Minireactor-based high-throughput temperature profiling for the optimization of microbial and enzymatic processes.

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Enzyme Technology

Background:

• Temperature is a critical parameter in bioprocesses, yet rapid determination of temperature-dependent kinetics remains challenging.
• Existing micro-bioreactor systems offer potential but face difficulties in precise temperature control.
• There is a clear need for high-throughput screening methods to analyze temperature-dependent process behavior.

Purpose of the Study:

• To develop and evaluate a novel system for high-throughput temperature profiling of bioprocesses.
• To enable precise temperature control and real-time monitoring in microtiter plate formats.
• To facilitate the optimization of microbial and enzymatic reactions at the micro-scale.

Main Methods:

• A unique system combining an optical on-line monitoring device with a customized temperature control unit for 96-well microtiter plates was developed.
• Fluorescence thermometry using Rhodamine B and Rhodamine 110 was employed for single-well resolved temperature measurement.
• The system was validated by determining temperature optima for microbial growth and recombinant protein production in Escherichia coli and Kluyveromyces lactis, and for a commercial cellulase enzyme activity.

Main Results:

• The developed system successfully achieved high-throughput temperature optimization for microbial and enzymatic systems in 200 μL volumes.
• Real-time monitoring provided extensive data output for microbial and enzymatic reactions.
• Temperature optima were successfully determined for selected microorganisms and enzymes, demonstrating the system's efficacy.

Conclusions:

• Microtiter plate-based high-throughput temperature profiling is an effective tool for characterizing temperature-dependent reaction processes.
• The system allows for rapid evaluation of diverse conditions, including microorganisms, enzymes, and media.
• The combination of simple temperature control and commercial on-line monitoring results in a user-friendly and efficient platform.