Metabolic flux analysis of Shewanella spp. reveals evolutionary robustness in central carbon metabolism

Shewanella spp. are a group of facultative anaerobic bacteria widely distributed in marine and freshwater environments. In this study, we profiled the central metabolic fluxes of eight recently sequenced Shewanella species grown under the same condition in minimal medium with [3‐13C] lactate. Although the tested Shewanella species had slightly different growth rates (0.23–0.29 h−1) and produced different amounts of acetate and pyruvate during early exponential growth (pseudo‐steady state), the relative intracellular metabolic flux distributions were remarkably similar. This result indicates that Shewanella species share similar regulation in regard to central carbon metabolic fluxes under steady growth conditions: the maintenance of metabolic robustness is not only evident in a single species under genetic perturbations (Fischer and Sauer, 2005; Nat Genet 37(6):636–640), but also observed through evolutionary related microbial species. This remarkable conservation of relative flux profiles through phylogenetic differences prompts us to introduce the concept of metabotype as an alternative scheme to classify microbial fluxomics. On the other hand, Shewanella spp. display flexibility in the relative flux profiles when switching their metabolism from consuming lactate to consuming pyruvate and acetate. Biotechnol. Bioeng. 2009;102: 1161–1169. © 2008 Wiley Periodicals, Inc.

[1]  Yinjie J. Tang,et al.  Pathway Confirmation and Flux Analysis of Central Metabolic Pathways in Desulfovibrio vulgaris Hildenborough using Gas Chromatography-Mass Spectrometry and Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry , 2006, Journal of bacteriology.

[2]  David E. Goldberg,et al.  Genetic Algorithms in Search Optimization and Machine Learning , 1988 .

[3]  R. K. Ursem Multi-objective Optimization using Evolutionary Algorithms , 2009 .

[4]  U. Sauer,et al.  Metabolic Flux Ratio Analysis of Genetic and Environmental Modulations of Escherichia coli Central Carbon Metabolism , 1999, Journal of bacteriology.

[5]  Kalyanmoy Deb,et al.  Simulated Binary Crossover for Continuous Search Space , 1995, Complex Syst..

[6]  Yinjie J. Tang,et al.  Shewanella oneidensis MR-1 Fluxome under Various Oxygen Conditions , 2006, Applied and Environmental Microbiology.

[7]  Sunwon Park,et al.  MetaFluxNet, a program package for metabolic pathway construction and analysis, and its use in large-scale metabolic flux analysis of Escherichia coli. , 2003, Genome informatics. International Conference on Genome Informatics.

[8]  D C White,et al.  Polyphasic taxonomy of the genus Shewanella and description of Shewanella oneidensis sp. nov. , 1999, International journal of systematic bacteriology.

[9]  Kazuyuki Shimizu,et al.  An improved method for statistical analysis of metabolic flux analysis using isotopomer mapping matrices with analytical expressions. , 2003, Journal of biotechnology.

[10]  K. Shimizu,et al.  Metabolic flux analysis of Escherichia coli K12 grown on 13C-labeled acetate and glucose using GC-MS and powerful flux calculation method. , 2003, Journal of biotechnology.

[11]  Wolfgang Wiechert,et al.  New tools for mass isotopomer data evaluation in 13C flux analysis: Mass isotope correction, data consistency checking, and precursor relationships , 2004, Biotechnology and bioengineering.

[12]  Yinjie J. Tang,et al.  A kinetic model describing Shewanella oneidensis MR‐1 growth, substrate consumption, and product secretion , 2007, Biotechnology and bioengineering.

[13]  Grigoriy E. Pinchuk,et al.  Towards environmental systems biology of Shewanella , 2008, Nature Reviews Microbiology.

[14]  William H. Press,et al.  Numerical Recipes in Fortran 77 , 1992 .

[15]  W. Wiechert,et al.  Bidirectional reaction steps in metabolic networks: III. Explicit solution and analysis of isotopomer labeling systems. , 1999, Biotechnology and bioengineering.

[16]  U. Sauer,et al.  Large-scale in vivo flux analysis shows rigidity and suboptimal performance of Bacillus subtilis metabolism , 2005, Nature Genetics.

[17]  Gregory Stephanopoulos,et al.  Accurate assessment of amino acid mass isotopomer distributions for metabolic flux analysis. , 2007, Analytical chemistry.

[18]  Kalyanmoy Deb,et al.  Real-coded Genetic Algorithms with Simulated Binary Crossover: Studies on Multimodal and Multiobjective Problems , 1995, Complex Syst..

[19]  Yinjie J. Tang,et al.  Evaluation of the effects of various culture conditions on Cr(VI) reduction by Shewanella oneidensis MR‐1 in a novel high‐throughput mini‐bioreactor , 2006, Biotechnology and bioengineering.

[20]  U. Sauer,et al.  A Novel Metabolic Cycle Catalyzes Glucose Oxidation and Anaplerosis in Hungry Escherichia coli* , 2003, Journal of Biological Chemistry.

[21]  U. Sauer,et al.  Impact of Global Transcriptional Regulation by ArcA, ArcB, Cra, Crp, Cya, Fnr, and Mlc on Glucose Catabolism in Escherichia coli , 2005, Journal of bacteriology.

[22]  Katherine H. Huang,et al.  The MicrobesOnline Web site for comparative genomics. , 2005, Genome research.

[23]  U. Sauer,et al.  Systematic evaluation of objective functions for predicting intracellular fluxes in Escherichia coli , 2007, Molecular systems biology.

[24]  D. Goldberg,et al.  Modeling tournament selection with replacement using apparent added noise , 2001 .

[25]  James M. Tiedje,et al.  Shewanella—the environmentally versatile genome , 2002, Nature Biotechnology.

[26]  U. Sauer,et al.  Article number: 62 REVIEW Metabolic networks in motion: 13 C-based flux analysis , 2022 .

[27]  U. Sauer,et al.  Large-scale 13C-flux analysis reveals mechanistic principles of metabolic network robustness to null mutations in yeast , 2005, Genome Biology.