Metabolic response of Geobacter sulfurreducens towards electron donor/acceptor variation

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Abstract

Background: Geobacter sulfurreducens is capable of coupling the complete oxidation of organic compounds to iron reduction. The metabolic response of G. sulfurreducens towards variations in electron donors (acetate, hydrogen) and acceptors (Fe(III), fumarate) was investigated via 13C-based metabolic flux analysis. We examined the 13C-labeling patterns of proteinogenic amino acids obtained from G. sulfurreducens cultured with 13C-acetate.Results: Using 13C-based metabolic flux analysis, we observed that donor and acceptor variations gave rise to differences in gluconeogenetic initiation, tricarboxylic acid cycle activity, and amino acid biosynthesis pathways. Culturing G. sulfurreducens cells with Fe(III) as the electron acceptor and acetate as the electron donor resulted in pyruvate as the primary carbon source for gluconeogenesis. When fumarate was provided as the electron acceptor and acetate as the electron donor, the flux analysis suggested that fumarate served as both an electron acceptor and, in conjunction with acetate, a carbon source. Growth on fumarate and acetate resulted in the initiation of gluconeogenesis by phosphoenolpyruvate carboxykinase and a slightly elevated flux through the oxidative tricarboxylic acid cycle as compared to growth with Fe(III) as the electron acceptor. In addition, the direction of net flux between acetyl-CoA and pyruvate was reversed during growth on fumarate relative to Fe(III), while growth in the presence of Fe(III) and acetate which provided hydrogen as an electron donor, resulted in decreased flux through the tricarboxylic acid cycle.Conclusions: We gained detailed insight into the metabolism of G. sulfurreducens cells under various electron donor/acceptor conditions using 13C-based metabolic flux analysis. Our results can be used for the development of G. sulfurreducens as a chassis for a variety of applications including bioremediation and renewable biofuel production. © 2010 Yang et al; licensee BioMed Central Ltd.

Figures

  • Table 1 Cultivation conditions for Geobacter sulfurreducens to study intracellular metabolism with different electron donor/acceptor conditions
  • Figure 1 Comparisons of amino acid carbon mass isotopomer distributions between the experiments listed in Table 1. The straight line corresponds to a one-to-one (y = x) line. R: correlation coefficient; D: Euclidian distance.
  • Table 2 Abbreviations and EC numbers of the enzymes considered in the central metabolic network of G. sulfurreducens for 13C metabolic flux analysis
  • Figure 2 In vivo flux estimates (95% confidence intervals given for the flux mean estimates) for the central metabolic network of G. sulfurreducens, corresponding to E1, E2, E3, and E4 (Table 1). All fluxes are normalized to acetate uptake (ACKr, PTAr). Gray double-headed arrows indicate bidirectional reactions, and the black arrowa with plus signs correspond to the net direction of a positive value for the bidirectional reactions. Unidirectional reactions were indicated by gray single-headed arrows. The subscript “ex“ denotes extracellular metabolites. The abbreviations (pathway enzymes) are listed in Table 2, and precursor demand for biomass synthesis (precursors in yellow color) in Table 4.
  • Figure 3 Relative flux distributions (95% confidence interval) at key branch points of the central metabolic network for different experimental conditions of E1, E2, E3, and E4 (Table 1).
  • Table 3 Specific fluxes (fluxes normalized by the biomass yield estimate: (mmolmetablite/h)(gbiomass/h)-1) in the central metabolic pathways of the experiments listed in Table 1.
  • Table 4 Precursor demand of G. sulfurreducens from the intermediary metabolites involved in the central metabolic pathways in Figure 2
  • Figure 4 95% confidence intervals given for key TCA fluxes (νi) in the fumarate cultures (E3, E4), normalized by fumarate uptake. νnet = ν2 - ν3.

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Yang, T. H., Coppi, M. V., Lovley, D. R., & Sun, J. (2010). Metabolic response of Geobacter sulfurreducens towards electron donor/acceptor variation. Microbial Cell Factories, 9. https://doi.org/10.1186/1475-2859-9-90

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