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(13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H.
Metabolites. 2015 Sep 02; 5(3):455-74.M

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.

Authors+Show Affiliations

Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. s.ostermann@fz-juelich.de.Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. j.richhardt@fz-juelich.de.Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. st.bringer-meyer@fz-juelich.de.Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. m.bott@fz-juelich.de.Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. w.wiechert@fz-juelich.de.Institute of Bio- and Geosciences-IBG-1: Biotechnology, Leo-Brandt-Straβe, 52428 Jülich, Germany. m.oldiges@fz-juelich.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

26404385

Citation

Ostermann, Steffen, et al. "(13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter Oxydans 621H." Metabolites, vol. 5, no. 3, 2015, pp. 455-74.
Ostermann S, Richhardt J, Bringer S, et al. (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. Metabolites. 2015;5(3):455-74.
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 S, et al. (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter Oxydans 621H. Metabolites. 2015 Sep 2;5(3):455-74. PubMed PMID: 26404385.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - (13)C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. AU - Ostermann,Steffen, AU - Richhardt,Janine, AU - Bringer,Stephanie, AU - Bott,Michael, AU - Wiechert,Wolfgang, AU - Oldiges,Marco, Y1 - 2015/09/02/ PY - 2015/07/10/received PY - 2015/08/20/revised PY - 2015/08/24/accepted PY - 2015/9/26/entrez PY - 2015/9/26/pubmed PY - 2015/9/26/medline KW - 13C tracer KW - Entner–Doudoroff pathway KW - glucose KW - microtiter cultivation KW - pentose phosphate pathway SP - 455 EP - 74 JF - Metabolites VL - 5 IS - 3 N2 - 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. SN - 2218-1989 UR - https://www.unboundmedicine.com/medline/citation/26404385/_13_C_Tracers_for_Glucose_Degrading_Pathway_Discrimination_in_Gluconobacter_oxydans_621H_ L2 - https://www.mdpi.com/resolver?pii=metabo5030455 DB - PRIME DP - Unbound Medicine ER -
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