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MARIE SKLODOWSKA-CURIE ACTIONS

Co-funding of regional, national and international programmes (COFUND)

DOC2AMU THESIS PROJECT 2018 CALL FOR APPLICATIONS

Integrative study of the energy metabolism of H2 in Desulfovibrio

fructosovorans: functional and molecular characterization of Hnd

hydrogenase

1. GENERAL INFORMATION

Call 2018-27

Topic Climate change

Keywords Hydrogen, hydrogenase, energy metabolism, omics, bacteria

2. THESIS DIRECTOR(S), RESEARCH UNITS AND DOCTORAL SCHOOLS

Thesis director Marie-Thérèse GIUDICI-ORTICONI

Research Unit Bioénergétique et Ingénierie des Protéines

Doctoral school ED 062 - Sciences de la Vie et de la Santé

Thesis co-director Laetitia SHINTU

Research Unit Institut des Sciences Moléculaires de Marseille

Doctoral school ED 250 - Sciences Chimiques

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MARIE SKLODOWSKA-CURIE ACTIONS

Co-funding of regional, national and international programmes (COFUND)

DOC2AMU THESIS PROJECT 2018 CALL FOR APPLICATIONS

Integrative study of the energy metabolism of H2 in Desulfovibrio fructosovorans:

functional and molecular characterization of Hnd hydrogenase

1. DESCRIPTION OF THE PHD THESIS PROJECT

1.1 OBJECTIVES OF THE PROJECT BASED ON THE CURRENT STATE OF THE ART

Background: In a context of global warming and depletion of fossil fuel reserves, the development of

renewable non-polluting energy is an absolute necessity. H2 constitutes the most likely environmentally

friendly future fuel because the only byproduct of H2 combustion is water, although to date it cannot be

produced using any "green" methods. An option would be to produce bio-H2 using microorganisms. One of the

most potentially useful ways to use hydrogen would be in electric cars equipped with fuel cells that would

convert H2 into electricity. However, the development of fuel cells is considerably limited due to the low

availability and the cost of the platinum that constitutes the catalyst for the conversion reaction of H2 into

protons and electrons. In this context, an innovative area of research is the development of biofuel cells using

hydrogenases as biological catalyst for H2 oxidation. The precise understanding of the molecular mechanism of

hydrogenases and of the microbial H2 metabolism at the fundamental level is thus an essential step towards

applications for the use of hydrogenases and bioproduction of H2.

Hydrogenases are complex metalloenzymes that catalyze the reversible oxidation of H2 at a bimetallic

(FeFe or NiFe) active site. These enzymes are very diverse and widespread in archaea and bacteria, but only

few representatives of these enzymes are deeply characterized. Exploration of the biodiversity of hydrogenases

has shown that these enzymes are involved in energy metabolism and can play a major physiological role in

energy conservation. In addition to the functional diversity of these enzymes, there is a great structural

variability, which is based on the subunit composition but also on the cofactor content (1).

H2 plays an important role in the energy metabolism of sulfate reducers belonging to the genus

Desulfovibrio. The versatility of the Desulfovibrio H2 metabolism is due to a complex hydrogenase system

composed by several different enzymes, which varies considerably from one Desulfovibrio species to another.

The precise physiological role of these enzymes has not been yet elucidated. The biological system used as a

model in this project is the sulfate-reducing anaerobic bacterium Desulfovibrio fructosovorans. Its hydrogenase

content is exceptional since its genome encodes 6 hydrogenases (2 NiFe and 4 FeFe) with different subunit

composition and cellular localization. Studies have been carried out to determine their physiological role within

the cellular energy metabolism. However, their multiplicity makes it difficult to establish their role since

compensation phenomena can come into play when the genes coding for one enzyme are deleted.

