1.

Metabolic Flux Analysis

Metabolic Flux Analysis (MFA) is a very useful feature in the field of metabolic
engineering27,28. As we have seen in the case of genome scale reconstruction, similarly flux
analysis methods have been developed for various organisms. The flux analysis techniques
have been used to study simple microbial systems such as Escherichia coli29, to complex
eukaryotic organisms such as Saccharomyces cervisiae30, mammalian cells31 as well as
plants32.

Labelling experiments have been widely used to study the biochemical reaction network of
organisms that have the potential to be used as cell factories for chemical production33,34. Use
of stable isotopes of Carbon, nitrogen etc along with mass spectrometry techniques has led to
better understanding of the reaction mechanisms. Liquid chromatography, gas
chromatography and Nuclear magnetic resonance (NMR) have been employed to detect the
isotoplogues of the metabolites. The labelling pattern of the metabolites at a reasonably long-
time frame or a steady state reveals the rate at which the reactions are proceeding. Following
are the various techniques that have been used to obtain the intracellular fluxes from the
extracellular data obtained.