Understanding the conformational and dynamic changes of biomacromolecular complexes in different states, such as the membrane protein photoreceptor-transducer complex NpSRII/NpHtrII, is a key step to gaining insight into the functional mechanism of these important classes of protein complexes, since ~30 % of the human proteome are membrane proteins, yet they are largely underrepresented in terms of structural information with <1 % of all structures in the protein data bank. Hence for the development of methods suitable to study the conformation and dynamics of such complexes there is a strong demand and a vast potential field of applications. Here we combined method development at the interface between biomolecular simulations and model-based analysis of EPR- and fluorescence spectroscopic data with application studies using state-of-the-art spectroscopic techniques in conjunction with site-directed spin- or fluorescence labeling. In an initial benchmark study on the rigid globular protein complex Rpo4/7, we compared experimental inter fluorescence label distances or spin label distance distributions to a variety of predicted inter label distances based on molecular dynamics simulations, Monte Carlo sampling and a discrete rotamer library analysis. We found that while for the molecular dynamics simulations with explicit solvent considerable sampling challenges have to be overcome to reproduce the experimentally observed inter label distance distributions, the Monte Carlo sampling performed well when compared to the experimental data and was computationally less demanding. Significantly more efficient and equally accurate for our examples was the so-called rotamer library analysis available for the spin labels since it relies on a pre-calculated set of rotational isomers. In general, predictions for the mean distances were in agreement within the error margins while distribution shapes were more challenging to reproduce. Overall this study shows a positive evaluation for the assessed tools and the developed simulation protocols as well as their potential applications. Using the combination of EPR and fluorescence spectroscopy for distance determination we studied the structural influence of RNA binding on Rpo4/7, and showed that the protein complex stays conformationally rigid and thereby serves as a guiding rail for the nascent RNA chain that leaves the RNA polymerase along the Rpo4/7 RNA binding interface. To enhance the interpretation of experimentally determined changes of conformation and dynamics in protein complexes and to discuss the observed changes in terms of structural information, we built models of the two transcription factors TFE and the Spt4/5 complex, as well as of Argonaute, a 713 amino acid four-domain protein nuclease from Methanocaldococcus jannaschii. These structural models not only allowed a more accurate planning of fluorescence or EPR labeling experiments, but also the models enabled the discussion of the experimental data in structural terms. Based on such an initial structure further computational analysis techniques may be applied to identify putative structural changes or dynamic modes. This was shown for the histidine transporter HisQMP2, where we combined normal mode analysis to model protein flexibility with the rotamer library analysis to screen for possible conformational changes in comparison to experimental inter spin distance data. The most prominent agreement with one mode led to a working hypothesis of a conformational change and provides the basis for validation in future experiments. Due to the inherent synergy effects, we applied a combined experimental and simulation approach for the EPR-based distance determination in the globular DNA-binding protein LexA to probe conformation and dynamics of the N-terminal DNA-binding domains with respect to the C-terminal domains within the LexA homodimer. While the C-terminal dimerization domains exhibit a well-defined conformation that proved to be independent of DNA-binding, large-scale changes in conformation and dynamics were detected for the N-terminal domains. They were only found in a defined conformation when bound to DNA while in its absence a large rotational freedom of the entire N-terminal domains contributed to the conformational ensemble. Combined with a biochemical characterization of the autocatalytic cleavage of LexA, our data explains how LexA induces the SOS response after DNA damage or under latent antibiotic stress. We further studied the membrane photoreceptor-transducer complex NpSRII/NpHtrII that governs the light-dependent swimming behavior in Natronomonas pharaonis by a two-component signaling system. This system comprises extraordinary features of sensitivity, signal amplification, integration and transducer cooperativity, yet the molecular details of these features are poorly understood, as is signal propagation itself. By combining time-resolved cw EPR spectroscopy of NpSRII/NpHtrII variants spin labeled in the HAMP1 domain with time-resolved optical absorbance spectroscopy to report on the receptor signaling state, we found a tight kinetic coupling of receptor and transducer during the relaxation back to the ground state and hence a prolonged activation period, that with ~500 - ~700 ms is sufficiently long to cause phosphorylation bursts of the cognate kinase CheA. This explains signal amplification already on the level of the NpSRII/NpHtrII dimers. We further determined the transient difference spectra