engleski

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 of therapeutically- valuable iminocyclitols is discussed herein. A two-enzyme system consisting of chloroperoxidase (CPO) and D-fructose-6-phosphate aldolase (FSA) was evaluated for the synthesis of a precursor of D-fagomine from a β-amino alcohol (N-Cbz-3- aminopropanol). Both the oxidation and aldol addition reactions were investigated to elucidate the reaction mechanism, prepare suitable stabilized biocatalysts, and find the optimal conditions for its application. Also, the inactivation of both enzymes and many other side reactions (the oxidation of the intermediate amino aldehyde, an aldehyde-peroxide reaction, etc.), are discussed and analyzed in detail. An in-depth, systematic, step-by-step kinetic modeling of all the reactions was proposed as an approach for a better understanding of the system, and a possible strategy to optimize it. The model was able to accurately describe the system and predict the outcome at different experimental conditions. The inactivation of FSA caused by CPO-catalyzed reactions was identified as the many bottleneck in the reaction. A two- step reaction approach and the use of immobilized enzymes derivatives on magnetic nanoparticle clusters and functionalized agarose carriers increased the stability of FSA, and thus a higher product yield was obtained with a 577-fold higher preFagomine formation per mol of enzyme.

D-fagomine ; kinetic modeling ; enzyme cascade ; peroxidase ; aldolase

nije evidentirano