Supplementary Tables for: Resolution of the ABA Biosynthesis Controversy: Discovery of a Dihydroxylating Terpene Synthase and its Convergent Evolution

Supplementary tables S1–S9 for the manuscript: "Resolution of the ABA Biosynthesis Controversy: Discovery of a Dihydroxylating Terpene Synthase and its Convergent Evolution". This dataset contains the following supplementary information: Table S1: Distribution of the BcStc family among the Botrytis genera with NCBI ID. Table S2: List of key homologs of BcStc5 identified in other fungi. Table S3: Detailed results of the ConSurf analysis for BcStc5, including conservation scores and functional/structural predictions for each residue. Table S4: Calculated binding affinity (ΔG) and root mean square deviation (RMSD) values for the top five docking models of FDP in the BcStc5 active site. Table S5: Complete list of protein-protein interactions and associated STRING scores for the supercluster networks: (A) Dw1 and (B) T4. Table S6: Functional annotation and enrichment analysis (GO terms, KEGG pathways, and STRING clusters) for the supercluster proteins: (A) B. cinerea DW1 and (B) B. cinerea T4. Table S7: Functional Conservation of the Metabolic Supercluster in B. cinerea Strains T4 and DW1. Table S8: Experimental design matrix and observed responses for the DoE (Design of Experiments) study. Table S9: Bacterial Strains and Plasmids Utilized in This Study. Abstract: Fungal terpene biosynthesis is a vital source of bioactive metabolites. Here, we elucidate the function of BeStc5, a Botrytis cinerea terpene synthase previously linked to abscisic acid (ABA) biosynthesis. Using bioinformatics and heterologous expression, we demonstrate that BcStc5 catalyzes the formation of (4S, 5S, 7R, 10S)-4B,10a-eudesmane-5ẞ,11-diol (1), a dihydroxylated sesquiterpenoid not previously reported in fungi. Mechanistic modeling suggests BcStc5 employs a novel, P450-independent dihydroxylation strategy. We further established a scalable bioproduction platform in E. coli, achieving titers >550 mg/L. This work resolves a long-standing biosynthetic controversy, characterizing BcStc5 as an eudesmanediol synthase rather than the ABA cyclase, settling the controversy over this protein.