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The fungal pathogen  Penicillium marneffei  produces melanin-like pigment  in vitro . The synthetic pathway of melanin and its possible influence in the protective yeast cells surviving within macrophage cells are not known. In this work,  P. marneffei  produced brown black pigment in the presence of L-DOPA and black particles were extracted from yeast cells treated with proteolytic enzymes, denaturant and concentrated hot acid. Kojic acid inhibited the brown-black pigment production of  P. marneffei  yeast grown on brain heart infusion agar. Transmitting electron microscopy showed spherical granular electron-dense particles with an average diameter of 100 nm in a beaded arrangement in the innermost cell wall. Electron-paramagnetic resonance revealed that the black particles contain a stable free radical compound. The UV-visible and Fourier transform infrared spectra of particles extracted from  P. marneffei  and synthetic DOPA-melanin showed a high degree of similarity. Melanized yeast cells decreased phagocytosis by macrophage cells and increased resistance to intracellular digestion  in vitro . These results indicate that  P. marneffei  can synthesize DOPA-melanin or melanin-like compounds  in vitro  and suggest that the DOPA-melanin pathway is associated with cell wall structure and enhances the resistance to phagocytosis by macrophages.

Detection of DOPA-Melanin in the Dimorphic Fungal Pathogen Penicillium marneffei and Its Effect on Macrophage Phagocytosis In Vitro