World Journal of Microbiology and Biotechnology

, Volume 24, Issue 10, pp 2125–2131 | Cite as

Isolation and characterization of melanin pigment from Pleurotus cystidiosus (telomorph of Antromycopsis macrocarpa)

  • P. Selvakumar
  • S. Rajasekar
  • K. Periasamy
  • N. Raaman
Original Paper


Melanins are enigmatic pigments that are produced by a wide variety of microorganisms including several species of bacteria and fungi. For more than 40 years, fungi have been known to produce pigments called melanins. Melanin pigment production by mushrooms was not intensively studied. The present study was carried out on isolation and characterization of melanin from an edible mushroom Pleurotus cystidiosus var. formosensis. The mushroom produced dark mucous mass of hyaline arthrospores on mycelium. The coremia exclusively produced dikaryotic arthrospores with the remnant of a clamp connection. Continuous cell extension and division in the coremium stipe supplied cells for arthroconidiation at the coremium apex, which is surrounded by a liquid droplet (coremioliquid). The black coloured coremea (conidia) were produced by Antromycopsis macrocarpa (anamorph of P. cystidiosus) when cultured on potato dextrose agar medium. The agar plate was incubated at continuous light illumination for high amount of pigment (coremea) production. The slimy layer of the coremea was extracted and partially purified by alkaline and acid treatment. The black pigment was confirmed as melanin based on UV, IR and EPR spectra apart from chemical analysis. This is the first report on characterization of melanin obtained from Pleurotus cystidiosus var. formosensis.


Basidiomycetes Mushroom Pleurotus cystidiosus Antromycopsis macrocarpa Melanin Pigment 



Thanks are due to Prof. N. Anand, Director, Centre for Advanced Studies in Botany, University of Madras for laboratory facilities and Dr. K. Victor Babu and Dr. J. Subramanian, Chemical Physics Laboratory, Central Leather Research Institute, Chennai for EPR spectral analysis.


