Microliter-scale parallel cultivation system

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 547888200

The research at the Institute of Molecular Microbiology and Biotechnology (IMMB) focuses on the optimization of microbial processes and strains for the production of biobased materials and chemicals, especially biobased and biodegradable exopolysaccharides (EPS) (AG Schmid), bacterial cell-cell interactions and metabolism (AG Phillip), and the central carbon metabolism in bacteria and archaea (AG Berg). AG Schmid uses in vivo genetic engineering to develop chassis organisms and to generate strain variants, that produce various EPS and biobased chemicals. Based on growth and product formation under reproducible and controllable conditions, strain-specific optimization of the biosynthesis pathways is performed to increase productivity or modify products. Due to the high energy- and thus oxygen-dependent biosynthesis of polysaccharides, many EPS can only be produced under optimal conditions in bioreactors. The strain portfolio for this approach includes mainly Gram-negative and Gram-positive bacteria, but also filamentous fungi. The AG Phillip studies novel metabolic pathways and interspecies and interdomain metabolic interactions in microbial communities via cross-feeding. The growth of the individual strains in co-cultures can be measured by the expression of different fluorescent proteins, which is a great advantage for this research. AG Berg studies carbon dioxide fixation pathways in anaerobic, mainly thermophilic bacteria and archaea. The cultivation of these microorganisms requires a system that limits the evaporation at high temperatures and can maintain anaerobic conditions. The requested microliter-scale parallel cultivation system fulfils all of the above requirements, parallel cultivation (up to 48), stable temperature control, gas mixing, exhaust gas analysis and headspace control, as well as fluorescence measurements, and the possibility of online biomass and dissolved oxygen measurement. By integration of a pipetting robot, the system is equipped for automated optimization of cultivation media, which is a very powerful tool to increase growth rates and product titers, and a fundamental step for the translation into larger production scales. In addition, the ability to perform targeted and automated adaptive laboratory evolution (aALE) will massively extend our methodical capabilities to develop optimized strains for process optimization in our existing 1 parallel bioreactor system. Due to the large number of cultivation vessels and the high degree of automation, the microliter scale parallel cultivation system is the perfect system for design of experiments (DoE) and ALE approaches, as well as for high-throughput evaluation of microbial growth, protein expression (fluorescence) and media optimization. The system is the perfect high-throughput complement to the existing fermentation capabilities from 1 L - 500 L and will massively contribute to the scientific success of the research groups at the IMMB.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Paralleles Kultivierungssystem im Mikrolitermaßstab
• Instrumentation Group: 3520 Bakterien-Zuchtgeräte, Fermenter
• Applicant Institution: Universität Münster
• Leader: Professor Dr.-Ing. Jochen Schmid