Metabolic fluxes and beyond—systems biology understanding and engineering of microbial metabolism

The recent years have seen tremendous progress towards the understanding of microbial metabolism on a higher level of the entire functional system. Hereby, huge achievements including the sequencing of complete genomes and efficient post-genomic approaches provide the basis for a new, fascinating era of research—analysis of metabolic and regulatory properties on a global scale. Metabolic flux (fluxome) analysis displays the first systems oriented approach to unravel the physiology of microorganisms since it combines experimental data with metabolic network models and allows determining absolute fluxes through larger networks of central carbon metabolism. Hereby, fluxes are of central importance for systems level understanding because they fundamentally represent the cellular phenotype as integrated output of the cellular components, i.e. genes, transcripts, proteins, and metabolites. A currently emerging and promising area of research in systems biology and systems metabolic engineering is therefore the integration of fluxome data in multi-omics studies to unravel the multiple layers of control that superimpose the flux network and enable its optimal operation under different environmental conditions.

[1]  V. Wendisch,et al.  Lysine and glutamate production by Corynebacterium glutamicum on glucose, fructose and sucrose: roles of malic enzyme and fructose-1,6-bisphosphatase. , 2005, Metabolic engineering.

[2]  W. Wiechert 13C metabolic flux analysis. , 2001, Metabolic engineering.

[3]  U. Sauer,et al.  GC‐MS Analysis of Amino Acids Rapidly Provides Rich Information for Isotopomer Balancing , 2000, Biotechnology progress.

[4]  Christoph Wittmann,et al.  Amplified Expression of Fructose 1,6-Bisphosphatase in Corynebacterium glutamicum Increases In Vivo Flux through the Pentose Phosphate Pathway and Lysine Production on Different Carbon Sources , 2005, Applied and Environmental Microbiology.

[5]  G. Stephanopoulos,et al.  Systematic quantification of complex metabolic flux networks using stable isotopes and mass spectrometry. , 2003, European journal of biochemistry.

[6]  C. Wittmann,et al.  Metabolic Flux Analysis in Corynebacterium glutamicum , 2005 .

[7]  Christoph Wittmann,et al.  Theoretical aspects of 13C metabolic flux analysis with sole quantification of carbon dioxide labeling , 2005, Comput. Biol. Chem..

[8]  Christoph Wittmann,et al.  Systems level engineering of Corynebacterium glutamicum – Reprogramming translational efficiency for superior production , 2010 .

[9]  C. Wittmann,et al.  Respirometric 13C flux analysis, Part I: design, construction and validation of a novel multiple reactor system using on-line membrane inlet mass spectrometry. , 2006, Metabolic engineering.

[10]  E. Tsalikian,et al.  Increased leucine flux in short-term fasted human subjects: evidence for increased proteolysis. , 1984, The American journal of physiology.

[11]  A. D. de Graaf,et al.  Quantitative Determination of Metabolic Fluxes during Coutilization of Two Carbon Sources: Comparative Analyses withCorynebacterium glutamicum during Growth on Acetate and/or Glucose , 2000, Journal of bacteriology.

[12]  Uwe Sauer,et al.  Metabolic-flux and network analysis in fourteen hemiascomycetous yeasts. , 2005, FEMS yeast research.

[13]  H Sahm,et al.  Response of the central metabolism in Corynebacterium glutamicum to the use of an NADH-dependent glutamate dehydrogenase. , 1999, Metabolic engineering.

[14]  U. Sauer,et al.  Getting Closer to the Whole Picture , 2007, Science.

[15]  Suteaki Shioya,et al.  Study on roles of anaplerotic pathways in glutamate overproduction of Corynebacterium glutamicum by metabolic flux analysis , 2007, Microbial cell factories.

[16]  J. Nielsen,et al.  Metabolic network analysis. A powerful tool in metabolic engineering. , 2000, Advances in biochemical engineering/biotechnology.

[17]  Lars M. Blank,et al.  Metabolic flux distributions: genetic information, computational predictions, and experimental validation , 2010, Applied Microbiology and Biotechnology.

[18]  Pei Yee Ho,et al.  Multiple High-Throughput Analyses Monitor the Response of E. coli to Perturbations , 2007, Science.

[19]  Christoph Wittmann,et al.  Metabolic Fluxes in Corynebacterium glutamicum during Lysine Production with Sucrose as Carbon Source , 2004, Applied and Environmental Microbiology.

[20]  C. Des Rosiers,et al.  Myocardial phenotyping using isotopomer analysis of metabolic fluxes. , 2005, Biochemical Society transactions.

[21]  C. Wittmann,et al.  Metabolic flux analysis using mass spectrometry. , 2002, Advances in biochemical engineering/biotechnology.