from the time-resolved EPR data that show local differences in dynamics and steric restrictions upon light-activation. Comparing these experimentally observed differences to predictions confirms the assumed two-state structural model and shows this transition between the two states for a single HAMP domain in a light-dependent manner. Additionally, our approach integrates a dynamic view into the model, since the two states are shown to exhibit different local dynamics in a fashion described previously as a competing model for signaling by dynamic differences based on biochemical studies. Here we show unification of the two models into one congruent description encompassing a transition between the two previously suggested states by concerted structural and dynamic changes. In an independent analysis using all-atom and coarse grained molecular dynamics of the NpSRII/NpHtrII complex in the minimal unit that can exert kinase control, the trimer of receptor-transducer dimers, we revealed a distinct dynamical pattern encoded in the primary sequence of the coiled-coil heptad-repeats. Upon receptor activation, these segments alter their dynamics in a concerted fashion with regions such as HAMP1 and the adaptation region becoming more compact, while HAMP2 and the tip become more dynamic, leading to dynamic and to limited structural changes at the CheA-kinase binding sites. Together with an extensive validation against experimental data, these findings suggest the altered dynamics as the mechanism for signal propagation along the extended coiled-coil structure of NpHtrII. This working model, that explains the current body of experimental data, allows for further refinement by all-atom molecular dynamics and provides a basis to devise future experiments for validation. The presented studies outline the versatile methodology of combined experimental and simulation approaches to analyze the conformation and dynamics of biomacromolecules including membrane protein complexes.

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Вещества, обладающие антирадикальной активностью (АРА) играют ведущую роль в системе защиты организма от свободных радикалов. Общие недостатки известных способов оценки АРА выражаются в том, что измерения проводятся непрямыми методами, что затрудняет количественно оценивать содержание веществ с антирадикальными свойствами. Результаты измерений представляются в относительных единицах, в качестве эталонных веществ чаще всего используются: тролокс, галловая и аскорбиновая кислоты, проявляющие разную АРА, что не позволяет сравнивать результаты между собой. Таким образом, существует необходимость в разработке прямого, безэталонного метода оценки антирадикальной активности. Одним из потенциально возможных решений данной проблемы является метод ЭПР-спектроскопии на модели 2,2-дифенил-1-пикрилгидразил. Цель и задачи исследования. Исследование безэталонным методом оценки антирадикальной активности природных и синтетических антиоксидантов на модели 2,2-дифенил-1-пикрилгидразил с использованием ЭПР-спектроскопии. Отработка методики определения антирадикальной активности на модели ДФПГ методом ЭПР-спектроскопии. Получение и анализ значений антирадикальной активности для ряда веществ с антирадикальными свойствами. Сравнение с результатами полученными методом потенциометрического определения антиоксидантной активности с использованием медиаторной системы. Полученные результаты. Установлены оптимальные условия для проведения анализа. Проведено исследование - новым, безэталонным методом оценки антирадикальной активности, ряда природных и синтетических антиоксидантов. Установлены значения антирадикальной активности ряда природных и синтетических антиоксидантов. Рассмотрены различные варианты механизмов ингибирования свободных радикалов и их зависимость от структуры антиоксиданта. / Substances with anti-radical activity (ARA) play a leading role in the body's defense system against free radicals. General disadvantages of the known methods of evaluating the APA expressed in the fact that measurements are carried out by indirect methods, making it difficult to quantify the content of substances with antiradical properties. The measurement results are presented in relative units, as reference substances are most often used: Trolox, ascorbic and gallic acid, exhibit different ARA, which does not allow to compare the results with each other. Thus, there is a need to develop direct, reference-free method of evaluating anti-radical activity. One possible solution to this potential problem is the method of EPR model 2,2-diphenyl-1-picrylhydrazyl. The purpose and objectives of the study. Study standardless method for assessing the antiradical activity of natural and synthetic antioxidants model 2,2-diphenyl-1-picrylhydrazyl using EPR spectroscopy. Testing methods for determining the anti-radical activity in the DPPH model EPR spectroscopy. Obtaining and analyzing anti-radical activity values for a number of substances with antiradical properties. Comparison with the results obtained by potentiometric determination of antioxidant activity using a mediator system Results. The optimum conditions for analysis. The research - the new reference-free method of evaluating anti-radical activity, a number of natural and synthetic antioxidants. Established values antiradical activity of natural and synthetic antioxidants. Various embodiments of the mechanisms of inhibition of free radicals and their antioxidant dependency structure.

