Biosynthesis of the alpha-D-Mannosidase Inhibitor (-)-Swainsonine

(-)-Swainsonine is a polyhydroxylated indolizidine alkaloid with potent inhibitory activity against α-D-mannosidases. In this work, we successfully reconstituted swainsonine biosynthetic pathway both in vivo and in vitro. Our study unveiled a surprising epimerization mechanism involving an imine reductase (SwnN) and two non-heme iron and α-ketoglutarate-dependent oxygenases (SwnH2 and SwnH1). SwnH2 is a multifunctional enzyme responsible for both C2-hydroxylation and amine desaturation, while SwnN completes the oxidation-reduction sequence to achieve the net redox neutral epimerization. Notably, SwnN exhibits substrate-dependent stereospecificity and the stereochemical outcome of SwnN-catalyzed iminium reduction is determined by SwnH1-catalyzed C8-hydroxylation. We also discovered that an