Abstract
BACKGROUND: During the last decade, enormous research efforts have been directed at identifying potent microorganisms as sustainable green cell factories for eco-friendly pigments. Talaromyces atroroseus has recently been shown to excrete large amounts of azaphilone mycotoxin-free red pigment mixture comprising some known coloring components together with many uncharacterized metabolites. In this study, a new Talaromyces atroroseus isolate was identified via sequencing of the fragment of the nuclear ribosomal gene cluster containing internal transcribed spacers and 5.8S rRNA gene. The parameters that affected the level of pigment production were optimized in uncommon static conditions of culture and genetic improvement, via γ-irradiation, to improve pigment yield. Moreover, chemical characterization using LC/MS and skin safety test of the target pigment mixture were precisely conducted to maximize its benefits as a natural and safe red pigment for wool fabrics. RESULTS: Molecular identification via the sequencing of the ITS of the rDNA encoding gene cluster revealed that the fungal isolate TRP-NRC was T. atroroseus TRP-NRC (deposited in GenBank under accession number MW282329). In the static conditions of culture, pigment production was dramatically enhanced to 27.36 g/L in an optimum yeast malt peptone medium of 2% mannitol at pH 2-4.5 and 30 °C for 7 days of incubation. Under exposure to a 400-Gy γ-radiation dose, pigment yield was increased to a 3-fold level higher than that recorded for the wild type. Based on the inter-simple sequence repeats (ISSR), as a molecular marker tool, the wild-type T. atroroseus TRP-NRC strain and its mutants were discriminated. The UHPLC/HRESI-MS analytical tool characterized 60 metabolites, including many unknown molecules, at appropriate concentrations. It is worthy to note that four mitorubrin derivatives were identified for the first time in T. atroroseus, i.e., mitorubrinolamine acetate, dihydro-PP-O, mitorobrinal, and mitorubrinol. The range of irritation indexes (0-0.1) demonstrated an adequate skin safety after the direct local application of the pigment mixture. Finally, the pigment mixture exhibited a remarkably good dyeing ability in wool fabrics, with high-fastness properties. CONCLUSIONS: Because of its sustainable and economic production, the target red pigment mixture may be applied in the future in textile, food, cosmetics, or different pharmaceutical industries after extensive conventional safety and toxicity studies, which are currently under consideration.