Tracing Carbon Dioxide Metabolism Using 13c NMR in the Anaerobic Thermophile, Clostridium thermocellum
Additional Funding Sources
The project described was supported by the EPSCoR Research Infrastructure through the National Science Foundation under Award No. 1738785.
Abstract
Metabolites are small molecular compounds that are found in various stages of the metabolic processes within an organism. Understanding the complexity of these molecules in solution will allow further insight on how a cell sustains life. Molecular compounds such as glucose and sodium pyruvate can be isotopically labeled with C-13 to trace metabolic flux within a living cell using nuclear magnetic resonance (NMR) spectroscopy. 1-D NMR spectra can then be collected to observe specific chemical shifts of known C-13 labeled metabolites in living cells. However, these chemical shifts are subject to change in solutions with different solutes or pH. Solution conditions were tested at different pH values (from 7.2-5.0) and two temperatures (25 °C and 55 °C) to form a greater database of known chemical shifts that, in future work, will be used against unknown metabolites. This NMR spectra can be accurately replicated as each chemical shift is derived from its corresponding metabolite. The aim of this specific research is to compile an NMR database of known C-13 labeled metabolites in varying solution conditions.
Tracing Carbon Dioxide Metabolism Using 13c NMR in the Anaerobic Thermophile, Clostridium thermocellum
Metabolites are small molecular compounds that are found in various stages of the metabolic processes within an organism. Understanding the complexity of these molecules in solution will allow further insight on how a cell sustains life. Molecular compounds such as glucose and sodium pyruvate can be isotopically labeled with C-13 to trace metabolic flux within a living cell using nuclear magnetic resonance (NMR) spectroscopy. 1-D NMR spectra can then be collected to observe specific chemical shifts of known C-13 labeled metabolites in living cells. However, these chemical shifts are subject to change in solutions with different solutes or pH. Solution conditions were tested at different pH values (from 7.2-5.0) and two temperatures (25 °C and 55 °C) to form a greater database of known chemical shifts that, in future work, will be used against unknown metabolites. This NMR spectra can be accurately replicated as each chemical shift is derived from its corresponding metabolite. The aim of this specific research is to compile an NMR database of known C-13 labeled metabolites in varying solution conditions.
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