Multi-reaction kinetic modeling for the peroxidase–aldolase cascade synthesis of a D-fagomine precursor

The feasibility of a peroxidase–aldolase cascade reaction for the synthesis therapeutically-valuable iminocyclitols is discussed herein. A two-enzyme system consisting chloroperoxidase (CPO) and D-fructose-6-phosphate aldolase (FSA) was evaluated D-fagomine precursor (preFagomine) from N-Cbz-3-aminopropanol. An in-depth, systematic, step-by-step kinetic modeling seven reactions two inactivation decays proposed to elucidate mechanism, prepare suitable stabilized biocatalysts, find optimal conditions its application. model described accurately data predicted outcome at different experimental conditions. FSA caused by CPO identified as main bottleneck in reaction. two-step approach use immobilized enzymes on magnetic nanoparticle clusters functionalized agarose carriers increased stability FSA, with an 1839-fold higher preFagomine formation per mol enzyme comparison one-pot using soluble enzymes.