Scaling-up with the eyes of the microbe - Sky

• Applicant: Professor Dr.-Ing. Ralf Takors
• Subject Area: Biological Process Engineering
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 569559247

The successful establishment of new biomanufacturing processes is an essential step towards a sustainable circular economy based on renewable resources. Unfortunately, experience shows that the implementation of promising ideas from the laboratory into large-scale applications is a major hurdle. Too often this is not successfully overcome. Too many very good approaches do not make the leap into implementation, which is often due to a lack of fundamental understanding of the underlying mechanisms. This is where the Sky research project comes in. Sky aims to develop robust Escherichia coli chassis that exhibit maximum performance even under the heterogeneous cultivation conditions that prevail on a production scale. This approach not only ensures the successful transfer from the laboratory to production, but also opens up the possibility of realizing new, large-volume production processes with less technical effort. Both support the successful establishment of sustainable biomanufacturing as a pillar of the circular bioeconomy. To achieve this, Sky is pursuing a synergistic approach of microbial strain construction and engineering scale-down. Both disciplines are anchored at the Institute of Bioprocess Engineering (IBVT). Based on already existing, very extensive data on the transcriptional response of E. coli to large-volume stress, comprehensive genome scale modeling will provide specifications for a targeted genome reduction. These are implemented using molecular biology. It should be emphasized that no genome-reduced strain should exhibit growth deficits or additional substrate requirements. This means that the concept developed for the E. coli RM214 strain, successfully developed at the IBVT, is being pursued for the stringent response deficient strain E. coli SR and rigorously implemented. In addition, intracellular possibilities of carbohydrate supply are also implemented in the strains for performing in zones of substrate deficiency. The performance data of such strains will be evaluated in newly constructed scale-down bioreactors. The focus is on the engineering design and realization of the single multi-compartment bioreactor (SMCB) for microorganisms. The latter was recently successfully developed by the applicant for the scale-down of mammalian cell processes. Real data from a 600 m³ bubble column, collected and modeled by the applicant's working group, are used as the basis for the scale-down. This SMCB bioreactor serves as a test system for the parallel work on targeted genome reduction in the synergistic Sky project.

• DFG Programme: Research Grants