Exploration of the Out-of-Phase Phenomenon in Shake Flasks by CFD Calculations of Volumetric Power Input, kLa Value and Shear Rate at Elevated Viscosity.

Culture broth with secreted macromolecules and culture broth of filamentous fungi showing disperse growth exhibit elevated viscosity, usually with shear-thinning flow behavior. High viscosity, however, poses a serious challenge in the production and research of these compounds and organisms. It commonly causes insufficient mixing and oxygen transfer in large- and small-scale bioreactors. Computational Fluid dynamics (CFD) has been proven to be a valuable tool for the computation of important bioprocess parameters. The published literature for small-scale shaken bioreactors, especially shake flasks, however, almost exclusively focuses on water-like viscosity. In this paper, a previously published CFD model for 250 mL shake flasks was used to simulate experiments at high viscosities of up to 100 mPa·s. Compared to experimental data, the CFD model accurately predicted the liquid distribution and computed the volumetric power input with a deviation of less than 7% and the kLa value within a factor of two, compared to the kLa correlation from Henzler and Schedel. Furthermore, a novel approach to compute the shear rate was tested. Lastly, new insights into the out-of-phase phenomenon were gained. The presented data confirms the usefulness of the already established critical phase numbers of 0.91 and 1.26, while underlying the fundamentally smooth transition from in-phase to out-of-phase operating conditions.