NPtagM: A Tailoring Enzyme Genome Mining Toolkit and Its Application in Terpenoid P450s from Phytopathogenic Fungi

Terpenoids derived from phytopathogenic fungi are major participants in interactions among microorganisms, plants, and animals. The modifications catalyzed by cytochrome P450s significantly influence the structural and bioactivity diversity of the terpenoids. To conduct genome mining of P450s in pathogenic fungi, in this study, we developed a new software called Natural Products Tailoring Enzymes Genome Mining (NPtagM). By optimizing the workflow and gene prediction software, NPtagM demonstrated a 3-fold increase in the number of predicted P450s and an 8-fold reduction in runtime compared to antiSMASH. We then used it to extract 1189 dereplicated terpenoid P450s from our in-house fungal genomes. Using a sequence similarity network analysis, we identified a family that potentially produced eremophilane-type sesquiterpenoids. The heterologous expression in Aspergillus oryzae resulted in the production of two new and four known eremophilanes. Our results highlight the potential of NPtagM in genome mining for tailoring enzymes from phytopathogenic fungi.