For the first time, an integrated multi-enzyme cascade reaction in a two-phase system is scaled up to miniplant-scale, mathematically modeled, and optimized through computer-aided process development methods in an effort towards reducing the gap between the academic research in biotechnology and the industrial application of enzyme-based processes identified in a significant number of literature reviews. After characterization in laboratory scale, the reference reaction system is implemented to a miniplant-scale reactor setup and mathematically modeled in Aspen Custom Modeler. Validated with independent experimental data, the process model is then used for mathematical process optimization.