13C Metabolic Flux Analysis (MFA)
- Post by: wangikar_8khp2z
- 11:25AM Apr 11, 2018
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The intracellular reaction rate analysis can provide a system level phenotypic endpoint for biological systems. This however cannot be extracted from the static measurements of individual protein or metabolite concentrations.
Figure 1. Representative data to show 13C isotopic enrichment in 3-phosphoglycerate.
Flux analysis requires metabolic model based interpretation of dynamic patterns of 13C-isotopic labelling of intermediate metabolites. Needless to say, the recursive data fitting exercise depends heavily on precise and quantitative data from mass spectrometry. Apart from the strength in modelling, researchers in this lab have developed state of the art experimental techniques to conduct 13C-labelling followed by the acquisition and analysis of mass spectrometric data using an AB Sciex LC-MS/MS instrument available in house. The computationally and experimentally challenging isotopic non-stationary approach has been perfected for 13C MFA to obtain fluxes that are significantly better resolved compared to those obtained in the stationary 13C MFA approach.
Figure 2. Flux map of cyanobacteria
Broader Impact- A single genetic perturbation can lead to sweeping changes in overall cellular metabolism. The researchers of this lab have demonstrated through their studies of a glycogen knockout mutant that there are cascading effects as a result of this mutation leading to an overall slowdown of gluconeogenesis pathway in the phototrophic metabolism of cyanobacteria. The excess carbon was found to be successfully redirected to desired products of heptadecane, pentadecane and 2,3-butanediol. This approach can be used for metabolic engineering based production of desired products. The approach can also be used to assess metabolic effects of candidate molecules during drug discovery process and will be complementary to the conventional toxicity assays.