MetaToul – Metabolomics-fluxomics

Its mission

The platform, specialised in the functional analysis of metabolic networks, designs, develops and provides metanalytical approaches in the field of metabolomics and fluxomics.

Key words

Metabolomics, Fluxomics, Mass Spectrometry, Nuclear Magnetic Resonance, High-throughput, Metabolic Networks

Person in charge

Scientific manager

PORTAIS Jean-Charles –

Operational and technical managers

BELLVERT Floriant –

PEYRIGA Lindsay –

Team numbers in 2017: 11

Teachers and researchers: 1 Engineers: 7 Assistant engineers and technicians : 2 Post-doctoral and doctoral students: 1

Thematic axes

Functional analysis of cellular metabolism (prokaryote and eukaryote) Robotisation and High-throughput Fluxomics

Main activities

• Development and applications of complementary NMR and MS methods for the analysis of metabolic networks
• Targeted quantitative metabolomics
• Identification of metabolic pathways by isotopic labelling (13C-metabolomics)
• Measurement of metabolic flux (13C-Fluxomics)
• Modelling of metabolic systems (calculation of flux, etc.)

Research models

The Platform is open to all the research models with high expertise on cellular models (prokaryote or eukaryote).

Fields of application and target products

The MetaToul platform provides its support in the functional analysis of the metabolism in many fields of application. Principally, these are:

• Microbial and plant biotechnologies
• Systems biology
• Cancerology
• Human health
• Host-microorganism interactions
• Sustainable development

Technologies, techniques, specific tools

Mass spectrometry

• Triple Quadripole 4000QTrap (AB Sciex): ESI ionisation source, can be coupled to a HPLC U3000 (Thermo) fitted with a UV detector or coupled to an ion chromatography system (Dionex ICS 5000+)
• LTQ Orbitrap Velos (Thermo): ESI ionisation source, can be coupled to a HPLC U3000 (Thermo) fitted with a UV detector or coupled to an ion chromatography system (Dionex ICS 5000+)
• QTof Xevo G2-XS (Waters): ESI ionisation source, coupled to a UPLC Acquity
• Q Exactive+ (Thermo): ESI source, can be coupled to a UHPLC Vanquish (Thermo) fitted with a UV detector or coupled to an ion chromatography system (Dionex ICS 5000+)

Nuclear Magnetic Resonance

• Spectrometer Ascend III 800 MHz (Bruker): cryoprobe 5 mm ATMA QPCI 1H-13C-31P-15N, Z-gradient, microcryoprobe 1.7 mm ATMA QTCI 1H-13C-31P, Z-gradient, refrigerated autosampler SampleJet
• Spectrometer Avance II 500 MHz (Bruker): probe BBI H-BB-D Z-Grd 5mm (BTO), probe TXI S2 H-P/C Z-Grd 5mm ATMA (BTO), probe BBO SB BB-H-D 10mm (BVT), probe BBI S2 H-BB-D 8mm (BVT), TXO SB P/C-H 15mm (BTO), LC-RMN SEI 3mm gradient Z ATMA (BTO), ambient temperature autosampler SampleXPress

Robotics and automation

• High-throughput Fluxomic robot (TECAN): this robot, jointly designed by the staff of the platform, the MetaSys team from the LISBP, and the TECAN company, enables parallel microorganism cultivation with substrates enriched with 13C for fluxomics. This robot enables the fully controlled cultivation of 48 bioreactors in parallel as well as automated sampling for metabolomics and fluxomics. This device is unique in the world, custom-made for the needs of the platform.
• Sample preparation robot (TECAN Evo200): Pipetting robot enabling automated sample preparation for mass spectrometry or NMR analyses. SPE module enabling pre-treatment of samples before injection or analysis.

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Significant publications and patents

• Pierre Millard, Edern Cahoreau, Maud Heuillet, Jean-Charles Portais, and Guy Lippens : 15N-NMR-Based Approach for Amino Acids-Based 13C-Metabolic Flux Analysis of Metabolism, Anal Chem, 2017
• L Poulain, P Sujobert, F Zylbersztejn, S Barreau, L Stuani, M Lambert, T L Palama, V Chesnais, R Birsen, F Vergez, T Farge, C Chenevier-Gobeaux, M Fraisse, F Bouillaud, C Debeissat, O Herault, C Récher, C Lacombe, M Fontenay, P Mayeux, T T Maciel, J-C Portais, J-E Sarry, J Tamburini, D Bouscary and N Chapuis : High mTORC1 activity drives glycolysis addiction and sensitivity to G6PD inhibition in acute myeloid leukemia cells, Leunkemia 2017
• Farge Thomas, Saland Estelle, Fabienne de Toni, Nesrine Aroua, Moshen Hosseini, Perry Robin, Claudie Bosc, Mayumi Sugita, Lucille Stuani, Marine Fraisse, Sarah Scotland, Clément Larrue, Héléna Boutzen, Virginie Féliu, Marie-Laure Nicolau-Travers, Stephanie Cassant-Sourdy, Nicolas Broin, Marion David, Nizar Serhan, Audrey Sarry, Suzanne Tavitian, Tony Kaoma, Laurent Vallar, Jason Iacovoni, Laetitia K. Linares, Camille Montersino, Remy Castellano, Emmanuel Griessinger, Yves Collette, Olivier Duchamp, Yara Barreira, Pierre Hirsch, Tony Palama, Lara Gales, Francois Delhommeau, Barbara H. Garmy-Susini, Jean-Charles Portais, Vergez Francois, Mary Selak, Gwenn Danet-Desnoyers, Martin Carroll, Christian Récher and Jean-Emmanuel Sarry : Chemotherapy Resistant Human Acute Myeloid Leukemia Cells are Not Enriched for Leukemic Stem Cells but Require Oxidative Metabolism, Cancer Discovery, 2017
• Teresa Pérez-Berezo, Julien Pujo, Patricia Martin, Pauline Le Faouder, Jean-Marie Galano, Alexandre Guy, Claude Knauf, Jean Claude Tabet, Sophie Tronnet, Frederick Barreau, Maud Heuillet, Gilles Dietrich, Justine Bertrand-Michel, Thierry Durand, Eric Oswald, and Nicolas Cenac : Identification  of an analgesic lipopeptide produced by the probiotic Escherichia coli strain Nissle 1917, Nature Communication, 2017
• Stéphanie Heux, Cécilia Bergès, Pierre Millard, Jean-Charles Portais and Fabien Létisse : Recent advances in high-throughput 13C-fluxomics, COB, 2016

Involvement in TWB precompetitive projects

• SYNTHOL: Valorisation SYNTHétique du méthanOL par E. coli
• ROCOCO: The Reverse tricarboxylic acid cycle (rTCA), a promising and flexible biological CO2 Capture pathway