U.S. Department of Energy Office of Biological and Environmental Research

BER Research Highlights


3D NMR Method Enhances Analyses of Metabolic Networks in Cells
Published: February 05, 2016
Posted: June 16, 2016

New technique dramatically reduces data acquisition time and amplifies metabolite identification.

The Science
More rapid and improved estimates of metabolic fluxes in cells are possible thanks to a new technique that combines 13C-metabolic flux analysis (13C-MFA) with non-uniform sampling nuclear magnetic resonance (NMR) spectroscopy data.

The Impact
Key insights into metabolic networks provided by the new technique could be used for synthetic biology-based efforts to modify living systems for production of metabolites or other products of interest, such as biofuels or fine chemicals.

Summary
The use of 13C-MFA can provide key insights into the metabolic networks of microbial cells that are used for producting biofuels or valuable chemicals. This technique can be combined with either NMR spectrometry or mass spectrometry to infer metabolic fluxes within cells based on the characteristic rearrangement of 13C tracers through metabolic pathways. However, position-specific 13C-labeling of metabolites has been particularly difficult to obtain using conventional NMR or mass spectrometry techniques, hindering accurate estimations of metabolic fluxes. To overcome this problem, researchers from the Department of Energy’s Environmental Molecular Sciences Laboratory (EMSL; a national scientific user facility), Washington State University, Duke University Medical Center, and Miami University developed a new technique that combines 13C-MFA with non-uniform sampling (NUS), which dramatically reduces the time required to collect high-resolution NMR data. NUS techniques acquire only a subset of NMR data points and use sophisticated reconstruction methods that ultimately allow extraction of complete sets of chemical shift information. Using EMSL’s 600 MHz and 800 MHz NMR spectrometers, the research team demonstrated that their approach provides detailed information about position-specific labeling patterns that can be incorporated into metabolic flux models. By enabling more accurate estimations of metabolic fluxes in complex biological systems, the new technique could shed light on environmental nutrient cycling and enhance synthetic biology-based engineering efforts to modify living systems for production of metabolites or other products of interest, such as biofuels or fine chemicals.

BER PM Contact
Paul Bayer, SC-23.1, 301-903-5324

PI Contact
Patrick Reardon, EMSL, 509-371-7673, Patrick.Reardon@pnnl.gov

Funding
This work was supported by the U.S. Department of Energy’s Office of Science, Office of Biological and Environmental Research, including support of EMSL, an Office of Science user facility; a William Wiley distinguished postdoctoral fellowship from EMSL; and an EMSL intramural research project entitled “Development of an Integrated EMSL Mass Spectrometry and Nuclear Magnetic Resonance Metabolic Flux Analysis Capability in Support of Systems Biology: Test Application for Biofuels Production.”

Publication
Reardon, P. N., C. L. Marean-Reardon, M. A. Bukovec, B. E. Coggins, and N. G. Isern. 2016. “3D TOCSY-HSQC NMR for Metabolic Flux Analysis Using Non-Uniform Sampling,” Analytical Chemistry 88(5), 2825-31. DOI: 10.1021/acs.analchem.5b04535. (Reference link) .

Related Links
EMSL News

Topic Areas:

  • Research Area: DOE Environmental Molecular Sciences Laboratory (EMSL)
  • Research Area: Genomic Analysis and Systems Biology
  • Research Area: Microbes and Communities
  • Research Area: Sustainable Biofuels and Bioproducts
  • Research Area: Biosystems Design

Division: SC-23.1 Climate and Environmental Sciences Division, BER

 

BER supports basic research and scientific user facilities to advance DOE missions in energy and environment. More about BER

Recent Highlights

Aug 24, 2019
New Approach for Studying How Microbes Influence Their Environment
A diverse group of scientists suggests a common framework and targeting of known microbial processes [more...]

Aug 08, 2019
Nutrient-Hungry Peatland Microbes Reduce Carbon Loss Under Warmer Conditions
Enzyme production in peatlands reduces carbon lost to respiration under future high temperatures. [more...]

Aug 05, 2019
Amazon Forest Response to CO2 Fertilization Dependent on Plant Phosphorus Acquisition
AmazonFACE Model Intercomparison. The Science Plant growth is dependent on the availabi [more...]

Jul 29, 2019
A Slippery Slope: Soil Carbon Destabilization
Carbon gain or loss depends on the balance between competing biological, chemical, and physical reac [more...]

Jul 15, 2019
Field Evaluation of Gas Analyzers for Measuring Ecosystem Fluxes
How gas analyzer type and correction method impact measured fluxes. The Science A side- [more...]

List all highlights (possible long download time)