World Journal of Environmental Engineering
ISSN (Print): 2372-3076 ISSN (Online): 2372-3084 Website: Editor-in-chief: Apply for this position
Open Access
Journal Browser
World Journal of Environmental Engineering. 2019, 6(1), 1-6
DOI: 10.12691/wjee-6-1-1
Open AccessArticle

Purification of Phycocyanin from Chaohu Algae by Various Salting out Methods

Kolawole B. H. E Boni1, , Fayu Zhang1, Wu Kang1 and Jiaquan Wang1

1Hefei University of Technology

Pub. Date: January 14, 2019

Cite this paper:
Kolawole B. H. E Boni, Fayu Zhang, Wu Kang and Jiaquan Wang. Purification of Phycocyanin from Chaohu Algae by Various Salting out Methods. World Journal of Environmental Engineering. 2019; 6(1):1-6. doi: 10.12691/wjee-6-1-1


This research was carried out to extract and purify phycocyanin from blue algae in Chaohu Lake by utilizing salting out methods. Thus a two-step salting out was performed using Ammonium sulphate, Triammonium citrate, Sodium citrate and Sodium sulfate. The phycocyanin and the impurity solution collected at each phase were subjected to analysis by using UV-Vis spectrophotometer to identify the optimal dose of ammonium sulfate, triammonium citrate, sodium citrate and sodium sulfate. The optimal molar concentration for Ammonium sulphate, Triammonium citrate, Sodium citrate and Sodium sulfate in the first and second salting out process were 1.0 mol/L and 1.7mol/L for Ammonium sulphate (NH4)2SO4, 0.7, mol/L and 1.3 mol/L for Triammonium citrate C6H17N3O7, 0.5mol/L and 0.9 mol/L for Sodium citrate C6H5Na3O7, 1.0mol/L and 1.3 mol/L for Sodium sulfate Na2S04. After the two-step salting out processes, it was observed that higher molar concentrations can remove impurities in large quantities, and both purity and yield was greatly increased. The results indicate a purity of phycocyanin above 2.0 with a phycocyanin recovery relatively high. This was carried out to affirm and estimate if the result from the salt use will vary from a previous experiment carried out on the same river by the authors with three (3) different kinds of salt K3C6H5O7H2O, C6H5O7(NH4)3 and (NH4)2SO4 as compared to the four (4) salt compound used in this experiment.

