FluxProbe - Using biocatalysis for probing the functional space of single microbial catalysts

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Whole-cell biocatalysts are powerful tools for organic synthesis due to their unmatched chiral and positional selectivity. But what is the contribution of the minimal catalytic unit, the single cell, to biocatalysis? Discrete rates for reactivity of individual cells, as well as mechanisms that control cellular functionality, are still largely unknown due to a lack of suitable analytics. Project 7 will establish a chip-based analytical framework for quantifying biocatalyst performance at the single cell level. By integrating microfluidic bioreactor technologies and miniaturized analytics for mass spectrometry of biocatalytic products, the conditions-dependent catalytic performance of single cells in terms will be evaluated quantitatively for the first time. However, catalytic products will not only be quantified, but also metabolically enriched with stable isotopes to serve as a metabolic probe (“flux probe”). With the power of mass spectrometry, the isotope labelling in the product molecules will reveal the catalytic network operation of the cell and finally enable connecting metabolic status with reactivity and extracellular reaction conditions. Key experiments performed in collaboration with the WG Belder already validated the feasibility of this concept.