This project focuses on the FeFe hydrogenase Hnd of D. fructosovorans. Until now, all attempts to

purify and characterize Hnd have never been successful due to its very small amount in the cell (2). However,

our first results show that it is possible to purify a recombinant form of the enzyme. It is a soluble cytoplasmic

enzyme constituted of 4 different subunits and predicted to contain 8 FeS clusters as well as a binding site for a

flavin mononucleotide (FMN) and for NAD(P) that was previously described as an NADP-reducing hydrogenase

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(3). Moreover, Hnd shares a high sequence similarity with hydrogenases belonging to a new class of enzymes

that carry out flavin-based electron bifurcation. This newly discovered process could be regarded as a third

mode of energy conservation in addition to electron transport phosphorylation and substrate level

phosphorylation (4). These electron-bifurcating hydrogenases, which have recently been preliminarily

described in bacteria (5, 6), present the unique feature of using 2 electron donors (or acceptors depending on

the direction of the reaction) simultaneously, which are a ferredoxin, a small electron transfer protein, and

NAD (or NADP). These enzymes carry out the energy coupling of an exergonic reaction (the oxidation of

ferredoxin to produce H2) with an endergonic reaction (the oxidation of NAD(P)H). The molecular mechanism

of flavin-based electron bifurcation remains unexplained.

Objectives: The aim of our research is to understand the H2 metabolism and more broadly the energy

metabolism of our model bacteria D. fructosovorans. It is now well known that “omics” techniques represent a

cutting-edge tool for the characterization of metabolism modulations. Recent studies have also shown the

importance of multi-omics approaches for deciphering enzyme activity and for the identification of affected

metabolic pathways (7). In this context, the combination of in vitro biochemical and physico-chemical studies

and integrative systemic approaches (i.e. transcriptomics, proteomics and metabolomics) should enable us to

understand further the functions of Hnd hydrogenase and, in particular, to highlight how it may use the

electron bifurcation mechanism that was only recently described as the third energy conservation mechanism

in micro-organisms. Therefore, this doctoral project has a dual objective: 1- to understand the physiological

role of the Hnd hydrogenase in the energy metabolism of D. fructosovorans and 2- to characterize this

hydrogenase at the molecular level.

Understanding these mechanisms at the fundamental level will make it possible to use this hydrogenase for

applications such as biofuel cells. Moreover, the mechanism of electron bifurcation by the hydrogenase Hnd

should allow the bacterium a better conservation of energy and thus reduced consumption of ATP. The

production of bio-H2 could then be considered, in fermentation conditions, by an interesting process that

consumes less energy.

In addition, the student will acquire training at the interface between biochemistry/microbiology and chemistry

and expertise in the field of third-generation biofuels, a job-creating sector.

(1)-Vignais P, Billoud B (2007) Chem. Rev., 107: 4206.

(2)- de Luca G et al. (1998) Biochem. Biophys. Res. Commun., 248: 591.

(3)- Malki S et al. (1995) J. Bacteriol. 177: 2628.

(4)- Buckel W and Thauer RK (2013) BBA Bioernergetics, 1827: 94.

(5)- Schut GJ and Adams MWW (2009) J. Bacteriol. 191: 4451.

(6)- Schuchmann K and Müller V (2012) J. Biol. Chem. 287: 31165.

(7)- Prosser et al. (2014) EMBO Rep. 15: 657

1.2 METHODOLOGY

1- Metabolic role of Hnd hydrogenase:

The role of Hnd hydrogenase in the physiology of D. fructosovorans will be studied using cell batches

grown under different growth conditions. First, we will carry out physiological studies of the wild-type strain

under a respiratory condition (with sulfate as terminal electron acceptor), a fermentation condition (without

sulfate) and a mixed condition (with a limiting amount of sulfate) since our first results show that hnd

expression is different in these three conditions. The WT strain will then be compared to the hydrogenase

mutant strain (Δhnd) (7) under the same conditions.

