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Isolation, characterization and antioxidative activity of C-phycocyanin from Limnothrix sp. strain 37-2-1

Gantar, Miroslav, Simović, Dragan, Djilas, Sonja, Gonzalez, Walter W., Miksovska, Jaroslava
Journal of biotechnology 2012 v.159 no.1-2 pp. 21-26
Spirulina, absorbance, activated carbon, ammonium sulfate, anti-inflammatory activity, antioxidant activity, antioxidants, biomass, cells, chemical precipitation, chitosan, color, filtration, fluorescence, free radicals, freshwater, habitats, ion exchange chromatography, molecular weight, photosynthesis, phycocyanin, protein quaternary structure, purification methods, rutin, screening, Florida
C-phycocyanin (C-PC) is a blue colored accessory photosynthetic pigment found in cyanobacteria. Some of the medicinal properties of Spirulina have been attributed to this pigment, which includes anticancer, antioxidant, and anti-inflammatory activity. We have screened cyanobacteria isolated from freshwater habitats in Florida for their high content of C-PC. Of 125 strains tested, one filamentous strain identified as Limnothrix sp. was selected for further research. This strain produced 18% C-PC of total dry biomass. Here we describe a simple method for obtaining C-PC of high purity without the use of ion exchange chromatography. The procedure is based on pigment precipitation from the cell lysate with an appropriate concentration of ammonium sulfate, then purification with activated carbon and chitosan, followed by a sample concentration using tangential flow filtration. We have shown that when the lower concentration of ammonium sulfate was used, C-PC with higher purity index was recovered. Characterization of C-PC from Limnothrix showed that it had an absorbance maximum at 620nm and fluorescence at 639nm. The molecular mass of intact C-PC was estimated to be ∼50kDa with α and β subunits forming dimmers. When C-PC content per unit biomass was compared to that of marketed Spirulina powder, we found that Limnothrix was superior. C-phycocyanin from Limnothrix had an antioxidative activity on DPPH free radicals similar to that found in a natural antioxidant – rutin.