Properties and Functions of Melanin Pigment from Klebsiella sp. GSK

  • Sajjan, Shrishailnath S. (Department of Biochemistry, Gulbarga University) ;
  • Anjaneya, O (Department of Biochemistry, Gulbarga University) ;
  • Kulkarni, Guruprasad B. (Department of Biochemistry, Gulbarga University) ;
  • Nayak, Anand S. (Department of Biochemistry, Gulbarga University) ;
  • Mashetty, Suresh B. (Department of Biochemistry, Gulbarga University) ;
  • Karegoudar, T.B. (Department of Biochemistry, Gulbarga University)
  • Received : 2012.10.10
  • Accepted : 2012.12.15
  • Published : 2013.03.28


Purified melanin pigment from Klebsiella sp. GSK was characterized by thermogravimetric, differential thermal, X-ray diffraction and elemental analysis. This melanin pigment is structurally amorphous in nature. It is thermally stable up to $300^{\circ}C$ and emits a strong exothermic peak at $700^{\circ}C$. Its carbon, hydrogen and nitrogen composition is 47.9%, 6.9% and 12.0%, respectively. It was used to scavenge metal ions and free radicals. After immobilizing the pigment and using it to adsorb copper and lead ions, the metal ion adsorption capacity was evaluated by atomic absorption spectroscopy (AAS) and the identity of melanin functional groups involved in the binding of metal ions was determined by Fourier transform infrared (FT-IR) spectroscopy. Batch adsorption studies showed that 169 mg/g of copper and 280 mg/g of lead were adsorbed onto melanin-alginate beads. The metal ion adsorption capacity of the melanin-alginate beads was relatively significant compared to alginate beads. The metal ion desorption capacity of HCl was greater (81.5% and 99% for copper and lead, respectively) than that of EDTA (80% and 71% for copper and lead, respectively). The ability of the melanin pigment to scavenge free radicals was evaluated by inhibition of the oxidation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and was shown to be about 74% and 98%, respectively, compared with standard antioxidants.




  1. Albuquerque, J. E., C. Giacomantonio, A. G. White, and P.Meredith. 2006. Study of optical properties of electropolymerized melanin films by photopyroelectric spectroscopy Eur. Biophys. J. 35: 190-195.
  2. Averyanov, A. A., V. P. Lapikova, G. G. Petelina, and V. G. Dzhavakhiya. 1986. Prevention by fungal melanins of photodynamic spore damage. Izv. Akad. Nauk SSSR 4: 541-549.
  3. Baig, T. H., A. E. Garcia, K. J. Tiemann, and Gardea-Torresdey. 1999. Adsorption of heavy metal ions by the biomass of Solanum elaeagnifolium (Silver leaf night-shade). Proceedings of the 1999 Conference on Hazardous Waste Research.
  4. Baraldi, P., R. Capelletti, P. R. Crippa, and N. Romeo. 1979. Electrical characteristics and electret behavior of melanine. J. Electrochem. Soc. 126: 1207-1212.
  5. Bridelli, M. G. 1998. Self-assembly of melanin studied by laser light scattering. Biophys. Chem. 73: 227-239.
  6. Byrnes, B. J., R. L. Ryan, and M. Pazirandeh. 1997. Comparison of ion-exchange resins and biosorbents for the removal of heavy metals from plating factory wastewater. Environ. Sci. Technol. 31: 2910-2914.
  7. Casadevall, A., A. Nakouzi, P. R. Crippa, and M. Eisner. 2012. Fungal melanins differ in planar stacking distances. PLoS ONE 7: e30299.
  8. Deziderio, S. N., C. A. Brunello, M. I. N. Da Silva, M. A. Cotta, and C. F. O. Graeff. 2004. Thin films of synthetic melanin. J. Non-Crystal. Sol. 338-340: 634-638.