In order to characterize the complexity of this biological system, combined transcriptomic, proteomic

and metabolomic analyses will be carried out. A comparative transcriptomic approach using the RNA-Seq

method will be used to demonstrate the fine gene expression under controlled growth conditions (respiratory,

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fermentative and mixed conditions) in the WT and the mutant strains, as well as the downstream regulation

mechanisms within the bacterium. The information provided at gene level will be completed and compared at

protein level by a mass spectrometry-based labelled quantitative proteomic method (isobaric tags for relative

quantification or quantitative targeted proteomics). Finally, the WT and mutant metabolism variations will be

followed using NMR-based metabolomic analysis. Intra-cellular metabolic profiles of the strains will be

acquired using High-Resolution Magic Angle Spinning NMR spectroscopy (HRMAS NMR) that allows the direct

analysis of the cells without preliminary extraction steps. The excreted metabolites will also be detected

through the liquid-state NMR analysis of the culture media under each condition. Quantification of the

discriminant metabolites will also be performed. It is becoming clear that any single "omics" approach may not

be sufficient to characterize the complexity of biological systems. The objective, herein, is to simultaneously

exploit the data resulting from the different "omics" approaches to enhance our understanding of the H2

metabolism and more broadly the energy metabolism in D. fructosovorans as well as to determine the role of

Hnd hydrogenase by comparing the WT strain with the Δhnd mutant strain.

2- Molecular characterization of Hnd hydrogenase:

A multidisciplinary approach (biochemistry, molecular biology, spectroscopy, electrochemistry) will be

conducted to determine whether Hnd is an electron-bifurcating hydrogenase and which are the redox

partner(s) of the enzyme. For this purpose, we will greatly benefit from the presence of platforms labeled by

AMU in the Institute (plateformes technologiques AMU: EPR, Marseille Protéomique, Biomasse et

biohydrogène). We have already cloned the genes encoding Hnd and a recombinant form of the hydrogenase is

homologously produced and easily purified, which is a strong pre-requisite to this project. In the same way,

mutants will eventually be produced and purified.

In addition, the three ferredoxins of the bacterium, which may be potential redox partners of Hnd, will also be

cloned and purified to test electron bifurcation. Ferredoxins are small, acidic, FeS-containing proteins involved

in electron transfer. Hnd being predicted to be an electron-bifurcating hydrogenase, it must therefore reduce

two electron acceptors in synergy, which are probably the NADP (8) and a ferredoxin. The specificity of the

ferredoxin used in the Hnd reaction will be determined.

Redox centers of Hnd will be characterized (EPR, FTIR, HPLC, fluorescence, UV-visible spectroscopy...), in

particular the number and the nature of the flavin cofactor(s) will be determined. H2 oxidation or production

activity of the purified enzyme will be measured in the presence of O2, which is usually a strong inhibitor of

FeFe hydrogenases. Preliminary results show that Hnd, unlike the large majority of other FeFe hydrogenases,

exhibits high activity even when it is purified in the presence of O2. The O2-tolerance, combined with the high

stability of the enzyme, makes Hnd an excellent candidate for use in biotechnological applications such as H2

bioproduction or biofuel cells.

(7)- Malki S et al. (1997) Arch. Microbiol., 167: 38.

(8)- Malki S et al. (1995) J. Bacteriol. 177: 2628.

1.3 WORK PLAN

The PhD student will start first by the determination of the role of Hnd in the metabolism of the

bacterium (metabolic phenotyping and metabolomic analysis). He/she will then, during the second year,

prepare the samples for proteomic (total protein fraction of the WT and Δhnd strains grown under different

conditions) and transcriptomic analyses (total mRNA preparation for RNA-Seq analysis). The proteomic analysis

will be implemented in collaboration with the "plateforme protéomique de l'Institut de Microbiologie de la

Méditerranée" (R. lebrun), CNRS, Marseille, and the transcriptomic analysis will be outsourced. The

transcriptomic data will be processed by the bioinformatic group of the "Laboratoire de Chimie Bactérienne",

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IMM, CNRS, Marseille (E. Talla). During the last year, after the omics analyses will be completed, the PhD

student will have in charge to integrate the data from the three omic analyses. The molecular characterisation

of Hnd will start six month after the beginning of the thesis by the cloning and the purification of the three

ferredoxins of D. fructosovorans, which are potential redox partners of Hnd. This molecular characterisation of

the hydrogenase will be continued until almost the end of the thesis. The last months of the thesis will be

devoted to the writing of the thesis manuscript.

