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59 results on '"Stéphane Massou"'

1. Mechanochemical Synthesis and Biological Evaluation of Novel Isoniazid Derivatives with Potent Antitubercular Activity

Author: Paulo F. M. Oliveira, Brigitte Guidetti, Alain Chamayou, Christiane André-Barrès, Jan Madacki, Jana Korduláková, Giorgia Mori, Beatrice Silvia Orena, Laurent Roberto Chiarelli, Maria Rosalia Pasca, Christian Lherbet, Chantal Carayon, Stéphane Massou, Michel Baron, and Michel Baltas
Subjects: Mycobacterium tuberculosis, mechanochemistry, hydrazone, Organic chemistry, QD241-441
Abstract: A series of isoniazid derivatives bearing a phenolic or heteroaromatic coupled frame were obtained by mechanochemical means. Their pH stability and their structural (conformer/isomer) analysis were checked. The activity of prepared derivatives against Mycobacterium tuberculosis cell growth was evaluated. Some compounds such as phenolic hydrazine 1a and almost all heteroaromatic ones, especially 2, 5 and 7, are more active than isoniazid, and their activity against some M. tuberculosis MDR clinical isolates was determined. Compounds 1a and 7 present a selectivity index >1400 evaluated on MRC5 human fibroblast cells. The mechanism of action of selected hydrazones was demonstrated to block mycolic acid synthesis due to InhA inhibition inside the mycobacterial cell.
Published: 2017
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2. Gelation Mechanism Revealed in Organometallic Gels: Prevalence of van der Waals Interactions on Oligomerization by Coordination Chemistry

Author: Yinping Wang, Yannick Coppel, Juliette Fitremann, Stéphane Massou, Christophe Mingotaud, Myrtil L. Kahn, Interactions moléculaires et réactivité chimique et photochimique (IMRCP), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Fédération de Recherche Fluides, Energie, Réacteurs, Matériaux et Transferts (FERMAT), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), IDeAS - Interfaces Dynamiques et Assemblages Stimulables (IDeAS), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie de Toulouse (ICT), Laboratoire de chimie de coordination (LCC), and Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [CHIM]Chemical Sciences, Physical and Theoretical Chemistry, Atomic and Molecular Physics, and Optics
Abstract: International audience
Published: 2023
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3. Cordiasecosides G-J, 9,10-Seco-29-norcycloartane glycosides isolated from Cordia lutea and their antibacterial activities

Author: Inés Castro, Christiane André-Barrès, Nicolas Fabre, Stéphane Massou, Michel Sauvain, Denis Castillo Pareja, Valérie Jullian, Pharmacochimie et Biologie pour le Développement (PHARMA-DEV), Institut de Recherche pour le Développement (IRD)-Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Laboratorios de Investigación y Desarrollo (LID), Universidad Peruana Cayetano Heredia (UPCH), Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and Financement récurrent des institutions d'affiliation des auteurs,IRD : Bourse de thèse ARTSIRD : Laboratoire Mixte International LAVI
Subjects: Pharmacology, Cordia, Helicobacter pylori, Cordia lutea, Molecular Structure, Secocycloartane, Plant Extracts, [SDV]Life Sciences [q-bio], Traditional medicine, General Medicine, Microbial Sensitivity Tests, traditional medicine, Boraginaceae, Anti-Bacterial Agents, Antibacterial, antibacterial, Drug Discovery, [CHIM]Chemical Sciences, secocycloartane, Glycosides, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; Four undescribed secocycloartane monoglycosides (1-4) were isolated from an ethanolic extract of the dry flowers of Cordia lutea Lam. Their structural assignment is based on NMR and MS analysis. Their stereochemistry is confirmed by molecular modelling studies using DFT-NMR calculations done for compound 3. In vitro antibacterial activity of the four compounds was moderate on Helicobacter pylori (MIC = 15.6 µg/mL), and much weaker on Staphylococcus aureus, Pseudomonas aeruginosa or Escherichia coli (MIC > 125 µg/mL). Toxicity evaluated against RAW 264.7 cells was weak (IC50 values ranging from 24 to 41 µM i.e 15 to 24 µg/mL), but in the same range as anti-Helicobacter activity.
Published: 2021
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4. Cyclometalated gold(III) complexes : noticeable differences between (N,C) and (P,C) ligands in migratory insertion

Author: Sonia Mallet-Ladeira, Didier Bourissou, Karinne Miqueu, Xavi Ribas, E. Daiann Sosa Carrizo, Abderrahmane Amgoune, Teodor Parella, Jordi Serra, Stéphane Massou, Pau Font, Institut de Química Computacional i Catàlisi and Departament de Química, Universitat de Girona (IQCC), Universitat de Girona (UdG), Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM), Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de chimie de coordination (LCC), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, NMR Service, Universitat Autònoma de Barcelona (UAB), Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Spanish MINECO (CTQ2016-77989-P and CTQ2015-64436-P) and Generalitat de Catalunya (2017 SGR 264), European Project: 277801,EC:FP7:ERC,ERC-2011-StG_20101014,SUSCATCU3(2011), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), and Ministerio de Economía y Competitividad (Espanya)
Subjects: Ethylene, Química organometàl·lica, Or, 010402 general chemistry, 01 natural sciences, Medicinal chemistry, Catalysis, chemistry.chemical_compound, Catàlisi, Reactivity (chemistry), [CHIM.COOR]Chemical Sciences/Coordination chemistry, Alkyl, chemistry.chemical_classification, Olefin fiber, 010405 organic chemistry, Ligand, Migratory insertion, Cationic polymerization, General Chemistry, 0104 chemical sciences, chemistry, Organometallic chemistry, Reaction mechanisms (Chemistry), Mecanismes de reacció (Química), Gold
Abstract: Gold(III) complexes are garnering increasing interest for opto-electronic, therapeutic and catalytic applications. But so far, very little is known about the factors controlling their reactivity and the very influence of the ancillary ligand. This article reports the first comprehensive study on this topic. The reactivity of a cationic (N,C) gold(iii) complex, namely 1A, towards ethylene has been thoroughly studied and compared with that of the related (P,C) complex 1C. A cationic gold(iii) complex 5A resulting from double insertion of ethylene was selectively obtained. Complex 5A was found to be remarkably stable. It was trapped with chloride and fully characterized. In marked contrast to that observed with 1C, no β-H elimination or linear-to-branched rearrangement of the alkyl chain occurred with 1A. The energy profile for the reactions of 1A with ethylene has been comprehensively investigated computationally, and the influence of the ancillary ligand has been precisely delineated. Because nitrogen is a weaker donor than carbon (and phosphorus), the (N,C) ligand is very electronically dissymmetric, much more than the (P,C) ligand. This makes the two reactive sites at gold quite different, which noticeably influences the competition between migratory insertion and β-H elimination, and actually changes the outcome of the olefin insertion at gold. This study provides valuable insight into the influence of ancillary ligands on gold(iii) reactivity, something critical to further develop Au(iii) and Au(i)/Au(iii) catalysis Financial support from the Centre National de la Recherche Scientique and the Universit´e de Toulouse, the European Research Council (Starting Grant Project ERC-2011-StG277801), Spanish MINECO (CTQ2016-77989-P and CTQ2015-64436-P) and Generalitat de Catalunya (2017 SGR 264) is gratefully acknowledged
Published: 2021
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5. Synthesis of a Stable N‐Hetero ‐ Rh I ‐Metallacyclic Silanone

Author: Norio Nakata, Kazuki Nakaya, Tsuyoshi Kato, Stéphane Massou, Shintaro Takahashi, Maria Frutos, Antoine Baceiredo, Vicenç Branchadell, Nathalie Saffon-Merceron, Daisuke Hashizume, Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), and Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Steric effects, Bicyclic molecule, 010405 organic chemistry, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Silanone, Substituent, General Medicine, General Chemistry, [CHIM.INOR]Chemical Sciences/Inorganic chemistry, 010402 general chemistry, Photochemistry, 01 natural sciences, Catalysis, 0104 chemical sciences, 3. Good health, chemistry.chemical_compound, Silanol, Reagent, Intramolecular force, [CHIM]Chemical Sciences, [CHIM.COOR]Chemical Sciences/Coordination chemistry, ComputingMilieux_MISCELLANEOUS
Abstract: A novel N-hetero-RhI -metallacyclic silanone 2 has been synthesized. The silanone 2, showing an extremely large dimerization energy (ΔG=+86.2 kcal mol-1 ), displays considerable stability and persists in solution up to 60 °C. Above 120 °C, an intramolecular Csp3 -H insertion occurs slowly over a period of two weeks leading to the bicyclic silanol 5. The exceptional stability of 2, related to the unusual electronic and steric effects of RhI -substituent, should allow for a more profound study and understanding of these new species. Furthermore, the metallacyclic silanone 2 presents two reactive centers (Si=O and Rh), which can be involved depending upon the nature of reagents. Of particular interest, the reaction with H2 starts with the hydrogenation of RhI center leading to the corresponding RhIII -dihydride complex 7 and it undergoes a cis/trans-isomerization via a particular mechanism, demonstrating that addition-elimination processes can also happen for silanones just like for their carbon analogues!
Published: 2020
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6. Synthesis of a Stable N-Hetero-Rh

Author: Shintaro, Takahashi, Kazuki, Nakaya, María, Frutos, Antoine, Baceiredo, Nathalie, Saffon-Merceron, Stéphane, Massou, Norio, Nakata, Daisuke, Hashizume, Vicenç, Branchadell, and Tsuyoshi, Kato
Abstract: A novel N-hetero-Rh
Published: 2020

7. Evidence for genuine hydrogen bonding in gold(I) complexes

Author: Karinne Miqueu, Didier Bourissou, Abderrahmane Amgoune, Stéphane Massou, E. Daiann Sosa Carrizo, Sonia Mallet-Ladeira, Mathilde Rigoulet, Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM), Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de chimie de coordination (LCC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Université Fédérale Toulouse Midi-Pyrénées, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), and Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: chemistry.chemical_classification, Multidisciplinary, 010405 organic chemistry, Hydrogen bond, Cationic polymerization, Solid-state, gold, hydrogen bonding, 010402 general chemistry, 01 natural sciences, relativistic effects, 0104 chemical sciences, Characterization (materials science), Crystallography, non-covalent interactions, chemistry, Physical Sciences, [CHIM]Chemical Sciences, Non-covalent interactions, Relativistic quantum chemistry, Proton acceptor
Abstract: The ability of gold to act as proton acceptor and participate in hydrogen bonding remains an open question. Here, we report the synthesis and characterization of cationic gold(I) complexes featuring ditopic phosphine-ammonium (P,NH + ) ligands. In addition to the presence of short Au∙∙∙H contacts in the solid state, the presence of Au∙∙∙H–N hydrogen bonds was inferred by NMR and IR spectroscopies. The bonding situation was extensively analyzed computationally. All features were consistent with the presence of three-center four-electron attractive interactions combining electrostatic and orbital components. The role of relativistic effects was examined, and the analysis is extended to other recently described gold(I) complexes.
Published: 2018
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8. Cyclic (Amino)(Phosphonium Bora-Ylide)Silanone: A Remarkable Room-Temperature-Persistent Silanone

Author: Daisuke Hashizume, Vicenç Branchadell, Isabel Alvarado-Beltran, Tsuyoshi Kato, Nathalie Saffon-Merceron, Stéphane Massou, Antoine Baceiredo, and Alfredo Rosas-Sánchez
Subjects: chemistry.chemical_classification, Silicon, 010405 organic chemistry, Silanone, Substituent, chemistry.chemical_element, General Chemistry, General Medicine, Photochemistry, 010402 general chemistry, 01 natural sciences, Catalysis, 0104 chemical sciences, chemistry.chemical_compound, chemistry, Ylide, Polymer chemistry, Borylene, Lewis acids and bases, Phosphonium
Abstract: Silanone substituted by bulky amino- and phosphonium bora-ylide substituents has been isolated in crystalline form. Thanks to the exceptionally strong electron-donating phosphonium bora-ylide substituent, the life-time at room temperature of silanone 2 is dramatically extended (t1/2 = 4 days) compared to the related (amino)(phosphonium-ylide)silanone (t1/2 = 5 hours), allowing easier manipulation and its use as precursor of new valuable silicon compounds. The interaction of silanone with a weak Lewis acid such as MgBr2 increases further its stability (no degradation after 3 weeks at room temperature).
Published: 2017
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9. Exceptionally Strong Electron-Donating Ability of Bora-Ylide Substituent vis-à-vis Silylene and Silylium Ion

