Isolation and characterization of thermostable phycocyanin from Galdieria sulphuraria

Myounghoon Moon; Sanjiv K. Mishra; Chul Woong Kim; William I. Suh; Min S. Park; Ji -Won Yang

Korean Journal of Chemical Engineering, Vol.31, No.3, 490-495, March, 2014

DOI10.1007/s11814-013-0239-9 Export Citation

Isolation and characterization of thermostable phycocyanin from Galdieria sulphuraria

Myounghoon Moon¹, Sanjiv K. Mishra², Chul Woong Kim¹, William I. Suh², Min S. Park^{1, 2}, and Ji -Won Yang^{1, 2, †}

¹Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea
²Advanced Biomass R&D Center, #2502 Building W1-3, KAIST, 291 Daehak-ro, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea

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Phycocyanin is a highly valuable pigmented protein synthesized by several species of cyanobacteria and red alga. In this study we demonstrate the production of thermostable phycocyanin from the unicellular red alga Galdieria sulphuraria. Phycocyanin was extracted by repeated freeze-thaw cycles and purified in a two-step process using ammonium sulfate fractionation, at 25% and 50% concentrations. Purified phycocyanin exhibited maximum absorbance at 620 nm, and the purity ratio (A620/A280) was found to be greater than 4. The recovery efficiency of phycocyanin from the crude extract was above 80%. In total, approximately 19 milligram pure phycocyanin was obtained from 3 g of wet cell mass of Galdieria sp. Subunits α and β of the protein were separated by SDS-PAGE and analyzed by MALDITOF mass spectrometry for identification, which confirmed that the isolated protein is phycocyanin. The molecular weight of α and β subunits of phycocyanin was found to be 17.6 and 18.4 kDa, respectively.

Keywords:Galdieria sulphuraria;Red Algae;Value-added Product;Phycocyanin;Purification

[References]

Berns DS, MacColl R, Chem. Rev., 89, 4 (1989)
Eriksen NT, Appl. Microbiol. Biotechnol., 80(1), 1 (2008)
Chaiklahan R, Chirasuwan N, Bunnag B, Process Biochem., 47, 4 (2012)
Grossman AR, Schaefer MR, Chiang GG, Collier JL, Microbiol. Rev., 57, 3 (1993)
Ladero M, Santos A, Garcia-Ochoa F, Enzyme Microb. Technol., 38(1-2), 1 (2006)
Pang P, Koska J, Coad BR, Brooks DE, Haynes CA, Biotechnol. Bioeng., 90(1), 1 (2005)
Campbell D, Hurry V, Clarke AK, Gustafsson P, Oquist G, Microbiol. Mol. Biol. Rev., 62, 3 (1998)
Govindjee, Shevela D, Front. Plant Sci., 2, 1 (2011)
MacColl R, J. Struct. Biol., 124, 2 (1998)
Glazer A, J. Appl. Phycol., 6, 2 (1994)
Wedemayer GJ, Wemmer DE, Glazer AN, J. Biol. Chem., 266, 8 (1991)
O’Carra P, Murphy RF, Killilea SD, Biochem. J., 187, 2 (1980)
Samsonoff W, MacColl R, Arch. Microbiol., 176, 6 (2001)
Gross W, Schnarrenberger C, Plant Cell Physiol., 36, 4 (1995)
Stec B, Troxler RF, Teeter MM, Biophys. J., 76, 6 (1999)
Adir N, Dobrovetsky Y, Lerner N, J. Mol. Biol., 313, 1 (2001)
Yoshida A, Takagaki Y, Nishimune T, Biosci., Biotechnol., Biochem., 60, 1 (1996)
Brody SS, Brody M, Biochim. Biophys. Acta, 50, 2 (1961)
Graziani G, Schiavo S, Nicolai MA, Buono S, Fogliano V, Pinto G, et al., Food Funct., 4, 1 (2013)
Kronick MN, Grossman PD, Clin. Chem., 29, 9 (1983)
Li B, Gao MH, Zhang XC, Chu XM, Biotechnol. Appl. Biochem., 43, 3 (2006)
Chakdar H, Jadhav SD, Dhar DW, Pabbi S, J. Sci. Ind. Res., 71, 1 (2012)
Mishra SK, Shrivastav A, Mishra S, Process Biochem., 43, 4 (2008)
Spolaore P, Joannis-Cassan C, Duran E, Isambert A, J. Biosci. Bioeng., 101, 2 (2006)
Patel A, Mishra S, Pawar R, Ghosh PK, Protein Exp. Purif., 40, 2 (2005)
Mimuro M, Fuglistaller P, Rumbeli R, Zuber H, Biochim. Biophys. Acta, Bioenergy, 848, 2 (1986)
Lien TS, Wu ST, Yu ST, Too JR, Korean J. Chem. Eng., 24, 5 (2007)
Bennett A, Bogorad L, J. Cell Biol., 58, 2 (1973)
Mishra SK, Shrivastav A, Mishra S, Protein Expression Purif., 80, 2 (2011)
Shevchenko A, Wilm M, Vorm O, Mann M, Anal. Chem., 68, 5 (1996)
Soni B, Kalavadia B, Trivedi U, Madamwar D, Process Biochem., 41, 9 (2006)
Mishra SK, Shrivastav A, Pancha I, Jain D, Mishra S, Int. J. Biol. Macromol., 47, 5 (2010)
Patel A, Mishra S, Pawar R, Ghosh PK, Protein Exp. Purif., 40, 2 (2005)
Santiago-Santos MC, Ponce-Noyola T, Olvera-Ramirez R, Ortega-Lopez J, Canizares-Villanueva RO, Process Biochem., 39, 12 (2004)
Boussiba S, Richmond A, Arch. Microbiol., 120, 2 (1979)
MacColl R, Berns DS, Koven NL, Arch. Biochem. Biophys., 146, 2 (1971)
Olvera-Ramirez R, Coria-Cedillo M, Canizares-Villanueva RO, Martinez-Jeronimo F, Ponce-Noyola T, Rios-Leal E, Bioresour. Technol., 72, 2 (2000)
Chae KJ, Jang A, Yim SK, Kim IS, Bioresour. Technol., 99(1), 1 (2008)
Singh NK, Parmar A, Madamwar D, Bioresour. Technol., 100, 4 (2009)
Brock TD, Genetics, 146, 1 (1997)

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[10] Glazer A, J. Appl. Phycol., 6, 2 (1994)

[11] Wedemayer GJ, Wemmer DE, Glazer AN, J. Biol. Chem., 266, 8 (1991)

[12] O’Carra P, Murphy RF, Killilea SD, Biochem. J., 187, 2 (1980)

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[14] Gross W, Schnarrenberger C, Plant Cell Physiol., 36, 4 (1995)

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