  9. Fourest, E. and B. Volesky. 1997. Alginate properties and heavy metal biosorption by marine algae. Appl. Biochem. Biotechnol. 67: 215-226.
  10. Gomes, R. C., A. S. Mangrich, R. R. R. Coelho, and L. F. Linhares. 1996. Elemental, functional group and infrared spectroscopic analysis of actinomycete melanins from Brazilian soils. Biol. Fertil. Soils 21: 84-88.
  11. Gómez-Marín, A. M. and I. S. Carlos. 2010. Thermal and mass spectroscopic characterization of a sulphur-containing bacterial melanin from Bacillus subtilis. J. Non-Crystal. Sol. 356: 1576-1580.
  12. Hong, L. and J. D. Simon. 2006. Insight into the binding of divalent cations to Sepia eumelanin from IR absorption spectroscopy. Photochem. Photobiol. 82: 1265-1269.
  13. Howell, R. C., A. D. Schweitzer, A. Casadevall, and E. A. Dadachova. 2008. Chemosorption of radiometals of interest to nuclear medicine by synthetic melanins. Nucl. Med. Biol. 35: 353-357.
  14. Inglezakis, V. J., M. D. Loizidou, and H. P. Grigoropoulou. 2003. Ion exchange of $Pb^{2+}$, $Cu^{2+}$, $Fe^{3+}$, and $Cr^{3+}$ on natural clinoptilolite: selectivity determination and influence of acidity on metal uptake. J. Colloid Interface Sci. 261: 49-54.
  15. Ito, S. and K. Fujita. 1985. Microanalysis of eumelanin and pheomelanin in hair and melanomas by chemical degradation and liquid chromatography. Anal. Biochem. 144: 527-536.
  16. Koroleva, O. V., N. A. Kulikova, T. N. Alekseva, E. V. Stepanova, V. N. Davidchik, E. Y. Beliaeva, and E. A. Tsvetkova. 2007. A comparative characterization of fungal melanin and the huminlike substances synthesized by Cerrena maxima 0275. Appl. Biochem. Microbiol. 43: 61-67.
  17. Kurchenko, V. P., K. A. Mosse, and A. T. Pikulev. 1991. Abstracts of Papers, Vsesoyuznaya konferentsiya po radiobiologicheskim issledvoaniyam avarii na Cherno-byl'skoi AES, (All-Union Conf. on Radiobiological Studies of the Chernobyl Power Plant Accident). Minsk: Naukai Tekhnika. pp. 70-71.
  18. Lin, S. H. and R. S. Juang. 2002. Heavy metal removal from water by sorption using surfactant-modified montmorillonite. J. Hazard. Mater. B 92: 315-326.
  19. Liu, Y., L. Hong, V. R. Kempf, K. Wakamatsu, S. Ito, and J. D. Simon. 2004. Ion exchange and adsorption of Fe(III) by Sepia melanin. Pigment Cell Res. 17: 262-269.
  20. Liyana-Pathiranan, C. M. and F. Shahidi. 2005. Antioxidant activity of commercial soft and hard wheat (Triticum aestivum L) as affected by gastric pH conditions. J. Agr. Food Chem. 53: 2433-2440.
  21. Lyakh, S. P. 1981. Mikrobnyi melaninogenez i ego funktsii (Microbial Melaninogenesis: Its Function). Moscow: Nauka. pp. 224.
  22. Meredith, P. and T. Sarna. 2006. The physical and chemical properties of eumelanin. Pigment Cell Res. 19: 572-594.
  23. Nies, D. H. 1999. Microbial heavy metal resistance. Appl. Microbiol. Biotechnol. 51: 730-750.
  24. Oliveira, H. P., C. F. O. Graeff, C. A. Brunello, and E. M. Guerr. 2000. Electrochromic and conductivity properties: a comparative study between melanin-like/$V_2O_5$.$nH_2O $ and polyaniline/ $V_2O_5$.$nH_2O $ hybrid materials. J. Non-Crystal. Sol. 273: 193-197.
