NPtagM: A Tailoring\nEnzyme Genome Mining Toolkit and\nIts Application in Terpenoid P450s from Phytopathogenic Fungi

Terpenoids derived from phytopathogenic fungi are major\nparticipants\nin interactions among microorganisms, plants, and animals. The modifications\ncatalyzed by cytochrome P450s significantly influence the structural\nand bioactivity diversity of the terpenoids. To conduct genome mining\nof P450s in pathogenic fungi, in this study, we developed a new software\ncalled <u>N</u>atural <u>P</u>roducts <u>Ta</u>iloring Enzymes <u>G</u>enome <u>M</u>ining (NPtagM). By optimizing the workflow and gene\nprediction software, NPtagM demonstrated a 3-fold increase in the\nnumber of predicted P450s and an 8-fold reduction in runtime compared\nto antiSMASH. We then used it to extract 1189 dereplicated terpenoid\nP450s from our in-house fungal genomes. Using a sequence similarity\nnetwork analysis, we identified a family that potentially produced\neremophilane-type sesquiterpenoids. The heterologous expression in Aspergillus oryzae resulted in the production of\ntwo new and four known eremophilanes. Our results highlight the potential\nof NPtagM in genome mining for tailoring enzymes from phytopathogenic\nfungi.