Abstract. In this work, Evolutionary Algorithms (EAs) are used to achieve optimal feedforward control in a recombinant bacterial fed-batch fermentation process, that aims at producing a bio-pharmaceutical product. Three different aspects are the target of the optimization procedure: the feeding trajectory (the amount of substrate introduced in a bioreactor per time unit), the duration of the fermentation and the initial conditions of the process. A novel EA with variable size chromosomes and using real-valued representations is proposed that is capable of simultaneously optimizing the aforementioned aspects. Outstanding productivity levels were achieved and the results are validated by practice.
In the present work the characterisation by image analysis of anaerobic biomass under organic and hydraulic shocks was performed. The digester was fed with a synthetic substrate, containing 50% oleic acid (as COD). Organic and hydraulic shocks were performed by stepwise increasing the substrate concentration or by reducing the hydraulic retention time. In both cases the organic loading rate changed from 6 to 30 kg COD/m3.d. Hydraulic shock induced a fast decrease in the number of free filaments and in total filament length, which attained a minimum value 51 hours after beginning the shock. The initial filament values were not recovered 340 hours after the hydraulic shock. In the organic shock, the minimum values of these parameters were detected 200 hours after beginning the shock and initial values were recovered 840 hours after. During the hydraulic shock the methanogenic acetoclastic activity was directly correlated to the number and length of free filaments. This result suggests that filaments are predominantly acetoclastic bacteria, probably Methanosaeta.
The specific growth rate is one of the most important process variables characterizing the state of microorganisms during fermentations mainly because the biosynthesis of many products of interest is often related with the values assumed by this parameter. In the particular case of the fed-batch operation of Escherichia coli for the production of recombinant proteins, it is important to maintain the specific growth rate below a certain threshold in order to avoid the accumulation of acetic acid throughout the fermentation and, additionally, it is often argued that both pre-and the post-induction specific growth rates should be closely controlled in order to achieve maximum productivities of the desired recombinant protein. In a previous work the authors have developed and validated by simulations a strategy for the automatic control of the specific growth rate in E. coli fed-batch fermentations based on an asymptotic observer for biomass and on developed estimators for the specific growth rates. The main purpose of the present work was to implement experimentally the developed observer, estimator and controller in a real fed-batch fermentation process. For that purpose a data acquisition and control program was developed in LabVIEW that allows the acquisition of the necessary on line data (off gas and dissolved oxygen concentration and culture weight) and the calculation of the feeding rates using the developed equations. The feedforward-feedback controller developed was able to keep the culture growing in an exponential phase throughout the fermentation without accumulation of glucose and acetate.
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