Design and Application of Software Sensors in Batch and Fed-batch Cultivations during Recombinant Protein Expression in Escherichia coli

The aim of the current project was to develop software sensors which probe for important state parameters in batch and fed-batch cultivations and which enable to control processes in a more advanced and effective way.

Software sensors are a potent tool to improve biotechnological real time process monitoring and control. In the current project, algorithms for six partly novel, software sensors were established and tested in a microbial reactor system. Eight batch and two fed-batch runs were carried out with a recombinant Escherichia coli to investigate the suitability of the different software sensor models in diverse cultivation stages. Special respect was given to effects on the sensors after recombinant protein expression was initiated by addition of an inducer molecule…

Contents

1.Introduction
1.1Aim
1.2 Approach
1.3 Background
1.3.1General background
1.3.2 Software sensors
1.3.3 Industrial Application
1.4 Organisms and metabolites
1.4.1 Escherichia coli
1.4.2 Green Fluorescence protein
1.4.3 Vector system and induction strategy
1.4.4 Stress mediated proteins
1.5 Limitations of software sensing
2. Material and Methods
2.1 Strain
2.2 Medium
2.3 Cultivation conditions
2.4 Analyses
2.4.1 Online measurements
2.4.2 Offline measurements
3. Modeling and Theoretical Aspects
3.1 Nomenclature and definition of parameters
3.2 Estimation of biomass
3.3 Specific growth rate estimation
3.4 Specific glucose consumption
3.5 Specific production of acetate
3.6 Presentation and distribution of data
4. Results and Discussion
4.1 Estimation of biomass
4.2 Specific growth rate estimation
4.3 Specific glucose consumption
4.4 Specific production of acetate
4.5 Effects of IPTG pulse induction
5.Conclusions
6.Acknowledgements
7.References

Author: Warth, Benedikt

Source: Linköping University

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