  1. Bartnicki-Garcia S, Reyes E (1964) Chemistry of spore wall differentiation in Mucor rouxii. Arch Biochem Biophys 108:125–133. Medline doi: 10.1016/0003-9861(64)90363-7 CrossRefGoogle Scholar
  2. Bell AA, Wheeler MH (1986) Biosynthesis and functions of fungal melanins. Annu Rev Phytopath 24:411–451. doi: 10.1146/ CrossRefGoogle Scholar
  3. Bonner TG, Duncan A (1962) Infrared spectra of some melanins. Nature 194:1078–1079. Medline doi: 10.1038/1941078a0 CrossRefGoogle Scholar
  4. Butler MJ, Day AW (1988) Destruction of fungal melanins by ligninases of Phanerochaete chrysosporium and other white rot fungi. Internat J Plant Sci 159:989–995Google Scholar
  5. Butler MJ, Day AW (1998) Fungal melanins: a review. Can J Microbiol 44:1115–1136. doi: 10.1139/cjm-44-12-1115 CrossRefGoogle Scholar
  6. Butler MJ, Day AW, Henson JM, Money NP (2001) Pathogenic properties of fungal melanins. Mycologia 93:1–8. doi: 10.2307/3761599 CrossRefGoogle Scholar
  7. Capelari M (1999) First record of Antromycopsis macrocarpa for Brazil. Mycotaxon 72:101–105Google Scholar
  8. Casadevall A, Rosas AL, Nosanchuk JD (2000) Melanin and virulence in Cryptococcus neoformans. Curr Opin Microbiol 3: 354–358. Medline doi: 10.1016/S1369-5274(00)00103-X CrossRefGoogle Scholar
  9. Chet I, Henis Y, Mitchell R (1967) Chemical composition of hyphal and sclerotial walls of Sclerotium rolfsii Sacc. Can J Bot 13:137–141Google Scholar
  10. Cockell CS, Knowland J (1999) Ultraviolet radiation screening compounds. Biol Rev 74:311–345. Medline doi: 10.1017/S0006323199005356 CrossRefGoogle Scholar
  11. Cooke MC (1892) Handbook of Australian fungi. Williams & Norgate, LondonGoogle Scholar
  12. Daniel J (1938) Studies on multiple allelomorphic series in the house mouse. III. A spectrophotometric study of mouse melanin. J Genet 36:139–143Google Scholar
  13. Elliott ML (1995) Effect of melanin biosynthesis inhibiting compounds on Gaeumannomyces species. Mycologia 87:370–374. doi: 10.2307/3760835 CrossRefGoogle Scholar
  14. Ellis DH, Griffiths DA (1974) The location and analysis of melanins in the cell walls of some soil fungi. Can J Microbiol 20:1379–1386CrossRefGoogle Scholar
  15. Enochs WS, Nilges MJ, Swartz HW (1993) A standardized test for the identification and characterization of melanins using electron paramagnetic resonance (EPR) spectroscopy. Pigment Cell Res 6:91–99. Medline doi: 10.1111/j.1600-0749.1993.tb00587.x CrossRefGoogle Scholar
  16. Gadd GM (1982) Effects of media composition and light on colony differentiation and melanin synthesis in Microdochium bolleyi. Trans Br Mycol Soc 78:115–122CrossRefGoogle Scholar
  17. Gasowska B, Kafarski P, Wojtasek H (2004) Interaction of mushroom tyrosinase with aromatic amines, o-diamines and o-aminophenols. Biochim Biophys Acta 1673:170–177. MedlineGoogle Scholar
  18. Hearing VJ, Tsukamoto K (1991) Enzymatic control of pigmentation in mammals. FASEB J 5:2902–2909. MedlineGoogle Scholar
  19. Henson JM, Butler MJ, Day AW (1999) The dark side of the mycelium: melanins of phytopathogenic fungi. Annu Rev Phytopathol 37:447–471. Medline doi: 10.1146/annurev.phyto.37.1.447 CrossRefGoogle Scholar
  20. Howard RJ, Ferrari MA (1989) Role of melanin in appressorium function. Expt Mycol 13:403–418. doi: 10.1016/0147-5975(89)90036-4 CrossRefGoogle Scholar
  21. Ito S (1998) Advances in chemical analysis of melanins. In: Nordlund JJ, Boissy RE, Hearing VJ, King RA, Ortonne JP (eds) The pigmentary system: physiology and pathophysiology. Oxford University Press, New York, pp 439–450Google Scholar
  22. Jacobson ES (2000) Pathogenic roles for fungal melanins. Clin Microbiol Rev 13:708–717. Medline doi: 10.1128/CMR.13.4.708-717.2000 CrossRefGoogle Scholar
  23. Jacobson FW, Millot N (1953) Phenolases and melanogenesis in the coelomic fluid of the ethinoid Diadema antillarum philippi. Proc R Soc Lond B 141:231–247. MedlineGoogle Scholar
  24. Langfelder K, Streibel M, Jahn B, Haase G, Brakhage AA (2003) Biosynthesis of fungal melanins and their importance for human pathogenic fungi. Fungal Genet Biol 38: 143–158. Medline doi: 10.1016/S1087-1845(02)00526-1 CrossRefGoogle Scholar
  25. Mason HS (1948) The chemistry of melanin III. Mechanism of the oxidation of dihydroxyphenylalanine by tyrosinase. J Biol Chem 172:83–92Google Scholar
  26. Miller OK (1969) A new species of Pleurotus with a coremioid imperfect stage. Mycologia 61:887–893. doi: 10.2307/3757633 CrossRefGoogle Scholar