  25. Plonka, P. M. and M. Grabacka. 2006. Melanin synthesis in microorganisms-biotechnological and medical aspects. Acta. Biochim. Pol. 53: 429-443.
  26. Prota, G. 1992. Melanins and Melanogenesis. San Diego: Academic. pp. 1-290.
  27. Przemyslaw, M. P. and G. Maja. 2006. Melanin synthesis in microorganisms: Biotechnological and medical aspects. Acta. Biochem. Pol. 53: 429-443.
  28. Rafika, S., T. Djilali, B. Benchreit, and B. Ali. 2009. Adsorption of heavy metals (Cd, Zn and Pb) from water using keratin powder prepared from Algerien sheep hoofs. Eur. J. Sci. Res. 35: 416-425.
  29. Re, R., N. Pellegrini, A. Proteggente, A. Pannala, M. Yang, and C. Rice-Evans. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol. Med. 26: 1231-1237.
  30. Sajjan, S., G. Kulkarni, V. Yaligara, K. Lee, and T. B. Karegoudar. 2010. Purification and physiochemical characterization of melanin pigment from Klebsiella sp. GSK. J. Microbiol. Biotechnol. 20: 1513-1520.
  31. Schaeffer, P. 1953. A black mutant of Neurospora crassa. Mode of action of the mutant allele and action of light on melanogenesis. Arch. Biochem. Biophys. 47: 359-379.
  32. Schiewer, S. and Volesky, B. 1995. Modeling of the protonmetal ion exchange in biosorption. Environ. Sci. Technol. 29:3049-3058.
  33. Shiguo, C., X. Changhu, W. Jingfeng, F. Hui, W. Yuming, M. Qin, and W. Dongfeng. 2009. Adsorption of Pb(II) and Cd(II) by squid Ommastrephes bartrami melanin. Bioinorg. Chem. Appl. 2009: 901563.
  34. Simonovic, B., V. Vucelic, A. Hadzi-Pavlovic, K. Stepien, T. Wilczok, and D. Vucelic. 1990. Thermogravimetry and differential scanning calorimetry of natural and synthetic melanins. J. Thermal. Anal. 36: 2475-2482.
  35. Subianto, S. 2006. Electrochemical synthesis of melanin-like polyindolequinone. Thesis presented to Queensland University of Technology.
  36. Tu, Y., Y. Sun, Y. Tian, M. Xie, and J. Chen. 2009. Physicochemical characterization and antioxidant activity of melanin from the muscles of Taihe Black-bone silky fowl (Gallus gallus domesticus Brisson). Food Chem. 114: 1345-1350.
  37. Volesky, B. and Z. R. Holan. 1995. Biosorption of heavy metals. Biotechnol. Prog. 11: 235-250.
  38. Wang, M., J. Li, M. Rangarajan, Y. Shao, E. J. La Voie, T. Huang, and C. Ho. 1998. Antioxidative phenolic compounds from Sage (Salvia officinalis). J. Agr. Food Chem. 46: 4869-4873.
  39. Wuyep, P. A., A. G. Chuma, S. Awodi, and A. J. Nok. 2007. Biosorption of Cr, Mn, Fe, Ni, Cu and Pb metals from petroleum refinery effluent by calcium alginate immobilized mycelia of Polyporus squamosus. Sci. Res. Essay 2: 217-221.
  40. Yu, L., S. Haley, J. Perret, M. Harris, J. Wilson, and M. Qian. 2002. Free radical scavenging properties of wheat extracts. J. Agr. Food Chem. 50: 1619-1624.
  41. Zajac, G., J. M. Gallas, J. Cheng, M. Eisner, S. C. Moss, and E. Alvarado-Swaisgood. 1994. The fundamental unit of synthetic melanin: A verification by tunneling microscopy of X-ray scattering results. Biochem. Biophys. Acta. 1199: 271-278.