Abstract
13C label-based metabolic flux analysis is a powerful technique for the determination of intracellular reaction rates and is used in such different research fields as quantitative physiology, metabolic engineering, and systems biology. Metabolic fluxes can be determined at high quality using (pseudo)-steady-state cultures and advanced mathematical models for data interpretation. Here, we describe a protocol for parallel metabolic flux analysis that consists of downsized microbial (yeast) cultivation, miniaturized sample preparation, and semiautomated analytics and data evaluation. With this protocol dozens of metabolic flux analyses can be carried out in 1 week, thereby enabling for example the analysis of genetic and environmental perturbations on the operation of metabolic networks.
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Acknowledgement
This work was supported by the Cluster of Excellence “Tailor-Made Fuels from Biomass,” which is funded by the Excellence Initiative of the German federal and state governments to promote science and research at German universities.
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Ebert, B.E., Blank, L.M. (2014). Successful Downsizing for High-Throughput 13C-MFA Applications. In: Krömer, J., Nielsen, L., Blank, L. (eds) Metabolic Flux Analysis. Methods in Molecular Biology, vol 1191. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1170-7_8
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DOI: https://doi.org/10.1007/978-1-4939-1170-7_8
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Online ISBN: 978-1-4939-1170-7
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