Incorporating metabolic flux ratios into constraint-based flux analysis by using artificial metabolites and converging ratio determinants.
One of the well-established approaches for the quantitative characterization of large-scale underdetermined metabolic network is constraint-based flux analysis, which quantifies intracellular metabolic fluxes to characterize the metabolic status. The system is typically underdetermined, and thus usually is solved by linear programming with the measured external fluxes as constraints. Thus, the intracellular flux distribution calculated may not represent the true values. (13)C-constrained flux analysis allows more accurate determination of internal fluxes, but is currently limited to relatively small metabolic networks due to the requirement of complicated mathematical formulation and limited parameters available. Here, we report a strategy of employing such partial information obtained from the (13)C-labeling experiments as additional constraints during the constraint-based flux analysis. A new methodology employing artificial metabolites and converging ratio determinants (CRDs) was developed for improving constraint-based flux analysis. The CRDs were determined based on the metabolic flux ratios obtained from (13)C-labeling experiments, and were incorporated into the mass balance equations for the artificial metabolites. These new mass balance equations were used as additional constraints during the constraint-based flux analysis with genome-scale E. coli metabolic model, which allowed more accurate determination of intracellular metabolic fluxes.
Choi
Hyung Seok
HS
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, Institute for the BioCentury, Yuseong-Gu, Daejeon, Republic of Korea.
Kim
Tae Yong
TY
Lee
Dong-Yup
DY
Lee
Sang Yup
SY
D001709:Biotechnology / Q000379:methods; D002247:Carbon Isotopes; D004926:Escherichia coli / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D007700:Kinetics; D008660:Metabolism; D008954:Models, Biological; D008962:Models, Theoretical
D002247:Carbon Isotopes