Figure : Tasks schedule

1.4 SUPERVISORS AND RESEARCH GROUPS DESCRIPTION

BIP, UMR 7281, AMU-CNRS, Marseille: BIP laboratory studies focus on the diversity of energy metabolisms of

micro-organisms and their applications in the field of bioenergy and environment. The hydrogen metabolism is

one of the major thematic axes of the laboratory. The objectives of our research are to better understand the

universality of hydrogenases in the microbial world. Among our several microbial model organisms,

Desulfovibrio fructosovorans has a complex hydrogenase system composed of 6 different enzymes. Why such a

redundancy of hydrogenases in the same organism? Some of these enzymes have been particularly well

studied at the molecular level in the laboratory but others are still completely unknown. Our aim is to elucidate

the precise physiological role as well as the molecular mechanism of these enzymes in Desulfovibrio.

Project's supervisor: Marie-Thérèse Giudici ORTICONI: M-T Giudici's group has characterized the main

metabolic chains of organisms involving metalloenzymes, complexes and supercomplexes, and now studying

their dynamics and interactions under different conditions. New strategies of immobilization and encapsulation

of suitable biocatalysts for biotechnological devices are being developed, especially for new energy sources

(biomass, biogas, biohydrogen). The immobilization of bacterial extremophilic enzymes on electrode supports

for efficient enzymatic reactions is studied, i.e. catalytic oxidation of H2 by hydrogenases. New multi-resistant

hydrogenases are characterized using multidisciplinary, functional approaches. Giudici’s group is the first to

have developed the technology of H2/O2 biofuel cell in France, which was highlighted this year by the CNRS and

awarded by the French Agency for Environment and Energy Management (ADEME). She has initiated an

innovative axis of ecological engineering, which consists in the microbial study of bacterial consortia involved in

biohydrogen production. The objective is to establish the parameters governing the metabolic networks in view

of biotechnological applications.

ISM2, UMR 7313, AMU-CNRS, Marseille: The Institute of Molecular Sciences of Marseille (iSm2) is a

multidisciplinary research center acting mainly at the interface between Chemistry and Biology. This structure,

built around three main structures, Aix-Marseille University, the CNRS and the Ecole Centrale de Marseille,

presents an architecture based on clusters of skills powered by four research teams (Chiroscience, Stereo,

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CTOM and Biosciences). The specificities and scientific objectives of the Institute aim at answering major

scientific problems in the fields of chirality, catalysis, chemical models and mechanisms, natural products,

supramolecular assemblies, of living chemistry and NMR-based metabolomics at the fundamental and applied

levels. The NMR group belongs to the Biosciences team and develops NMR methodologies and applications for

the analysis of complex mixtures.

Project's co-supervisor: Laetitia SHINTU: Dr L. Shintu is an assistant professor who is part of the Biosciences

team. LS is a Metabolomics expert since 2002 and one of the pioneers who used High Resolution Magic Angle

Spinning NMR spectroscopy for metabolomics purpose. LS is currently working on the use of NMR-based

metabolomics as a tool for the diagnosis or the classification of different types of cancer (thyroid, pancreas,

lymphomas, sarcomas…). She is also involved in projects that aim at highlighting the metabolic impact of

dysbiosis in mice. In parallel, she develops NMR methods such as slow-MAS NMR that allows the HRMAS NMR

analysis of fragile tissues at low spinning speed preserving their physiological integrity.

