ChBE Seminar Series: Igor Libourel
Tuesday, October 14, 2014
11:00 a.m.-12:15 p.m.
Room 2108, Chemical and Nuclear Engineering Bldg.
Professor Jeffery Klauda
Towards Massive Parallel 13C Metabolic Flux Analysis
Department of Plant Biology
College of Biological Sciences
University of Minnesota
13C metabolic flux analysis (MFA) has become the experimental method of choice to investigate the cellular metabolism of microbes, cell cultures and plant seeds. Conventional steady-state MFA utilizes isotopic labeling measurements of amino acids obtained from protein hydrolysates. By hydrolyzing protein, spatial and temporal information associated with proteins is lost. By utilizing peptides instead of amino acids for MFA, flux maps may be associated with the specific conditions associated with peptides of a given protein used for MFA. With this in mind, we investigated the suitability of peptide mass distributions (PMDs) for flux analysis, and compared PMD-based flux estimates to values obtained through a conventional GC-MS measurement-based approach. The peptide-based flux maps were of comparable precision, and suffered less from the presence of unlabeled biomass in plant seed-derived protein. The developed technology enables large-scale parallel flux analysis that can be applied to flux-imaging of tissues, MFA of microbial communities, and screening of mutant libraries.