Google Scholar
- Stewart DE, Farmer FH (1984) Extraction, identification, and quantitation of phycobiliprotein pigments from phototrophic plankton. Limnol Oceanogr 29:392–397Google Scholar
- Wyman M (1992) An in vivo method for the estimation of phycoerythrin concentration in marine cyanobacteria (Synechococcus spp.). Limnol Oceanogr 37:1300–1306Google Scholar