Author: Antoine Baceiredo, Alfredo Rosas-Sánchez, Isabel Alvarado-Beltran, Tsuyoshi Kato, Nathalie Saffon-Merceron, Vicenç Branchadell, and Stéphane Massou
Subjects: chemistry.chemical_classification, Ligand, 010405 organic chemistry, Substituent, Silylene, chemistry.chemical_element, General Chemistry, General Medicine, Photochemistry, 010402 general chemistry, 01 natural sciences, Catalysis, Coordination complex, 0104 chemical sciences, chemistry.chemical_compound, chemistry, Ylide, Silylium ion, Phosphonium, Boron
Abstract: Electropositive boron-based substituent (phosphonium bora-ylide) with an exceptionally strong π- and σ-electron donating character dramatically increases the stability of a new type of N-heterocyclic silylene 2 featuring amino- and bora-ylide-substituents. Moreover, the related silylium ion 4 and transition-metal–silylene complexes, with trigonal-planar geometries around the silicon center, are also well stabilized. Therefore, the N,B-heterocyclic silylene 2 can be used as a strongly electron-donating innocent ligand in coordination chemistry similarly to N-heterocyclic carbenes.
Published: 2017
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10. A Stable N-Hetero -Rh -Metallacyclic Silylene

Author: Shintaro Takahashi, Norio Nakata, Vicenç Branchadell, Tsuyoshi Kato, Stéphane Massou, Antoine Baceiredo, Daisuke Hashizume, Nathalie Saffon-Merceron, Ekaterina V. Bellan, Laboratoire de chimie de coordination (LCC), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), RIKEN - Institute of Physical and Chemical Research [Japon] (RIKEN), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Institut de Chimie de Toulouse (ICT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UPS), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UPS), and RIKEN
Subjects: Materials science, 010405 organic chemistry, Silylene, Tetrahedral molecular geometry, chemistry.chemical_element, General Chemistry, General Medicine, 010402 general chemistry, 01 natural sciences, Catalysis, 3. Good health, 0104 chemical sciences, Rhodium, Crystallography, chemistry.chemical_compound, chemistry, Atom, Single bond, Moiety, [CHIM]Chemical Sciences, Derivative (chemistry), ComputingMilieux_MISCELLANEOUS
Abstract: A cyclic (amino)metal-substituted dicoordinated silylene derivative has been synthesized and fully characterized. Of particular interest is that the N-hetero-RhI -metallacyclic silylene exhibits a distorted tetrahedral geometry around the rhodium atom and a considerably shortened Si-Rh bond (2.138 A) compared to classical Si-Rh single bonds (ca. 2.30-2.35 A). A theoretical investigation reveals that the geometrical deviation around the rhodium center from the classical square-planar to a tetrahedral geometry increases the π-donating and σ-accepting character of the rhodium atom, thereby efficiently stabilizing the silylene moiety.
Published: 2019
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11. A sterically congested 1,2-diphosphino-1'-boryl-ferrocene: synthesis, characterization and coordination to platinum

Author: Stéphane Massou, Patrice Renaut, Hélène Cattey, Julien Roger, Ghenwa Bouhadir, Paul Fleurat-Lessard, Jean-Cyrille Hierso, Didier Bourissou, Emmanuel Lerayer, Nadine Pirio, Maxime Boudjelel, Institut de Chimie Moléculaire de l'Université de Bourgogne [Dijon] (ICMUB), Université de Bourgogne (UB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Bourgogne (UB)-Institut de Chimie du CNRS (INC), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)
Subjects: Steric effects, 010405 organic chemistry, Chemistry, Ligand, chemistry.chemical_element, 010402 general chemistry, 01 natural sciences, Nmr data, 3. Good health, 0104 chemical sciences, Inorganic Chemistry, chemistry.chemical_compound, Ferrocene, Polymer chemistry, [CHIM]Chemical Sciences, Moiety, Chelation, Lewis acids and bases, Platinum
Abstract: International audience; A new class of tritopic ferrocene-based ambiphilic compound has been prepared by assembling diphosphino- and boryl-substituted cyclopentadienides at iron. The presence of five sterically demanding substituents on the ferrocene platform induces conformation constraints, as apparent from XRD and NMR data, but does not prevent chelating coordination to platinum. The Lewis acid moiety is pendant in both the free ligand and the platinum complex.
Published: 2019
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12. Genome-scale reconstruction and system level investigation of the metabolic network of Methylobacterium extorquens AM1.

Author: Rémi Peyraud, Kathrin Schneider, Patrick Kiefer, Stéphane Massou, Julia A. Vorholt, and Jean-Charles Portais
Published: 2011
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13. Room-Temperature Functionalization of N2 to Borylamine at a Molybdenum Complex

Author: Qian Liao, Stéphane Massou, Nathalie Saffon-Merceron, Noel Nebra, Soukaina Bennaamane, Maria F. Espada, Marie Fustier-Boutignon, Nicolas Mézailles, Eric Clot, Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC), Institut Charles Gerhardt Montpellier - Institut de Chimie Moléculaire et des Matériaux de Montpellier (ICGM ICMMM), and Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut de Chimie du CNRS (INC)
Subjects: 010405 organic chemistry, Chemistry, Intermolecular force, chemistry.chemical_element, Boranes, General Medicine, General Chemistry, [CHIM.CATA]Chemical Sciences/Catalysis, Borane, Hydride ligands, 010402 general chemistry, 01 natural sciences, Catalysis, 3. Good health, 0104 chemical sciences, Metal, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, chemistry.chemical_compound, Molybdenum, visual_art, Polymer chemistry, visual_art.visual_art_medium, Surface modification, [PHYS.PHYS.PHYS-CHEM-PH]Physics [physics]/Physics [physics]/Chemical Physics [physics.chem-ph], ComputingMilieux_MISCELLANEOUS
Abstract: Intermolecular, stepwise functionalization by BH bonds of a (triphosphine)MoIV -nitrido complex generated by N2 splitting is reported. The imido-hydride and di-hydride-amido MoIV complexes have been isolated and characterized. Addition of PinBH to the [Mo(H)2 (N(BPin)2 )]+ complex at room temperature results in the liberation of borylamines from the metal center.
Published: 2018
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14. Room-Temperature Functionalization of N

Author: Maria F, Espada, Soukaina, Bennaamane, Qian, Liao, Nathalie, Saffon-Merceron, Stéphane, Massou, Eric, Clot, Noel, Nebra, Marie, Fustier-Boutignon, and Nicolas, Mézailles
Abstract: Intermolecular, stepwise functionalization by BH bonds of a (triphosphine)Mo
Published: 2018

15. Catalytic Dehydrogenation of (Di)Amine-Boranes with a Geometrically Constrained Phosphine-Borane Lewis Pair

Author: Didier Bourissou, Karinne Miqueu, E. Daiann Sosa Carrizo, Sonia Ladeira, Stéphane Massou, Maxime Boudjelel, Ghenwa Bouhadir, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM), Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), and ANR-15-CE07-0003,COLAMET,Cooperativity between Lewis acids and transition metals(2015)
Subjects: small molecule activation, Boranes, phosphine-borane, Borane, 010402 general chemistry, Lewis pair, 01 natural sciences, Medicinal chemistry, Catalysis, Ring strain, chemistry.chemical_compound, amine-borane, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Dehydrogenation, Electron pair, 010405 organic chemistry, General Chemistry, [CHIM.MATE]Chemical Sciences/Material chemistry, 0104 chemical sciences, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, metal-free catalysis, [CHIM.POLY]Chemical Sciences/Polymers, chemistry, dehydrogenation, Amine gas treating, Phosphine
Abstract: International audience; The o-phenylene bridged phosphine-borane iPr2P(o-C6H4)B(Fxyl)2 2 was prepared. Despite ring strain, it adopts a closed form, as substantiated by NMR, XRD, and DFT analyses. However, the corresponding open form is only slightly higher in energy. The dormant Lewis pair 2 proved to efficiently catalyze the dehydrogenation of a variety of amine- and diamine-boranes under mild conditions. The corresponding phosphonium-borate iPr2PH(o-C6H4)BH(Fxyl)2 3 was authenticated as a key intermediate of these dehydrogenation reactions. The propensity of 3 to release H2 plays a major role in the catalytic turnover
Published: 2018
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16. Isotopic profiling of 13C-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy

Author: Edern Cahoreau, Jane Hubert, Stéphane Massou, Frédéric Bringaud, Jean-Charles Portais, Lindsay Peyriga, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Centre de résonance magnétique des systèmes biologiques (CRMSB), Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB), Résonance magnétique des systèmes biologiques (RMSB), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), MetaToul FluxoMet (TBI-MetaToul), MetaboHUB-MetaToul, MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)
Subjects: C-13, [SDV]Life Sciences [q-bio], Biophysics, Analytical chemistry, H-1 NMR, 010402 general chemistry, Mass spectrometry, 01 natural sciences, Biochemistry, J-resolved spectroscopy, GLUTAMATE, 03 medical and health sciences, Nuclear magnetic resonance, Metabolic flux analysis, 2D NMR, METABOLIC FLUX ANALYSIS, Spectroscopy, Molecular Biology, 030304 developmental biology, J resolved, 0303 health sciences, Isotopic profiling, STABILITY, Chemistry, MIXTURES, MASS-SPECTROMETRY, Cell Biology, Nuclear magnetic resonance spectroscopy, QUANTITATIVE-ANALYSIS, 0104 chemical sciences, Intermediate complexity, Heteronuclear molecule, ESCHERICHIA-COLI, NMR-SPECTROSCOPY, C-13 enrichment, Two-dimensional nuclear magnetic resonance spectroscopy, Carbon labeling experiment, LABELING EXPERIMENTS
Abstract: The use of two-dimensional heteronuclear J-resolved (2D H-JRES) nuclear magnetic resonance (NMR) spectroscopy for fast and reliable measurement of isotopic patterns from C-13-enriched compounds resulting from carbon labeling experiments was evaluated. Its use with biological samples of increasing complexity showed that 2D H-JRES spectroscopy is suitable for high-throughput isotopic profiling of any kind of labeled samples. Moreover, the method enabled accurate quantification of C-13 enrichments and, thus, can be used for metabolic flux analysis. The excellent trade-off between reduced experimental time and the number of measurable isotopic data makes 2D H-JRES NMR a promising approach for high-throughput flux analysis of samples of intermediate complexity. (C) 2012 Elsevier Inc. All rights reserved.
Published: 2012
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17. 95 Mo NMR: a useful tool for structural studies in solution

Author: Helena Teruel, José A. Brito, Stéphane Massou, and Montserrat Gómez
Subjects: chemistry.chemical_classification, Oxygen transfer, Ligand, Stereochemistry, General Chemistry, HEXA, Coordination complex, Metal, Crystallography, chemistry, visual_art, visual_art.visual_art_medium, General Materials Science, Reactivity (chemistry), Bimetallic strip, Electronic density
Abstract: Oxomolybdenum(VI) complexes containing diverse ligands from an electronic and topological point of view have been analysed by means of (95)Mo NMR in solution with the purpose of using this technique as a tool to study their coordination chemistry and reactivity. The relationship between the electronic density on the metal tuned by the electron-donor ability of the coordinated ligands and the (95)Mo chemical shift has been proved for mono- and bimetallic complexes showing a hexa- or hepta-coordination around the metal centre. The different origins of the signal broadening (associated either to the symmetry of the metallic polyhedron or to the presence of isomers or to the ligand de-coordination) have been also considered to rationalise the obtained data.
Published: 2009
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18. Germylenes and Stannylenes with Chelating Anilido–Imine Ligands: Syntheses, Structures and Reactivity