  27. Moncalvo JM (1995) Pleurotus cystidiosus var. formosensis var. nov. an unusual Pleurotus collection of subgenus Coremiopleurotus from Taiwan. Mycol Res 99:1479–1482CrossRefGoogle Scholar
  28. Natarajan K, Raman N (1984) Occurrence of Pleurotus cystidiosus in India. Curr Sci 53:658–659Google Scholar
  29. Nosanchuk JD, Casadevall A (2003) The contribution of melanin to microbial pathogenesis. Cell Microbiol 5:203–223. Medline doi: 10.1046/j.1462-5814.2003.00268.x CrossRefGoogle Scholar
  30. Petersen RH, Nicholl DBG, Hughes KW (1997) Mating systems of some putative polypore-agaric relatives. Plant Syst Evol 201:135–158. doi: 10.1007/BF00984386 CrossRefGoogle Scholar
  31. Pollack FG, Miller OK (1976) Antromycopsis broussonetiae found to be the name of the imperfect state of Pleurotus cystidiosus. Mem New York Bot Gard 28:174–178Google Scholar
  32. Prota G (1992) Melanins and melanogenesis. Academic Press, New York, pp 1–290Google Scholar
  33. Raper HS (1928) The aerobic oxidases. Physiol Rev 8:245–282Google Scholar
  34. Ravishankar JP, Muruganandam V, Suryanarayanan TS (1995) Isolation and characterization of melanin from a marine fungus. Botanica Marina 38:413–416CrossRefGoogle Scholar
  35. Rescigno A, Sollai F, Pisu B, Rinaldi A, Sanjust E (2002) Tyrosinase inhibition: general and applied aspects. J Enzyme Inhib Med Chem 17: 207–218. Medline doi: 10.1080/14756360210000010923 CrossRefGoogle Scholar
  36. Rizner TL, Wheeler MH (2003) Melanin biosynthesis in the fungus Curvularia lunata (teleomorph: Cochliobolus lunatus). Can J Microbiol 49:110–119. Medline doi: 10.1139/w03-016 CrossRefGoogle Scholar
  37. Sanchez-Ferrer A, Rodriguez-Lopez JN, Garcia-Canovas F, Garcia-Carmona F (1995) Tyrosinase: a comprehensive review of its mechanism. Biochim Biophys Acta 1247:1–11. MedlineGoogle Scholar
  38. Sealy RC, Felix CC, Hyde JS, Swartz MH (1980) Structure and reactivity of melanins influence of free radicals and metal ions. In: Pryor WA (ed) Free radicals in biology, vol IV. Academic Press, New York, pp 209–259Google Scholar
  39. Segedin BP, Buchanan PK, Wilkie JP (1995) Studies in the Agaricales of New Zealand: new species, new records and renamed species of Pleurotus (Pleurotaceae). Aust Syst Bot 8:453–482. doi: 10.1071/SB9950453 CrossRefGoogle Scholar
  40. Soler-Rivas C, Jolivet S, Arpin N, Olivier JM, Wihers HJ (1999) Biochemical and physiological aspects of brown blotch disease of Agaricus bisporus. FEMS Microbiol Rev 23:591–614. Medline doi: 10.1111/j.1574-6976.1999.tb00415.x CrossRefGoogle Scholar
  41. Stalpers JA, Seifert KA, Samson RA (1991) A revision of the genera Antromycopsis, Sclerostilbum, and Tilachlidiopsis (Hyphomycetes). Can J Bot 69:6–15Google Scholar
  42. Suryanarayanan TS, Ravishankar JP, Venkatesan G, Murali TS (2004) Characterization of the melanin pigment of a cosmopolitan fungal endophyte. Mycol Res 108(8):974–978. Medline doi: 10.1017/S0953756204000619 CrossRefGoogle Scholar
  43. Thomas M (1955) Modern methods of plant analysis. In: Paech K, Tracey MV (eds) Melanin, vol 4. Springer, Verlag, Berlin, pp 661–675Google Scholar
  44. Toussaint O, Lerch K (1987) Catalytic oxidation of 2-aminophenols and ortho hydroxylation of aromatic amines by tyrosinase. Biochemistry 26: 8567–8571. Medline doi: 10.1021/bi00400a011 CrossRefGoogle Scholar
  45. Truong BN, Okazaki K, Hitoshi Neda HS, Fukiharu T, Le Akira Suzuki XT (2006) Dikaryotic arthroconidiation of Pleurotus subgenus Coremiopleurotus. Mycoscience 47:84–90. doi: 10.1007/s10267-005-0276-2 CrossRefGoogle Scholar
  46. Wheeler MH, Bell AA (1988) Melanins and their importance in pathogenic fungi. In: McGinnis MR (ed) Current topics in medical mycology, vol 2. Springer-Verlag, New York, pp 338–387Google Scholar
  47. Zervakis G (1998) Mating competence and biological species within the subgenus Coremiopleurotus. Mycologia 90:1063–1074. doi: 10.2307/3761281 CrossRefGoogle Scholar
  48. Zervakis G, Dimou D, Balis C (1992) First record of the natural occurrence in Europe of the basidiomycete Pleurotus cystidiosus on a new host. Mycol Res 96:874–876Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • P. Selvakumar
    • 1
  • S. Rajasekar
    • 1
  • K. Periasamy
    • 1
  • N. Raaman
    • 1
  1. 1.Fungal Biotechnology, Natural Products and Tissue Culture Laboratory, Centre for Advanced Studies in BotanyUniversity of MadrasChennaiIndia

Personalised recommendations