High-Throughput Stationary ¹³C-Based Metabolic Flux Analysis of Pichia pastoris Strains

¹³C-based Metabolic Flux Analysis (¹³C-MFA) is an experimental approach that allows the quantification of the intracellular reaction rates (fluxes) that has become a valuable tool in the field of metabolic systems engineering. Rapid advances in the field of synthetic biology are leading to the generation of large numbers of engineered strains for the same research project. Thereby, the use of high-throughput cultivation and analytical platforms is required to test and characterize these sets of strains and extract meaningful information for further strain engineering cycles. Here, a detailed protocol for the characterization of the fluxome of a set of Komagataella phaffii strains using a high-throughput cultivation platform is described.