Author: Annie Castel, Ali Mcheik, Heinz Gornitzka, Stéphane Massou, Pierre Rivière, Tayssir Hamieh, and Nadia Katir
Subjects: 010405 organic chemistry, chemistry.chemical_element, Germanium, 010402 general chemistry, 01 natural sciences, Imine ligands, Cycloaddition, 0104 chemical sciences, Inorganic Chemistry, chemistry, Polymer chemistry, Molecule, Organic chemistry, Reactivity (chemistry), Lithium, Chelation, Tin
Abstract: Germylenes and stannylenes with anilido–imine ligands were synthesized in high yields by the reaction of group 14 dichlorides (Cl2Ge·dioxane, Cl2Sn) and the lithium salts of the ligands. Complexes were characterized by 1H and 13C NMR spectroscopy, and the molecular structure of methoxygermylene was determined by X-ray crystallography. These data indicate the formation of N,N′-chelate complexes having three-coordinate germanium or tin atoms and quasiplanar six-membered heterocycles. In the germanium series, cycloaddition reactions with o-quinone were performed, and the structures of the cycloadducts were determined by X-ray structure analyses. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Published: 2008
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19. Molybdenum(VI)-catalysed olefin epoxidation: Structure and reactivity study

Author: Montserrat Gómez, Stéphane Massou, José A. Brito, Helena Teruel, and Guillermo Muller
Subjects: Olefin fiber, Chiral ligand, chemistry.chemical_element, Epoxide, Medicinal chemistry, Catalysis, Kinetic resolution, Inorganic Chemistry, chemistry.chemical_compound, chemistry, Molybdenum, Cyclooctene, Materials Chemistry, Organic chemistry, Physical and Theoretical Chemistry, Selectivity
Abstract: Dioxo and oxoperoxo molybdenum(VI) complexes, I – III , containing the non-labile bidentated oxazolinyl-pyridine ligand 1 , were used as catalytic precursors in the epoxidation of cyclooctene and ( R )-limonene, to explore the nature of the catalytic species. The high diastereoselectivity showed by I and II towards limonene epoxide formation ( trans / cis - 8 = 4/1) could be justified by the kinetic resolution observed to give the double epoxide ( 9 ), but also by the olefin coordination to the metal centre due to the presence of a labile ligand (isothiocyanate). 95 Mo NMR monitoring experiments of complex II in the presence of ( R )-limonene, together with conductivity measurements, showed the formation of an ionic metallic species (1:1 electrolyte). These results point to a bimetallic species where one of the metal atoms is coordinated to the olefin by the dissociation of the isothiocyanate group, remaining coordinated the spectator chiral ligand, which is the responsible of the selectivity observed.
Published: 2008
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20. Phosphavinylidene(oxo)phosphorane MesP(O)=C=PMes: A Diphosphaallene Featuring λ 5 σ 3 ‐ and λ 3 σ 2 ‐Phosphorus Atoms

Author: Heinz Gornitzka, Ioan Silaghi-Dumitrescu, Raluca Septelean, Jean Escudié, Stéphane Massou, Henri Ranaivonjatovo, Luminita Silaghi-Dumitrescu, and Gabriela Nemes
Subjects: chemistry.chemical_classification, Double bond, Chemistry, Stereochemistry, Coordination number, Phosphorus, Oxide, chemistry.chemical_element, Phosphorane, Medicinal chemistry, Inorganic Chemistry, chemistry.chemical_compound, Hydrolysis, Intramolecular force, Reactivity (chemistry), Methylene
Abstract: Bis(2,4,6-tri-tert-butylphenyl)phosphavinylidene(oxo)phosphorane (1) (Mes* = 2,4,6-tri-tert-butylphenyl), the first 1λ5σ3,3λ3σ2-diphosphaallene, has been synthesised by addition of the phosphaalkenyllithium Mes*P=C(Li)Cl to dichlorophosphane oxide Mes*P(O)Cl2. Compound 1 is stable in the solid-state but undergoes a slow intramolecular cyclisation in solution by addition of a C–H bond of an ortho-tert-butyl group to the σ3-P=C double bond. Water adds chemoselectively and regioselectively to the λ5σ3-P=C bond, which appears to be more reactive than the λ3σ2-P=C bond. This leads to the hydrolysis compound 8 which has been structurally characterised. The new bis(phosphane oxide)methylene 9 formed from the successive cyclisation of anortho-tert-butyl group to the λ5σ3-P=C bond and addition of water to the λ3σ2-P=C bond has also been structurally characterised. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)
Published: 2006
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21. Isolation and Antimalarial Activity of Alkaloids fromPseudoxandra cuspidata

Author: Stéphane Massou, Florence Souard, Geneviève Bourdy, Séverine Maurel, Claude Moulis, Vincent Roumy, Nicolas Fabre, and Alexis Valentin
Subjects: Stereochemistry, Plasmodium falciparum, Annonaceae, Pharmaceutical Science, Pharmacognosy, Analytical Chemistry, Antimalarials, chemistry.chemical_compound, Alkaloids, Cell Line, Tumor, Toxicity Tests, Drug Discovery, Animals, Humans, Cytotoxicity, Nuclear Magnetic Resonance, Biomolecular, Pharmacology, biology, Alkaloid, Organic Chemistry, biology.organism_classification, Rodiasine, Complementary and alternative medicine, chemistry, visual_art, Plant Bark, visual_art.visual_art_medium, Molecular Medicine, Bark, Pseudoxandra cuspidata
Abstract: A novel and very unusual azaanthracene alkaloid, 1-aza-7,8,9,10-tetramethoxy-4-methyl-2-oxo-1,2-dihydroanthracene ( 1) and a new diastereoisomer of the bis-benzylisoquinoline alkaloid rodiasine, 1 S,1' R-rodiasine ( 2), as well as the alkaloids O-methylpunjabine ( 3) and O-methylmoschatoline ( 4) have been isolated from Pseudoxandra cuspidata bark, used in French Guiana as an antimalarial. Their structures were elucidated by spectroscopic analyses, especially 2D-NMR techniques (ADEQUATE and NOESY). We found that the antimalarial activity of this bark was mostly due to bis-benzylisoquinoline 1 S,1' R-rodiasine ( 2) (IC (50)= 1 microM) also displaying a low cytotoxicity.
Published: 2006
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22. 13C-NMR Study of the Ruthenium(II)-Catalyzed Degradation of Labeled G-Factor Endoperoxides: Proposal of a Likely Biogenetic Pathway and Evidence for CO2 Release

Author: Michel Baltas, Christiane André-Barrès, and Stéphane Massou
Subjects: chemistry.chemical_classification, Carbon Isotopes, Magnetic Resonance Spectroscopy, Chemistry, Stereochemistry, g-factor, chemistry.chemical_element, Bioengineering, General Chemistry, General Medicine, Nuclear magnetic resonance spectroscopy, Carbon Dioxide, Carbon-13 NMR, Biochemistry, Catalysis, Ruthenium, Peroxides, Electron transfer, Enzyme, Plant Growth Regulators, Molecular Medicine, Degradation (geology), Molecular Biology
Abstract: G-factor endoperoxides are natural products acting as phytohormones and contribute to root inhibition and frost resistance in Eucalyptus grandis and other Myrtaceae. Several (Z)- and (E)-configured cyclopentane-l,3-dione metabolites of G-factors have been found in some Eucalyptus extracts. Interestingly, these probable metabolites are identical to the products obtained by Ru II -catalyzed reduction of G-factors. In the present work, we, thus, studied the mechanism involved in the formation of these compounds by means of in-depth NMR distribution analysis of two differently 13 C-labeled G 3 -factors. The observed CO 2 release is underlined, and a comparison with the corresponding Fe H -induced degradation of G-factor endoperoxides is made. Also, the type of electron transfer proposed in the two processes is discussed and, tentatively, connected to the role of endoperoxides in plants.
Published: 2006
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23. Synthesis, Structures, and Reactivity of Mono- and Bis(ferrocenyl)-Substituted Group 14 Metallocenes

Author: Ali Mcheik, Stéphane Massou, Mounia Joudat, Heinz Gornitzka, Annie Castel, Pierre Rivière, Fabien Delpech, and Alix Sournia-Saquet
Subjects: Catechol, Stereochemistry, Organic Chemistry, chemistry.chemical_element, Crystal structure, Medicinal chemistry, Cycloaddition, Inorganic Chemistry, chemistry.chemical_compound, chemistry, Cyclopentadienyl complex, Electrophile, Moiety, Reactivity (chemistry), Lithium, Physical and Theoretical Chemistry
Abstract: New mono- and bis(ferrocenyl)-substituted germanocenes and stannocenes have been synthesized in good yield by the reaction between group 14 dichlorides (Cl2Ge·dioxane, Cl2Sn) and the lithium salts of the corresponding mono- and bis(cyclopentadienyl)-substituted ferrocenes. The X-ray crystal structure analyses of germanocenes 3a and 4a reveal a bent sandwich structure with an angle between the cyclopentadienyl planes of 45.4° and 40.9° for 3a and 4a, respectively. Mass spectrometry and electrochemical studies show the stabilization of the generated transient cations by ferrocenyl groups, particularly in the germanium series. Their reactions with electrophilic reagents such as catechol and cycloaddition with o-quinone also are reported and show, in the latter case, an increase of the specific reactivity of the dimethylcyclopentadienyl moiety.
Published: 2004
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24. Heterocyclisation of ferrocenyl- and benzenetricarbonyl chromium-cyclopentadienes with o-quinone. Evidence for a SET route

Author: Jacques Rouzaud, Fabien Delpech, Stéphane Massou, Annie Castel, Heinz Gornitzka, Pierre Rivière, and Mounia Joudat
Subjects: Chemistry, Stereochemistry, chemistry.chemical_element, Carbon-13 NMR, Medicinal chemistry, Cycloaddition, Ion, Inorganic Chemistry, Chromium, Stereospecificity, Transition metal, Cyclopentadienyl complex, Radical ion, Materials Chemistry, Physical and Theoretical Chemistry
Abstract: A series of cyclopentadienyl ligands substituted by unsaturated systems including various transition metal groups ((C5H5)2Fe, PhCr(CO)3) were reacted with 3,5-di-tert-butyl orthoquinone producing 1,4-benzodioxines in high yields. These cycloaddition reactions were stereospecific. 1H and 13C NMR studies and X-ray structural analysis are in agreement with the exclusive formation of “endo” cycloadducts. Moreover, an ESR study shows the transient formation of both the radical anion of the o-quinone and the radical cation of the dienic system indicating an alternative single electron transfer pathway.
Published: 2004
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25. A 1H NMR study of the specificity of α-l-arabinofuranosidases on natural and unnatural substrates

Author: Vinciane Borsenberger, Emmie Dornez, Stéphane Massou, Marie-Laure Desrousseaux, Michael J. O’Donohue, Régis Fauré, Claire Dumon, Maija Tenkanen, Christophe M. Courtin, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Lab Food Chem & Biochem, Université Catholique de Louvain = Catholic University of Louvain (UCL), Department of Food and Environmental Sciences, University of Helsinki, Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Université Catholique de Louvain (UCL), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and Helsingin yliopisto = Helsingfors universitet = University of Helsinki
Subjects: chemistry.chemical_classification, 0303 health sciences, 030306 microbiology, Chemistry, Chromogenic, Stereochemistry, di-arabinofuranosylated substrates, screening, Biophysics, food and beverages, enzymatic hydrolysis, specificity, Biochemistry, Xylan, 03 medical and health sciences, Hydrolysis, Enzyme, Enzymatic hydrolysis, Proton NMR, arabinoxylan arabinofuranohydrolases, Glycoside hydrolase, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Selectivity, Molecular Biology, 030304 developmental biology
Abstract: Background The detailed characterization of arabinoxylan-active enzymes, such as double-substituted xylan arabinofuranosidase activity, is still a challenging topic. Ad hoc chromogenic substrates are useful tools and can reveal subtle differences in enzymatic behavior. In this study, enzyme selectivity on natural substrates has been compared with enzyme selectivity towards aryl-glycosides. This has proven to be a suitable approach to understand how artificial substrates can be used to characterize arabinoxylan-active α- l -arabinofuranosidases (Abfs). Methods Real-time NMR using a range of artificial chromogenic, synthetic pseudo-natural and natural substrates was employed to determine the hydrolytic abilities and specificity of different Abfs. Results The way in which synthetic di-arabinofuranosylated substrates are hydrolyzed by Abfs mirrors the behavior of enzymes on natural arabinoxylo-oligosaccharide (AXOS). Family GH43 Abfs that are strictly specific for mono-substituted d -xylosyl moieties (AXH-m) do not hydrolyze synthetic di-arabinofuranosylated substrates, while those specific for di-substituted moieties (AXH-d) remove a single l -arabinofuranosyl ( l -Ara f ) group. GH51 Abfs, which are supposedly AXH-m enzymes, can release l -Ara f from disubstituted d -xylosyl moieties, when these are non-reducing terminal groups. Conclusions and general significance The present study reveals that although the activity of Abfs on artificial substrates can be quite different from that displayed on natural substrates, enzyme specificity is well conserved. This implies that carefully chosen artificial substrates bearing di-arabinofuranosyl d -xylosyl moieties are convenient tools to probe selectivity in new Abfs. Moreover, this study has further clarified the relative promiscuity of GH51 Abfs, which can apparently hydrolyze terminal disubstitutions in AXOS, albeit less efficiently than mono-substituted motifs.
Published: 2014
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26. Hydrolytic pathway of 5-fluorouracil in aqueous solutions for clinical use