[22]  H. Sahm,et al.  Pyruvate carboxylase is a major bottleneck for glutamate and lysine production by Corynebacterium glutamicum. , 2001, Journal of molecular microbiology and biotechnology.

[23]  Wolfgang Wiechert,et al.  Computational tools for isotopically instationary 13C labeling experiments under metabolic steady state conditions. , 2006, Metabolic engineering.

[24]  C. Wittmann,et al.  Physiological response of Corynebacterium glutamicum to oxidative stress induced by deletion of the transcriptional repressor McbR. , 2008, Microbiology.

[25]  T. Szyperski Biosynthetically directed fractional 13C-labeling of proteinogenic amino acids. An efficient analytical tool to investigate intermediary metabolism. , 1995, European journal of biochemistry.

[26]  T. Terwilliger,et al.  Rapid gas chromatographic-mass spectrometric analysis of [15N]urea: application to human metabolic studies. , 1981, Clinica chimica acta; international journal of clinical chemistry.

[27]  Jens Nielsen,et al.  Antibiotic Overproduction in Streptomyces coelicolor A3(2) Mediated by Phosphofructokinase Deletion* , 2008, Journal of Biological Chemistry.

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

[29]  Christoph Wittmann,et al.  Fluxome analysis using GC-MS , 2007, Microbial cell factories.

[30]  Christoph Wittmann,et al.  Response of fluxome and metabolome to temperature-induced recombinant protein synthesis in Escherichia coli. , 2007, Journal of biotechnology.

[31]  E. Heinzle,et al.  Mass spectrometry for metabolic flux analysis. , 1999, Biotechnology and bioengineering.

[32]  K. Hyung-Min,et al.  Deregulation of aspartokinase by single nucleotide exchange leads to global flux rearrangement in the central metabolism of Corynebacterium glutamicum , 2006 .

[33]  H Sahm,et al.  Characterization of the phosphoenolpyruvate carboxykinase gene from Corynebacterium glutamicum and significance of the enzyme for growth and amino acid production. , 2001, Journal of molecular microbiology and biotechnology.

[34]  Chikara Furusawa,et al.  Effect of odhA overexpression and odhA antisense RNA expression on Tween-40-triggered glutamate production by Corynebacterium glutamicum , 2009, Applied Microbiology and Biotechnology.

[35]  Masato Ikeda,et al.  A novel gnd mutation leading to increased L-lysine production in Corynebacterium glutamicum. , 2005, FEMS microbiology letters.

[36]  L. Quek,et al.  OpenFLUX: efficient modelling software for 13C-based metabolic flux analysis , 2009, Microbial cell factories.

[37]  Uwe Sauer,et al.  Different Biochemical Mechanisms Ensure Network-Wide Balancing of Reducing Equivalents in Microbial Metabolism , 2009, Journal of bacteriology.

[38]  Bernhard O Palsson,et al.  Latent Pathway Activation and Increased Pathway Capacity Enable Escherichia coli Adaptation to Loss of Key Metabolic Enzymes* , 2006, Journal of Biological Chemistry.

[39]  Hyun Uk Kim,et al.  Metabolic engineering of microorganisms: general strategies and drug production. , 2009, Drug discovery today.

[40]  Jens Nielsen,et al.  Metabolic Network Analysis , 1999 .

[41]  J. Villadsen,et al.  Modeling isotopomer distributions in biochemical networks using isotopomer mapping matrices. , 1997, Biotechnology and bioengineering.

[42]  H. Mori,et al.  Systematic phenome analysis of Escherichia coli multiple-knockout mutants reveals hidden reactions in central carbon metabolism , 2009, Molecular systems biology.

[43]  Uwe Sauer,et al.  TCA cycle activity in Saccharomyces cerevisiae is a function of the environmentally determined specific growth and glucose uptake rates. , 2004, Microbiology.

[44]  Nicola Zamboni,et al.  Novel biological insights through metabolomics and 13C-flux analysis. , 2009, Current opinion in microbiology.

[45]  A. D. de Graaf,et al.  Response of the central metabolism of Corynebacterium glutamicum to different flux burdens. , 1997, Biotechnology and bioengineering.

[46]  Stephan Hans,et al.  Metabolic phenotype of phosphoglucose isomerase mutants of Corynebacterium glutamicum. , 2003, Journal of biotechnology.

[47]  C. Wittmann,et al.  Investigation of the central carbon metabolism of Sorangium cellulosum: metabolic network reconstruction and quantification of pathway fluxes. , 2009, Journal of microbiology and biotechnology.

