Advances in analysis of microbial metabolic fluxes via (13)C isotopic labeling.
Metabolic flux analysis via (13)C labeling ((13)C MFA) quantitatively tracks metabolic pathway activity and determines overall enzymatic function in cells. Three core techniques are necessary for (13)C MFA: (1) a steady state cell culture in a defined medium with labeled-carbon substrates; (2) precise measurements of the labeling pattern of targeted metabolites; and (3) evaluation of the data sets obtained from mass spectrometry measurements with a computer model to calculate the metabolic fluxes. In this review, we summarize recent advances in the (13)C-flux analysis technologies, including mini-bioreactor usage for tracer experiments, isotopomer analysis of metabolites via high resolution mass spectrometry (such as GC-MS, LC-MS, or FT-ICR), high performance and large-scale isotopomer modeling programs for flux analysis, and the integration of fluxomics with other functional genomics studies. It will be shown that there is a significant value for (13)C-based metabolic flux analysis in many biological research fields.
Tang
Yinjie J
YJ
Joint Bio-Energy Institute, Emeryville, CA 94608, USA.
Martin
Hector Garcia
HG
Myers
Samuel
S
Rodriguez
Sarah
S
Baidoo
Edward E K
EE
Keasling
Jay D
JD
D018407:Bacterial Physiological Phenomena; D001426:Bacterial Proteins / Q000378:metabolism*; D019149:Bioreactors / Q000382:microbiology*; D002244:Carbon / Q000378:metabolism*; D002247:Carbon Isotopes / Q000032:analysis / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods / Q000639:trends; D013058:Mass Spectrometry / Q000379:methods* / Q000639:trends*; D009097:Multienzyme Complexes / Q000378:metabolism*
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D009097:Multienzyme Complexes; D002244:Carbon