Author: Myriam Malet-Martino, Joëlle Azéma, Stéphane Massou, Robert Martino, Rémi Legay, Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées
Subjects: Tris, Oxazolidine, Magnetic Resonance Spectroscopy, Double bond, Chemistry, Pharmaceutical, Clinical Biochemistry, Pharmaceutical Science, 030226 pharmacology & pharmacy, Aldehyde, Medicinal chemistry, Analytical Chemistry, 03 medical and health sciences, chemistry.chemical_compound, Hydrolysis, 0302 clinical medicine, Drug Discovery, Organic chemistry, [CHIM]Chemical Sciences, Sodium Hydroxide, Reactivity (chemistry), Uracil, Spectroscopy, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, Fluoroacetaldehyde, Aqueous solution, Hydrogen-Ion Concentration, Solutions, chemistry, 030220 oncology & carcinogenesis, Fluorouracil
Abstract: The purpose of the study was to investigate the degradation pathway of 5-fluorouracil (FU) in the situation of commercial formulations for clinical use, namely FU dissolved in sodium hydroxide (NaOH) solutions or Tris buffer at pH 8.5-9. Combination of data from (19)F, (1)H and (13)C NMR and in some cases MS led to the identification of 8 and 13 FU degradation products in NaOH and Tris solutions respectively. In FU NaOH solutions, the first stage of FU degradation is a stereoselective hydration of the C5-C6 double bond leading to 5,6-dihydro-5-fluoro-6-hydroxyuracil, the cis stereoisomer being predominant relative to the trans. The second stage involves either a defluorination step with formation of fluoride ion and 5-hydroxyuracil or the cleavage of the N3-C4 bond giving the two diastereoisomeric 2-fluoro-3-hydroxy-3-ureidopropanoic acids. The subsequent N1-C6 bond breakdown of these compounds releases urea and 2-fluoro-3-oxopropanoic acid (FOPA) which in turn losses easily carbon dioxide leading to the formation of fluoroacetaldehyde (Facet). The degradation pathway in FU-Tris solutions is identical, except that Tris reacts with the aldehydes FOPA and Facet to form oxazolidine adducts stable at pH 8.5 but in equilibrium with the aldehyde forms at physiological pH, whereas the high reactivity of free aldehydes leads to numerous unidentified degradation compounds all in very low amounts. The FOPA diastereoisomeric adducts react with Facet to form four diastereoisomeric fused bicyclic five-membered ring compounds. Facet and FOPA are highly cardiotoxic. In Tris formulations, they are trapped as stable oxazolidine adducts which release the free aldehydes at physiological pH thus explaining the higher cardiotoxicity of FU in Tris solutions compared to that of FU in NaOH solutions.
Published: 2014
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27. Synthesis, NMR characterization and pharmacological evaluation of ligands derived from diprenorphine for central opioid receptors imaging

Author: Pierre Tisnès, François Bourrel, Yann Prigent, Stéphane Massou, Michel Bergon, Michel Baltas, Jean-Paul Esquere, and Mathieu Tafani
Subjects: Chemistry, Stereochemistry, Organic Chemistry, Alcohol, Carbon-13 NMR, chemistry.chemical_compound, Acetylation, Spect imaging, medicine, Reactivity (chemistry), Physical and Theoretical Chemistry, Receptor, Diprenorphine, Two-dimensional nuclear magnetic resonance spectroscopy, medicine.drug
Abstract: The aim of this work was to explore the synthesis of a tosylated derivative of diprenorphine (DPN) able to be radiolabeled with either fluorine-18 or iodine-123, making it suitable for PET or SPECT imaging studies of central opioid receptors, respectively. The strategy was based on the reactivity of the C-19 alcohol tertiary function. As an unexpected deacetylation of the phenolic function of the diprenorphine occurred, the prosthetic group reacted with the deprotected C-3 phenolic function instead of the C-19 alcohol group. UV spectroscopy and 1H and 13C NMR studies provided good evidence for the 3-phenolic substituted diprenorphine structure. Thorough 2D NMR experiments such as 1H–1H COSY, 1H–1H TOCSY, 1H–13C HMQC, 1H–13C HMBC and 1H–1H NOESY allowed us to assign fully 3-O-[(Z)-4-fluorobut-2-enyl] diprenorphine (10a) and gave us an unambiguous proof of the C-3 prosthetic group position. In vitro binding studies showed low affinity for both fluoro and iodo derivatives of diprenorphine, Ki = 0.31 ± 0.05 and 0.09 ± 0.03 µM, respectively, for mouse brain membranes, these inhibition constants also being in agreement with a 3-phenolic substituted structure. Copyright © 2001 John Wiley & Sons, Ltd.
Published: 2001
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28. [Untitled]

Author: V. Puech, Stéphane Massou, Pascal Demange, Marielle Tropis, Alain Milon, Nic D. Lindley, and Franck Talmont
Subjects: chemistry.chemical_classification, biology, biology.organism_classification, Biochemistry, Yeast, Pichia pastoris, Amino acid, chemistry.chemical_compound, Membrane, chemistry, Biosynthesis, Membrane protein, Methanol, Heterologous expression, Spectroscopy
Abstract: Methylotrophic yeast has previously been shown to be an excellent system for the cost-effective production of perdeuterated biomass and for the heterologous expression of membrane receptors. A protocol for the expression of 85% deuterated, functional human μ-opiate receptor was established. For partially deuterated biomass, deuteration level and distribution were determined for fatty acids, amino acids and carbohydrates. It was shown that prior to biosynthesis of lipids and amino acids (and of carbohydrates, to a lower extent), exchange occurs between water and methanol hydrogen atoms, so that 80%–90% randomly deuterated biomass and over-expressed proteins may be obtained using only deuterated water.
Published: 1999
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29. NMR analyses of deuterated phospholipids isolated from Pichia angusta

Author: Alain Milon, Nic D. Lindley, Stéphane Massou, Marielle Tropis, and Sophie Augé
Subjects: chemistry.chemical_compound, Deuterium, Chemistry, Pichia angusta, Stereochemistry, Lipid composition, Phospholipid, Phosphatidyl inositol, Phosphatidylinositol, Biochemistry
Abstract: La composition en phospholipides de levures methylotrophes ayant pousse sur des milieux de cultures hydrogenes et deuteries a ete determinee par RMN du proton et du phosphore 31. L'utilisation d'un solvant d'affinement a permis d'obtenir des largeurs de raies inferieures a 2 Hz et de resoudre toutes les classes de phospholipides. Ils sont ensuite identifies par leur deplacement chimique et par des correlations phosphore - proton specifiques (experiences HMQC-HOHAHA gradients). Cette approche a permis une analyse qualitative et quantitative de melanges de phospholipides directement apres extraction au chloroforme-methanol. La composition en phospholipides est profondement modifiee lors de la croissance en milieu perdeuterie ou l'on observe un lipide majoritaire, le phosphatidyl Inositol (PI), au lieu d'un melange PC, PS, PI en milieu hydrogene.
Published: 1998
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30. A novel platform for automated high-throughput fluxome profiling of metabolic variants

Author: Jean-Charles Portais, Stéphane Massou, Serguei Sokol, Stéphanie Heux, Juliette Poinot, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), and Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MICROBIAL CULTIVATIONS, [SDV]Life Sciences [q-bio], CENTRAL CARBON METABOLISM, Metabolic network, Bioengineering, High-throughput, Computational biology, Biology, Applied Microbiology and Biotechnology, Peptide Mapping, ISOTOPE LABELING EXPERIMENTS, Metabolic engineering, Automation, QUANTITATIVE 2D NMR, INFORMATION-CONTENT, Escherichia coli, Profiling (information science), Fluxomics, MAMMALIAN-CELL CULTURES, business.industry, Escherichia coli Proteins, Gene Expression Profiling, Equipment Design, Robotics, Flux calculation, NMR, Metabolic Flux Analysis, Data flow diagram, Equipment Failure Analysis, Systems Integration, Workflow, Robotic systems, Biochemistry, CORYNEBACTERIUM-GLUTAMICUM, Statistical analysis, ESCHERICHIA-COLI, Batch Cell Culture Techniques, Mutation, Metabolome, CLOSTRIDIUM-ACETOBUTYLICUM, business, ZWF GENE-KNOCKOUT, Biotechnology
Abstract: Advances in metabolic engineering are enabling the creation of a large number of cell factories. However, high throughput platforms do not yet exist for rapidly analyzing the metabolic network of the engineered cells. To fill the gap, we developed an integrated solution for fluxome profiling of large sets of biological systems and conditions. This platform combines a robotic system for C-13-labelling experiments and sampling of labelled material with NMR-based isotopic fingerprinting and automated data interpretation. As a proof-of-concept, this workflow was applied to discriminate between Escherichia coli mutants with gradual expression of the glucose-6-phosphate dehydrogenase. Metabolic variants were clearly discriminated while pathways that support metabolic flexibility towards modulation of a single enzyme were elucidating. By directly connecting the data flow between cell cultivation and flux quantification, considerable advances in throughput, robustness, release of resources and screening capacity were achieved. This will undoubtedly facilitate the development of efficient cell factories. (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
Published: 2014
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31. Isotopic studies of metabolic systems by mass spectrometry: using pascal's triangle to produce biological standards with fully controlled labeling patterns

Author: Fabien Létisse, Jean-Charles Portais, Stéphane Massou, Pierre Millard, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), INRA (program CJS), European Union (267196), European Project: 267196,EC:FP7:PEOPLE,FP7-PEOPLE-2010-COFUND,AGREENSKILLS(2012), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA)
Subjects: Quality Control, [SDV]Life Sciences [q-bio], Analytical chemistry, INTRACELLULAR METABOLITES, Mass spectrometry, 01 natural sciences, Mass Spectrometry, Pichia, Analytical Chemistry, 03 medical and health sciences, Metabolic flux analysis, Escherichia coli, Isotopologue, 030304 developmental biology, FRAGMENT, 0303 health sciences, Isotope, integumentary system, Chemistry, EXTRACTS, 010401 analytical chemistry, Ms analysis, Reference Standards, FLUX ANALYSIS, 0104 chemical sciences, Metabolism, Isotopic ratio outlier analysis, Isotope Labeling, GC-MS, Biological system, Quantitative analysis (chemistry), Algorithms
Abstract: Mass spectrometry (MS) is widely used for isotopic studies of metabolism in which detailed information about biochemical processes is obtained from the analysis of isotope incorporation into metabolites. The biological value of such experiments is dependent on the accuracy of the isotopic measurements. Using MS, isotopologue distributions are measured from the quantitative analysis of isotopic clusters. These measurements are prone to various biases, which can occur during the experimental workflow and/or MS analysis. The lack of relevant standards limits investigations of the quality of the measured isotopologue distributions. To meet that need, we developed a complete theoretical and experimental framework for the biological production of metabolites with fully controlled and predictable labeling patterns. This strategy is valid for different isotopes and different types of metabolisms and organisms, and was applied to two model microorganisms, Pichia augusta and Escherichia coli, cultivated on (13)C-labeled methanol and acetate as sole carbon source, respectively. The isotopic composition of the substrates was designed to obtain samples in which the isotopologue distribution of all the metabolites should give the binomial coefficients found in Pascal's triangle. The strategy was validated on a liquid chromatography-tandem mass spectrometry (LC-MS/MS) platform by quantifying the complete isotopologue distributions of different intracellular metabolites, which were in close agreement with predictions. This strategy can be used to evaluate entire experimental workflows (from sampling to data processing) or different analytical platforms in the context of isotope labeling experiments.
Published: 2014
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32. Extending the Structural Diversity of alpha-Flavonoid Glycosides with Engineered Glucansucrases