2. 3I DIMENSIONS AND OTHER ASPECTS OF THE PROJECT

2.1 INTERDISCIPLINARY DIMENSION

Proposed role of the supervisor (M.-T. Giudici-Orticoni), BIP, AMU-CNRS, Marseille:

The BIP laboratory, focusing on the diversity of energy metabolisms in microorganisms with a

multidisciplinary approach, is a leader in the field of hydrogenases, biohydrogen production and biofuel cells.

By a combination of a biological (biochemistry, molecular biology, physiology) and a physico-

chemical/biophysical (electrochemistry, spectroscopies) approach, it studies the functional and structural

properties of hydrogenases in bacteria. The precise understanding of the mechanism of these enzymes is

important for both fundamental and applied research. This interdisciplinary strategy has already been used by

the BIP laboratory for the study of another hydrogenase of D. fructosovorans and has given spectacular results

on the molecular mechanism of [NiFe] hydrogenases. It will be used in the framework of this project to initiate

the biochemical and physico-chemical characterization of a hydrogenase that bifurcates electrons, flavin-based

electron bifurcation being a recently discovered mechanism whose molecular bases remain to be elucidated. In

addition to the molecular characterization of the enzyme, the BIP laboratory will also be in charge of the

metabolic phenotyping of the strains and the integration of the "omic" data to reconstruct the metabolic

pathways involved in the energy metabolism of the bacterium.

Proposed role of the co-supervisor (L. Shintu), ISM2, AMU-CNRS-CM, Marseille:

NMR-based metabolomics is the one of the main resarch area of the NMR group of ISM2. The group owns two

NMR spectrometers that allow the analysis of both liquid and semi-solid samples, as well as all the hardwares

and softwares that are necessary to perform metabolomic analysis. The co-supervisor L. Shintu will be in charge

of the training of the student for NMR spectroscopy (sample preparation, spectrum acquisition, data

processing, metabolite identification) and for statistical treatment of the NMR data (supervised and

unsupervised multivariate statistical analysis, use of the statistical software SIMCA-P, Matlab). Connections

between affected metabolites will also be performed using dedicated softwares (Cytoscape, metabolanalyst) in

order to provide hypotheses on the metabolic pathways involved in Hnd activity.

2.2 INTERSECTORAL DIMENSION:

Dr M. Maillot, President of MS_Nutrition, will participate to this project as a non-academic partner.

The start-up, created in 2014, has a unique expertise in modeling and statistics. It will enable the integration

and statistical analysis of the “omics” data that will be collected during the first 2 years of the project. The

future PhD student will be welcomed and accommodated in the laboratory of MS Nutrition for 3-4 months

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where training will be provided in data processing and filtering, multi-omics statistical analyses, and the

interpretation of results.

This project is in agreement with the SRI-S3 objective related to the energetic transition since it aims at

understanding how to produce renewable energy using biofuel cells.

2.2 INTERNATIONAL DIMENSION:

The molecular characterization of the hydrogenase Hnd will be done in collaboration with the group of

A. de Lacey from the "Laboratory de Bioelectrocatalysis", Instituto de Biocatalisis y Petroleochimica, CSIC,

Madrid, Spain. This group pioneered the use of Fourier-transform InfraRed spectroscopy (FTIR) for the study of

hydrogenases. These enzymes have CO and CN ligands at the active site, whose vibrations report on the

electronic structure of the active site. A. de Lacey has a long history of collaboration on hydrogenases with the

BIP laboratory.

The PhD student will attend several international conferences to present the results obtained during the thesis,

among them: -the 12th

International Hydrogenase Conference, Lisbon (Portugal), April 2019.

