CFD-guided scaling of Pseudomonas putida fermentation

Abstract

This study evaluated the scale-up of Pseudomonas putida fed-batch fermentation from a 2 L benchtop-scale stirred bioreactor to a 200 L pilot-scale tank by using a validated computational fluid dynamic (CFD) model. One of the major concerns in this fermentation process is the potential reduction in mixing quality with increasing scale, leading to lower yield or product quality. For a low-risk scale-up, a multiphase Euler-Euler CFD model was developed that simulated the hydrodynamics of the fed-batch system at various filling volumes, representing different stages of the fermentation process. The model was validated using experimental data of mixing time and mass transfer coefficient. The hydrodynamic model was then coupled with a Monod kinetic model of P. putida ‘s fermentation. Response surface methodology was used to generate a performance map of the pilot bioreactor at various aeration, agitation, and bioreactor filling volumes. The study considered different established scale-up approaches, such as constant tip speed and aeration rate across scales, constant $k_{L}a$, as well as constant power to unit of liquid volume (P/V). The performance of the bioreactor was assessed, and the optimum operating ranges of the input parameters were obtained at different stages of the fermentation. Using performance map the possibility of substrate and oxygen gradient formation, and the gradient severity inside the pilot bioreactor at different working volumes were evaluated.