Author: Pierre Monsan, Emmanuelle Cambon, Stéphane Massou, Isabelle André, Sandrine Morel, François Lefoulon, Sandra Pizzut-Serin, Yannick Malbert, Magali Remaud-Simeon, Sandrine Laguerre, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), SERVIER, and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Glycosylation, TRANSGLYCOSYLATION, glycosylation, Stereochemistry, [SDV]Life Sciences [q-bio], Mutant, Flavonoid, enzymes, water, BIOLOGICAL-ACTIVITIES, Catalysis, Inorganic Chemistry, chemistry.chemical_compound, GLUCOSYLATION, [SDV.IDA]Life Sciences [q-bio]/Food engineering, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, Physical and Theoretical Chemistry, CHEMOENZYMATIC SYNTHESIS, NEISSERIA-POLYSACCHAREA, ACCEPTOR REACTIONS, chemistry.chemical_classification, Flavonoid glycosides, ACTIVE-SITE, Organic Chemistry, fungi, protein engineering, Protein engineering, Bioavailability, AMYLOSUCRASE, carbohydrates (lipids), Enzyme, antioxidants, chemistry, Biochemistry, STEAROTHERMOPHILUS MALTOGENIC AMYLASE, LEUCONOSTOC-MESENTEROIDES, Luteolin
Abstract: International audience; Flavonoids constitute an important class of bioactive molecules, the physicochemical properties of which can be modulated by glucosylation. A structurally guided approach has been used to isolate glucansucrases modified in their acceptor-binding site and specialized for luteolin glucosylation. Of a small-size library, we isolate mutants showing up to an 8-fold increase in flavonoid conversion rate over that observed with the parental enzyme. Di- and triglucosylated luteolin derivatives never described before have been obtained. They exhibit 282- and 17708-fold increases in water solubility, respectively, and are protected from oxidation by the glucosylation reaction. Molecular docking enables insight into the product specificity of the best mutants. These results demonstrate that atransglucosylase engineering is a powerful means to generate highly specific catalysts for flavonoid glucosylation and deliver new structural scaffolds with increased bioavailability and high relevance for therapeutic applications.
Published: 2014
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33. Sampling of intracellular metabolites for stationary and non-stationary C-13 metabolic flux analysis in [i]Escherichia coli[/i]

Author: Christoph Wittmann, Stéphane Massou, Pierre Millard, Jean-Charles Portais, Fabien Létisse, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Université Fédérale Toulouse Midi-Pyrénées, Saarland University [Saarbrücken], INRA (CJS), European Project: 267196, Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and Université de Toulouse (UT)
Subjects: 0106 biological sciences, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], Biophysics, C-13-labeling experiments, Mass isotopomer, 01 natural sciences, Biochemistry, Models, Biological, 03 medical and health sciences, Isotopes, 010608 biotechnology, Metabolic flux analysis, Sampling procedures, Extracellular, Metabolomics, Isotopologue, Molecular Biology, 030304 developmental biology, 0303 health sciences, Carbon Isotopes, Quenching (fluorescence), Chromatography, Escherichia coli K12, Chemistry, Sampling (statistics), Cell Biology, Metabolism, Solvent, Isotope Labeling, Flux (metabolism)
Abstract: The analysis of metabolic intermediates is a rich source of isotopic information for (13)C metabolic flux analysis ((13)C-MFA) and extends the range of its applications. The sampling of labeled metabolic intermediates is particularly important to obtain reliable isotopic information. The assessment of the different sampling procedures commonly used to generate such data, therefore, is crucial. In this work, we thoroughly evaluated several sampling procedures for stationary and non-stationary (13)C-MFA using Escherichia coli. We first analyzed the efficiency of these procedures for quenching metabolism and found that procedures based on cold or boiling solvents are reliable, in contrast to fast filtration, which is not. We also showed that separating the cells from the broth is not necessary in isotopic stationary state conditions. On the other hand, we demonstrated that the presence of metabolic intermediates outside the cells strongly affects the transient isotopic data monitored during non-stationary (13)C-labeling experiments. Meaningful isotopic data can be obtained by recovering intracellular labeled metabolites from pellets of cells centrifuged in cold solvent. We showed that if the intracellular pools are not separated from the extracellular ones, accurate flux maps can be established provided that the contribution of exogenous compounds is taken into account in the metabolic flux model.
Published: 2014
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34. Structurally original oxathioethers macrocycles containing an exocyclic double-bond: synthesis, characterization, reactivity, and coordination

Author: Marc Vedrenne, Alina Saponar, David Madec, Stéphane Massou, Gabriela Nemes, Nathalie Saffon, Guillaume Carel, Annie Castel, Ghassoub Rima, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Babes-Bolyai University [Cluj-Napoca] (UBB), Centre National de la Recherche Scientifique (CNRS), Universite de Toulouse, Agence Nationale pour la Recherche ANR-08-CSOG-00, and PCCE 129/2008
Subjects: palladium(ii), Double bond, Stereochemistry, Polyether chain, Ether, Biochemistry, Sandwich-type compounds, thiacrown, chemistry.chemical_compound, ionophores, Thioether, Drug Discovery, [CHIM.CRIS]Chemical Sciences/Cristallography, Reactivity (chemistry), Hydroxymethyl, Oxathioethers, chemistry.chemical_classification, Bicyclic molecule, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, benzothiacrown ethers, thia-crown-ethers, polyether sulfide syntheses, NMR, Hydroboration, chemistry, Coordination, Benzyl group, derivatives, Macrocycles, ring, crystal-structures, metal-complexes
Abstract: International audience; New oxathioethers macrocycles have been synthesized and characterized. Each macrocycle consists in structurally defined ether and thioether moieties and an exocyclic double-bond (2a-c) or a hydroxymethyl group (3a-c). Macrocycles (2a-c) have been synthesized by reaction of dianions of thioethers dials (1a-c) with 3-chloro-2-chloromethylprop-1-ene. Their hydroboration/oxidation led to corresponding primary alcohols (3a-c). Structures of compounds (2b) and (3a) have been determined by X-ray diffraction. The reactivity of the hydroxyl group allowed the preparation of oxathioethers macrocycles bearing a polyether chain or a benzyl group (4a,b) and the synthesis of new bicyclic sandwich-type compounds (5a,b). The ability of these functionalized macrocycles to coordinate to palladium has been investigated.
Published: 2014
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35. A ¹H NMR study of the specificity of α-l-arabinofuranosidases on natural and unnatural substrates

Author: Vinciane, Borsenberger, Emmie, Dornez, Marie-Laure, Desrousseaux, Stéphane, Massou, Maija, Tenkanen, Christophe M, Courtin, Claire, Dumon, Michael J, O'Donohue, and Régis, Fauré
Subjects: Bacterial Proteins, Glycoside Hydrolases, Oligosaccharides, Bacillus, Nuclear Magnetic Resonance, Biomolecular, Substrate Specificity
Abstract: The detailed characterization of arabinoxylan-active enzymes, such as double-substituted xylan arabinofuranosidase activity, is still a challenging topic. Ad hoc chromogenic substrates are useful tools and can reveal subtle differences in enzymatic behavior. In this study, enzyme selectivity on natural substrates has been compared with enzyme selectivity towards aryl-glycosides. This has proven to be a suitable approach to understand how artificial substrates can be used to characterize arabinoxylan-active α-l-arabinofuranosidases (Abfs).Real-time NMR using a range of artificial chromogenic, synthetic pseudo-natural and natural substrates was employed to determine the hydrolytic abilities and specificity of different Abfs.The way in which synthetic di-arabinofuranosylated substrates are hydrolyzed by Abfs mirrors the behavior of enzymes on natural arabinoxylo-oligosaccharide (AXOS). Family GH43 Abfs that are strictly specific for mono-substituted d-xylosyl moieties (AXH-m) do not hydrolyze synthetic di-arabinofuranosylated substrates, while those specific for di-substituted moieties (AXH-d) remove a single l-arabinofuranosyl (l-Araf) group. GH51 Abfs, which are supposedly AXH-m enzymes, can release l-Araf from disubstituted d-xylosyl moieties, when these are non-reducing terminal groups.The present study reveals that although the activity of Abfs on artificial substrates can be quite different from that displayed on natural substrates, enzyme specificity is well conserved. This implies that carefully chosen artificial substrates bearing di-arabinofuranosyl d-xylosyl moieties are convenient tools to probe selectivity in new Abfs. Moreover, this study has further clarified the relative promiscuity of GH51 Abfs, which can apparently hydrolyze terminal disubstitutions in AXOS, albeit less efficiently than mono-substituted motifs.
Published: 2013

36. Fast Spatially Encoded 3D NMR Strategies for C-13-Based Metabolic Flux Analysis

Author: Benoît Charrier, Patrick Giraudeau, Renaud Boisseau, Stéphane Massou, Jean-Charles Portais, Serge Akoka, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), 'Agence Nationale de la Recherche' for young researchers (ANR) [2010-JCJC-0804-01], Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), and Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
Subjects: Magnetic Resonance Spectroscopy, Time Factors, [SDV]Life Sciences [q-bio], Analytical chemistry, ULTRAFAST 2D NMR, Measure (mathematics), Spectral line, Homonuclear molecule, 030218 nuclear medicine & medical imaging, Analytical Chemistry, 03 medical and health sciences, 0302 clinical medicine, Dimension (vector space), Metabolic flux analysis, Escherichia coli, SPECTRA, NUCLEAR-MAGNETIC-RESONANCE, PHYSIOLOGY, 030304 developmental biology, SENSITIVITY LOSSES, Coupling, 0303 health sciences, Carbon Isotopes, Chemistry, Nuclear magnetic resonance spectroscopy, SINGLE-SCAN, Metabolic Flux Analysis, J-RESOLVED SPECTROSCOPY, INTERMEDIARY METABOLISM, C-13 ISOTOPOMER ANALYSIS, Biological system, Two-dimensional nuclear magnetic resonance spectroscopy, ENRICHMENTS
Abstract: The measurement of site-specific (13)C enrichments in complex mixtures of (13)C-labeled metabolites is a powerful tool for metabolic flux analysis. One of the main methods to measure such enrichments is homonuclear (1)H 2D NMR. However, the major limitation of this technique is the acquisition time, which can amount to a few hours. This drawback was recently overcome by the design of fast COSY experiments for measuring specific (13)C-enrichments, based on single-scan 2D NMR. However, these experiments are still limited by overlaps because of(1)H-(13)C splittings, thus limiting the metabolic information accessible for complex biological mixtures. To circumvent this limitation, we propose to tilt the (1)H-(13)C coupling into a third dimension via fast-hybrid 3D NMR methods combining the speed of ultrafast 2D NMR with the high resolution of conventional methods. Two strategies are described that allow the acquisition of a complete 3D J-resolved-COSY spectrum in 12 min (for concentrations as low as 10 mM). The analytical potentialities of both methods are evaluated on a series of (13)C-enriched glucose samples and on a biomass hydrolyzate obtained from Escherichia coli cells. Once optimized, the two complementary experiments lead to a trueness and a precision of a few percent and an excellent linearity. The advantages and drawbacks of these approaches are discussed and their potentialities are highlighted.
Published: 2013
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37. Hydrogenation processes at the surface of ruthenium nanoparticles: A NMR study

