GLUCOSE IS TAKEN UP BY GALACTOSE PERMEASE IN METABOLIC ENGINEERED ESCHERICHIA COLI TO PRODUCE SUCCINATE
Keywords:
Succinate, E. coli, PTS systems, GalP, metabolic flux analysisAbstract
E. coli strain KJ073 was previously developed to produce succinate by a combination of metabolicengineering and metabolic evolution. The strain produces succinate as a major product underanaerobic fermentation with high titer and yield. Metabolic Flux Analysis (MFA) results revealedthat about 70% of PEP (phosphoenolpyruvate) flux from glucose generated during glycolysis flowedto a succinate production route in strain KJ073. This event is not normally found in E. coli wild typesince almost the entire PEP pool flows through PYK (pyruvate kinase) to generate ATP for growthand maintenance rather than flowing through PPC (pyruvate carboxylase) since this latter pathwaydoes not produce ATP. However, strain KJ073 has been reported to show an increase of PCK (PEPcarboxykinase) activity but a decrease in PPC activity. It is worth noting that the production ofOAA (oxaloacetate) by PEP carboxylation via PCK produces an extra mole of ATP. In addition, theresults from gene deletion analysis showed that galP- mutants of KJ073 exhibited severely lowglucose consumption rates, while the deletions of genes involving in the classical PEP(phosphoenolpyruvate)-dependent PTS transporters (ptsG and manX) did not affect the rates ofglucose consumption in the strains. This result could confirm that the activation of GalP (galactosepermease) for glucose transporting in strain KJ073 would enhance the succinate production bypreserving PEP pool size in which PEP is not used for phosphorylating glucose. The strain gainedthe benefit in which the elevated PEP level was used for OAA production simultaneously withincreasing ATP production through PCK.
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