Preliminary Purification of C-Phycocyanin through the Foam Fractionation

Document Type : Regular Article

Authors

Department of Chemical Engineering, Ker. C., Islamic Azad University, Kermanshah, Iran

10.22034/ijche.2026.579517.1592
Abstract
Objective: C-Phycocyanin (C-PC) is a blue-colored protein antioxidant produced in the microalgae Spirulina platensis (S. platensis) that is used in the food, cosmetic and pharmaceutical industries. The purification methods are often time-consuming and expensive. In this study, Foam fractionation (FF) was used as a simple, rapid, cost-effective, and environmentally friendly method for the purification of C-PC.
Methods: C-PC pigment solution was extracted from dry S. plantensis biomass in phosphate buffer. FF method for the purification of C-PC was optimized using the response surface methodology (RSM). A UV-Vis spectrophotometer was used to determine the identifying absorbance peaks of the C-PC solutions.
Results: The RSM obtained an optimal setting at a pH = 6, aeration rate of 3.5 vvm, and operation time of 21 min for achievement of the highest purification fold (PF), purity index (PI), and C-PC recovery percentage (R) of about 1.56 ± 0.02, 0.59 ± 0.02, and 45.59 ± 0.72%, respectively.
Conclusion: In FF method there are no additional chemicals, also it is fast and has low operating costs, which make it an attractive method.