Author: Stéphane Massou, Emmanuelle Teuma, Montserrat Gómez, Bruno Chaudret, Isabelle Favier, P. Lavedan, Karine Philippot, Laboratoire Hétérochimie Fondamentale et Appliquée (LHFA), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Institut de Chimie de Toulouse (ICT-FR 2599), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire de chimie de coordination (LCC), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de physique et chimie des nano-objets (LPCNO), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie de Toulouse (ICT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut de Chimie de Toulouse (ICT), Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), and Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], chemistry.chemical_element, Arenes, NOE effects, 010402 general chemistry, Photochemistry, 01 natural sciences, Catalysis, Ruthenium, Styrene, chemistry.chemical_compound, Adsorption, Surface reactivity, Desorption, Pyridine, [SDV.IDA]Life Sciences [q-bio]/Food engineering, Molecule, Reactivity (chemistry), [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, 010405 organic chemistry, DOSY NMR, General Chemistry, Nuclear magnetic resonance spectroscopy, 0104 chemical sciences, chemistry, Nanoparticles, Hydrogenation, Hydrogen transfer
Abstract: 1022-5528; The reactivity of ruthenium nanoparticles stabilized by 4-(3-phenylpropyl)pyridine in hydrogen transfer and hydrogenation processes was monitored by NMR spectroscopy. Unsaturated substrates such as styrene, 4-vinylpyridine and 4-phenyl-but-3-en-2-one were used as model molecules to investigate the surface properties of nanoparticles by a combination of NMR studies. Interestingly, the hydrides present at the metallic surface after nanoparticles synthesis are selectively transferred to vinylic groups without reducing the aromatic rings, under dihydrogen-free atmosphere. DOSY and NOE NMR experiments permitted to propose a way of interaction of the organic compounds at the metallic surface. In particular, the coordination of the substrate could be evidenced for 4-vinylpyridine and 4-ethylpyridine but not for styrene derivatives. Curved double arrows represent magnetization exchanges. Straight arrows represent adsorption/desorption phenomena.
Published: 2013
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38. Isotopic profiling of ¹³C-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy

Author: Edern, Cahoreau, Lindsay, Peyriga, Jane, Hubert, Frédéric, Bringaud, Stéphane, Massou, and Jean-Charles, Portais
Subjects: Carbon Isotopes, Trypanosoma brucei brucei, Escherichia coli, Amino Acids, Complex Mixtures, Nuclear Magnetic Resonance, Biomolecular, High-Throughput Screening Assays
Abstract: The use of two-dimensional heteronuclear J-resolved (2D H-JRES) nuclear magnetic resonance (NMR) spectroscopy for fast and reliable measurement of isotopic patterns from ¹³C-enriched compounds resulting from carbon labeling experiments was evaluated. Its use with biological samples of increasing complexity showed that 2D H-JRES spectroscopy is suitable for high-throughput isotopic profiling of any kind of labeled samples. Moreover, the method enabled accurate quantification of ¹³C enrichments and, thus, can be used for metabolic flux analysis. The excellent trade-off between reduced experimental time and the number of measurable isotopic data makes 2D H-JRES NMR a promising approach for high-throughput flux analysis of samples of intermediate complexity.
Published: 2012

39. Photobioreactor design for isotopic non-stationary 13C-metabolic flux analysis (INST 13C-MFA) under photoautotrophic conditions

Author: Lindsay Peyriga, Jeremy Pruvost, Guillaume Cogne, Emmanuel Dechandol, Olivier Gonçalves, Jack Legrand, Jean-Charles Portais, Fabienne Le Grand, Arnaud Martzolff, Edern Cahoreau, Stéphane Massou, Laboratoire de génie des procédés - environnement - agroalimentaire (GEPEA), Université de Nantes (UN)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), French National Research Agency project ALGOMICS [ANR-08-BIOE-002], FP7 European project [SOLAR-H2], Region Midi-Pyrenees, European Regional Development Fund (ERDF), French Ministry for Higher Education Research, SICOVAL, IBiSa, Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Université Bretagne Loire (UBL)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Bioprocédés Appliqués aux Microalgues (GEPEA-BAM), Institut Universitaire de Technologie - Nantes (IUT Nantes), Université de Nantes (UN)-Université de Nantes (UN)-Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Institut Universitaire de Technologie Saint-Nazaire (IUT Saint-Nazaire), Université de Nantes (UN)-Ecole Polytechnique de l'Université de Nantes (EPUN), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Institut Universitaire de Technologie - La Roche-sur-Yon (IUT La Roche-sur-Yon), Université de Nantes (UN)-Institut Universitaire de Technologie - Nantes (IUT Nantes), Université de Nantes (UN), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), ANR-08-BIOE-0002,ALGOMICS,ETUDES GLOBALES DE LA CONVERSION ET DU STOCKAGE DE L'ENERGIE CHEZ LES MICROALGUES(2008), Université de Nantes (UN)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL)-IMT Atlantique (IMT Atlantique), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), MetaToul FluxoMet (TBI-MetaToul), MetaboHUB-MetaToul, MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université de Nantes (UN)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Universitaire Européen de la Mer (IUEM), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Université de Brest (UBO)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS), Bioprocédés Appliqués aux Microalgues ( GEPEA-BAM ), Laboratoire de génie des procédés - environnement - agroalimentaire ( GEPEA ), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique ( ONIRIS ) -Université Bretagne Loire ( UBL ) -IMT Atlantique Bretagne-Pays de la Loire ( IMT Atlantique ) -Université de Nantes ( UN ) -Centre National de la Recherche Scientifique ( CNRS ) -Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique ( ONIRIS ) -Université Bretagne Loire ( UBL ) -IMT Atlantique Bretagne-Pays de la Loire ( IMT Atlantique ) -Université de Nantes ( UN ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés ( LISBP ), Institut National de la Recherche Agronomique ( INRA ) -Institut National des Sciences Appliquées - Toulouse ( INSA Toulouse ), Institut National des Sciences Appliquées ( INSA ) -Institut National des Sciences Appliquées ( INSA ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire des Sciences de l'Environnement Marin (LEMAR) ( LEMAR ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Brest ( UBO ) -Institut Français de Recherche pour l'Exploitation de la Mer ( IFREMER ) -Institut Universitaire Européen de la Mer ( IUEM ), Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées, Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Université Bretagne Loire (UBL)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), and Institut de Recherche pour le Développement (IRD)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0106 biological sciences, TORUS PHOTOBIOREACTOR, carbon labeling experiment, [SDV]Life Sciences [q-bio], Analytical chemistry, 01 natural sciences, 7. Clean energy, Applied Microbiology and Biotechnology, Isotopomers, law.invention, Photobioreactors, law, Metabolic flux analysis, Dissolved organic carbon, MICROORGANISMS, Biomass, [ CHIM.GENI ] Chemical Sciences/Chemical engineering, ComputingMilieux_MISCELLANEOUS, Carbon Isotopes, 0303 health sciences, Chemistry, Systems Biology, Fatty Acids, Hydrogen-Ion Concentration, isotopomer analysis, Biotechnology, Photobioreactor, Bioengineering, CHLAMYDOMONAS-REINHARDTII, MICROALGAE, 03 medical and health sciences, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, photobioreactor, 010608 biotechnology, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, photoautotrophic metabolism, SHORT-TIME-SCALE, Filtration, 030304 developmental biology, Chromatography, Reproducibility of Results, Substrate (chemistry), MASS-SPECTROMETRY, Liquid nitrogen, METABOLOME ANALYSIS, isotopic non-stationary 13C-metabolic flux analysis, isotopic non-stationary 13 C-metabolic flux analysis, [ CHIM.ANAL ] Chemical Sciences/Analytical chemistry, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, Flux (metabolism), Chlamydomonas reinhardtii, LABELING EXPERIMENTS
Abstract: International audience; Adaptive metabolic behavior of photoautotrophic microorganisms toward genetic and environmental perturbations can be interpreted in a quantitative depiction of carbon flow through a biochemical reaction network using isotopic non-stationary 13C-metabolic flux analysis (INST 13C-MFA). To evaluate 13C-metabolic flux maps for Chlamydomonas reinhardtii, an original experimental framework was designed allowing rapid, reliable collection of high-quality isotopomer data against time. It involved (i) a short-time 13C labeling injection device based on mixing control in a torus-shaped photobioreactor with plug-flow hydrodynamics allowing a sudden step-change in the 13C proportion in the substrate feed and (ii) a rapid sampling procedure using an automatic fast filtration method coupled to a manual rapid liquid nitrogen quenching step. 13C-substrate labeling enrichment was controlled through the total dissolved inorganic carbon concentration in the pulsed solution. First results were obtained from steady-state continuous culture measurements allowing the characterization of the kinetics of label incorporation into light-limited growing cells cultivated in a photobioreactor operating at the maximal biomass productivity for an incident photon flux density of 200 mu molm-2s-1. 13C label incorporation was measured for 21 intracellular metabolites using IC-MS/MS in 58 samples collected across a labeling experiment duration of 7 min. The fastest labeling rate was observed for 2/3-phosphoglycerate with an apparent isotopic stationary state reached after 300s. The labeling rate was consistent with the optimized mixing time of about 4.9s inside the reactor and the shortest reliable sampling period assessed at 5s. Biotechnol. Bioeng. 2012; 109: 30303040. (C) 2012 Wiley Periodicals, Inc.
Published: 2012
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40. A Versatile and Colorful Screening Tool for the Identification of Arabinofuranose-Acting Enzymes

Author: Fernando Ferreira, Annick Pollet, Marie-Laure Desrousseaux, Christophe M. Courtin, Michael J. O’Donohue, Vinciane Borsenberger, Stéphane Massou, Emmie Dornez, Régis Fauré, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Lab Food Chem & Biochem, Université Catholique de Louvain = Catholic University of Louvain (UCL), Region Midi-Pyrenees [DAER-Recherche 10008500, 07009817], Agence Nationale de la Recherche (CP2D PENTOVAL), European Regional Development Fund, SICOVAL, Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), and Université Catholique de Louvain (UCL)
Subjects: [SDV]Life Sciences [q-bio], Chromogenic Substrates, Catechols, carbohydrates, Bifidobacterium adolescentis, Biology, 010402 general chemistry, ARABINOXYLAN, 01 natural sciences, Biochemistry, high-throughput screening, 03 medical and health sciences, chromogenic substrates, nitrocatechol, Escherichia coli, Glycoside hydrolase, Screening tool, Biorefining, ALPHA-L-ARABINOFURANOSIDASES, Molecular Biology, 030304 developmental biology, Enzyme Assays, chemistry.chemical_classification, 0303 health sciences, Hydrolysis, Organic Chemistry, Nitro Compounds, BIFIDOBACTERIUM-ADOLESCENTIS, Arabinose, 0104 chemical sciences, OLIGOSACCHARIDES, Enzyme, chemistry, RESIDUES, Molecular Medicine, glycoside hydrolases, Identification (biology), Colorimetry
Abstract: because they provide quick and easy detectionof enzyme activity at the microbial colony level and are com-patible with automated strategies. Arabinofuranose-acting gly-coside hydrolases (a-l-arabinofuranosidases and related arabi-nofuranose hydrolases) are often components of naturalenzyme systems that break down plant cell walls, specificallyby targeting polysaccharides such as arabinoxylans. Thesepolymers are widespread and abundant in industrial and agri-cultural residues, such as fruit pulp and straw (Scheme 1A). Ac-cordingly, increasing attention is now being given to arabino-furanose hydrolases, because these can contribute to the de-velopment of enzyme-based biorefining processes and pavethe way to more sustainable industrial strategies.
Published: 2012
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41. UFJCOSY: A Fast 3D NMR Method for Measuring Isotopic Enrichments in Complex Samples

