Genome-scale metabolic network modeling results in minimal interventions that cooperatively force carbon flux towards malonyl-CoA
Malonyl-coenzyme A is an important precursor metabolite for the biosynthesis of polyketides, flavonoids and biofuels. However, malonyl-CoA naturally synthesized in microorganisms is consumed for the production of fatty acids and phospholipids leaving only a small amount available for the production...
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Veröffentlicht in: | Metabolic engineering 2011-09, Vol.13 (5), p.578-587 |
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Sprache: | eng |
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Zusammenfassung: | Malonyl-coenzyme A is an important precursor metabolite for the biosynthesis of polyketides, flavonoids and biofuels. However, malonyl-CoA naturally synthesized in microorganisms is consumed for the production of fatty acids and phospholipids leaving only a small amount available for the production of other metabolic targets in recombinant biosynthesis. Here we present an integrated computational and experimental approach aimed at improving the intracellular availability of malonyl-CoA in
Escherichia coli. We used a customized version of the recently developed OptForce methodology to predict a minimal set of genetic interventions that guarantee a prespecified yield of malonyl-CoA in
E. coli strain BL21 Star™. In order to validate the model predictions, we have successfully constructed an
E. coli recombinant strain that exhibits a 4-fold increase in the levels of intracellular malonyl-CoA compared to the wild type strain. Furthermore, we demonstrate the potential of this
E. coli strain for the production of plant-specific secondary metabolites naringenin (474
mg/L) with the highest yield ever achieved in a lab-scale fermentation process. Combined effect of the genetic interventions was found to be synergistic based on a developed analysis method that correlates genetic modification to cell phenotype, specifically the identified knockout targets (Δ
fumC and Δ
sucC) and overexpression targets (ACC, PGK, GAPD and PDH) can cooperatively force carbon flux towards malonyl-CoA. The presented strategy can also be readily expanded for the production of other malonyl-CoA-derived compounds like polyketides and biofuels.
► Metabolic network modeling identified minimal set of genetic interventions leading to improved intracellular malonyl-CoA. ► Engineered strain exhibits 5.6-fold increase in flavanone production. ► Combined effect of the genetic interventions was found to be synergistic based on an analysis correlating genetic modifications to cell phenotype. ► Same strategy can be applied to the production of other malonyl-CoA-derived compounds. |
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ISSN: | 1096-7176 1096-7184 |
DOI: | 10.1016/j.ymben.2011.06.008 |