Enzyme-Mediated Exponential Glucose Release: A Model-Based Strategy for Continuous Defined Fed-Batch in Small-Scale...

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Synthetic Biology

Background:

• Miniaturized cultivation systems boost experimental throughput but often lack precise feeding capabilities for fed-batch processes.
• Enzyme-mediated glucose release from starch derivatives offers continuous supply but lacks precise control over feed rates.
• Implementing controlled feeding strategies, like exponential feeding, in small-scale systems remains challenging.

Purpose of the Study:

• To develop a model-based approach for precise control of enzyme-mediated glucose release in cultivations.
• To enable defined continuous fed-batch processes in small-scale bioprocess development.
• To overcome limitations in achieving accurate feeding in miniaturized bioreactors.

Main Methods:

• Integrated an existing mathematical model into a computational framework.
• Calculated setpoints for enzyme additions to control glucose release.
• Utilized a robotic facility with parallel mini-bioreactors for Escherichia coli cultivations.

Main Results:

• Demonstrated precise control over enzyme-mediated glucose release.
• Successfully maintained pre-defined exponential growth rates during Escherichia coli cultivations.
• Showcased the adaptability of the approach for manual applications.

Conclusions:

• The proposed model-based approach enables straightforward and robust implementation of defined continuous fed-batch processes in small-scale systems.
• This method overcomes previous limitations in achieving accurate and controlled feeding in miniaturized bioreactors.
• Enhances the utility of miniaturized systems for bioprocess development and experimental throughput.