Biotechnological Production and Characterization of Extracellular Melanin by Streptomyces nashvillensis

Melanins are pigments employed in food, cosmetic, and textile industries, manufactured by extraction from cuttlefishes. Their biotechnological production by Streptomycetes, instead, has been poorly investigated so far. In this paper, for the first time, the strain Streptomyces nashvillensis DSM 40314 was tested as an extracellular melanin producer by investigating the influence of diverse temperatures (26, 28, and 30 C-degrees) and pH values (6.0 and 7.0) on bacterial growth, melanin production, and on the activity of the secreted tyrosinase, the first enzyme of the pigment biosynthetic pathway. In physiological 96-h shake flask experiments, the optimal growth parameters resulted to be 28(degrees)C and pH 7.0, at which a maximum biomass of 8.4 +/- 0.5 gcdw/L, a melanin concentration of 0.74 +/- 0.01 g/L (yield on biomass of 0.09 +/- 0.01 g/gcdw and productivity of 0.008 +/- 0.001 g/L/h), and a final tyrosinase activity of 10.1 +/- 0.1 U/mL were reached. The produced pigment was purified from the broth supernatant with a two-step purification process (75.0 +/- ; 2.0% of purity with 65.0 +/- 5.0% of recovery) and tested for its chemical, antioxidant, and photoprotective properties. Finally, characterization by UV-visible and FT-IR spectroscopy, elemental analyses, and mono- and bi-dimensional NMR suggested the eumelanin-like nature of the pigment.