The outcome of chemical reactions proceeding via radical pair (RP) intermediates can be influenced by the magnitude and direction of applied magnetic fields, even for interaction strengths far smaller than the thermal energy. Sensitivity to Earth-strength magnetic fields has been suggested as a biophysical mechanism of animal magnetoreception and this thesis is concerned with simulations of the effects of such weak magnetic fields on RP reaction yields. State-space restriction techniques previously used in the simulation of NMR spectra are here applied to RPs. Methods for improving the efficiency of Liouville-space spin dynamics calculations are presented along with a procedure to form operators directly into a reduced state-space. These are implemented in the spin dynamics software Spinach. Entanglement is shown to be a crucial ingredient for the observation of a low field effect on RP reaction yields in some cases. It is also observed that many chemically plausible initial states possess an inherent directionality which may be a useful source of anisotropy in RP reactions. The nature of the radical species involved in magnetoreception is investigated theoretically. It has been shown that European Robins are disorientated by weak radio-frequency (RF) fields at the frequency corresponding to the Zeeman splitting of a free electron. The potential role of superoxide and dioxygen is investigated and the anisotropic reaction yield in the presence of a RF-field, without a static field, is calculated. Magnetic field effect data for Escherichia coli photolyase and Arabidopsis thaliana cryptochrome 1, both expected to be magnetically sensitive, are satisfactorily modelled only when singlet-triplet dephasing is included. With a view to increasing the reaction yield anisotropy of a RP magnetoreceptor, a brief study of the amplification of the magnetic field experienced by a RP from nearby magnetite particles is presented. Finally in a digression from RPs, Spinach is used to determine the states expected to be immune from relaxation and therefore long-lived in NMR experiments on multi-spin systems.

L’étude des transitions structurales dans les protéines est d’un intérêt crucial car ces transformations sont impliquées dans de nombreux processus biologiques essentiels. De tels phénomènes structuraux peuvent être à l’origine de propriétés remarquables dans les protéines flexibles ou désordonnées, propriétés difficilement accessibles par les techniques structurales usuelles. Le marquage de spin couplé à la spectroscopie de résonance paramagnétique électronique (RPE) est une technique bien adaptée pour l’étude de ces transitions structurales. L’insertion d’un radical nitroxyde sur une cystéine, naturelle ou introduite par mutagenèse dirigée, située à un endroit clé de la protéine permet d’obtenir des informations locales sur les changements structuraux éventuels provoqués par l’ajout d’un partenaire.Cette technique a été appliquée à deux systèmes biologiques comportant un degré de flexibilité différent. La flexibilité de la protéine chaperon NarJ, intervenant dans la biogenèse du complexe Nitrate Réductase de la bactérie Escherichia coli, a été étudiée en présence de son peptide partenaire. Ces études ont permis d’une part de déterminer le site d’interaction et d’autre part, de montrer que l’association des deux partenaires entraîne un verrouillage dans une conformation préférentielle de NarJ. Le deuxième sujet d’étude est la protéine CP12 de Chlamydomonas reinhardtii, intervenant dans la régulation d’un complexe supramoléculaire du cycle de Calvin. La CP12 s’apparente à une protéine intrinsèquement désordonnée, ayant la particularité de posséder des cystéines naturelles et fonctionnelles. Le marquage classique a permis de mettre en évidence un nouveau rôle de son partenaire et de montrer que la CP12 garde un caractère désordonné dans le complexe. Par ailleurs, cette protéine a servi de système d’étude pour développer une nouvelle stratégie de marquage sur Tyrosine et démontrer sa faisabilité. / The study of structural transitions in proteins is of crucial interest because these transformations are involved in many biological processes. Such structural phenomena can be the source of remarkable properties in flexible or disordered proteins, properties hardly accessible by conventional structural techniques. Site-directed spin labeling combined with electron paramagnetic resonance spectroscopy (EPR) is a technique well suited for the study of these structural transitions. The insertion of a nitroxide reagent on a cysteine, natural or introduced by site-directed mutagenesis, located in a key position of a protein provides local information on possible structural changes induced by the addition of a partner. This technique was applied on two biological systems with a different degree of flexibility. The flexibility of NarJ, a chaperon protein involved in the biogenesis of the complex nitrate reductase of Escherichia coli was studied in the presence of its peptide partner. These studies enabled us to determine the interaction site and to show that the association of the two partners induced a locked conformation of NarJ. The second system is the CP12 protein of Chlamydomonas reinhardtii, involved in the regulation of a supramolecular complex of the Calvin cycle. CP12 shares some similarities with the intrinsically disordered protein but having natural and functional cysteines. The conventional labeling allowed us to highlight a new role of its partner and to demonstrate that CP12 remains disordered in the complex. Moreover, this protein was used as a model system to develop a new labeling strategy on tyrosine and to demonstrate its feasibility.

