Extraction and Separation of Phycocyanin from Spirulina using Aqueous Two-Phase Systems of Ionic Liquid and Salt

Xifeng Zhang¹, Fenqin Zhang¹, Guanghong Luo^2, , Shenghui Yang² and Danxia Wang²

¹The College of Agriculture and Biotechnology (CAB), Hexi University, Zhangye, P.R. China

²Kaiyuan Bio-tech Development Center, Hexi University, Zhangye, P.R. China

Cite this paper:
Xifeng Zhang, Fenqin Zhang, Guanghong Luo, Shenghui Yang and Danxia Wang. Extraction and Separation of Phycocyanin from Spirulina using Aqueous Two-Phase Systems of Ionic Liquid and Salt. Journal of Food and Nutrition Research. 2015; 3(1):15-19. doi: 10.12691/jfnr-3-1-3

Abstract

To explore a new and simple rapid extraction and purification technique for phycocyanin, an ionic liquids(ILs)-based aqueous two-phase system(ATPS) was developed for the purification of phycocyanin from Spirulina extracts. Effects of various process parameters such as the concentrations of [Bmim]Cl, the concentrations of KH₂PO₄, the concentrations of crude phycocyanin, the system pH and the temperature on partitioning of phycocyanin were evaluated. The obtained data indicated that phycocyanin was preferentially partitioned into the ILs-rich phase and the ATPS composed of 23% (w/w) [Bmim]Cland 29% (w/w) KH₂PO₄ at 30°C and pH 7.0 showed good selectivity on phycocyanin. Under the optimum conditions, phycocyanin with a purity of 3.98 and yield of about 90.23 % was obtained. Therefore, ILs-based ATPS was an effective method for partitioning and recovery of phycocyanin from Spirulina extracts.

