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PNNL Metabolomics Measurement and Validation Development for Renewable Energy Research


EMSL Project ID
16092

Abstract

We propose the in-situ/live analysis of microbes inside of a controlled bioreactor. Further, we will determine the effects of existing extraction methods by comparing quantitative NMR metabolite profiles measured both before and after sample extraction. Finally, we will employ this in-situ/live analysis method to study the metabolism of Thermotoga lettingae, a hydrogen-producing anaerobic bacterium that could be exploited for biological hydrogen production.

A self-contained chemostat bioreactor will be developed capable of controlling the cell cultivation environment (pH, pO2, nutrient concentrations, etc.) inside of the strong field of a NMR superconducting magnet. NMR, a key metabolomics technology, will be performed to continuously and non-invasively monitor the bioreactor contents, providing time-resolved metabolite concentrations, cell densities, intracellular pH and related information in a known, user-controlled culture environment. (The estimated detectable metabolite concentration threshold is 1 ?M.) The resulting metabolomics capability will be compatible with and complementary to existing high-throughput genomics and proteomics methods at PNNL, thus suitable for comprehensive integrated systems biology studies.

Project Details

Project type
Capability Research
Start Date
2005-10-01
End Date
2007-05-21
Status
Closed

Team

Principal Investigator

Paul Majors
Institution
Washington State University

Team Members

Johannes Scholten
Institution
Pacific Northwest National Laboratory

Jeffrey Mclean
Institution
University of Washington

Related Publications

Majors PD, JS McLean and JCM Scholten (2008). " NMR bioreactor development for live in-situ microbial functional analysis," Journal of Magnetic Resonance 192(1): 159-166, doi:10.1016/j.jmr.2008.02.014.