Chromatographic and spectroscopic characterization of phycocyanin and its subunits purified from Anabaena variabilis CCC421
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Phycocyanin, a high value pigment was purified from diazotrophic cyanobacteria Anabaena variabilis CCC421 using a strategy involving ammonium sulfate precipitation, dialysis and anion exchange chromatography using DEAE-cellulose column. 36% phycocyanin with a purity of 2.75 was recovered finally after anion exchange chromatography. Purified phycocyanin was found to contain 2 subunits of 17 and 18 kDa which were identified as α and (β subunits by SDS-PAGE and MALDI-TOF. HPLC method using a C5 column coupled with fluorescence or photodiode-based detection was also developed to separate and detect the A. variabilis CCC421 phycocyanin subunits. The fluorescence method was more sensitive than photodiode one. The purified phycocyanin from A. variabilis CCC421 as well as its subunits was characterized with respect to absorption and IR spectra. Spectral characterization of the subunits revealed that α and β-subunits contained one and two phycocyanobilin groups as chromophores, respectively.
KeywordsPCBs Apply Biochemistry Anabaena DEAE Cellulose Anion Exchange Chromatography
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