No presente trabalho foram estudados os espectros de RPE de um radical nitróxido, 4-hidroxi-2,2,6,6-tetrametilpiperidina-l-oxil (TEMPOL, 11), introduzido como impureza em uma matriz diamagnética, 4-hidroxi-2,2,6,6-tetrametilpiperidina (I), de estrutura conhecida. O uso do radical deuterado, 4-hidroxi-2,2,6,6 -tetrametilpiperidina-d17-l-oxil (PD-TEMPOL, 111) mostrou um aumento considerável na resolução dos espectros para a maioria das orientações do campo magnético. A interação do grupo paramagnético N-O com os prótons vizinhos foram estudadas e os tensores superhiperfinos de dois prótons fortemente acoplados foram determinados. Um desses prótons forma ponte de hidrogênio com o grupo N-O e a distância estimada é 2,09ª&#176. Os autovalores do tensor são: -5,2; 5,4 e -5,1 gauss. O segundo próton está mais distanciado do radical e provavelmente não forma ponte com o radical. Os valores nas direções principais são: 2,0; -1,7;e -2,7 gauss. Com a finalidade de estudar o envolvimento dos prótons na relação eletrônica dos radicais foram estimados os tempos de relaxação T1 e T2 no intervalo de temperatura de -160 a 25&#176C através do método de saturação continua. Foram obtidos os valores em diversas orientações para os radicais protonado (II) e deuterado - (III) para compará-los. Os valores de T2 obtidos são praticamente constante no intervalo de temperatura onde foram medidos e sistematicamente são maiores para o radical deuterado. O tempo de relaxação spin-rede, T1 é dependente da orientação para ambos os radicais. Em geral é obtida urna dependência linear de T1-1 versus T que foi explicada considerando um mecanismo de relaxação por tunelamento dos protons vizinhos entre dois poços de potenciais. Para o radical protonado é observada uma descontinuidade na curva de T1-1 versu T na região de -60&#176C; para o radical deuterado - isto é observado somente para algumas orientações. Este comporta mento poderia ser explicado como devido o congelamento da vibração das pontes de hidrogênio em torno desta temperatura. Um alagamento das linhas de RPE é observado para muitas orientações a partir de -100D&#176C que foi interpretado como devido o congelamento da rotação dos grupos metilas, do radical no caso (II) e da matriz no caso (I) / In the present work the ESR spectra of a nitroxide radical, 4-hydroxi-2,2,6,6-tetramethylpiperidine-l-oxyl (TANOL, lI), introduced as impurity in a diamagnetic host, 4-hydroxi-2, 2,6,6-tetramethylpiperidine (1), of know structure, has been investigated. The use of the deuterated radical, 4-hydroxi-2,2, 6,6-tetramethylpiperidine-d17-1-oxyl (PD-TANOL, 111) showed an improved resolution in the ESR spectra for most orientations of the magnetic field. The interactions of protons in the neighbourhood of the paramagnetic group N-O were studied and the superhyperfine tensors for the two strongly compled protons were determined one of these protons is hydrogen bonded to the N-O and the distance is estimated to be 2,09ª&#176. The superhyperfine tensor has principal values -5.2; +5.4; -5.1 gauss. The second proton is further away from the electron and probably is not hydrogen bonded. The principal values for this proton are 2.0; -1.7 and -2,7 gauss. In order to study the involvment of the protons in the electronic relaxation of the radicaIs, the relaxation times T1 and T2 were estimated in the temperature range -160&#176C 25&#176C for many orientations and comparing data for the protonated and deuterated radicaIs II and III, using the continuous wave saturation method. It is shown that the relaxation time T2 is practically constant in the temperature range studied and that it is sistematically greater for the deuterated radical implying narrower lines. The spin-lattice relaxation time TI is dependent upon the orientation of the magnetic field for both radicaIs. In general a linear temperature dependence of T1-1 versus T is obtained and a mechanism of tunelling of protons between close potential walls in used to explain this behavior. In the case of the protonated radical for many orientations of the magnetic field a break is observed in the T1-1 versus T curve around -60&#176C for the deuterated radical this is observed only for fewer orientations. This behaviour could be explained as due to the freezing of hydrogen bonds vibration near this temperature. For many orientations it is also observed that below -100&#176C a broadening of the ESR lines occurs which is probably re1ated to the freezing of the rotations of the methyl groups of the radical, in the case of II, and of the hast I in the case of radical III

