Analysis across scales: Integrated insights in metabolism and economic feasibility of recombinant protein production from CO2 and H2

Efficient microbial production processes are essential for sustainable biomanufacturing,yet optimizing strain performance and bioreactor conditions remains challenging due tothe complex interplay between cellular metabolism and process parameters. Processesinvolving hydrogen-oxidizing bacteria (HOB), such as Xanthobacter sp. SoF1, are par-ticularly challenging due to limitations in gas–liquid mass transfer throughout the reac-tor.This study leverages constraint-based models of microbial metabolism to gain a deeperunderstanding of food-grade heterologous protein production by Xanthobacter sp.SoF1, predict cellular responses under bioreactor conditions, and assess how strainand process design choices influence performance and cost-effectiveness.Metabolic modeling is combined with simulations of various process conditions to iden-tify metabolic bottlenecks and key response patterns. Furthermore, a techno-economicassessment is used to identify target productivities and titers for an economically feasi-ble production process. The effect of different production scenarios on the strain perfor-mance and the economic feasibility serves as a basis for decisions in the envisionedbioprocess. By systematically mapping metabolic responses to process variations, thiswork provides a framework to connect the different scales affecting bioprocesses. Theresult of this framework are actionable insights for designing more efficient and cost-ef-fective microbial production systems. Furthermore, it guides rational decision-making instrain development and bioprocess optimization.