Keywords:
Ionic liquid Aqueous two-phase system phycocyanin Purification

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	G.G. Choi, M.S. Bae, C.Y. Ahn, H.M. Oh, Induction of axenic culture of Arthrospira (Spirulina) platensis based on antibiotic sensitivity of contaminating bacteria, Biotechnol. Lett. 30 (2008) 87-92.
[2]	M.G. Sajilata, R.S. Singhal, M.Y. Kamat, Fractionation of lipids and purification of gamma-linolenic acid (GLA) from Spirulina platensis, Food Chem. 109 (2008) 580-586.
[3]	T. Gireesh, A. Jayadeep, K.N. Rajasekharan, V.P. Menon, M. Vairamany, G. Tang, P.P. Nair, P.R. Sudhakaran, Production of deuterated beta-carotene by metabolic labelling of Spirulina platensis, Biotechnol. Lett. 23 (2001) 447-449.
[4]	H.B. Chen, J.Y. Wu, C.F. Wang, C.C. Fu, C.J. Shieh, C.I. Chen, C.Y. Wang, Y.C. Liu, Modeling on chlorophyll a and phycocyanin production by Spirulina platensis under various light-emitting diodes, Biochem. Eng. J. 53 (2010) 52-56.
[5]	T.Silveira, J.F.M.Burkert, J.A.V.Costa, C.A.V.Burkert, S.J.Kalil, Opti mization of phycocyanin extraction from Spirulina platensis using factorial design, Bioresour. Technol. 98 (2007) 1629-1634.
[6]	M.C.Santiago-Santos,T.Ponce-Noyola,R.Olvera-Ramirez, J.Ortega-Lopez, R.O. Canizares-Villanueva, Extraction and purification of phycocyanin from Calothrix sp, Process Biochem. 39 (2004) 2047-2052.
[7]	G. Patil, K. Raghavarao, Aqueous two phase extraction for purification of C-phycocyanin, Biochem. Eng. J. 34 (2007) 156-164.
[8]	V.B. Bhat, K.M. Madyastha, C-Phycocyanin: a potent peroxyl radical scavenger in vivo and in vitro, Biochem. Biophys. Res. Commun. 275 (2000) 20-25.
[9]	M.C. Reddy, J. Subliashini, S.V.K. Mahipal, V.B. Bhat, P.S. Reddy, G. Kiranmai, K.M. Madyastha, P. Reddanna, C-Phycocyanin, a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW 264.7 macrophages, Biochem. Biophys. Res. Commun. 304 (2003) 385-392.
[10]	M.G. de Morais, J.A.V. Costa, Carbon dioxide fixation by Chlorella kessleri, C-vulgaris, Scenedesmus obliquus and Spirulina sp cultivated in flasks and vertical tubular photobioreactors, Biotechnol. Lett. 29 (2007) 1349-1352.
[11]	C.Y. Wang, C.C. Fu, Y.C. Liu, Effects of using light-emitting diodes on the cultivation of Spirulina platensis, Biochem. Eng. J. 37 (2007) 21-25.
[12]	S.G. Yan, L.P. Zhu, H.N. Su, X.Y. Zhang, X.L. Chen, B.C. Zhou, Y.Z. Zhang, Single-step chromatography for simultaneous purification of C-phycocyanin and allophycocyanin with high purity and recovery from Spirulina (Arthrospira) platensis, J. Appl. Phycol. 23 (2011) 1-6.
[13]	Boussiba, S., & Richmond,A.E. (1979). Isolation and characterization of phycocyanins from the blue-green alga Spirulina platensis. Archives of Microbiology, 120, 155-159.
[14]	Ranjitha, K., & Kaushik, B. D. (2005). Purification of phycobiliproteins from Nostoc muscorum. Journal of Scientific and Industrial Research, 64, 372-375.
[15]	Eriksen, N. T. (2008).Production of phycocyanin-a pigment with applications in biology, biotechnology, foods and medicine. Applied Microbiology and Biotechnology, 80, 1-14.
[16]	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. Journal of Applied Phycology, 23, 1-6.
[17]	Tchernov, A. A., Minkova, K. M., Houbavenska, N. B., & Kovacheva, N. G. (1999). Purification of phycobiliproteins from Nostoc sp. by aminohexyl-sepharose chromatography. Journal of Biotechnology, 69, 69-73.
[18]	Aguilar, O., & Rito-Palomares, M. (2010). Aqueous two‐phase systems strategies for the recovery and characterization of biological products from plants. Journal of the Science of Food and Agriculture, 90 (9), 1385-1392.
[19]	S. Dreyer, P. Salim, U. Kragl, Driving forces of protein partitioning in an ionic liquid-based aqueous two-phase system, Biochem. Eng. J. 46 (2009) 176-185.
[20]	J.A.P. Coutinho, C.M.S.S. Neves, S.P.M. Ventura, M.G. Freire, I.M. Marrucho, Evaluation of cation influence on the formation and extraction capability of ionic-liquid-based aqueous biphasic systems, J. Phys. Chem. B 113 (2009) 5194-5199.
[21]	Y.C. Pei, J.J. Wang, K. Wu, X.P. Xuan, X.J. Lu, Ionic liquid-based aqueous two-phase extraction of selected proteins, Sep. Purif. Technol. 64 (2009) 288-295.
[22]	H.B. Chen, J.Y. Wu, C.F. Wang, C.C. Fu, C.J. Shieh, C.I. Chen, C.Y. Wang, Y.C. Liu, Modeling on chlorophyll a and phycocyanin production by Spirulina platensis under various light-emitting diodes, Biochem. Eng. J. 53 (2010) 52-56.
[23]	Boussiba S,Richmond A E(1979)Isolation and purification of phycocyanins from the blue green algae Spirulina platensis. Arch Microbiol 120: 155-159.
[24]	Deutscher M (1990) Guide to protein purification, vol 182. Methods in enzymology. Academic Press, New York.
[25]	novak U, Pohar a, Plazl l, Znidarsic-Plazl P (2012) Ionic liquid-based aqueous two-phase extraction within a microchannel system. Sep Purif Technol 97: 172-178.
[26]	Ma CH, liu TT, Yang l, Zu Yg, Chen XQ, Zhang l, Zhang Y,Chao CJ (2011) Ionic liquid-based microwave-assisted extraction of essential oil and biphenyl cyclooctene lignans from Schisandra chinensis Baill fruits. J Chromatogr a 1218: 8573-8580.
[27]	M.G. Freire, A.F.M. Cláudio, J.M.M. Araújo, J.A.P. Coutinbo, I.M. Marrucho, J.N.C.Lopes, L.P.N. Rebelo, Aqueous biphasic systems: a boost brought about byusing ionic liquids, Chem. Soc. Rev. 41 (2012) 4966-4995.
[28]	F.Z. Deng, D.F. Guo, Extraction separation of bovine serum albumin in ionic liquid aqueous two-phase system, Chin. J. Anal. Chem. 34 (2006) 1451-1453.