Global Metabolomics and Isotopically-Labeled Metabolomics for Metabolic Flux Analysis
Metabolomics is a key aspect of an integrated systems biology approach to understanding engineered and basal strains under different conditions to enable learning that leads to the next set of Designs for DBTL.
Global metabolomics is the derivatization GC-MS analysis of the intracellular metabolome. The extracellular metabolites can be analyzed separately to assess substrate utilization and secretion of target molecules as well as side-products.
13C Metabolic Flux Analysis (MFA) involves the use of 13C labeled substrates and subsequent metabolomics and metabolic modeling analysis discussed in the Modeling and Machine Learning Capability to determine fluxes through pathways.
LC-MS and NMR methods are techniques complementary to our standard GC-MS metabolomics analysis. They are useful for examining large hydrophilic molecules and can be utilized for verification of structures of complex or novel intermediates or target molecules.
The Agile BioFoundry has three GC-MS instruments devoted to metabolomics and one LC-MS, as well as high resolution instruments for isotopically labelled metabolite assignments, enabling 13C-MFA, and multiple NMR instruments of moderate to high fields with diverse probes available.
Preference for this activity is ~50 mg of wet cell weight per sample, quadruplicate biological replicates, and sample sets of 48-60 though twice that number can be accommodated.
- Lawrence Berkeley National Laboratory
- Pacific Northwest National Laboratory