cyanobacteria phycocyanin purity salting-out yield

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Kibria, G., (2016). Blue-green algal/cyanobacterial blooms (BGA), climate change and BGA impacts on water quality, fish kills, recreation, crops, seafood, livestock, wild animals and humans, 1-7.
[2]  Devendra; Dolly; Sunil; Neeraj, and Suresh, (2014). Extraction and purification of C-phycocyanin from Spirulinaplatensis (CCC540) Indian J. Plant Physiol., 19: 184-188.
[3]  Reis, A.; Mendes, A.; Lobo-Fernandes, H.; Empis, J.A.; Novais, J.M., (1998). Production, extraction and purification of phycobiliproteins from Nostoc sp. Bioresour. Technol. 66: 181-187.
[4]  Sonani, R.R.; Singh, N.K.; Kumar, J.; Thakar, D.; Madamwar, D., (2014). Concurrent purification and antioxidant activity of phycobiliproteins from Lyngbya sp. A09DM: an antioxidant and anti-aging potential of phycoerythrin in Caenorhabditiselegans. Process Biochem., 49:1757-1766.
[5]  Bermejo, R.; Acién, F.G.; Ibanez, M.J.; Fernandez, J.M.; Molina, E.; Alvarez-Pez, J.M., (2003). Preparative purification of B-phycoerythrin from the microalga Porphyridiumcruentum by expanded-bed adsorption chromatography. J. Chromatogr. B. 790: 317-325.
[6]  Khattar, J.I.S.; Kaur, S.; Kaushal, S.; Singh, Y.; Singh, D.P.; Rana, S.; Gulati, A., (2015). Hyperproduction of phycobiliproteins by the cyanobacterium Anabaenafertilissima PUPCCC 410.5 under optimised culture conditions. Algal Res., 12: 463-469.
[7]  Singh, N.K.; Parmar, A.; Madamwar, D., (2009). Optimisation of medium components for increased production of C-phycocyanin from Phormidiumceylanicum and its purification by single step process. Bioresour. Technol., 100: 1663-1669.
[8]  Klotz, A.V.; Glazer, A.N.; Bishop, J.E.; Nagy, J.O.; Rapoport, H., (1986). Phycobiliprotein-bilin linkage diversity. II. Structural studies on A and D-ring-linkedphycoerythrobilins. J. Biol. Chem., 261: 6797-6805.
[9]  Apt, K.E.; Collier, J.L.; Grossman, A.R., (1995). Evolution of the phycobiliproteins. J.Mol. Biol., 248: 79-96.
[10]  Glazer, A.N.; Fang, S.; Brown, D.M.,(1973). Spectroscopic properties of C-phycocyanin and of its a and b subunits. J. Biol. Chem., 248: 5679-5685.
[11]  Sekar, S.; Chandramohan, M., (2008). Phycobiliproteins as a commodity: trends in applied research, patents and commercialization. J. Appl. Phycol., 20:113-136.
[12]  Qureshi, M.A.; Garlich, J.D., & Kidd, M.T., (1996). Dietary Spirulinaplatensis enhances humoral and cell-mediated immune functions in chickens. Immunopharmacol. Immunotoxicol., 18: 465-476.
[13]  Romay, C.; Gonzalez, R., (2000). Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals. J. Pharm. Pharmacol., 52:367-368.
[14]  Cherng, S.C.; Cheng, S.N.; Atarn C.T.C., (2007). Anti-inflammatory activity of C-phycocyanin in lipopolysaccharide-stimulated RAW264. 7 macrophages. Life Sci., 81:1431-1435.
[15]  Eriksen, N.T., (2008). Production of phycocyanina pigment with applications in biology, biotechnology, foods and medicine. Appl. Microbiol. Biotechnol., 80: 1-14.
[16]  Chaiklahan, R.; Chirasuwan, N.; Loha, V.; Tia, S.; Bunnag, B., (2011). Separation and purification of phycocyanin from Spirulinasp. using a membrane process. Bioresour. Technol., 102:7159-7164.
[17]  Kuddus, M.; Singh, P.; Thomas, G.; Al-Hazimi, A., (2013). Recent developments in production and biotechnological applications of C-phycocyanin. Biomed. Res. Int., 1-10.
[18]  Roman, R.B.; Alvarez-Pez, J.M.; Acien Fernandez, F.G.; Grima, E.M., (2002). Recovery of pure B-PE from the microalgae Porphyriumcruenlum. J. Biotechnol., 93: 73-85.
[19]  Rassano, R.; Ungaro, N.; D’Ambariso, A.; Liuzzi, G.M.; Riccio, P., (2003). Extractingandpurifying RPE frommediterranean red algaeCorallinaelongata Ellis andSolander. J. Biotechnol., 101: 289-293.
[20]  Vonshak, A., (1997). Spirulinaplatensis (Arthospira): Physiology, cell biology and biotechnology. Taylor & Francis, London.
[21]  Boussiba, S.;& Richmond, A. E., (1979). Isolation and characterization of phycocyanins from the blue-green alga Spirulinaplatensis. Arch. Microbiol., 120: 155-159.
[22]  Hirata, T.; Tanaka, M.; Ooike, M.; Tsunomura, T.; Sakaguchi, M., (2000). Antioxidant activities of phycocyanobilin prepared from Spirulinaplatensis. J. Appl. Phycol., 12: 435-439.
[23]  Minkova, K. M.; Tchernov, A.A.; Tchorbadjieva, M.I.; Fournadjieva, S.T.; Antova, R.E.; Busheva, M.C., (2003). Purification of C-phycocyanin from Spirulina (Arthrospira) fusiformis. J. Biotechnol., 102: 55-59.
[24]  Yan, S.; Zhu, L.; Su, H.; Zhang, X.; Chen, X.; Zhou, B.; Zhang, Y., (2011). Single-step chromatography for simultaneous purification of C-phycocyanin and allophycocyanin with high purity and recovery from Spirulina (Arthrospira) platensis. J. Appl. Phycol., 23: 1-6.
[25]  Ferreira, H.A.; Torres, D.P.V., and Juarez-Oropeza, M.A., (2010). Hepatoprotective effects of Spirulinamaximain patients with non-alcoholic fatty liver disease: a case series. J. Med. Case Rep., 4: 1-5.
[26]  Gantar, M.;Simovic, D.; Djils, S.; Gonzalez, W.W., and Miksovska, J., (2012). Isolation, characterization and antioxidative activity of C-phycocyanin from Limnothrixsp. Strain 37-2-1. J. Biotechnol., 159: 21-26.
[27]  Deng, R., and Chow, T.J., (2010). Hypolipidemic, antioxidant and anti-inflammatory activities of microalgae Spirulina. Cardiovasc. Ther., 28: 33-45.
[28]  Reddy, M.C.; Subhashini, J.; Mahipal, S.V.K.; Bhat, V.B.; Reddy, P.S.; Kiranmai, G.; Madyastha, K.M.; Reddanna, P., (2003). C-Phycocyanin,a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW 264.7 macrophages. Biochem. Biophys. Res. Commun., 304: 385-392.
[29]  Soni, B.;Kalavadia, B.; Trivedi, U.; Madamwar, D., (2006). Extraction, purification and characterization of phycocyanin from Oscillatoria quadripunctulata Isolated from the rocky shores of Bet-Dwarka, Gujarat, India. Process Biochem., 41: 2017-2023.
[30]  Niu, J.F.; Wang, G.C.; Tseng, C.K., (2006). Method for Large-Scale Isolation and Purification of R-Phycoerytrin Red Alga Polysiphoniaurceolata Grev. Protein Expression Purif., 49: 23-31.
[31]  Moraes, C.; Sala, L.; Cerveira, G.P.;Kalil, S.J., (2011). C-Phycocyanin extraction from Spirulinaplatensis wet biomass. Braz. J. Chem. Eng., 28: 45-48.
[32]  Zhang, F.; Yu, J.; Zhang, L.; Sheng, J.; Wang, J., (2017). Ultraviolet-Visible absorption spectra characteristics analysis of cyanophycocyanin from Chaohu Lake during the purification process. J. Spectrosc. Spect. Anal., 37: 806-810.
[33]  Herrera, A.;Boussiba, S.;Hohlberg, A.; Napoleone, V., (1989). Recovery of c-phycocyanin from the cyanobacterium Spirulinamaxima. J. Appl. Phycol. 1: 325-331.
[34]  MacColl, R., (1998). Cyanobacterialphycobilisomes. J. Struct. Biol., 124: 311-334.
[35]  Yuan Meng Yuan. Zhang Fayu. Sheng Jing Meng. Lu Yi Nan. Wang Jiaquan (2016). Food science and Technology. 41. (5) 267-272.