Successful downsizing for high-throughput ¹³C-MFA applications

(13)C 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.