摘要Thermomyces dupontii harbors two P450 paralogs(P450S and P450L)in the gene cluster for the biosynthesis of prenylated indole alkaloids(PIAs)and correponding iron chelators with P450L assigned as one protein containing a CYP like domain fused with a FAD-binding domain-containing oxidore-ductase.Genetic manipulation and metabolic profile analysis indicated both P450S and P450L were involved in transforming simple PIAs to their corresponding iron chelators.Moreover,P450S is respon-sible for bolstering simple PIAs to complex PIAs,and P450L for reinforcing conjugating unsaturated systems in complex PIAs.Chemical investigation led to isolation and characterization of novel complex PIA metabolites with more oxidations.P450L also contributed to forming the third iron-chelating core in iron chelators.A series of iron bioassays and infrastructure analysis revealed that lack of these P450 genes caused strongly elevated Fe3+levels but attenuated Fe2+levels,together with abnormal mito-chondria in mycelia and lipid droplets and vacuoles in conidia.Phenotype analysis revealed that P450S and P450L facilitated fungal colony pigments,conidial formation and germination via bolstering conidiophores and cell walls in response to temperature reduction.