Purification and characterisation of a sulphur rich melanin from edible mushroom Termitomyces albuminosus Heim

Production, purification and characterisation of a black pigment from Termitomyces albuminosus as melanin is reported, for the first time, from shaken submerged culture condition using scanning electron microscopy (SEM), elemental analysis, ultraviolet-visible (UV-VIS), and Fourier transformed infrared spectroscopy (FTIR), electron paramagnetic resonance (EPR) and 13 C (CP/MAS) NMR spectra. SEM results on T. albuminosus revealed nanogranular nature of melanin nanoparticles within size range of 400-100 nm with fractal dimension D = 1.195-1.73. Elemental analysis of melanin indicated 54.6% C, 3.5% H, 2.4% N, 26.9% O, and 12% S. UV-VIS and FTIR spectra confirmed to the characteristic of melanin and were identical to the reference commercial sepia melanin. Further validation of the identity of pigment as melanin was achieved by EPR analysis. Termitomyces albuminosus melanin is postulated to be DOPA-type melanin confirmed by 13 C (CP/MAS) NMR spectral analysis showing chemical shift at 200-170 ppm carbonyl, 160-110 ppm aromatic region, and with high 40-30 ppm open chain aliphatic region. Chemical modification through oxidation and cysteinylation (Pheomelanin) is implied as indicated by relatively high sulphur content (12%).