Metabolic characterization of two white-rot fungi in diverse cultivation environments reveals biotechnologically relevant biosynthetic pathways

White-rot fungi stand out as highly efficient organisms for the mineralization of biopolymers, such as lignin and polysaccharides, into CO2 and H2O. Despite their significant potential for biotechnological applications, the intracellular metabolism of WRF remains largely overlooked. Building on recent findings regarding the utilization of lignin-related aromatic compounds as carbon sources by WRF, our objective was to gain further insights into these catabolic processes. For this purpose, two WRF, Trametes versicolor and Gelatoporia subvermispora were cultivated in different environments – agitation and static incubation modes and variations in the level of antioxidants – during the conversion of a lignin-related aromatic compound and cellobiose. To comprehensively assess their metabolic responses, we employed transcriptomics, proteomics, lipidomics, metabolomics, and microscopy analyses. The results underscore that both incubation mode and the presence of antioxidants impact sugars and aromatic catabolism, as well as lipid composition of the fungal mycelia. In addition, these analyses reveal biosynthetic pathways for the production of extracellular fatty acids and phenylpropanoids by these WRF, both products with relevance in biotechnological applications. Overall, we demonstrate that cultivation conditions play a crucial role in shaping bioprocesses that involve WRF and provide additional insights into carbon flux in nature. Overall design: Gelatoporia subvermispora and Trametes versicolor were cultured for 10 and 6 days, respectively, then treated with 4-hydroxybenzoate in the presence or absence of antioxidents, with or without agitation.