Author: Meerakhan Pathan, Patrick Giraudeau, Stéphane Massou, Serge Akoka, Edern Cahoreau, Jean-Charles Portais, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), French National Research Agency (ANR) [QUANTUM 2010-JCJC-0804-01, ANR 08-BIOE-002], Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), MetaToul FluxoMet (TBI-MetaToul), MetaboHUB-MetaToul, MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
Subjects: Deuterium NMR, Time Factors, [SDV]Life Sciences [q-bio], Analytical chemistry, Fluorine-19 NMR, Complex Mixtures, 010402 general chemistry, ULTRAFAST 2D NMR, 01 natural sciences, 03 medical and health sciences, Escherichia coli, SPECTRA, Phosphorus-31 NMR spectroscopy, Physical and Theoretical Chemistry, PHYSIOLOGY, Nuclear Magnetic Resonance, Biomolecular, isotopic labeling, 030304 developmental biology, SENSITIVITY LOSSES, 0303 health sciences, Carbon Isotopes, Alanine, Chemistry, MIXTURES, Pulse sequence, spatial encoding, Nuclear magnetic resonance spectroscopy, SINGLE-SCAN, Atomic and Molecular Physics, and Optics, J-RESOLVED SPECTROSCOPY, 0104 chemical sciences, Heteronuclear molecule, RESOLUTION, 3D NMR spectroscopy, Isotope Labeling, ultrafast NMR spectroscopy, INTERMEDIARY METABOLISM, PRINCIPLES, Proton NMR, Two-dimensional nuclear magnetic resonance spectroscopy, metabolism
Abstract: Methods based on C isotopic enrichment are common procedures for metabolite quantification and metabolic flux analysis in complex biological samples. 2D NMR spectroscopy is a powerful tool in this field, as it offers the capacity of measuring site-specific C enrichments in complex samples. However, existing methods relying on 2D NMR are hampered by numerous overlaps when complex mixtures are studied. Here, we propose a fast 3D NMR method, based on ultrafast spatially-encoded NMR, which gives unambiguous access to isotopic enrichments (IEs) in biological mixtures in a few minutes, an experiment duration roughly 60 times shorter than a conventional 3D NMR approach. The combination of NMR with C-labeling strategies is a powerful tool for studying metabolic fluxes in living systems. Biological samples are generally incubated with Clabeled substrates, and the metabolic fluxes in complex networks can be estimated from the labeling pattern of metabolites. As the isotope distribution is generally not uniform over a given molecule, analytical methods must be able to measure site-specific C enrichments (i.e. the C enrichment for each position of each molecule). The simplest procedure consists of integrating the C-satellite peaks on H NMR spectra, 13] however this approach is impractical due to the large overlap between peaks in complex mixtures. Several approaches based on 2D H NMR have been designed to circumvent this drawback. Homonuclear 2D spectra are recorded, where H C couplings are expressed in one dimension only, which reduces overlaps while permitting the determination of site-specific isotopic enrichments by extracting rows (parallel to the F2 dimension) from the 2D spectrum. Whereas this approach initially suffered from long experiment durations, we recently proposed a strategy to reduce the experiment time by several orders of magnitude, based on the ultrafast 2D NMR methodology. This acquisition strategy, relying on spatial encoding, makes it possible to record 2D NMR spectra in a single scan— or a few scans when signal averaging is necessary for sensitivity purposes. However, both conventional and ultrafast 2D NMR methods are still affected by signal overlaps due to the H C splitting (typically 100–200 Hz) in one of the two dimensions. Consequently, many isotopomers cannot be resolved by these methods, thus limiting the metabolic information accessible for complex biological mixtures. 14] In order to bypass this limitation, it would be interesting to tilt the H C splittings in a third dimension, thus avoiding overlaps caused by heteronuclear couplings while preserving the information on C enrichments. Such an acquisition strategy, however, would require the sampling of an additional spectral dimension, thus increasing the acquisition duration to several tens of hours, which would be highly impractical. This communication presents a new NMR acquisition strategy permitting the acquisition of 3D data in a few minutes, based on the ultrafast 2D NMR strategy developed by Frydman and described in recent papers. In the corresponding NMR pulse sequence (Figure 1), two dimensions are recorded in an ultrafast fashion, while the third one is recorded in a conventional way. Therefore, a full 3D spectrum can be recorded in the time generally required to obtain a conventional 2D spectrum. The pulse sequence starts like a conventional correlation spectroscopy (COSY) experiment, with a linear time incrementation to sample the H chemical shifts in the first dimension, including a 1808 C pulse in the middle of this period to refocus the heteronuclear couplings. The 908 coherence transfer pulse is then followed by a constant-time spatial encoding scheme, while the heteronuclear couplings are again refocused by a 1808 pulse. Finally, the information is decoded in an echo-planar-imaging fashion (EPI), and 1808 pulses are applied during the detection to refocus chemical shift interactions. Therefore, the H C splittings are detected in the third dimension, similarly to the ultrafast heteronuclear J-resolved strategy that we have described recently. Following the acquisition, a processing specific to ultrafast acquisitions is applied (see Experimental Section). Figure 2a presents the 3D NMR spectrum obtained in 11 min on a model alanine sample with this pulse sequence. The H chemical shifts are expressed in dimensions F1 and F2, the conventionally sampled and spatially encoded dimensions, respectively. A F1F2 plane read from the 3D spectrum gives rise to a COSY-type correlation (Figure 2b), where the peaks are not broadened by heteronuclear couplings in spite of the C enrichment, contrary to the 2D methods previously published. The heteronuclear couplings are obtained in di[a] Dr. P. Giraudeau, M. Pathan, Prof. S. Akoka EBSI team, CEISAM UMR 6230, Universit de Nantes, CNRS B.P. 92208, 2 rue de la Houssini re, 44322 Nantes Cedex 03 (France) E-mail : patrick.giraudeau@univ-nantes.fr [b] E. Cahoreau, Dr. S. Massou, Prof. J.-C. Portais Universit de Toulouse; INSA, UPS, INP 135 Avenue de Rangueil, 31077 Toulouse (France) [c] E. Cahoreau, Dr. S. Massou, Prof. J.-C. Portais INRA, UMR792 Ing nierie des Syst mes Biologiques et des Proc d s 31400 Toulouse (France) [d] E. Cahoreau, Dr. S. Massou, Prof. J.-C. Portais CNRS, UMR5504, 31400 Toulouse (France) [**] UFJCOSY: Ultrafast J-Resolved Correlation Spectroscopy
Published: 2012
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42. Genome-scale reconstruction and system level investigation of the metabolic network of Methylobacterium extorquens AM1

Author: Kathrin Schneider, Julia A. Vorholt, Stéphane Massou, Patrick Kiefer, Jean-Charles Portais, Rémi Peyraud, Institute of Microbiology, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), We thank Philipp Christen for cultivation of M. extorquens AM1 in bioreactors. We thank Birgit Roth Zgraggen of the Functional Genomics Center Zurich for performing amino acid quantification. This work was supported by ETH Zurich, Research Grant ETH-25 08-2. The Swiss Academy of Engineering Science (SATW) and the Centre Francais pour l'Accueil et les Echanges Internationaux (Egide) supported the work with a travel grant (Germaine de Stael program). The work carried out at the LISBP (Toulouse, France) was supported by the Region Midi-Pyrenees, the European Regional Development Fund (ERDF), the French Ministry for Higher Education & Research, the SICOVAL, and the Reseau RMN Midi-Pyrenees., Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and Portais, Jean-Charles
Subjects: genome, bacterial, [SDV]Life Sciences [q-bio], physiological, Genome scale, Metabolic network, adaptation, Genome, Metabolic Network, Structural Biology, adaptation, physiological, [SDV.IDA]Life Sciences [q-bio]/Food engineering, lcsh:QH301-705.5, 0303 health sciences, Applied Mathematics, systems biology, Computer Science Applications, Methylotrophic Bacterium, models, biological, methylobacterium extorquens, metabolic networks and pathways, mathematical and computational biology, Modeling and Simulation, Methylobacterium extorquens, bactérie méthylotrophe, Research Article, Glyoxylate, Glyoxylate Cycle, Central Carbon Metabolism, Systems biology, Genomics, Computational biology, Biology, models, 03 medical and health sciences, Modelling and Simulation, System level, bacterial, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, genome, Molecular Biology, 030304 developmental biology, Facultative, 030306 microbiology, business.industry, biology.organism_classification, Biotechnology, lcsh:Biology (General), réseau métabolique, business, biological
Abstract: Background Methylotrophic microorganisms are playing a key role in biogeochemical processes - especially the global carbon cycle - and have gained interest for biotechnological purposes. Significant progress was made in the recent years in the biochemistry, genetics, genomics, and physiology of methylotrophic bacteria, showing that methylotrophy is much more widespread and versatile than initially assumed. Despite such progress, system-level description of the methylotrophic metabolism is currently lacking, and much remains to understand regarding the network-scale organization and properties of methylotrophy, and how the methylotrophic capacity emerges from this organization, especially in facultative organisms. Results In this work, we report on the integrated, system-level investigation of the metabolic network of the facultative methylotroph Methylobacterium extorquens AM1, a valuable model of methylotrophic bacteria. The genome-scale metabolic network of the bacterium was reconstructed and contains 1139 reactions and 977 metabolites. The sub-network operating upon methylotrophic growth was identified from both in silico and experimental investigations, and 13C-fluxomics was applied to measure the distribution of metabolic fluxes under such conditions. The core metabolism has a highly unusual topology, in which the unique enzymes that catalyse the key steps of C1 assimilation are tightly connected by several, large metabolic cycles (serine cycle, ethylmalonyl-CoA pathway, TCA cycle, anaplerotic processes). The entire set of reactions must operate as a unique process to achieve C1 assimilation, but was shown to be structurally fragile based on network analysis. This observation suggests that in nature a strong pressure of selection must exist to maintain the methylotrophic capability. Nevertheless, substantial substrate cycling could be measured within C2/C3/C4 inter-conversions, indicating that the metabolic network is highly versatile around a flexible backbone of central reactions that allows rapid switching to multi-carbon sources. Conclusions This work emphasizes that the metabolism of M. extorquens AM1 is adapted to its lifestyle not only in terms of enzymatic equipment, but also in terms of network-level structure and regulation. It suggests that the metabolism of the bacterium has evolved both structurally and functionally to an efficient but transitory utilization of methanol. Besides, this work provides a basis for metabolic engineering to convert methanol into value-added products., BMC Systems Biology, 5, ISSN:1752-0509
Published: 2011
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43. Ultrafast Quantitative 2D NMR: An Efficient Tool for the Measurement of Specific Isotopic Enrichments in Complex Biological Mixtures

Author: Stéphane Massou, Yoann Robin, Edern Cahoreau, Serge Akoka, Jean-Charles Portais, Patrick Giraudeau, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire Charles Coulomb (L2C), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Region Midi-Pyrenees, European Regional Development Fund (ERDF), French Ministry for Higher Education Research, SICOVAL, reseau RMN Midi-Pyrenees, Agence Nationale de la Recherch [ANR-08-BIO-002-02], Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), MetaToul FluxoMet (TBI-MetaToul), MetaboHUB-MetaToul, MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)
Subjects: Protein Hydrolysates, ISOTOPOMERS, Analytical chemistry, 010402 general chemistry, 01 natural sciences, Homonuclear molecule, Analytical Chemistry, Escherichia coli, SPECTRA, [CHIM]Chemical Sciences, AMINO-ACIDS, Biomass, Single scan, NUCLEAR-MAGNETIC-RESONANCE, C-13 NMR, Nuclear Magnetic Resonance, Biomolecular, Fluxomics, SENSITIVITY LOSSES, Carbon Isotopes, SPECTROSCOPY, 010405 organic chemistry, Chemistry, Linearity, Limiting, SINGLE-SCAN, 0104 chemical sciences, RESOLUTION, INTERMEDIARY METABOLISM, Biological system, Two-dimensional nuclear magnetic resonance spectroscopy, Ultrashort pulse
Abstract: International audience; Two-dimensional nuclear magnetic resonance (2D NMR) is a promising tool for studying metabolic fluxes by measuring C-13-enrichments in complex mixtures of C-13-labeled metabolites. However, the methods reported so far are hampered by very long acquisition durations limiting the use of 2D NMR as a quantitative tool for fluxomics. In this paper, we propose a new approach for measuring specific C-13-enrichments in a very fast way, by using new experiments based on ultrafast 2D NMR. Two homonudear 2D experiments (ultrafast COSY and zTOCSY) are proposed to measure C-13-enrichments in a single scan. Their advantages and limitations are discussed, and their high analytical potentialities are highlighted. Both methods are characterized by an accuracy of 1-2%, an average precision of 3%, and an excellent linearity. The analytical performance is equivalent or better than any of the conventional methods previously reported. The two ultrafast experiments are applied to the measurement of C-13-enrichments on a biomass hydrolyzate, showing the first known application of ultrafast 2D NMR to a real biological extract. The experiment duration is divided by 200 compared to the conventional methods, while preserving 80% of the quantitative information. This new approach opens new perspectives of application for fluxomics and metabonomics.
Published: 2011
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44. NMR-Based Structural Glycomics for High-Throughput Screening of Carbohydrate-Active Enzyme Specificity

