Biosynthesis of the trichothecene 3-acetyldeoxynivalenol. Identification of the oxygenation steps after isotrichodermin

Abstract

Upon feeding an excess of the substrate isotrichodermin, five tricyclic metabolites accumulated in Fusarium culmorum cultures. These compounds were also identified as transient intermediates of trichothecene biosynthesis by kinetic pulse labeling. Their structures were characterized by spectroscopic techniques (1H NMR, 13C NMR, 2H NMR, and nuclear Overhauser effect difference experiments) as: 1, 15-deacylcalonectrin; 2, calonectrin; 3, 7-hydroxyisotrichodermin; 4, 8-hydroxyisotrichodermin; and 5, 7, hydroxycalonectrin. Four of these metabolites (1-4) were rigorously proven to be biosynthetic precursors to 3-acetyldeoxynivalenol. Indeed, their deuteriated derivatives were shown to be incorporated very efficiently into 3-acetyldeoxynivalenol by 2H-NMR. In addition, our experimental data suggests that the first oxygenation step after isotrichodermin is at C-15, producing 15-deacylcalonectrin.