This project is funded by the „Bridge to Industry” program of SystemsX.ch
The design and optimization of microorganisms that overproduce a certain compound or biopharmaceutical is one of the most common applications of current biotechnology. Major advances in particular in metabolomic steady state and transient analysis technologies allow unprecedented insights into the systems-level structure of metabolic networks. However, these methods are experimentally and theoretically very laborious and frequently still fail to pinpoint specific cellular targets, the modification of which should improve pathway performance.
Here, we propose to limit the analysis to a relevant subsystem – one specific pathway, including the comprehensive set of substrates, intermediates, and products and the corresponding enzymes - and furthermore to analyze the functionality of the subsystem in vitro instead of in vivo as it is typical for current flux determination methods. By analyzing in vitro, we gain an exceptional degree of freedom in the design of subsystem perturbations, including the possibility to systematically perturb compound concentrations to identify fundamental limitations and maximum potential of the pathway in question and to carry out precise additions of single or multiple purified pathway enzymes to discover more favorable pathway compositions. By reducing the size of the analyzed system, we allow a drastic shortening of analysis time, ultimately allowing highly time-resolved on-line analysis of system behavior under transient conditions.
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