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Effect of carbon sources differing in oxidation state and transport route on succinate production in metabolically engineered Escherichia coli

  • Henry Lin
  • George N. Bennett
  • Ka-Yiu San
Original Paper

Abstract

In mixed-acid fermentation, succinate synthesis requires one mole of phosphoenolpyruvate (PEP), one mole of CO2, and two moles of NADH for every mole of succinate to be formed. Different carbon sources with different properties were used to address these requirements. Sorbitol generates one more mole of NADH than glucose. Fermentation of sorbitol was shown in this study (and by others) to produce significantly more succinate than fermentation of glucose, due to increased NADH availability. Xylose fermentation conserves the intracellular PEP pool, since its transport does not require the phosphotransferase system normally used for glucose transport. The extra PEP can then be assimilated in the succinate pathway to improve production. In this study, fermentation of xylose did yield higher succinate production than glucose fermentation. Subsequent inactivation of the acetate and lactate pathways was performed to study metabolite redistribution and the effect on succinate production. With the acetate pathway inactivated, significant carbon flux shifted toward lactate rather than succinate. When both acetate and lactate pathways were inactivated, succinate yield ultimately increased with a concomitant increase in ethanol yield.

Keywords

Metabolic engineering Glucose phosphotransferase system NADH/NAD+ ratio Succinate production Pathway deletion 

Notes

Acknowledgements

The authors would like to thank Dr. Jean Vidal for providing the Sorghum pepc plasmid (pKK313). This work was supported by grants from the National Science Foundation (BES-0222691, BES-0000303). H.L. was supported by a training grant from the National Science Foundation (DGE0114264).

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Copyright information

© Society for Industrial Microbiology 2005

Authors and Affiliations

  1. 1.Department of Bioengineering MS 142Rice UniversityHoustonUSA
  2. 2.Department of Biochemistry and Cell Biology MS 140Rice UniversityHoustonUSA
  3. 3.Department of Chemical EngineeringRice UniversityHoustonUSA

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