Elucidation of metabolic pathways in glycogen-accumulating organisms with in vivo 13 C nuclear magnetic resonance

Paulo C. Lemos, Yu Dai, Zhiguo Yuan, Jurg Keller, Helena Santos, Maria A. M. Reis

Research output: Contribution to journalArticle

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Abstract

Glycogen-accumulating organisms (GAOs) are found in enhanced biological phosphorus removal systems where they compete with polyphosphate-accumulating organisms for external carbon substrates. 13 C nuclear magnetic resonance ( 13 C-NMR) was used to elucidate the metabolic pathways operating in an enriched GAO culture dominated by two known GAOs (81.2%). The experiments consisted of adding 13 C-acetate (labelled on position 1 or 2) to the culture under anaerobic conditions, and operating the culture through a cycle consisting of an anaerobic, an aerobic and a further anaerobic phase. The carbon transformations over the cycle were monitored using in vivo 13 C-NMR. The two-carbon moieties in hydroxybutyrate and hydroxyvalerate were derived from acetate, while the propionyl precursor of hydroxyvalerate was primarily derived from glycogen, with only a small fraction originating from acetate. Comparison of the labelling patterns in hydroxyvalerate at the end of the first and the second anaerobic periods in pulse experiments with 2- 13 C-acetate showed that the Entner-Doudoroff (ED) pathway was used for the breakdown of glycogen. This conclusion was further supported by the labelling pattern on glycogen observed in the pulse experiments with 1- 13 C-acetate, which can only be explained by the operation of ED with recycling of pyruvate and glyceraldehyde 3-phosphate via gluconeogenesis. The activity of the ED pathway for glycogen degradation by GAOs is demonstrated here for the first time. In addition, the decarboxylating part of the tricarboxylic acid cycle was confirmed to operate also under anaerobic conditions.

Original languageEnglish
Pages (from-to)2694-2706
Number of pages13
JournalEnvironmental Microbiology
Volume9
Issue number11
DOIs
Publication statusPublished - 1 Nov 2007

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Metabolic Networks and Pathways
Glycogen
glycogen
nuclear magnetic resonance
biochemical pathways
nuclear magnetic resonance spectroscopy
acetate
Magnetic Resonance Spectroscopy
organisms
Acetates
acetates
anoxic conditions
carbon
Carbon
anaerobic conditions
experiment
glyceraldehyde 3-phosphate
Glyceraldehyde 3-Phosphate
Hydroxybutyrates
Polyphosphates

Cite this

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abstract = "Glycogen-accumulating organisms (GAOs) are found in enhanced biological phosphorus removal systems where they compete with polyphosphate-accumulating organisms for external carbon substrates. 13 C nuclear magnetic resonance ( 13 C-NMR) was used to elucidate the metabolic pathways operating in an enriched GAO culture dominated by two known GAOs (81.2{\%}). The experiments consisted of adding 13 C-acetate (labelled on position 1 or 2) to the culture under anaerobic conditions, and operating the culture through a cycle consisting of an anaerobic, an aerobic and a further anaerobic phase. The carbon transformations over the cycle were monitored using in vivo 13 C-NMR. The two-carbon moieties in hydroxybutyrate and hydroxyvalerate were derived from acetate, while the propionyl precursor of hydroxyvalerate was primarily derived from glycogen, with only a small fraction originating from acetate. Comparison of the labelling patterns in hydroxyvalerate at the end of the first and the second anaerobic periods in pulse experiments with 2- 13 C-acetate showed that the Entner-Doudoroff (ED) pathway was used for the breakdown of glycogen. This conclusion was further supported by the labelling pattern on glycogen observed in the pulse experiments with 1- 13 C-acetate, which can only be explained by the operation of ED with recycling of pyruvate and glyceraldehyde 3-phosphate via gluconeogenesis. The activity of the ED pathway for glycogen degradation by GAOs is demonstrated here for the first time. In addition, the decarboxylating part of the tricarboxylic acid cycle was confirmed to operate also under anaerobic conditions.",
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Elucidation of metabolic pathways in glycogen-accumulating organisms with in vivo 13 C nuclear magnetic resonance. / Lemos, Paulo C.; Dai, Yu; Yuan, Zhiguo; Keller, Jurg; Santos, Helena; Reis, Maria A. M.

In: Environmental Microbiology, Vol. 9, No. 11, 01.11.2007, p. 2694-2706.

Research output: Contribution to journalArticle

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