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Appl Environ Microbiol, January 1998, p. 74-81, Vol. 64, No. 1
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

¹³C and ¹H Nuclear Magnetic Resonance Study of Glycogen Futile Cycling in Strains of the Genus Fibrobacter

Christelle Matheron,¹ Anne-Marie Delort,^1,* Geneviève Gaudet,^2,3 Evelyne Forano,^3,* and Tibor Liptaj^1,

Laboratoire de Synthèse, Electrosynthèse et Etude de Systèmes à Intérêt Biologique, UMR 6504-Centre National de la Recherche Scientifique,¹ and Centre Universitaire des Sciences et Techniques,² Université Blaise-Pascal, 63177 Abière, and Laboratoire de Microbiologie, Institut National de la Recherche Agronomique, Centre de Recherches de Clermont-Ferrand-Theix, 63122 Saint-Genès-Champanelle,³ France

Received 30 May 1997/Accepted 19 September 1997

We investigated the carbon metabolism of three strains of Fibrobacter succinogenes and one strain of Fibrobacter intestinalis. The four strains produced the same amounts of the metabolites succinate, acetate, and formate in approximately the same ratio (3.7/1/0.3). The four strains similarly stored glycogen during all growth phases, and the glycogen-to-protein ratio was close to 0.6 during the exponential growth phase. ¹³C nuclear magnetic resonance (NMR) analysis of [1-¹³C]glucose utilization by resting cells of the four strains revealed a reversal of glycolysis at the triose phosphate level and the same metabolic pathways. Glycogen futile cycling was demonstrated by ¹³C NMR by following the simultaneous metabolism of labeled [¹³C]glycogen and exogenous unlabeled glucose. The isotopic dilutions of the CH₂ of succinate and the CH₃ of acetate when the resting cells were metabolizing [1-¹³C]glucose and unlabeled glycogen were precisely quantified by using ¹³C-filtered spin-echo difference ¹H NMR spectroscopy. The measured isotopic dilutions were not the same for succinate and acetate; in the case of succinate, the dilutions reflected only the contribution of glycogen futile cycling, while in the case of acetate, another mechanism was also involved. Results obtained in complementary experiments are consistent with reversal of the succinate synthesis pathway. Our results indicated that for all of the strains, from 12 to 16% of the glucose entering the metabolic pathway originated from prestored glycogen. Although genetically diverse, the four Fibrobacter strains studied had very similar carbon metabolism characteristics.

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^* Corresponding author. Mailing address for Anne-Marie Delort: Laboratoire de Synthèse, Electrosynthèse et Etude de Systèmes à Intérêt Biologique, UMR 6504-CNRS, Université Blaise-Pascal, 63177, Aubière, France. Phone: (33) 04 73 40 77 14. Fax: (33) 04 73 40 77 17. E-mail: amdelort{at}chimtp.univ-bpclermont.fr. Mailing address for Evelyne Forano: Laboratoire de Microbiologie, Institut National de la Recherche Agronomique, Centre de Recherches de Clermont-Ferrand-Theix, 63122 Saint-Genès-Champanelle, France. Phone: (33) 04 73 62 42 48. Fax: (33) 04 73 62 45 81. E-mail: forano{at}clermont.inra.fr.

Present address: Central Laboratories, Slovak Technical University, 81237 Bratislava, Slovakia.

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