[48]  Jean-Charles Portais,et al.  Demonstration of the ethylmalonyl-CoA pathway by using 13C metabolomics , 2009, Proceedings of the National Academy of Sciences.

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

[50]  S. Lee,et al.  Application of systems biology for bioprocess development. , 2008, Trends in biotechnology.

[51]  Kazuyuki Shimizu,et al.  Metabolic flux analysis based on 13C-labeling experiments and integration of the information with gene and protein expression patterns. , 2004, Advances in biochemical engineering/biotechnology.

[52]  Christoph Wittmann,et al.  Metabolic flux engineering of L-lysine production in Corynebacterium glutamicum--over expression and modification of G6P dehydrogenase. , 2007, Journal of biotechnology.

[53]  Christoph Wittmann,et al.  Comparative Metabolic Flux Analysis of Lysine-Producing Corynebacterium glutamicum Cultured on Glucose or Fructose , 2004, Applied and Environmental Microbiology.

[54]  Christoph Wittmann,et al.  Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum , 2008, Microbial cell factories.

[55]  U. Sauer,et al.  Systems biology of microbial metabolism. , 2010, Current opinion in microbiology.

[56]  U. Sauer,et al.  Experimental Identification and Quantification of Glucose Metabolism in Seven Bacterial Species , 2005, Journal of bacteriology.

[57]  Yohei Yamada,et al.  Metabolic flux analysis in biotechnology processes , 2008, Biotechnology Letters.

[58]  C. Wittmann,et al.  Sampling for metabolome analysis of microorganisms. , 2007, Analytical chemistry.

[59]  U. Sauer,et al.  Metabolic flux profiling of Escherichia coli mutants in central carbon metabolism using GC-MS. , 2003, European journal of biochemistry.

[60]  Karl Hult,et al.  The distribution of the NADPH regenerating mannitol cycle among fungal species , 1980, Archives of Microbiology.

[61]  Uwe Sauer,et al.  The PEP-pyruvate-oxaloacetate node as the switch point for carbon flux distribution in bacteria. , 2005, FEMS microbiology reviews.

[62]  J K Kelleher,et al.  Flux estimation using isotopic tracers: common ground for metabolic physiology and metabolic engineering. , 2001, Metabolic engineering.

[63]  Christoph Wittmann,et al.  Analysis and engineering of metabolic pathway fluxes in Corynebacterium glutamicum. , 2010, Advances in biochemical engineering/biotechnology.

[64]  L. Eggeling,et al.  Handbook of Corynebacterium glutamicum , 2005 .

[65]  Takashi Gojobori,et al.  Comparative study of flux redistribution of metabolic pathway in glutamate production by two coryneform bacteria. , 2005, Metabolic engineering.

[66]  Annik Nanchen,et al.  Nonlinear Dependency of Intracellular Fluxes on Growth Rate in Miniaturized Continuous Cultures of Escherichia coli , 2006, Applied and Environmental Microbiology.

[67]  Andrew R. Joyce,et al.  Metabolic Characterization of Escherichia coli Strains Adapted to Growth on Lactate , 2007, Applied and Environmental Microbiology.

[68]  P. Verheijen,et al.  Cumulative bondomers: a new concept in flux analysis from 2D [13C,1H] COSY NMR data. , 2002, Biotechnology and bioengineering.

[69]  C. Wittmann,et al.  Application of MALDI-TOF MS to lysine-producing Corynebacterium glutamicum: a novel approach for metabolic flux analysis. , 2001, European journal of biochemistry.

[70]  U. Sauer,et al.  High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints. , 2004, Analytical biochemistry.

[71]  S. Lloyd,et al.  A critical perspective of the use of (13)C-isotopomer analysis by GCMS and NMR as applied to cardiac metabolism. , 2004, Metabolic engineering.

[72]  W Wiechert,et al.  A universal framework for 13C metabolic flux analysis. , 2001, Metabolic engineering.

[73]  H. Shimizu,et al.  Precise metabolic flux analysis of coryneform bacteria by gas chromatography-mass spectrometry and verification by nuclear magnetic resonance. , 2006, Journal of bioscience and bioengineering.

[74]  A. D. de Graaf,et al.  Flux partitioning in the split pathway of lysine synthesis in Corynebacterium glutamicum. Quantification by 13C- and 1H-NMR spectroscopy. , 1993, European journal of biochemistry.

[75]  Christoph Wittmann,et al.  Metabolic Engineering of the Tricarboxylic Acid Cycle for Improved Lysine Production by Corynebacterium glutamicum , 2009, Applied and Environmental Microbiology.

