Online estimation of changing metabolic capacities in continuous Corynebacterium glutamicum cultivations growing on a complex sugar mixture.

Model-based state estimators enable online monitoring of bioprocesses and, thereby, quantitative process understanding during running operations. During prolonged continuous bioprocesses strain physiology is affected by selection pressure. This can cause time-variable metabolic capacities that lead to a considerable model-plant mismatch reducing monitoring performance if model parameters are not adapted accordingly. Variability of metabolic capacities therefore needs to be integrated in the in silico representation of a process using model-based monitoring approaches. To enable online monitoring of multiple concentrations as well as metabolic capacities during continuous bioprocessing of spent sulfite liquor with Corynebacterium glutamicum, this study presents a particle filtering framework that takes account of parametric variability. Physiological parameters are continuously adapted by Bayesian inference, using noninvasive off-gas measurements. Additional information on current parameter importance is derived from time-resolved sensitivity analysis. Experimental results show that the presented framework enables accurate online monitoring of long-term culture dynamics, whereas state estimation without parameter adaption failed to quantify substrate metabolization and growth capacities under conditions of high selection pressure. Online estimated metabolic capacities are further deployed for multiobjective optimization to identify time-variable optimal operating points. Thereby, the presented monitoring system forms a basis for adaptive control during continuous bioprocessing of lignocellulosic by-product streams.

Noninvasive online monitoring of Corynebacterium glutamicum fed-batch bioprocesses subject to spent sulfite liquor raw material uncertainty.

In this study the use of a particle filter algorithm to monitor Corynebacterium glutamicum fed-batch bioprocesses with uncertain raw material input composition is shown. The designed monitoring system consists of a dynamic model describing biomass growth on spent sulfite liquor. Based on particle filtering, model simulations are aligned with continuously and noninvasively measured carbon evolution and oxygen uptake rates, giving an estimate of the most probable culture state. Applied on two validation experiments, culture states were accurately estimated during batch and fed-batch operations with root mean square errors below 1.1 g L-1 for biomass, 0.6 g L-1 for multiple substrate concentrations and 0.01 g g-1 h-1 for biomass specific substrate uptake rates. Additionally, upon fed-batch start uncertain feedstock concentrations were corrected by the estimator without the need of any additional measurements. This provides a solid basis towards a more robust operation of bioprocesses utilizing lignocellulosic side streams.