Keywords

Subjects

[1]   Athiyappan KD, Routray W, Paramasivan B. Phycocyanin from Spirulina: A comprehensive review on cultivation, extraction, purification, and its application in food and allied industries. Food and Humanity. 2024 May 1;2:100235. https://doi.org/10.1016/j.foohum.2024.100235.
[2] Liang Y, Kaczmarek MB, Kasprzak AK, Tang J, Shah MM, Jin P, Klepacz-Smółka A, Cheng JJ, Ledakowicz S, Daroch M. Thermosynechococcaceae as a source of thermostable C-phycocyanins: Properties and molecular insights. Algal Research. 2018 Nov 1;35:223-35. https://doi.org/10.1016/j.algal.2018.08.037.
[3] Patil G, Chethana S, Sridevi AS, Raghavarao KS. Method to obtain C-phycocyanin of high purity. Journal of chromatography A. 2006 Sep 15;1127(1-2):76-8. https://doi.org/10.1016/j.chroma.2006.05.073.
[4] Figueira FD, Moraes CC, Kalil SJ. C-phycocyanin purification: multiple processes for different applications. Brazilian Journal of Chemical Engineering. 2018;35(3):1117-28.https://doi.org/10.1590/0104-6632.20180353s20170160.
[5]   Moraes CC, Kalil SJ. Strategy for a protein purification design using C-phycocyanin extract. Bioresource technology. 2009 Nov 1;100(21):5312-7. http://doi:10.1016/j.biortech.2009.05.026.
[6]   Khazi MI, Demirel Z, Liaqat F, Dalay MC. Analytical grade purification of phycocyanin from cyanobacteria. InBiofuels from Algae: Methods and Protocols 2018 Nov 28 (pp. 173-179). New York, NY: Springer New York.https://doi.org/10.1007/7651_2018_202.
[7]   Phong WN, Show PL, Chow YH, Ling TC. Recovery of biotechnological products using aqueous two phase systems. Journal of bioscience and bioengineering. 2018 Sep 1;126(3):273-81.https://doi.org/10.1016/j.jbiosc.2018.03.005.
[8] Burghoff B. Foam fractionation applications. Journal of Biotechnology. 2012 Oct 15;161(2):126-37. https://doi:10.1016/j.jbiotec.2012.03.008.
[9]   Sochacki M, Michorczyk P, Vogt O. Foam fractionation as an efficient method for the separation and recovery of surfactants and surface-inactive agents: state of the art. ACS omega. 2024 Dec 31;10(1):55-75. https://doi.org/10.1021/acsomega.4c08413.
[10] Spolaore P, Joannis-Cassan C, Duran E, Isambert A. Commercial applications of microalgae. Journal of bioscience and bioengineering. 2006 Feb 1;101(2):87-96. https://doi.org/10.1263/jbb.101.87.
[11] Keshavarzi B, Krause T, Sikandar S, Schwarzenberger K, Eckert K, Ansorge-Schumacher MB, Heitkam S. Protein enrichment by foam fractionation: experiment and modeling. Chemical Engineering Science. 2022 Jul 20;256:117715. https://doi.org/10.1016/j.ces.2022.117715
[12] Antecka A, Klepacz-Smółka A, Szeląg R, Pietrzyk D, Ledakowicz S. Comparison of three methods for thermostable C-phycocyanin separation and purification. Chemical Engineering and Processing-Process Intensification. 2022 Jan 1;171:108563. https://doi.org/10.1016/j.cep.2021.108563.
[13] Chen L, Hu N, Zhao C, Sun X, Han R, Lv Y, Zhang Z. High-efficiency foam fractionation of anthocyanin from perilla leaves using surfactant-free active Al2O3 nanoparticle as collector and frother: Performance and mechanism. Food chemistry. 2023 Nov 30;427:136708. http://doi:10.1016/j.foodchem.2023.136708.
[14] Krause T, Keshavarzi B, Heitkam S, Ansorge-Schumacher MB. Foam fractionation Tags (F-Tags) enabling surfactant-free, activity-preserving recovery of enzymes. Applied Microbiology and Biotechnology. 2024 Dec;108(1):140.https://doi.org/10.1007/s00253-023-12837-1.
[15] Khuri AI, Mukhopadhyay S. Response surface methodology. Wiley interdisciplinary reviews: Computational statistics. 2010 Mar;2(2):128-49. https:// doi.org/10.1002/wics.73.
[16] Fabre JF, Niangoran NG, Gaignard C, Buso D, Mouloungui Z, Valentin R. Extraction, purification and stability of C-phycocyanin from Arthrospira platensis. European food research and technology. 2022 Jun;248(6):1583-99. http://doi.org/10.1007/s00217-022-03987-z.
[17] Athiyappan KD, Routray W, Paramasivan B. Phycocyanin from Spirulina: A comprehensive review on cultivation, extraction, purification, and its application in food and allied industries. Food and Humanity. 2024 May 1;2:100235. https://doi.org/10.1016/j.foohum.2024.100235.
[18] Chaiklahan R, Chirasuwan N, Loha V, Tia S, Bunnag B. Separation and purification of phycocyanin from Spirulina sp. using a membrane process. Bioresource technology. 2011 Jul 1;102(14):7159-64. http://doi:10.1016/j.biortech.2011.04.067.
[19] Hu D, Xu R, Jin Y, Sun S, Ye J, Wu J, Dai Z, Shen JW, Lu Y. Green and sustainable extraction of phycocyanin from Spirulina platensis by temperature-sensitive polymer-based aqueous two-phase system and mechanism study. Bioresource Technology. 2024 Sep 1;407:131142. https://doi.org/10.1016/j.biortech.2024.131142.
[20] Hailing PJ, Walstra P. Protein‐stabilized foams and emulsions. Critical Reviews in Food Science & Nutrition. 1981 Oct 1;15(2):155-203. https://doi.org/10.1080/10408398109527315.
[21]  Kumar S, Brooks MS. Enrichment and recovery of pea (Pisum sativum L.) proteins using foam fractionation for simultaneous enhancement of their functional properties. Separation and Purification Technology. 2025 Aug 30;364:132578. https://doi.org/10.1016/j.seppur.2025.132578.
[22] Aksay S, Mazza G. Optimization of protein recovery by foam separation using response surface methodology. Journal of food engineering. 2007 Mar 1;79(2):598-606.https://doi.org/10.1016/j.jfoodeng.2006.02.024.
[23] Narayan AV, Raghavarao KS. Extraction and Purification of C-Phycocyanin from Spirulina Platensis Employing Aqueous Two Phase Systems. International Journal of Food Engineering. 2007 Oct 1;3(4). http://doi.org/10.2202/1556-3758.1105.
[24] Deng B, Schroën K, de Ruiter J. Effects of dynamic adsorption on bubble formation and coalescence in partitioned-EDGE devices. Journal of Colloid and Interface Science. 2021 Nov 15;602:316-24.https://doi.org/10.1016/j.jcis.2021.06.014.
[25] Wang Y, Wang S, Li R, Wang Y, Xiang Q, Li K, Bai Y. Effects of combined treatment with ultrasound and pH shifting on foaming properties of chickpea protein isolate. Food Hydrocolloids. 2022 Mar 1;124:107351. https://doi.org/10.1016/j.foodhyd.2021.107351.
[26] Chew KW, Chia SR, Lee SY, Zhu L, Show PL. Enhanced microalgal protein extraction and purification using sustainable microwave-assisted multiphase partitioning technique. Chemical Engineering Journal. 2019 Jul 1;367:1-8. https://doi:10.1016/j.cej.2019.02.131
[27] Merz J, Schembecker G, Riemer S, Nimtz M, Zorn H. Purification and identification of a novel cutinase from Coprinopsis cinerea by adsorptive bubble separation. Separation and Purification Technology. 2009 Sep 15;69(1):57-62.https://doi.org/10.1016/j.seppur.2009.06.021.
[28] Ebrahimi A, Pazuki G, Mozaffarian M, Ahsaie FG, Abedini H. Separation and purification of C-phycocyanin from spirulina platensis using aqueous two-phase systems based on triblock thermosensitive copolymers. Food and Bioprocess Technology. 2023 Nov;16(11):2582-97.  doi.org/10.1007/s11947-023-03057-6.
 
 
 
 
 

Articles in Press, Accepted Manuscript
Available Online from 06 July 2026