“BioBreak – Enzymes for Biomass Breakdown
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Plant biomass is regarded as the most important source of renewable raw material for the bioeconomic conversion of energy, as well as production of fuels and chemicals. To date, the biotechnological utilization of plant biomass combines physical, chemical, and biocatalytic processes. However, substantial costs still prevent a thorough replacement of oil-based feedstocks. As the processing of the rather complex biomass requires many steps and therefore the use of a variety of different enzymes, it is necessary to produce those enzymes on large-scale and at low costs.
Therefore, the “BioBreak” project aims at evaluating two well-known bacterial expression strains, namely Escherichia coli and Bacillus subtilis, for the production and secretion of biomass-degrading enzymes to establish viable alternatives to existing fungal systems. Within the group of Prof. Büchs the optimization of the fermentation and the scale-up from microtiter plate to large-scale production will be carried out.
In order to save time and money, a thorough characterization of both microbial systems will be carried out in microtiter plates by using the BioLector® device. With the help of fluorescent reporter proteins, the product formation and other important process parameters (dissolved oxygen tension, biomass and NADH formation) can be monitored and compared online. After optimizing essential parameters like inducer concentration, time of induction, temperature optimum and media composition, a first scale-up of promising candidates into shake-flask scale will be performed.
Using the RAMOS®-device, promising candidates will be further characterized and suitable fed-batch conditions for high-level protein production will be determined by combining the RAMOS®-technique with newly developed shake flasks that allow feeding of different compounds. Subsequently, the fermentation will be transferred into standard laboratory fermenters.
Finally, high cell density cultivation will be carried out to achieve high enzyme concentrations. By applying a unique pressurized fermenter available at the chair of biochemical engineering, gas/liquid mass transfer is not a limiting issue anymore and enzyme production can finally be carried out at large-scale (50 L).
Participating Core Groups
Coordinator: Prof. Dr. Karl-Erich Jaeger, Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf
Prof. Dr. Lutz Schmitt, Institute of Biochemistry I, Heinrich-Heine-University Düsseldorf
Prof. Dr. Antje Spieß, Enzyme Process Technology (AVT.EPT), RWTH Aachen University, Aachen
Partners from industry
AB Enzymes GmbH, Feldbergstrasse 78, 64293 Darmstadt, Germany
Gefördert durch: BioSC (Bioeconomy Science Center)