Bioprocess engg. & technology

msbo309

Experiment 3

Aim of the Experiment

Data Analysis

a) Introduction to Metabolic Flux Analysis (MFA).

Principle

Metabolic flux analysis (MFA) is an experimental fluxomics technique used to examine production and consumption rates of metabolites in a biological system. At an intracellular level, it allows for the quantification of metabolites, thereby elucidating the central metabolism of the cell. Employing stoichiometric models of metabolism and mass spectrometry methods with isotopic mass resolution, the transfer of moieties containing isotopic tracers from one metabolite into another can be elucidated, and information about the metabolic network thus derived. Metabolic flux analysis (MFA) has many applications such as determining the limits on the ability of a biological system in producing a biochemical such as ethanoland predicting the response to gene additions or knockouts.

Metabolic flux analysis may use ¹³C to determine pathway fluxes, NMR techniques and mass spectrometry.

Thermodynamics-Based Metabolic Flux Analysis

Thermodynamics-Based Metabolic Flux Analysis (TMFA) is a specialized type of metabolic flux analysis which utilizes linear thermodynamic constraints in addition to mass balance constraints to generate thermo dynamically feasible fluxes and metabolite activity profiles. TMFA takes into consideration only pathways and fluxes that are feasible by using the Gibbs free energy change of the reactions and activities of the metabolites that are part of the model.