Author: Olivier Saurel, Magali Remaud-Simeon, Jean-Charles Portais, Gabrielle Potocki-Véronèse, Romain Irague, Pierre Monsan, Alain Milon, Claire Moulis, Stéphane Massou, Unité mixte de service Toulouse White Biotechnology, Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Region Midi-Pyrenees, European Regional Development Fund, Institut National de la Recherche Agronomique (INRA), European Structural funds, Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT), and Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Magnetic Resonance Spectroscopy, High-throughput screening, [SDV]Life Sciences [q-bio], Oligosaccharides, ALPHA-D-GLUCANS, 01 natural sciences, Analytical Chemistry, Substrate Specificity, Glycomics, 03 medical and health sciences, Gene bank, [SDV.IDA]Life Sciences [q-bio]/Food engineering, TOOL, Genomic library, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, Carbohydrate composition, 030304 developmental biology, Enzyme Assays, Gene Library, chemistry.chemical_classification, 0303 health sciences, SPECTROSCOPY, CONSTRUCTION, 010405 organic chemistry, Chemistry, Glycosyltransferases, Stereoisomerism, Carbohydrate, LIBRARIES, 0104 chemical sciences, Enzymes, Enzyme, Biochemistry, Mutation, Biocatalysis, Carbohydrate Metabolism, Active enzyme
Abstract: International audience; We report here the development of a straightforward, sensitive, and quantitative NMR-based method for high-throughput characterization of carbohydrate structure and screening of carbohydrate active enzyme (CAZyme) specificity. Automated assays starting from gene library expression to carbohydrate structure determination directly from crude reaction media have been established and successfully used to screen a library of 4032 CAZymes obtained by combinatorial engineering, at a rate of 480 enzyme variants per day. This allowed one to accurately discriminate 303 enzyme variants with altered specificity. The results demonstrate the potential of high-throughput NMR technology in novel biocatalysts in glycomics, to mine artificial and natural enzyme diversity for
Published: 2011
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45. Carboxyfluorescein fluorescence experiments

Author: Stéphane Massou, Renaud Albigot, and Michel Prats
Subjects: Biochemistry
Published: 2000
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46. Demonstration of the ethylmalonyl-CoA pathway by using 13C metabolomics

Author: Philipp Christen, Stéphane Massou, Patrick Kiefer, Jean-Charles Portais, Rémi Peyraud, and Julia A. Vorholt
Subjects: Time Factors, Glyoxylate cycle, Glycine, Metabolic network, Mass Spectrometry, Serine, chemistry.chemical_compound, Metabolomics, Isomerism, Methylmalonyl-CoA, Methylobacterium extorquens, Carbon Isotopes, Multidisciplinary, biology, Methanol, Glyoxylates, Assimilation (biology), Esters, Biological Sciences, biology.organism_classification, Kinetics, chemistry, Biochemistry, Isotope Labeling, Methylotroph, Acyl Coenzyme A, Metabolic Networks and Pathways, Chromatography, Liquid
Abstract: The assimilation of one-carbon (C1) compounds, such as methanol, by serine cycle methylotrophs requires the continuous regeneration of glyoxylate. Instead of the glyoxylate cycle, this process is achieved by a not yet established pathway where CoA thioesters are known to play a key role. We applied state-of-the-art metabolomics and 13 C metabolomics strategies to demonstrate how glyoxylate is generated during methylotrophic growth in the isocitrate lyase-negative methylotroph Methylobacterium extorquens AM1. High-resolution mass spectrometry showed the presence of CoA thioesters specific to the recently proposed ethylmalonyl-CoA pathway. The operation of this pathway was demonstrated by short-term 13 C-labeling experiments, which allowed determination of the sequence of reactions from the order of label incorporation into the different CoA derivatives. Analysis of 13 C positional enrichment in glycine by NMR was consistent with the predicted labeling pattern as a result of the operation of the ethylmalonyl-CoA pathway and the unique operation of the latter for glyoxylate generation during growth on methanol. The results also revealed that 2 molecules of glyoxylate were regenerated in this process. This work provides a complete pathway for methanol assimilation in the model methylotroph M. extorquens AM1 and represents an important step toward the determination of the overall topology of its metabolic network. The operation of the ethylmalonyl-CoA pathway in M. extorquens AM1 has major implications for the physiology of these methylotrophs and their role in nature, and it also provides a common ground for C1 and C2 compound assimilation in isocitrate lyase-negative bacteria.
Published: 2009

47. Response of the central metabolism of Escherichia coli to modified expression of the gene encoding the glucose-6-phosphate dehydrogenase

Author: Nic D. Lindley, Philippe Soucaille, Jean-Charles Portais, Jens O. Krömer, Patrick Kiefer, Stéphane Massou, Cécile Nicolas, Christoph Wittmann, Fabien Létisse, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire réactivité et chimie des solides (LRCS), Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS), Institute of Microbiology, University of Milan, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Saarland University [Saarbrücken], Institute of Systems Biotechnology, Saarland University, Laboratoire réactivité et chimie des solides - UMR CNRS 7314 (LRCS), Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Picardie Jules Verne (UPJV)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Università degli Studi di Milano = University of Milan (UNIMI), and Université de Toulouse (UT)
Subjects: 0106 biological sciences, Anabolism, [SDV]Life Sciences [q-bio], Biophysics, Glucosephosphate Dehydrogenase, Pentose phosphate pathway, Biology, medicine.disease_cause, Models, Biological, 01 natural sciences, Biochemistry, Gas Chromatography-Mass Spectrometry, 13C-labellingexperiments, 03 medical and health sciences, chemistry.chemical_compound, Structural Biology, 010608 biotechnology, Genetics, medicine, Escherichia coli, Glucose-6-phosphate dehydrogenase, Computer Simulation, Molecular Biology, Gene, 030304 developmental biology, Mass isotopomer analysis, Fluxomics, 13C-labelling experiments, 0303 health sciences, Metabolic network, Gene Expression Regulation, Bacterial, Cell Biology, Metabolism, Citric acid cycle, chemistry, Genes, Bacterial, Flux (metabolism), Gene Deletion
Abstract: International audience; The deletion of thezwfgene encoding G6PDH activ-ity led to restructuring of the carbon flux through central metab-olism inEscherichia coli, though over-expression of this gene hadonly minor consequences for overall carbon flux. The modifiedcarbon flux seen in thezwfdeletion mutant enabled alternativeroutes of anabolic precursor formation and an adequate supplyof NADPH synthesis via a modified TCA cycle to be generatedso as to sustain growth rates comparable to the WT.Ó2007 Federation of European Biochemical Societies.Published by Elsevier B.V. All rights reserved.
Published: 2007
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48. NMR-based fluxomics: Quantitative 2D NMR methods for isotopomers analysis

Author: Jean-Charles Portais, Fabien Létisse, Stéphane Massou, Cécile Nicolas, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire réactivité et chimie des solides (LRCS), Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Laboratoire réactivité et chimie des solides - UMR CNRS 7314 (LRCS), Université de Picardie Jules Verne (UPJV)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
Subjects: [SDV]Life Sciences [q-bio], Analytical chemistry, Plant Science, Horticulture, 010402 general chemistry, 01 natural sciences, Biochemistry, Isotopomers, 03 medical and health sciences, Specific 13C-enrichments, Escherichia coli, Computer Simulation, Quantitative 2D NMR, Nuclear Magnetic Resonance, Biomolecular, Molecular Biology, Analysis method, Fluxomics, 030304 developmental biology, 13C-labelling experiments, Carbon Isotopes, 0303 health sciences, Alanine, Chemistry, 2D ZQF-TOCSY, 2D-COSY, General Medicine, 0104 chemical sciences, Biological system, Two-dimensional nuclear magnetic resonance spectroscopy
Abstract: International audience; We have investigated the reliability of 2D-COSY and 2D-TOCSY experiments to provide accurate measurements of (13)C-enrichments in complex mixtures of (13)C-labelled metabolites. This was done from both theoretical considerations and experimental investigations. The results showed that 2D-TOCSY but not 2D-COSY could provide accurate measurements of (13)C-enrichments, provided efficient zero-quantum filters were applied during the mixing period. This approach extends the range of NMR methods applicable in (13)C-labelling experiments and is suitable to investigating the dynamic behaviour of metabolic systems.
Published: 2007
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49. Application of 2D-TOCSY NMR to the measurement of specific 13C-enrichments in complex mixtures of 13C-labeled metabolites

Author: Jean-Charles Portais, Fabien Létisse, Stéphane Massou, Cécile Nicolas, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie du Développement de Marseille (IBDM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Aix Marseille Université (AMU)-Collège de France (CdF)-Centre National de la Recherche Scientifique (CNRS), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Magnetic Resonance Spectroscopy, 13C-labeling experiments, [SDV]Life Sciences [q-bio], Biomass, Bioengineering, medicine.disease_cause, Sensitivity and Specificity, 01 natural sciences, Applied Microbiology and Biotechnology, 03 medical and health sciences, Escherichia coli, medicine, Spectroscopy, 030304 developmental biology, Fluxomics, Carbon Isotopes, 0303 health sciences, Chromatography, Chemistry, Hydrolysis, 010401 analytical chemistry, 0104 chemical sciences, Glucose, Complex biological mixtures, Quantitative 2D-NMR, Biotechnology
Abstract: International audience; A 2D-NMR method based on zero-quantum filtered (ZQF-) TOtal Correlation SpectroscopY (TOCSY) was applied to measure 13C-enrichments in complex mixtures of 13C-labeled metabolites generated in carbon-labeling experiments. Using ZQF-TOCSY, morethan 30 13C-enrichments could be potentially measured from the analysis of a biomass hydrolyzate prepared from Escherichia coli cellsgrown on a mixture of 20% [U-13C]-glucose and 80% [1-13C]-glucose, without need for separation of metabolites. The method isapplicable to biomass hydrolyzates, cell extracts, and other complex biological samples. It is also applicable to any combination oflabeled substrates and provides a basis for examining non-steady-state conditions.
Published: 2007
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50. In vivo 31P and 1H HR-MAS NMR spectroscopy analysis of the unstarved Aporrectodea caliginosa (Lumbricidae)

Author: Myriam Malet-Martino, Stéphane Massou, Delphine Bon, Robert Martino, Franck Desmoulin, Guénola Pérès, Véronique Gilard, Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Structure Fédérative Toulousaine en Chimie Moléculaire (SFTCM), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Ecosystèmes, biodiversité, évolution [Rennes] (ECOBIO), Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Equipe 7 Inserm U1048, Institut des Maladies Métaboliques et Cardiovasculaires (I2MC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Rennes (UR)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR), Université de Rennes (UR)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Centre National de la Recherche Scientifique (CNRS), Briand, Valerie, Institut de Chimie de Toulouse (ICT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut Ecologie et Environnement (INEE), and Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], Soil Science, 010501 environmental sciences, 01 natural sciences, Microbiology, chemistry.chemical_compound, Nuclear magnetic resonance, Lombricine, In vivo, biology.animal, Drilosphere, Ecotoxicology, Lumbricidae, [SDU.ENVI]Sciences of the Universe [physics]/Continental interfaces, environment, ComputingMilieux_MISCELLANEOUS, 0105 earth and related environmental sciences, biology, Earthworm, 04 agricultural and veterinary sciences, Nuclear magnetic resonance spectroscopy, Metabolism, biology.organism_classification, [SDU.ENVI] Sciences of the Universe [physics]/Continental interfaces, environment, Earthworm . Ecotoxicology . HR-MAS NMR . Glyphosate . Metabolism, Biochemistry, chemistry, 040103 agronomy & agriculture, 0401 agriculture, forestry, and fisheries, Agronomy and Crop Science
Abstract: This study demonstrates that 31P and 1H high resolution-magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR), a recently developed NMR technique, can be applied to the in vivo analysis of metabolites from an unstarved earthworm. The endogeic earthworm Aporrectodea caliginosa was cut into lengths and then the different body parts (anterior, middle, posterior) with the gut content were analyzed. With the HR-MAS NMR, metabolites show well-resolved signals, whereas, with conventional NMR, spectra are highly dependent on the gut content. 31P HR-MAS NMR has been used to evaluate the effect of an acute exposure of the earthworm to glyphosate. Our observations support a low toxicity of the herbicide and suggest that glyphosate could be trapped in the cutaneous mucus. Earthworms could therefore play a role in horizontal dispersion and stabilization of glyphosate in the drilosphere. Phosphorylated metabolites, such as phospholombricine and lombricine, were clearly identified and their amount measured during experiments. The 1H HR-MAS NMR method offers the opportunity to measure, on the same sample and simultaneously, both the hydrosoluble metabolites and lipids. The data on lipid location and relative succinate concentration shed light on the physiological and metabolic functions of the different body parts of the earthworm.
Published: 2006
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