Kinetics Modeling of a Convergent Cascade Catalyzed by Monooxygenase–Alcohol Dehydrogenase Coupled Enzymes

A convergent cascade reaction coupling a cyclohexanone monooxygenase variant and an alcohol dehydrogenase to make ε-caprolactone from cyclohexanone and 1,6-hexanediol was characterized via progress curve analysis with two kinetic models developed iteratively. A chemical side reaction occurring with the utilized Tris buffer and consequent byproduct formations were considered in Model 2, which reduced the root-mean-square error (RMSE) values by half, compared to Model 1 (RMSE values of 13%–40%). The optimized model, Model 2, led us to simulate the cascade reaction including 22 kinetic parameters with a maximum RMSE value in the range of 10%–21%.