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Ostermann S, Richhardt J, Bringer S, et al. (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. Metabolites. 2015;5(3):455-74doi: 10.3390/metabo5030455.
Ostermann, S., Richhardt, J., Bringer, S., Bott, M., Wiechert, W., & Oldiges, M. (2015). (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. Metabolites, 5(3), 455-74. https://doi.org/10.3390/metabo5030455
Ostermann, Steffen, et al. "(13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H." Metabolites vol. 5,3 (2015): 455-74. doi: https://doi.org/10.3390/metabo5030455
Ostermann S, Richhardt J, Bringer S, Bott M, Wiechert W, Oldiges M. (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. Metabolites. 2015 Sep 02;5(3):455-74. doi: 10.3390/metabo5030455. PMID: 26404385; PMCID: PMC4588806.
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Metabolites. 2015 Sep 02;5(3):455-74. doi: 10.3390/metabo5030455.

(13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H.

Metabolites

Steffen Ostermann, Janine Richhardt, Stephanie Bringer, Michael Bott, Wolfgang Wiechert, Marco Oldiges

Affiliations

  1. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].
  2. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].
  3. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].
  4. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].
  5. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].
  6. Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany. [email protected].

PMID: 26404385 PMCID: PMC4588806 DOI: 10.3390/metabo5030455

Abstract

Gluconobacter oxydans 621H is used as an industrial production organism due to its exceptional ability to incompletely oxidize a great variety of carbohydrates in the periplasm. With glucose as the carbon source, up to 90% of the initial concentration is oxidized periplasmatically to gluconate and ketogluconates. Growth on glucose is biphasic and intracellular sugar catabolism proceeds via the Entner-Doudoroff pathway (EDP) and the pentose phosphate pathway (PPP). Here we studied the in vivo contributions of the two pathways to glucose catabolism on a microtiter scale. In our approach we applied specifically (13)C labeled glucose, whereby a labeling pattern in alanine was generated intracellularly. This method revealed a dynamic growth phase-dependent pathway activity with increased activity of EDP in the first and PPP in the second growth phase, respectively. Evidence for a growth phase-independent decarboxylation-carboxylation cycle around the pyruvate node was obtained from (13)C fragmentation patterns of alanine. For the first time, down-scaled microtiter plate cultivation together with (13)C-labeled substrate was applied for G. oxydans to elucidate pathway operation, exhibiting reasonable labeling costs and allowing for sufficient replicate experiments.

Keywords: 13C tracer; Entner–Doudoroff pathway; glucose; microtiter cultivation; pentose phosphate pathway

References

  1. J Bacteriol. 1995 May;177(10):2637-43 - PubMed
  2. Microbiol Mol Biol Rev. 2005 Mar;69(1):12-50 - PubMed
  3. Arch Biochem Biophys. 1968 Oct;128(1):184-93 - PubMed
  4. Nucleic Acids Res. 2000 Jan 1;28(1):27-30 - PubMed
  5. J Mol Microbiol Biotechnol. 2007;13(1-3):147-55 - PubMed
  6. Appl Microbiol Biotechnol. 2010 Jan;85(4):1025-31 - PubMed
  7. Adv Biochem Eng Biotechnol. 2005;92:145-72 - PubMed
  8. Appl Environ Microbiol. 2013 Apr;79(7):2336-48 - PubMed
  9. Appl Environ Microbiol. 2012 Oct;78(19):6975-86 - PubMed
  10. J Bacteriol. 1953 Apr;65(4):405-12 - PubMed
  11. J Gen Microbiol. 1976 Feb;92(2):304-10 - PubMed
  12. Crit Rev Biotechnol. 2008;28(2):101-24 - PubMed
  13. Biosci Biotechnol Biochem. 2010;74(5):1084-8 - PubMed
  14. Appl Environ Microbiol. 2010 Jul;76(13):4369-76 - PubMed
  15. J Biotechnol. 2002 Mar 14;94(1):37-63 - PubMed
  16. Microb Cell Fact. 2012 Oct 31;11:144 - PubMed
  17. J Biotechnol. 2007 Jan 30;128(1):93-111 - PubMed
  18. J Mol Microbiol Biotechnol. 2001 Jul;3(3):445-56 - PubMed
  19. Appl Microbiol Biotechnol. 2010 Oct;88(3):711-8 - PubMed
  20. Appl Microbiol Biotechnol. 2013 Mar;97(6):2521-30 - PubMed
  21. Protein Eng Des Sel. 2005 Jul;18(7):345-57 - PubMed
  22. Antonie Van Leeuwenhoek. 1981;47(4):289-96 - PubMed
  23. Metab Eng. 2002 Apr;4(2):159-69 - PubMed
  24. Curr Opin Plant Biol. 2003 Jun;6(3):236-46 - PubMed
  25. Nat Biotechnol. 2005 Feb;23(2):195-200 - PubMed
  26. J Bacteriol. 2010 Oct;192(20):5304-11 - PubMed
  27. J Gen Microbiol. 1973 Oct;78(2):319-29 - PubMed
  28. J Bacteriol. 1945 May;49(5):495-501 - PubMed
  29. Bioprocess Biosyst Eng. 2008 Apr;31(3):227-39 - PubMed
  30. Appl Microbiol Biotechnol. 2013 May;97(10):4315-23 - PubMed
  31. Anal Bioanal Chem. 2008 Aug;391(7):2663-72 - PubMed

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