Development of NMR Sparse Sampling Methods for Quantitative Metabolic Flux Analysis
EMSL Project ID
49418
Abstract
Quantitative metabolomics and metabolic flux analysis have emerged in recent years as valuable approaches for the study of carbon cycling in living organisms and in the environment, whether for understanding biological processes impacting soil or atmospheric carbon, for the development of engineered microorganisms for biofuels production, or for the understanding of cancer biology in medicine. EMSL has identified metabolic flux analysis as a specific target for the Biosystem Dynamics and Design science theme. This proposal addresses that theme by developing new nuclear magnetic resonance (NMR) tools for the study of metabolite mixtures and metabolite flux. Established sparse sampling methods for protein NMR will be adapted for use in the small-molecule experiments used to study metabolite mixtures, permitting 2-D experiments to be collected at extremely high resolution in short acquisition times, and making it possible to apply 3-D and 4-D spectroscopy in studies of metabolism. Experiments for tracing both carbon and nitrogen will be developed. These experiments will be put to work monitoring the metabolism of a microbial culture for an important synthetic biology model system in real time. The enhanced methods developed in this proposal will improve the ability to identify and quantitate the metabolites in a mixture, to trace isotopic labels, and to carry out real-time pulse-chase labeling experiments, and should be broadly applicable to BER/DoE-relevant systems in future work.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2016-10-01
End Date
2018-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members