Parallel labeling experiments for pathway elucidation and (13)C metabolic flux analysis.

Metabolic pathway models provide the foundation for quantitative studies of cellular physiology through the measurement of intracellular metabolic fluxes. For model organisms metabolic models are well established, with many manually curated genome-scale model reconstructions, gene knockout studies and stable-isotope tracing studies. However, for non-model organisms a similar level of knowledge is often lacking. Compartmentation of cellular metabolism in eukaryotic systems also presents significant challenges for quantitative (13)C-metabolic flux analysis ((13)C-MFA). Recently, innovative (13)C-MFA approaches have been developed based on parallel labeling experiments, the use of multiple isotopic tracers and integrated data analysis, that allow more rigorous validation of pathway models and improved quantification of metabolic fluxes. Applications of these approaches open new research directions in metabolic engineering, biotechnology and medicine.

[1]  Christian M. Metallo,et al.  Regulation of substrate utilization by the mitochondrial pyruvate carrier. , 2014, Molecular cell.

[2]  U. Sauer,et al.  Metabolism of Pentose Sugars in the Hyperthermophilic Archaea Sulfolobus solfataricus and Sulfolobus acidocaldarius , 2010, The Journal of Biological Chemistry.

[3]  Jie Zhang,et al.  Optimization of 13C isotopic tracers for metabolic flux analysis in mammalian cells. , 2012, Metabolic engineering.

[4]  Maciek R Antoniewicz,et al.  Parallel labeling experiments with [1,2-(13)C]glucose and [U-(13)C]glutamine provide new insights into CHO cell metabolism. , 2013, Metabolic engineering.

[5]  Maciek R Antoniewicz,et al.  Publishing 13C metabolic flux analysis studies: a review and future perspectives. , 2013, Metabolic engineering.

[6]  Scott B. Crown,et al.  Rational design of 13C-labeling experiments for metabolic flux analysis in mammalian cells , 2012, BMC Systems Biology.

[7]  Thomas M. Wasylenko,et al.  The oxidative pentose phosphate pathway is the primary source of NADPH for lipid overproduction from glucose in Yarrowia lipolytica. , 2015, Metabolic engineering.

[8]  Maciek R Antoniewicz,et al.  Resolving the TCA cycle and pentose-phosphate pathway of Clostridium acetobutylicum ATCC 824: Isotopomer analysis, in vitro activities and expression analysis. , 2011, Biotechnology journal.

[9]  Edward J. O'Brien,et al.  Computing the functional proteome: recent progress and future prospects for genome-scale models. , 2015, Current opinion in biotechnology.

[10]  Maciek R Antoniewicz,et al.  (13)C-metabolic flux analysis of co-cultures: A novel approach. , 2015, Metabolic engineering.

[11]  Gregory Stephanopoulos,et al.  Evaluation of regression models in metabolic physiology: predicting fluxes from isotopic data without knowledge of the pathway , 2006, Metabolomics.

[12]  Joerg M. Buescher,et al.  A roadmap for interpreting (13)C metabolite labeling patterns from cells. , 2015, Current opinion in biotechnology.

[13]  C. Maranas,et al.  Recent advances in the reconstruction of metabolic models and integration of omics data. , 2014, Current opinion in biotechnology.

[14]  M. Antoniewicz Methods and advances in metabolic flux analysis: a mini-review , 2015, Journal of Industrial Microbiology & Biotechnology.

[15]  Yinjie J. Tang,et al.  Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain , 2009, Biotechnology and bioengineering.

[16]  Maciek R Antoniewicz,et al.  Tandem mass spectrometry for measuring stable-isotope labeling. , 2013, Current opinion in biotechnology.

[17]  Yinjie J. Tang,et al.  Investigation of Carbon Metabolism in “Dehalococcoides ethenogenes” Strain 195 by Use of Isotopomer and Transcriptomic Analyses , 2009, Journal of bacteriology.

