Probing intracellular yeast metabolism with deuterium magnetic resonance spectroscopy

Metabolomics provides insights into biochemical dynamics, with nuclear magnetic resonance (NMR) being one of the few noninvasive techniques. However, traditional 1H NMR often suffers from overlapping signals from proteins and numerous metabolites. To address this, we employed deuterium labeling, reducing background interference and streamlining metabolic analysis. Deuterium magnetic resonance spectroscopy (DMRS) enables rapid, noninvasive measurement of metabolic flux without specialized equipment. In our study, we first measured T1, T2, and chemical shifts for 25 deuterium-labeled compounds in phosphate-buffered saline: parameters functional for optimal DMRS settings. Among the 25 deuterated compounds tested with food-grade baker’s yeast (Saccharomyces cerevisiae) as a model solution, we observed and tracked the real-time consumption of pyruvate, glucose, fumarate, acetone, and nicotinamide. We redirected yeast metabolism by (i) varying concentrations of added pyruvate and (ii) osmotic pressure by changing buffer density. This study underscores DMRS’s potential as a robust, versatile tool for dissecting metabolic transformations exemplified here with the convenient yeast cell systems active for hundreds of minutes under typical NMR observation conditions.