Applied Biochemistry and Microbiology

, Volume 50, Issue 1, pp 62–68 | Cite as

Chromatographic and spectroscopic characterization of phycocyanin and its subunits purified from Anabaena variabilis CCC421

  • H. Chakdar
  • S. Saha
  • S. PabbiEmail author


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.


PCBs Apply Biochemistry Anabaena DEAE Cellulose Anion Exchange Chromatography 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Bogorad, L., Annu. Rev. Plant Physiol., 1975, vol. 26, no. 1, pp. 369–401.CrossRefGoogle Scholar
  2. 2.
    Glazer, A.N., Annu. Rev. Biophys. Biophys. Chem., 1985, vol. 14, no. 1, pp. 47–77.PubMedCrossRefGoogle Scholar
  3. 3.
    Glazer, A.N. and Bryant, D.A., Arch. Microbiol., 1975, vol. 104, no. 1, pp. 15–22.PubMedCrossRefGoogle Scholar
  4. 4.
    Oi, V.T., Glazer, A.N., and Stryer, L., J. Cell Biol., 1982, vol. 93, no. 3, pp. 981–986.PubMedCrossRefGoogle Scholar
  5. 5.
    Parks, D.R., Hardy, R.R., and Herzenberg, L.A., Cytometry, 1984, vol. 5, no. 2, pp. 159–168.PubMedCrossRefGoogle Scholar
  6. 6.
    Shapiro, H.M., Glazer, A.N., Christenson, L., Willians, J.M., and Strom, T.B., Cytometry, 1983, vol. 4, no. 3, pp. 276–279.PubMedCrossRefGoogle Scholar
  7. 7.
    Stec, B., Troxler, R.F., and Teeter, M.M., Biophys. J., 1999, vol. 76, no. 6, pp. 2912–2921.PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Boussiba, S. and Richmond, A.E., Arch. Microbiol., 1979, vol. 120, no. 2, pp. 155–179.CrossRefGoogle Scholar
  9. 9.
    Hirata, T., Tanaka, M., Ooike, M., Tsunomura, T., and Sakaguchi, M., J. Appl. Phycol., 2000, vol. 12, nos 3–5, pp. 435–439.CrossRefGoogle Scholar
  10. 10.
    Minkova, K.M., Tchernov, A.A., Tchorbadjieva, M.I., Fournadjieva, S.T., Antova, R.E., and Busheva, M.C., J. Biotechnol., 2003, vol. 102, no. 1, pp. 55–59.PubMedCrossRefGoogle Scholar
  11. 11.
    Abalde, J., Betancourt, L., Torres, E., Cid, A., and Barwell, C., Plant. Sci, 1998, vol. 136, no. 1, pp. 109–120.CrossRefGoogle Scholar
  12. 12.
    Gupta, A. and Sainis, J.K., J. Appl. Phycol., 2009, vol. 22, no. 3, pp. 231–233.CrossRefGoogle Scholar
  13. 13.
    Ranjitha, K. and Kaushik, B.D., J. Sci. Ind. Res., 2005, vol. 64, no. 5, pp. 372–375.Google Scholar
  14. 14.
    Reis, A., Mendes, A., Lobo-Fernandes, H., Empis, J.A., and Novais, J.M., Biores. Technol., 1998, vol. 66, no. 3, pp. 181–187.CrossRefGoogle Scholar
  15. 15.
    Tchernov, A.A., Minkova, K.M., Houbavenska, N.B., and Kovacheva, N.G., J. Biotechnol., 1999, vol. 69, no. 1, pp. 69–73.CrossRefGoogle Scholar
  16. 16.
    Bryant, D.A., Glazer, A.N., and Eiserling, F.A., Arch. Microbiol., 1976, vol. 110, no. 1, pp. 61–75.PubMedCrossRefGoogle Scholar
  17. 17.
    Santiago-Santos, M.C., Noyalo, P.T., and Olvera, R.R., Process. Biochem., 2004, vol. 39, no. 12, pp. 2047–2052.CrossRefGoogle Scholar
  18. 18.