[18]  Maciek R Antoniewicz,et al.  13C metabolic flux analysis: optimal design of isotopic labeling experiments. , 2013, Current opinion in biotechnology.

[19]  M. Antoniewicz,et al.  COMPLETE-MFA: complementary parallel labeling experiments technique for metabolic flux analysis. , 2013, Metabolic engineering.

[20]  Sang Yup Lee,et al.  Genome-scale reconstruction and in silico analysis of the Clostridium acetobutylicum ATCC 824 metabolic network , 2008, Applied Microbiology and Biotechnology.

[21]  Nathan D Price,et al.  Transparency in metabolic network reconstruction enables scalable biological discovery. , 2015, Current opinion in biotechnology.

[22]  Elmar Heinzle,et al.  Metabolic flux analysis in eukaryotes. , 2010, Current opinion in biotechnology.

[23]  Adam M. Feist,et al.  Tracing compartmentalized NADPH metabolism in the cytosol and mitochondria of mammalian cells. , 2014, Molecular cell.

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

[25]  Yinjie J. Tang,et al.  13C-MFA delineates the photomixotrophic metabolism of Synechocystis sp. PCC 6803 under light- and carbon-sufficient conditions. , 2014, Biotechnology journal.

[26]  Christian M. Metallo,et al.  Understanding metabolic regulation and its influence on cell physiology. , 2013, Molecular cell.

[27]  G. Stephanopoulos,et al.  Intracellular flux analysis in hybridomas using mass balances and in vitro 13C nmr , 1995, Biotechnology and bioengineering.

[28]  Maciek R Antoniewicz,et al.  Parallel labeling experiments validate Clostridium acetobutylicum metabolic network model for (13)C metabolic flux analysis. , 2014, Metabolic engineering.

[29]  Adam M. Feist,et al.  Evolution of E. coli on [U-13C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology , 2016, PloS one.

[30]  E. Papoutsakis,et al.  Genome‐scale model for Clostridium acetobutylicum: Part I. Metabolic network resolution and analysis , 2008, Biotechnology and bioengineering.

[31]  Jennifer L Reed,et al.  Refining metabolic models and accounting for regulatory effects. , 2014, Current opinion in biotechnology.

[32]  C. Wittmann,et al.  Large-Scale 13C Flux Profiling Reveals Conservation of the Entner-Doudoroff Pathway as a Glycolytic Strategy among Marine Bacteria That Use Glucose , 2015, Applied and Environmental Microbiology.

[33]  M. Antoniewicz,et al.  Parallel labeling experiments with [U-13C]glucose validate E. coli metabolic network model for 13C metabolic flux analysis. , 2012, Metabolic engineering.

[34]  Karsten Hiller,et al.  Profiling metabolic networks to study cancer metabolism. , 2013, Current opinion in biotechnology.

[35]  Scott B. Crown,et al.  Parallel labeling experiments and metabolic flux analysis: Past, present and future methodologies. , 2013, Metabolic engineering.

[36]  Christopher P. Long,et al.  Complete genome sequence, metabolic model construction and phenotypic characterization of Geobacillus LC300, an extremely thermophilic, fast growing, xylose-utilizing bacterium. , 2015, Metabolic engineering.

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

[38]  Björn H. Junker Flux analysis in plant metabolic networks: increasing throughput and coverage. , 2014, Current opinion in biotechnology.

[39]  Patrick F Suthers,et al.  Construction of an E. Coli genome‐scale atom mapping model for MFA calculations , 2011, Biotechnology and bioengineering.

[40]  Maciek R Antoniewicz,et al.  Selection of tracers for 13C-metabolic flux analysis using elementary metabolite units (EMU) basis vector methodology. , 2012, Metabolic engineering.

[41]  Chao Li,et al.  CeCaFDB: a curated database for the documentation, visualization and comparative analysis of central carbon metabolic flux distributions explored by 13C-fluxomics , 2014, Nucleic Acids Res..

