DOI: 10.5281/zenodo.13150702
title: Yeast solutions and hyperpolarization enable real-time observation of metabolized substrates even at natural abundance

description: This data corresponds to the following paper:

 

Title: Yeast solutions and hyperpolarization enable real-time observation of metabolized substrates even at natural abundance

Journal: Analytical Chemistry

Authors: Josh P. Peters, Charbel Assaf, Farhad Haj Mohamad, Eric Beitz, Sanjay Tiwari, Konrad Aden, Jan-Bernd Hövener and Andrey N. Pravdivtsev

 

The data is organized with respect to the subfigures in figure 2 to 6.

A description of acquisition parameters is provided in the "Acquisition parameters.xlsx" file for each dataset.

 

Figure 2:

- Overview of pyruvate metabolism in yeast cells using hyperpolarized and 13C labeled [1-13C]pyruvate. 

Hyperpolarized and thermally polarized spectra, if applicable, are included. 

 

Figure 3:

- Overview of pyruvate metabolism in yeast cells using co-hyperpolarized [1-13C] and [2-13C]pyruvate at an n.a. of 13C. 

Hyperpolarized and thermally polarized spectra, if applicable, are included.

 

Figure 4:

- Overview of fumarate metabolism in yeast cells using hyperpolarized and 13C labeled [1,4-13C2]fumarate

Hyperpolarized and thermally polarized spectra, if applicable, are included.

 

Figure 5:

- The fitted conversion exchange rate constants as a function of the yeast concentration. 

Hyperpolarized and thermally polarized spectra, if applicable, are included.

 

Figure 6:

- Metabolic data from pyruvate metabolism in yeast depending on the position of the yeast in the NMR tube. We 

Hyperpolarized and thermally polarized spectra, if applicable, are included.

 
description: Metabolism is one of the few instant indicators of changes in an organism’s state. Therefore, non-invasive tools to monitor metabo-lism are fundamental as they provide insights into in vivo biochemistry. NMR represents one of the gold standards for such in-sights by observing metabolites. Using nuclear spin hyperpolarization greatly increases the NMR sensitivity, enabling μmol/L sensi-tivity with a time resolution of about one second. However, a metabolic phantom with reproducible, extensive, and human-like metabolism is needed to progress research in this area. Using baker’s yeast as a convenient metabolic factory, we demonstrated in a single study that pyruvate and fumarate including n.a. substrates can be used with yeast as a phantom: we observed the production of ethanol, carbon dioxide, bicarbonate, lactate, alanine from pyruvate, malate, and oxaloacetate from fumarate. For observation, we hyperpolarized pyruvate and fumarate via the dissolution dynamic nuclear polarization (dDNP) technique to about 30% 13C polarization that is equivalent to 360,000 signal enhancement at 1 T and 310 K. Major metabolic pathways were observed using tracers at a natural abundance of 13C, demonstrating that isotope labeling is not always essential in vitro. Enriched [1-13C]pyruvate revealed minor lactate production, presumably via the D-lactate dehydrogenase (DLD) enzyme pathway, demon-strating the sensitivity gain using a dense yeast solution. We foresee that yeast as a metabolic factory can find application as an abundant MRI phantom standard to calibrate and optimize molecular MRI protocols. Our study highlights the potential of using hyperpolarization to probe the metabolism of yeast and other microorganisms even with naturally abundant substrates, offering valuable insights into their response to various stimuli such as drugs, treatment, nourishment, and genetic modification, thereby advancing drug development and our understanding of biochemical processes.
description: We acknowledge funding from German Federal Ministry of Education and Research (BMBF) within the framework of the e: Med research and funding concept (01ZX1915C), DFG (PR 1868/3-1, PR 1868/5-1, HO-4602/2-2, HO-4602/3, GRK2154-2019, EXC2167, FOR5042, TRR287). MOIN CC was founded by a grant from the European Regional Development Fund (ERDF) and the Zukunftsprogramm Wirtschaft of Schleswig-Holstein (Project no. 122-09-053).