Modeling the Glucose Concentration for the Recombinant E.coli Bioprocesses

This article presents the sophisticated-to-date carbon mass balance for fed-batch E.coli bioprocesses. The model originates from the distribution of carbon mass from glucose in biomass, off-gas, and hypothetical solutes. The suggested model complements Pirt's equation as a particular case scenario. The approach uses the linear relationship between biomass carbon content per carbon grams in glucose and average cell population age. The carbon balance brings two potential practical benefits. First, it has the potential to assess the type of cell metabolism pathway and to have a soft sensor for the concentration of dissolved products such as acetates. The measure of glucose concentration suggests another finding, assuring the reliance on off-gas information only. The paper introduces an average carbon content ratio in biomass and off-gas, with numerical values of 0.5 in growth-limiting experiments and 0.27 in nonlimiting ones, which may serve as a decision-making criterion for metabolic pathway detection in the future

Identification of Functional Bioprocess Model for Recombinant E. Coli Cultivation Process

The purpose of this study is to introduce an improved Luedeking–Piret model that represents a structurally simple biomass concentration approach. The developed routine provides acceptable accuracy when fitting experimental data that incorporate the target protein concentration of Escherichia coli culture BL21 (DE3) pET28a in fed-batch processes. This paper presents system identification, biomass, and product parameter fitting routines, starting from their roots of origin to the entropy-related development, characterized by robustness and simplicity. A single tuning coefficient allows for the selection of an optimization criterion that serves equally well for higher and lower biomass concentrations. The idea of the paper is to demonstrate that the use of fundamental knowledge can make the general model more common for technological use compared to a sophisticated artificial neural network. Experimental validation of the proposed model involved data analysis of six cultivation experiments compared to 19 experiments used for model fitting and parameter estimation

From Physics to Bioengineering: Microbial Cultivation Process Design and Feeding Rate Control Based on Relative Entropy Using Nuisance Time

For historic reasons, industrial knowledge of reproducibility and restrictions imposed by regulations, open-loop feeding control approaches dominate in industrial fed-batch cultivation processes. In this study, a generic gray box biomass modeling procedure uses relative entropy as a key to approach the posterior similarly to how prior distribution approaches the posterior distribution by the multivariate path of Lagrange multipliers, for which a description of a nuisance time is introduced. The ultimate purpose of this study was to develop a numerical semi-global convex optimization procedure that is dedicated to the calculation of feeding rate time profiles during the fed-batch cultivation processes. The proposed numerical semi-global convex optimization of relative entropy is neither restricted to the gray box model nor to the bioengineering application. From the bioengineering application perspective, the proposed bioprocess design technique has benefits for both the regular feed-forward control and the advanced adaptive control systems, in which the model for biomass growth prediction is compulsory. After identification of the gray box model parameters, the options and alternatives in controllable industrial biotechnological processes are described. The main aim of this work is to achieve high reproducibility, controllability, and desired process performance. Glucose concentration measurements, which were used for the development of the model, become unnecessary for the development of the desired microbial cultivation process

Adaptive control of the E. coli-specific growth rate in fed-batch cultivation based on oxygen uptake rate

In this study, an automatic control system is developed for the setpoint control of the cell biomass specific growth rate (SGR) in fed-batch cultivation processes. The feedback signal in the control system is obtained from the oxygen uptake rate (OUR) measurement-based SGR estimator. The OUR online measurements adapt the system controller to time-varying operating conditions. The developed approach of the PI controller adaptation is presented and discussed. The feasibility of the control system for tracking a desired biomass growth time profile is demonstrated with numerical simulations and fed-batch culture E.coli control experiments in a laboratory-scale bioreactor. The procedure was cross-validated with the open-loop digital twin SGR estimator, as well as with the adaptive control of the SGR, by tracking a desired setpoint time profile. The digital twin behavior statistically showed less of a bias when compared to SGR estimator performance. However, the adaptation—when using first principles—was outperformed 30 times by the model predictive controller in a robustness check scenario