La surexploitation des ressources de la planète est aujourd’hui une problématique de premier ordre, et fait du remplacement des sources non renouvelables, d'énergie et de matières premières, l’un des défis majeurs du XXIe siècle. Dans cet objectif, les lignines, par leur disponibilité et leur biocompatibilité, apparaissent comme l’une des alternatives aux ressources fossiles. C’est dans ce contexte que le Laboratoire PEIRENE a décidé de mener ce travail de thèse portant sur le développement de nouveaux matériaux photosensibles à base de lignines modifiées. Dans ce but, trois lignines d’origines différentes ont été acétylées. Leur étude par spectroscopie RPE a révélé que le blocage de leurs fonctions antioxydantes augmente considérablement la quantité d’espèces réactives de l’oxygène qu’elles sont capables de générer sous irradiation lumineuse, permettant ainsi d’envisager l’utilisation de ce biopolymère modifié dans de nombreux domaines tels que le traitement photodynamique antimicrobien. Afin de les rendre hydrodispersibles et d’élargir ainsi leur champ d’applications, ces matériaux aux propriétés prometteuses ont été mis sous forme de nanoparticules puis, leur comportement photosensible a été lui aussi évalué par spectroscopie RPE. Il a ainsi été démontré qu’une fois dispersées dans l’eau sous la forme de nanoparticules, les lignines acétylées étaient toujours capables de produire de l’oxygène singulet sous irradiation lumineuse. Cette activité, qui n’a pas encore été reportée dans la littérature à notre connaissance, reste cependant assez restreinte et nécessite donc d’être améliorée. Afin d’élargir le domaine du spectre solaire permettant leur activation, un photosensibilisateur a par ailleurs été associé à ces nano-objets par encapsulation et par greffage covalent. L’ensemble des résultats découlant de ces travaux permettent d’envisager le développement de systèmes à base de nanoparticules de lignines acétylées dans de nombreux domaines, notamment pharmaceutique et phytosanitaire. / The overexploitation of the planet's resources is nowadays a major problem and makes the replacement of non-renewable sources of energy and raw materials, one of the major challenges of the XXIe century. For this purpose, lignins, by their availability and their biocompatibility, appear as one of the alternatives to fossil resourcesIn this context, the PEIRENE Laboratory decided to carry out this PhD work on the development of new photosensitive materials based on modified lignins. For this purpose, three lignins from different origin were acetylated. Their study by EPR spectroscopy revealed that blocking their antioxidant functions considerably increases the quantity of reactive oxygen species they are able to generate under light irradiation. Thus it is possible to envisage the use of this modified biopolymer in many areas such as antimicrobial photodynamic therapy. In order to make them water-dispersible and thus to widen their field of applications, these materials with promising properties were put in the form of nanoparticles. Their photosensitive behavior has been also valuated by EPR spectroscopy. It has been demonstrated that once dispersed in water in the form of nanoparticles, the acetylated lignins were still capable of producing singlet oxygen under light irradiation. This activity, which has not yet been reported in the literature to our knowledge, however, remains quite limited and therefore needs to be improved. In order to widen the range of the solar spectrum allowing their activation, a photosensitizer has also been associated with these nano-objects by encapsulation and covalent grafting. The results of these studies make possible to envisage the development of systems based on acetylated lignins nanoparticles in in many field, in particular pharmaceutical and phytosanitary.