-the 21st

European bioenergetics conference, Marseille, 2020

3. RECENT PUBLICATIONS

Marie-Thérèse Giudici-Orticoni:

- Mazurenko I, Monsalve K, Rouhana J, Parent P, Laffon C, Goff AL, Szunerits S, Boukherroub R, Giudici-Orticoni MT, Mano N, Lojou E. (2017) How the Intricate Interactions between Carbon Nanotubes and Two Bilirubin Oxidases Control Direct and Mediated O2 Reduction. Energy & Environmental Science, 10, 1966-1982. DOI:10.1039/C7EE01830D

- Boughanemi S, Lyonnet L, Infossi P, Bauzan M ,Kosta A, Lignon S, Giudici-Orticoni M-T and Guiral M.(2016) Microbial oxidative sulfur metabolism: biochemical characterization of the membrane-bound heterodisulfide reductase-like complex of the bacterium Aquifex aeolicus FEMS Microbiology Letters, 363(15) doi: 10.1093/femsle/fnw156

- Benomar S, Ranava D, Cárdenas ML, Trably E, Rafrafi Y, Ducret A, Hamelin J, Lojou E, Steyer JP and Giudici-Orticoni MT. (2015) Nutritional stress induces exchange of cell material and energetic coupling between bacterial species, (2015) Nature comm. 10.1038/ncomms7283

- Castelle CJ, Roger M, Bauzan M, Brugna M, Lignon S, Nimtz M, Golyshina OV, Giudici-Orticoni MT, Guiral M (2015) The aerobic respiratory chain of the acidophilic archaeon Ferroplasma acidiphilum: A membrane-bound complex oxidizing ferrous iron. Biochimica et Biophysica Acta, 1847, 717–728.

Laetitia Shintu:

- Tranchida, F.; Rakotoniaina, Z. et al. (2017) Hepatic metabolic effects of Curcuma longa extract supplement in high-fructose and saturated fat fed rats. Sci. Rep., 7, 5880

- Tranchida F., Shintu* L., et al. (2015) Metabolomic and Lipidomic Analysis of Serum Samples following Curcuma longa Extract Supplementation in High-Fructose and Saturated Fat Fed Rats PLoS ONE, 10(8), e0135948

- Renault M, Shintu L, et al. (2013) Slow-spinning low-sideband HR-MAS NMR spectroscopy: delicate analysis of biological samples. Sci. Rep., vol 3, 3349

- Torregrossa L, Shintu L*, et al. (2012) Toward the Reliable Diagnosis of Indeterminate Thyroid Lesions: A HRMAS NMR-Based Metabolomics Case of Study, J. Proteome Res., 11(6), 3317-3325

- Shintu L, Baudoin R, et al. (2012) Metabolomics-on-a-Chip and Predictive Systems Toxicology in Microfluidic Bioartificial Organs, Anal. Chem., 84, 1840-1848

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4. EXPECTED PROFILE OF THE CANDIDATE

The candidate will be involved in the functional and the molecular characterization of the Hnd

hydrogenase in the sulfate-reducing bacterium Desulfovibrio fructosovorans. The objectives of this doctoral

project are to understand the physiological role of the Hnd hydrogenase in the energy metabolism of the

bacterium and to characterize this hydrogenase at the molecular level. The position requires a candidate at the

interface between biology and chemistry with a strong educational background in molecular biology, protein

biochemistry and microbiology. He/She will be in charge of the production, purification and mutagenesis of

proteins. For the characterization of the enzyme and the metabolomic analysis, knowledge of physico-chemical

and biophysical techniques, such as NMR, EPR or electrochemistry would be an advantage. Knowledge of

bioinformatics would be highly appreciated for the integration of the "omic' data, and a good knowledge in the

field of hydrogenase would be ideal. The PhD candidate must also have the capacity for interacting with

collaborators and communicating results at meetings and conferences. He/She will closely collaborate with

colleagues in other teams to bridge the multidisciplinary challenges.