[76]  Christoph Wittmann,et al.  Genealogy Profiling through Strain Improvement by Using Metabolic Network Analysis: Metabolic Flux Genealogy of Several Generations of Lysine-Producing Corynebacteria , 2002, Applied and Environmental Microbiology.

[77]  Weiwen Zhang,et al.  Integrating multiple 'omics' analysis for microbial biology: application and methodologies. , 2010, Microbiology.

[78]  B. Christensen,et al.  Isotopomer analysis using GC-MS. , 1999, Metabolic engineering.

[79]  E. Kimura,et al.  Altered Metabolic Flux due to Deletion of odhA causes l-Glutamate Overproduction in Corynebacterium glutamicum , 2006, Applied and Environmental Microbiology.

[80]  C. Wittmann,et al.  Bmc Microbiology , 2004 .

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

[82]  I. Nissim,et al.  Regulation of pool sizes and turnover rates of amino acids in humans: 15N-glycine and 15N-alanine single-dose experiments using gas chromatography-mass spectrometry analysis. , 1980, Metabolism: clinical and experimental.

[83]  Nicola Zamboni,et al.  FiatFlux – a software for metabolic flux analysis from 13C-glucose experiments , 2005, BMC Bioinformatics.

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

[85]  J. Nielsen,et al.  Network Identification and Flux Quantification in the Central Metabolism of Saccharomyces cerevisiae under Different Conditions of Glucose Repression , 2001, Journal of bacteriology.

[86]  S. Lee,et al.  Metabolic flux analysis and metabolic engineering of microorganisms. , 2008, Molecular bioSystems.

[87]  Shuichi Aiba,et al.  Identification of metabolic model: Citrate production from glucose by Candida lipolytica , 1979 .

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

[89]  G. Stephanopoulos,et al.  Elementary metabolite units (EMU): a novel framework for modeling isotopic distributions. , 2007, Metabolic engineering.

[90]  G. Stephanopoulos,et al.  Metabolic flux analysis in a nonstationary system: fed-batch fermentation of a high yielding strain of E. coli producing 1,3-propanediol. , 2007, Metabolic engineering.

[91]  C. Wittmann,et al.  Characterization of the metabolic shift between oxidative and fermentative growth in Saccharomyces cerevisiae by comparative 13C flux analysis , 2005, Microbial cell factories.

[92]  M. Ikeda Amino acid production processes. , 2003, Advances in biochemical engineering/biotechnology.

[93]  Jens Nielsen,et al.  Metabolic Network Analysis of Streptomyces tenebrarius, a Streptomyces Species with an Active Entner-Doudoroff Pathway , 2005, Applied and Environmental Microbiology.

[94]  Christoph Wittmann,et al.  Transcriptional and Metabolic Responses of Bacillus subtilis to the Availability of Organic Acids: Transcription Regulation Is Important but Not Sufficient To Account for Metabolic Adaptation , 2006, Applied and Environmental Microbiology.

[95]  Christoph Wittmann,et al.  Accumulation of Homolanthionine and Activation of a Novel Pathway for Isoleucine Biosynthesis in Corynebacterium glutamicum McbR Deletion Strains , 2006, Journal of bacteriology.

[96]  Elmar Heinzle,et al.  13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion-isotope ratio mass spectrometry. , 2010, Metabolic engineering.

[97]  Yinjie J. Tang,et al.  Metabolic flux analysis of Shewanella spp. reveals evolutionary robustness in central carbon metabolism , 2009, Biotechnology and bioengineering.

[98]  C. Wittmann,et al.  The l -Lysine Story: From Metabolic Pathways to Industrial Production , 2007 .

[99]  A. Yokota,et al.  Metabolic changes in a pyruvate kinase gene deletion mutant of Corynebacterium glutamicum ATCC 13032. , 2010, Metabolic engineering.

[100]  Ralf Takors,et al.  Metabolic flux analysis at ultra short time scale: isotopically non-stationary 13C labeling experiments. , 2007, Journal of biotechnology.

[101]  T. Szyperski Biosynthetically Directed Fractional 13C‐labeling of Proteinogenic Amino Acids , 1995 .

[102]  U. Sauer,et al.  Metabolic Flux Responses to Pyruvate Kinase Knockout in Escherichia coli , 2002, Journal of bacteriology.

[103]  C. Wittmann,et al.  In-Depth Profiling of Lysine-Producing Corynebacterium glutamicum by Combined Analysis of the Transcriptome, Metabolome, and Fluxome , 2004, Journal of bacteriology.

[104]  I. Nissim,et al.  Dynamic aspects of amino acid metabolism in alloxan-induced diabetes and insulin-treated rabbits: in vivo studies with 15N and gas chromatography-mass spectrometry. , 1986, Biochemical medicine and metabolic biology.