Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

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• 作者：Ruperto Bermejo (1) rbermejo@ujaen.es
  Amparo Ramos (1)
• 关键词：Expanded bed adsorption chromatography – ; Spirulina platensis – ; Phycobiliproteins – ; C ; phycocyanin – ; Preparative protein purification – ; Anion ; exchange chromatography
• 刊名：Chromatographia
• 出版年：2012
• 出版时间：March 2012
• 年：2012
• 卷：75
• 期：5-6
• 页码：195-204
• 全文大小：470.7 KB
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  9. Bermejo Rom谩n R, Alv谩rez-Pez JM, Aci茅n Fern谩ndez FG, Molina Grima E (2002) Recovery of pure B-phycoerythrin from the microalga Porphyridium cruentum. J Biotechnol 93(1):73–85
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  17. Bermejo R, Aci茅n FG, Ib谩帽ez MJ, Fern谩ndez JM, Molina E, Alvarez-Pez JM (2003) Preparative purification of B-phycoerythrin from the microalga Porphyridium cruentum by expanded-bed adsorption chromatography. J Chromatogr B Anal Technol Biomed Life Sci 790(1–2):317–325
  18. Bermejo R, Felipe MA, Talavera EM, Alvarez-Pez JM (2006) Expanded bed adsorption chromatography for recovery of phycocyanins from the microalga Spirulina platensis. Chromatographia 63(1–2):59–66
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  20. Ramos A, Aci茅n FG, Fern谩ndez-Sevilla JM, Gonz谩lez CV, Bermejo R (2010) Large scale isolation and purification of c-phycocyanin from the cyanobacteria Anabaena marina using expanded bed adsorption chromatography. J Chem Technol Biotechnol 85:783–792
  21. Ramos A, Aci茅n FG, Fern谩ndez-Sevilla JM, Gonz谩lez CV, Bermejo R (2011) Development of a process for large-scale purification of C-phycocyanin from Synechocystis aquatilis using expanded bed adsorption chromatography. J Chromatogr B 879:511–519
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• 作者单位：1. Department of Physical and Analytical Chemistry, E.P.S. of Linares, Ja茅n University, Linares, Ja茅n 23700, Spain
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Organic Chemistry
  Pharmacy
  Biochemistry
  Plant Sciences
  Measurement Science and Instrumentation
  
• 出版者：Vieweg Verlag
• ISSN：1612-1112

文摘

The use of expanded bed adsorption chromatography (EBAC) for pilot scale recovery of C-phycocyanin (C-PC) from the cyanobacteria Spirulina platensis is described. The description includes the whole procedure, from small-scale method optimization to pilot scale. Initially, the phycobiliproteins are extracted by osmotic shock and separated by applying the centrifuged cell suspension to an EBAC column, using Streamline-DEAE as adsorbent. Next, the PCs rich solution is eluted to finally be purified by packed bed chromatography using DEAE-cellulose. The EBAC operation was optimized on a small scale using a 15 mm internal diameter (ID) column. The optimal conditions were: a sample load of 0.91 mg C-PC mL−1 adsorbent, H/H ₀ = 2 and a sample viscosity of 1.084 mP. The EBAC process was then scaled up by increasing the column ID (15, 25, 40, 60 and 90 mm) and, finally, the process was developed at pilot scale using a 150 mm ID column obtaining a scale-up factor of 100. The yield of the EBAC step is in the range of 75–79% whatever the column diameter. To obtain pure C-PC, conventional chromatography with DEAE-cellulose was utilized. The overall yield of the purification process after all steps is 59%. The purification steps are monitored using SDS-PAGE and the purity of recovered C-PC is confirmed by absorption and emission spectroscopy. Results show that EBAC method is a scalable technology that allows large quantities of C-PC to be obtained without product loss, maintaining a high protein recovery while reducing both processing cost and times.