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Metabolites 2015, 5(3), 455-474; doi:10.3390/metabo5030455

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

Institute of Bio- and Geosciences—IBG-1: Biotechnology, Leo-Brandt-Straße, 52428 Jülich, Germany
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Academic Editor: Kazuyuki Shimizu
Received: 10 July 2015 / Revised: 20 August 2015 / Accepted: 24 August 2015 / Published: 2 September 2015
(This article belongs to the Special Issue Metabolic Flux Analysis)
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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 13C 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 13C fragmentation patterns of alanine. For the first time, down-scaled microtiter plate cultivation together with 13C-labeled substrate was applied for G. oxydans to elucidate pathway operation, exhibiting reasonable labeling costs and allowing for sufficient replicate experiments. View Full-Text
Keywords: microtiter cultivation; 13C tracer; pentose phosphate pathway; Entner–Doudoroff pathway; glucose microtiter cultivation; 13C tracer; pentose phosphate pathway; Entner–Doudoroff pathway; glucose
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Ostermann, S.; Richhardt, J.; Bringer, S.; Bott, M.; Wiechert, W.; Oldiges, M. 13C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H. Metabolites 2015, 5, 455-474.

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