Debottlenecking of an in vitro enzyme cascade using a combined model‐ and experiment‐based approach

Abstract

Designing a feasible in vitro enzyme cascade is a challenging task that is often preceded by optimization steps. Computational methods can increase the efficiency of the optimization process by providing a deeper insight into the system. In this study, bottlenecks of a farnesyl pyrophosphate-producing in vitro enzyme cascade were determined by a combination of in silico modeling and wet-lab experiments. ATP and the precursor-isomerizing enzyme isopentenyl diphosphate δ-isomerase were identified as limitations of the in vitro enzyme cascade.