A Clade II‐D Fungal Chimeric Diterpene Synthase from Colletotrichum gloeosporioides Produces Dolasta‐1(15),8‐diene

Abstract Based on a terpenoid overproduction platform in yeast for genome mining, a chimeric diterpene synthase from the endophytic fungus Colletotrichum gloeosporioides ES026 was characterized as the (5 R ,12 R ,14 S )‐dolasta‐1(15),8‐diene synthase. The absolute configuration was independently verified through the use of enantioselectively deuterated terpene precursors, which unequivocally established the predicted C1‐III‐IV cyclization mode for this first characterized clade II‐D enzyme. Extensive isotopic labeling experiments and isolation of the intermediate (1 R )‐δ‐araneosene supported the proposed cyclization mechanism.