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Metabolic flux and robustness analysis of glycerol metabolism in Klebsiella pneumoniae

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Abstract

The knowledge of the mechanism of flux distribution will benefit understanding cell physiology and regulation of metabolism. In this study, the measured fluxes obtained under steady-state conditions were used to estimate intracellular fluxes and identify the robustness of branch points of the anaerobic glycerol metabolism in Klebsiella pneumoniae for the production of 1,3-propanediol by metabolic flux analysis. The biomass concentration increased as NADH2/NAD+ decreased at low initial concentration and inversed at high initial glycerol concentration. The flux distribution revealed that the branch points of glycerol and dihydroxyacetonephosphate were rigid to the environmental conditions. However, the pyruvate and acetyl coenzyme A metabolisms gave cells the flexibility to regulate the energy and intermediate fluxes under various environmental conditions. Additionly, it was found that the formation rate of ethanol and the ratio of pyruvate dehydrogenase to pyruvate formate lyase appeared visible fluctuations at high glycerol uptake rate.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 20576018) and the grant from the Major State Basic Research Development Program of China (973 Program) (No.2007CB714306).

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Dalian University of Technology, Linggong Road 2, Dalian, 116023, People’s Republic of China

    Qingrui Zhang, Hu Teng, Yaqin Sun & Zhilong Xiu

  2. Department of Chemical Engineering, Qingdao University of Science and Technology, Zhengzhou Road 53, Qingdao, 266042, People’s Republic of China

    Qingrui Zhang

  3. Institute of Bioprocess and Biosystems Engineering, Technical University Hamburg-Harburg, Denickestr. 15, 21071, Hamburg, Germany

    Anping Zeng

Authors
  1. Qingrui Zhang
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  2. Hu Teng
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  3. Yaqin Sun
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  4. Zhilong Xiu
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Corresponding author

Correspondence to Zhilong Xiu.

Appendices

Appendix 1: List of metabolic reactions

  1. 1.

    Glycerol_ext → Glycerol

  2. 2.

    Glycerol → 3-HPA + H2O

  3. 3.

    3-HPA + NADH2 → 1,3-PD + NAD

  4. 4.

    Glycerol + NAD + ATP → DHAP + NADH2 + ADP

  5. 5.

    DHAP → GA3P

  6. 6.

    GA3P + NAD + PI + ADP → PEP + ATP + NADH2 + H2O

  7. 7.

    PEP + ADP → PYR + ATP

  8. 8.

    PYR + COA + NAD → ACCOA + CO2 + NADH2

  9. 9.

    PYR + COA → ACCOA + Formate

  10. 10.

    Formate → CO2 + H2

  11. 11.

    ACCOA + 2 NADH2 → Ethanol + 2 NAD + COA

  12. 12.

    ACCOA + ADP → AC + ATP + COA

  13. 13.

    2PYR → Acetoin + 2CO2

  14. 14.

    Acetoin + NADH2 → 2,3-BD + NAD

  15. 15.

    PYR + NADH2 → LAC + NAD

  16. 16.

    GA3P + DHAP + ADP → F6P + ATP

  17. 17.

    F6P → G6P

  18. 18.

    G6P + 2 NADP + H2O → R5P + CO2 + 2 NADPH

  19. 19.

    R5P → RIB5P

  20. 20.

    R5P → XYL5P

  21. 21.

    RIB5P + XYL5P → S7P + GA3P

  22. 22.

    S7P + GA3P → E4P + F6P

  23. 23.

    XYL5P + E4P → F6P + GA3P

  24. 24.

    ACCOA + OA → CIT + COA

  25. 25.

    CIT + NAD → AKG + NADH2 + CO2

  26. 26.

    OA + 2 NADH2 → SUCC + 2 NAD

  27. 27.

    PEP + ADP + CO2 → OA + ATP

  28. 28.

    0.35 G6P + 0.523 F6P + 0.952 RIB5P + 0.4075EP + 0.383 PEP + 3.13 PYR + 1.346AKG + 1.6 OA + 1.13 ACCOA + 16.3 ATP + 6.75 NAD + 17.8 NADPH → 1g Biomass + 16.3 ADP + 6.75 NADH2 + 17.8 NADP

  29. 29.

    PYR → PYR_ext

  30. 30.

    Formate → Formate_ext

  31. 31.

    CIT_ext → CIT

  32. 32.

    Acetoin → Acetoin_ext

  33. 33.

    CO2 → CO2_ext

Appendix 2: Summary of abbreviations for the list of metabolic reactions

Table 1

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Zhang, Q., Teng, H., Sun, Y. et al. Metabolic flux and robustness analysis of glycerol metabolism in Klebsiella pneumoniae . Bioprocess Biosyst Eng 31, 127–135 (2008). https://doi.org/10.1007/s00449-007-0155-7

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  • DOI: https://doi.org/10.1007/s00449-007-0155-7

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