Tuesday, July 31, 2007 - 10:15 AM
S62

A universal framework for high-throughput 13C flux analysis

Nicola Zamboni and Uwe Sauer. Institute of Molecular Systems Biology, ETH Zurich, Wolfgang Pauli Strasse 16, Zurich, 8093, Switzerland

Measurement of intracellular fluxes by means of stable isotopic tracers has maturated over the last decade to a quantitative and valuable workhorse for metabolic engineering. Developments were driven in two diverging directions by complementary philosophies: (i) the comprehensive and possibly detailed quantification of all reaction rates in a cell, and (ii) the targeted determination of local flux partitioning ratios by local analysis of specific labeling patterns. Nowadays, the latter approach is the method of choice in large scale flux studies because it requires less input information, it relies on a direct interpretation of labeling patterns, and, therefore, is more robust against measurement errors or ill-defined models.

Here we focus on the tools we have established for the purpose of high-throughput 13C flux analysis. These methods are applicable to a variety of microorganisms, are compatible with small scale cultivations, amenable to rapid and unsupervised computational analysis, and deliver quantitative information on carbon, energy, or redox metabolism. Unfortunately, their application is largely limited to cells grown in minimal media with single carbon sources, typically glucose.

The conceptually novel and generalized approach of synthetic flux ratios is presented, which overcomes this fundamental bottleneck and enables to identify and quantify calculable flux ratios at high-throughputs in any combination of isotopic tracer, biochemical reaction network, medium composition, and analytics.


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