    Soni, B., Kalavadia, B., Trivedi, U., and Madamwar, D., Process Biochem., 2006, vol. 41, no. 9, pp. 2017–2023.CrossRefGoogle Scholar
  19. 19.
    Soni, B., Trivedi, V., and Madamwar, D., Biores. Technol., 2008, vol. 99, no. 1, pp. 188–194.CrossRefGoogle Scholar
  20. 20.
    Colyer, C.L., Kinkade, C.S., Viskari, P.J., and Landers, J.P., Anal. Bioanal. Chem, 2005, vol. 382, no. 3, pp. 559–569.PubMedCrossRefGoogle Scholar
  21. 21.
    Swanson, R.N. and Glazer, A.N., Anal. Biochem., 1990, vol. 188, no. 2, pp. 295–299.PubMedCrossRefGoogle Scholar
  22. 22.
    Zolla, L. and Bianchetti, M., J. Chromatogr., A, 2001, vol. 912, no. 2, pp. 269–279.CrossRefGoogle Scholar
  23. 23.
    Zolla, L., Bianchetti, M., and Rinalducci, S., Eur. J. Biochem., 2002, vol. 269, no. 5, pp. 1534–1542.PubMedCrossRefGoogle Scholar
  24. 24.
    Stanier, R.Y., Kunisawa, R., Mandal, M., and Cohen-Bazire, G., Bacteriol. Rev., 1971, vol. 35, no. 2, pp. 171–205.PubMedCentralPubMedGoogle Scholar
  25. 25.
    Bennett, A. and Bogorad, L., J. Cell Biol., 1973, vol. 58, no. 2, pp. 419–435.PubMedCrossRefGoogle Scholar
  26. 26.
    Laemmli, U.K., Nature, 1970, vol. 227, no. 5259, pp. 680–685.PubMedCrossRefGoogle Scholar
  27. 27.
    Bringans, S., Eriksen, S., Kendrick, T., Gopalakrishnakone, P., Livk, A., Lock, R., and Lipscombe, R., Proteomics, 2008, vol. 8, no. 5, pp. 1081–1096.PubMedCrossRefGoogle Scholar
  28. 28.
    Benedetti, S., Rinalducci, S., Benvenuti, F., Francogli, S., Pagliarani, S., Giorgi, L., et al., J. Chromatogr. B, 2006, vol. 833, no. 1, pp. 12–18.CrossRefGoogle Scholar
  29. 29.
    Bermejo, R., Talavera, E.M., Alvarez-Pez, J.M., and Orte, J.C., J. Chromatogr., A, 1997, vol. 778, nos. 1–2, pp. 441–450.CrossRefGoogle Scholar
  30. 30.
    Patel, A., Mishra, S., Pawar, R., and Ghsoh, P.K., Protein Expres. Purif., 2005, vol. 40, no. 2, pp. 248–255.CrossRefGoogle Scholar
  31. 31.
    Chen, T., Wong, Y., and Zheng, W., Phytochemistry, 2006, vol. 67, no. 22, pp. 2424–2430.PubMedCrossRefGoogle Scholar
  32. 32.
    Ramos, A., Acien, F.G., Fernandez-Sevilla, J.M., Gonzalez, C., and Bermejo, R., J. Chromatogr. B, 2011, vol. 879, nos. 7–8, pp. 511–519.CrossRefGoogle Scholar
  33. 33.
    Glazer, A.N. and Cohen-Bazire, G., Proc. Nat. Acad. Sci. U.S.A., 1971, vol. 68, no. 7, pp. 1398–1401.CrossRefGoogle Scholar
  34. 34.
    Sudhakar, Babu T., Kumar, A., and Varma, A.K., Plant Physiol., 1991, vol. 95, no. 2, pp. 492–497.CrossRefGoogle Scholar

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© Pleiades Publishing, Inc. 2014

Authors and Affiliations

  1. 1.Centre for Conservation and Utilisation of Blue Green Algae, Division of MicrobiologyIndian Agricultural Research InstituteNew Delhi-12India
  2. 2.National Bureau of Agriculturally Important MicroorganismsKushmaur, Maunath BhanjanIndia
  3. 3.Division of Agricultural ChemicalsIndian Agricultural Research InstituteNew Delhi-12India

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