Early bioprocess development using Komagataella phaffii: application scenarios for the microTOM mini bioreactor...

Area of Science:

• Biotechnology
• Biochemical Engineering
• Microbial Physiology

Background:

• Developing novel bioprocesses involves extensive strain engineering and screening for optimal performance.
• Monitoring oxygen transfer rate (OTR) is crucial for aerobic bioprocesses to avoid oxygen limitation and assess metabolic activity.
• High-throughput methods for obtaining OTR data are essential for efficient bioprocess development.

Purpose of the Study:

• To evaluate the utility of the micro-scale Transfer rate Online Measurement (microTOM) system for rapid bioprocess development.
• To demonstrate the application of the microTOM system in high-throughput screening of microbial strains and optimization of cultivation conditions.
• To assess the system's capability in determining critical process parameters for scale-up.

Main Methods:

• Utilized a 96-well microtiter plate format with the microTOM system for online OTR monitoring.
• Tested the microbial host Komagataella phaffii.
• Investigated multiple strains, media formulations, carbon sources, antifoam agents, filling volumes, and temperatures.

Main Results:

• The microTOM system enabled high-throughput investigation of three strains in parallel.
• Methanol uptake and growth rates were estimated, providing data for process modeling and scale-up.
• Optimized screening conditions by assessing various media, carbon sources, antifoam agents, filling volumes, and temperatures.

Conclusions:

• The microTOM system significantly accelerates bioprocess development by enabling rapid, data-driven decision-making.
• It facilitates high-throughput determination of critical process parameters, previously not feasible.
• The system supports robust, data-driven bioprocess development for improved large-scale productivity.