Oosporein in Phlebia centrifuga: Prediction of Key Enzymes by a Combination of Multi-Omics and Molecular Dynamics.

The red bibenzoquinone oosporein, a promising biocontrol agent with potential to replace conventional pesticides in insect pest management in crops, was produced by the Basidiomycota Phlebia centrifuga P. Karst isolated from the Black Forest National Park. A submerged system was established, yielding up to 1.60 g L-1 oosporein in the culture supernatant upon supplementation with the key intermediate orsellinic acid, which strongly induced oosporein-specific biosynthetic genes. Using a multiomics approach, genes encoding enzymes for all necessary conversions were predicted, including a type I polyketide synthase, two monooxygenases, and a heme peroxidase. Enzymatic functions were investigated by extensive docking analyses and molecular dynamics simulations, ultimately leading to the prediction of the underlying biosynthetic pathway. In summary, a spore-free, high-yield, scalable production platform for oosporein was established, highlighting the potential of rare, protected fungal species as sources for valuable enzymes and bioactive secondary metabolites for efficient microbial biomanufacturing systems.