5. SUPERVISORS’ PROFILES

Dr. Marie-Thérèse Giudici-Orticoni is director of the Laboratory of Bioenergetics and Protein Engineering (BIP)

in Marseilles, director of the Mediterranean Institute of Microbiology (IMM, 5 laboratories, 8 technical

platforms and about 350 members), head of the biomass platform hydrogen CNRS Aix/Marseille University and

the Pôle competitivité Cap Énergies. MT Giudici-Orticoni obtained a PhD in enzymology at the University of Aix-

Marseille in 1991 and she got a research position at CNRS the same year, where she is currently research

director (DR1). In 2006 she created and is currently heading the team ‘Energetic metabolism of extremophiles

and bioenergy processes’ at the BIP-IMM. MT Giudici- Orticoni has expertise for implementation between

research, training, innovation and valorization. She is author of 90 publications in refereed journals and 3

patents, scientific coordinator for national and international programs, namely ANR Bioenergy, CNRS programs,

PACA call for applications, EC Marie Curie Research Fellow. She supervized and co-supervized 13 PhD, 6 PhD

students s during the last 5 years: Benomar Saïda (2009-2012, 100%, now postdoctorant in USA), Aussignargues

Clément (2009-2012, 50%, now post-doctorant in USA), Roger Magali (2011-2014, 50%, now postdoctorant in

United Kingdom), Ranava David (2012-2015, 100%, now postdoctorant in Toulouse), Backes Cassandra (2015-

2018, 100%, thesis currently being supervised), Zuili Lisa (2017-2020, 20%, thesis currently being supervised).

Dr L. Shintu is an assistant professor who is part of the Biosciences team. LS is a Metabolomics expert since

2002 and one of the pioneers who used High Resolution Magic Angle Spinning NMR spectroscopy for

metabolomics purpose. LS has worked in several fields (food science, environment, biomedicine) and published

more than 20 papers in the area of Metabolomics and NMR specroscopy. She was the PI of the ANR project

IMAPIST (2012-15) and was awarded by the CNRS-higher education chair program (2009-14). She supervised 2

PhD students (Jima N. Chandran (2010-2013) – now postdoctorant in Germany and Lamya Rezig (2012-2016)-

now engineer at Metabrain research), 3 research engineers and several bachelor and master students, as well

as 2 senior researchers through CNRS training workshop in NMR-based metabolomics. She is also in charge of

the “plateforme régionale de RMN métabolomique” supported by Cancéropôle PACA in 2016.

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AVIS DES DIRECTEURS DES LABORATOIRES CONCERNES PAR LE PROJET DE THESE

Avis du directeur du laboratoire du directeur de thèse, M. GIUDICI-ORTICONI

Marie-Thérèse

x Favorable □ Défavorable

Commentaires :

Ce projet de thèse s’inscrit dans la stratégie du laboratoire et permettra la compréhension d’un processus majeur de la bioénergétique. Par ailleurs la caractérisation d’une nouvelle hydrogénase permettra leur utilisation dans les biopiles, domaine dans lequel le laboratoire est leader

Fait à Marseille, le 08 janvier 2018

Signature

Laboratoire Nort – 1er étage Aile Bleue - Faculté de Médecine La Timone 27, bd jean Moulin - 13385 Marseille cedex 05

MS-Nutrition UMR NORT (C2VN) Faculté de Médecine de la Timone, 27, boulevard Jean Moulin 13385 Marseille Cedex 05 Tel: 04 91 32 45 94 RCS Marseille B 800 309 171 SIRET : 800 309 171 00018 TVA : FR 15 800 309 171 e-mail : contact@ms-nutrition.com

Marseille, le 01/08/2018

Letter of support from the non-academic partner

To whom it may concern,

Yours sincerely,

Matthieu Maillot Président de MS-Nutrition, matthieu.maillot@ms-nutrition.com

I, Dr Matthieu Maillot (the undersigned), President of MS-Nutrition (www.ms-nutrition.com), by this letter accept to participate as a non-academic partner in the project entitled “Integrative study of the energy metabolism of H2 in Desulfovibrio fructosovorans: functional and molecular characterization of Hnd hydrogenase”. My start-up’s expertise will enable the integration and statistical analysis of the “omics” data that will be collected during the first 2 years of the project. The future PhD student will be welcomed and accommodated in our laboratory for 3-4 months where training will be provided in data processing and filtering, multi-omics statistical analyses, and the interpretation of results.