GNPS2 molecular networking reveals metabolic diversity in fungi isolated from protease-rich fruits

Fungi are a rich source of chemical diversity for bioprospecting, yet most of their metabolic potential remains unexplored. This study investigated the metabolic profile of phytopathogenic fungi of the genus Fusarium, isolated from fruits with high protease content, such as papaya and pineapple. Comparative metabolomics was used to evaluate how culture substrates and co-cultivation with the endophyte Neofusicoccum ribis modulate secondary metabolite production. Molecular Networking analysis (GNPS2) revealed an impressive diversity of 173 annotated metabolites, including one previously undescribed compound. Metabolite production was highly dependent on the substrate and fungal strain, showing a strong correlation between phylogeny and metabolic profiles. Several extracts displayed significant inhibitory activity against the protease papain, with some co-cultivation combinations further enhancing this effect. These findings highlight how environmental conditions and microbial interactions activate silent biosynthetic pathways, confirming these fungi as promising sources for bioactive molecule discovery.