[42]  Bing Wu,et al.  Characterization of the Central Metabolic Pathways in Thermoanaerobacter sp. Strain X514 via Isotopomer-Assisted Metabolite Analysis , 2009, Applied and Environmental Microbiology.

[43]  M. Antoniewicz,et al.  Metabolic network reconstruction, growth characterization and 13C-metabolic flux analysis of the extremophile Thermus thermophilus HB8. , 2014, Metabolic engineering.

[44]  H. Shimizu,et al.  Flux analysis and metabolomics for systematic metabolic engineering of microorganisms. , 2013, Biotechnology advances.

[45]  Adam M. Feist,et al.  Next-generation genome-scale models for metabolic engineering. , 2015, Current opinion in biotechnology.

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

[47]  Maciek R Antoniewicz,et al.  Measuring complete isotopomer distribution of aspartate using gas chromatography/tandem mass spectrometry. , 2012, Analytical chemistry.

[48]  M. Jolicoeur,et al.  Unraveling the metabolism of HEK-293 cells using lactate isotopomer analysis , 2011, Bioprocess and biosystems engineering.

[49]  Christopher P. Long,et al.  Metabolic flux analysis of Escherichia coli knockouts: lessons from the Keio collection and future outlook. , 2014, Current opinion in biotechnology.

[50]  Xueyang Feng,et al.  Incomplete Wood–Ljungdahl pathway facilitates one-carbon metabolism in organohalide-respiring Dehalococcoides mccartyi , 2014, Proceedings of the National Academy of Sciences.

[51]  Yinjie J. Tang,et al.  Recent advances in mapping environmental microbial metabolisms through 13C isotopic fingerprints , 2012, Journal of The Royal Society Interface.

[52]  Jamey D. Young Metabolic flux rewiring in mammalian cell cultures. , 2013, Current opinion in biotechnology.

[53]  Xiao-Jiang Feng,et al.  Systems-Level Metabolic Flux Profiling Elucidates a Complete, Bifurcated Tricarboxylic Acid Cycle in Clostridium acetobutylicum , 2010, Journal of bacteriology.

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

[55]  M. Antoniewicz,et al.  (13)C metabolic flux analysis of the extremely thermophilic, fast growing, xylose-utilizing Geobacillus strain LC300. , 2016, Metabolic engineering.

[56]  Christopher P. Long,et al.  Integrated 13C-metabolic flux analysis of 14 parallel labeling experiments in Escherichia coli. , 2015, Metabolic engineering.

[57]  Xueyang Feng,et al.  Metabolic Flux Analysis of the Mixotrophic Metabolisms in the Green Sulfur Bacterium Chlorobaculum tepidum* , 2010, The Journal of Biological Chemistry.

[58]  T. Shlomi,et al.  Quantitative flux analysis reveals folate-dependent NADPH production , 2014, Nature.

[59]  M. Antoniewicz,et al.  Metabolic flux analysis of CHO cells at growth and non-growth phases using isotopic tracers and mass spectrometry. , 2011, Metabolic engineering.

[60]  Elmar Heinzle,et al.  Eukaryotic metabolism: Measuring compartment fluxes , 2011, Biotechnology journal.

[61]  Xueyang Feng,et al.  Carbon Flow of Heliobacteria Is Related More to Clostridia than to the Green Sulfur Bacteria* , 2010, The Journal of Biological Chemistry.

[62]  Xueyang Feng,et al.  Metabolic pathway determination and flux analysis in nonmodel microorganisms through 13C-isotope labeling. , 2012, Methods in molecular biology.

[63]  M. Antoniewicz,et al.  Towards dynamic metabolic flux analysis in CHO cell cultures , 2012, Biotechnology journal.

[64]  N. Kruger,et al.  Fluxes through plant metabolic networks: measurements, predictions, insights and challenges. , 2015, The Biochemical journal.