Related Experiment Video
Updated: Jun 1, 2026
06:24
Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Simple control of fed-batch processes for recombinant protein production with E. coli
Sebastian Schaepe1, Artur Kuprijanov, Mathias Aehle
1Institute of Biochemistry and Biotechnology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle, Saale, Germany. sebastian.schaepe@biochemtech.uni-halle.de
Biotechnology Letters
|June 1, 2011
Summary
A novel process control method enhances batch reproducibility for E. coli fermentations. This technique improves biomass concentration and growth rate, aiding recombinant protein production.
Area of Science:
- Biotechnology
- Biochemical Engineering
- Microbial Fermentation
Background:
- Achieving consistent batch-to-batch reproducibility in microbial fermentations is crucial for biopharmaceutical production.
- Current process control strategies for Escherichia coli (E. coli) fermentations can be limited in optimizing biomass concentration and growth rate profiles.
- Recombinant protein production in E. coli requires precise control over fermentation parameters.
Purpose of the Study:
- To introduce a simple yet effective process control technique for E. coli fermentations.
- To enhance batch-to-batch reproducibility in biomass concentration and specific biomass growth rate.
- To enable online estimation of oxygen mass transfer rates and k(L)a values.
Main Methods:
- Utilizing the difference between culture temperature (T_cult) and coolant temperature (T_coolin) from standard fermenter temperature controllers.
- Correcting the temperature difference for stirrer influences.
- Cumulating the corrected temperature difference to create a new process control variable.
Main Results:
- Demonstrated high batch-to-batch reproducibility in biomass concentration and specific biomass growth rate profiles.
- Successfully applied the novel control variable in E. coli fermentations for recombinant protein production.
- Enabled online estimation of oxygen mass transfer coefficients (k(L)a) as a byproduct of the control strategy.
Conclusions:
- The proposed simple process control technique significantly improves reproducibility in E. coli fermentations.
- This method offers a valuable tool for optimizing recombinant protein production processes.
- The ability to estimate oxygen mass transfer online provides additional process insights.
Related Concept Videos
01:27Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
01:23Fed-Batch Culture
Fed-batch culture is a widely used bioprocessing strategy combining aspects of batch culture with controlled substrate feeding to optimize cell growth and product formation. In this semi-closed system, nutrients are strategically added during fermentation, while the accumulated products and biomass remain within the bioreactor until the end of the operation. This controlled addition of substrates allows for better management of growth kinetics, nutrient limitation, and metabolite...
01:30Production of Pharmaceuticals
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
01:14Batch vs Continuous Culture
Fermentation is a foundational biotechnological process used to produce pharmaceuticals, biofuels, enzymes, and food additives. Among industrial strategies, batch and continuous fermentation are the two most widely applied. Although both rely on microbial conversion of substrates into desired products, they differ markedly in operation, productivity, and suitability for specific applications.Batch fermentation occurs in a closed system in which nutrient media and inoculum are added at the...
01:28Bioreactor Controls-I
Maintaining optimal conditions within fermenters is essential for maximizing microbial productivity and ensuring process efficiency. This lesson focuses on key parameters—temperature, foam, pH, carbon dioxide, oxygen, and pressure—and their precise measurement and control strategies in fermentation systems.Temperature ControlTemperature regulation is critical due to the exothermic nature of many fermentation processes. In small laboratory fermenters, temperature is commonly monitored using...
01:18Bioreactor Controls-II
In aerobic fermentations, oxygen is vital for microbial growth and metabolite production. Since air comprises only about 20% oxygen and the gas is poorly soluble in water—just 9 ppm at 20°C—supplying sufficient oxygen becomes a critical challenge, especially in high-demand processes like yeast growth or citric acid production. Even a fully saturated broth may offer only a few seconds of oxygen availability.To address this, sterile or scrubbed air is introduced into the fermentor via a sparger...