ABSTRACT
BACKGROUND Melanin production has been associated with virulence in various pathogenic fungi, including Fonsecaea pedrosoi, the major etiological agent for chromoblastomycosis, a subcutaneous fungal disease that occurs in South America. OBJECTIVE The aim of this study was to evaluate the effects of acid-basic extracted F. pedrosoi melanin particles and fungal cell ghosts obtained by Novozym 234 treatment on their ability to activate the human complement system. METHODS The ability of melanin particles and fungal cell ghosts to activate the human complement system was evaluated by complement consumption, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA). FINDINGS Unsensitised melanin particles and melanin ghosts presented complement consumption of 82.67 ± 2.08% and 96.04 ± 1.13%, respectively. Immunofluorescence assays revealed intense deposition of the C3 and C4 fragments on the surface of melanin particles and ghosts extracted from F. pedrosoi. Deposition of the C3, C4, and C5 fragments onto melanin samples and zymosan was confirmed by ELISA. Deposition of small amounts of C1q and C9 onto melanin samples and zymosan was detected by ELISA. CONCLUSION Fonsecaea pedrosoi melanin particles and fungal cell ghosts activated the complement system mainly through an alternative pathway.
Subject(s)
Humans , Ascomycota/chemistry , Complement Activation , Melanins/isolation & purification , Melanins/biosynthesis , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody TechniqueABSTRACT
Melanins are enigmatic pigments produced by a wide variety of microorganisms including bacteria and fungi. Here, we have isolated and characterized extracellular melanin from mushroom fungus, Schizophyllum commune. The extracellular dark pigment produced by the broth culture of S. commune, after 21 days of incubation was recovered by hot acid-alkali treatment. The melanin nature of the pigment was characterized by biochemical tests and further, confirmed by UV, IR, EPR, NMR and MALDI-TOF Mass Spectra. Extracellular melanin, at 100 µg/ml, showed significant antibacterial activity against Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae and Pseudomonas fluorescens and antifungal activity against Trichophyton simii and T. rubrum. At a concentration of 50 µg/ml, melanin showed high free radical scavenging activity of DPPH (2,2-diphenyl-1-picrylhydrazyl) indicating its antioxidant potential. It showed concentration dependent inhibition of cell proliferation of Human Epidermoid Larynx Carcinoma Cell Line (HEP-2). This study has demonstrated characterization of melanin from basidiomycetes mushroom fungus, Schizophyllum commune and its applications.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Antineoplastic Agents/pharmacology , Antioxidants/pharmacokinetics , Basidiomycota/chemistry , Electron Spin Resonance Spectroscopy/methods , Fungi , Melanins/biosynthesis , Melanins/isolation & purification , Melanins/pharmacokinetics , Melanins/metabolism , Schizophyllum/chemistry , Schizophyllum/classificationABSTRACT
Doenca de Jorge Lobo e uma doenca cronica granulomatosa causada por um fungo obrigatorio, em cuja parede celular demonstra-se a presenca de melanina constitutiva. Ao contrario, no P. brasiliensis, e em descricoes de outros ascomicetos filogeneticamente relacionados da familia Onygenaceae, a melanina nao e detectavel pela tecnica de coloracao histoquimica de Fontana-Masson