Mon objectif a consisté à élucider à l'échelle moléculaire le fonctionnement des systèmes enzymatiques complexes impliqués dans des processus de conversion d'énergie chez les êtres vivants. Je m'intéresse en particulier à la compréhension de deux étapes clés du fonctionnement commun à un grand nombre de ces systèmes, à savoir (i) les étapes d'interaction de ces complexes avec les quinones membranaires et (ii) les mécanismes catalytiques au niveau des sites actifs à molybdène. Le système modèle que j'étudie est la nitrate réductase A issue de la bactérie E. coli, en collaboration avec l'équipe du Dr. Axel Magalon (LCB, Marseille). Il permet la respiration anaérobie en catalysant la réduction du nitrate en nitrite et joue un rôle important dans le cycle biogéochimique de l'azote. Ma recherche vise en particulier à identifier les facteurs moléculaires qui permettent d'ajuster la réactivité de ces systèmes. Cela nécessite l'obtention d'informations structurales à l'échelle atomique sur ces complexes macromoléculaires. La stratégie utilisée a consisté dans un premier temps à générer des intermédiaires paramagnétiques clefs du fonctionnement de ces systèmes (radicaux semiquinones ou ion MoV). Puis j'ai caractérisé leurs propriétés rédox par potentiométrie suivie par spectroscopie RPE. Enfin, j'ai utilisé les techniques de spectroscopie RPE impulsionnelle à haute résolution, notamment la spectroscopie de corrélation des sous niveaux hyperfins (HYSCORE) pour sonder l'environnement magnétique local de ces intermédiaires à travers la détection des interactions nucléaires hyperfines et quadripolaires qui sont trop faibles pour être visibles par spectroscopie RPE classique. / The aim of my work is to elucidate at the molecular level the structure and the function of enzymes involved in energy conversion processes in living organisms. In particular, it is focused on the understanding of two important steps found in many of these systems, namely (i) their interaction with membrane quinones acting as electron/proton shuttles and (ii) the catalytic mechanism at the molybdenum active site. The nitrate reductase A (NarGHI) from the bacterium Escherichia coli is used as a model for these studies. This membrane-bound complex reduces nitrate into nitrite during anaerobic respiration and plays therefore an important role in the global nitrogen cycle. The goal of my research is mainly devoted to the identification of the molecular factors tuning the reactivity of this system at the two active sites. For this purpose, I mainly relied on the structural characterization of key paramagnetic intermediates e.g. semiquinone radicals or Mo(V) ion using electron paramagnetic resonance (EPR) spectroscopy in combination with rédox potentiometry. High resolution pulse EPR methods, especially Hyperfine Sublevel Correlation (HYSCORE) spectroscopy, were used to probe their local environment through the detection of hyperfine (and eventually quadrupole interactions) to nearby magnetic nuclei that are otherwise too weak to be measurable in conventional continuous wave EPR spectroscopy.

Metal-organic framework (MOF) compounds as a new class of porous coordination polymers consists of metal ions or clusters linked by organic molecules. They have gained recent interest because of their large surface areas and huge variety of the porous network structures. They exhibit interesting adsorption properties and therefore are potential candidates for various technical applications. In this work, continuous wave (cw) and pulsed electron paramagnetic resonance (EPR) methods such as pulsed electron-nuclear double resonance (ENDOR) and hyperfine sublevel correlation (HYSCORE) spectroscopy are applied to study metal-organic frameworks with respect to different aspects of their properties: The host-guest interactions between Cu2+ ions in [Cu3(btc)2]n (HKUST-1; btc: 1,3,5-benzenetricaboxylate) with adsorbed methanol (CH3OH), 13C enriched carbon monoxide and dioxide (13CO, 13CO2), hydrogen (H2), deuterium (D2) and mixed isotopic HD. In [Cu3(btc)2]n, the Cu2+ ions are connected to binuclear Cu/Cu paddle wheel units. Since the Cu2+ ions in the [Cu3(btc)2]n are antiferromagnetically coupled, the new compound [Cu2.97Zn0.03(btc)2]n is synthesized by isomorphous substitution containing about 1 % paramagnetic Cu/Zn paddle wheel units. The modified Cu/Zn paddle wheel units prove to be a very sensitive probe for the interactions with the adsorbed molecules. Secondly, the exchange interactions of antiferromagnetically coupled Cu/Cu paddle wheel units as well as additional inter-paddle wheel exchange interactions between the Cu/Cu pairs are studied in [Cu2(bdc)2(dabco)]n, a layered MOF with 1,4-benzenedicaboxylate (bdc) as linker and 1,4-diazabicyclo[2.2.2]octane (dabco) acting as pillars between the layers. In comparison to [Cu3(btc)2]n, the additional inter-paddle wheel exchange interactions are much easier disturbed by incorporation of Zn2+ ions into the framework structure. Third, the structural dynamics of the framework is investigated in the compound [Al(OH)(bdc)]n (MIL-53) which was isomorphously substituted by V(III)/V(IV) species. The 51V hyperfine structure revealed to be sensitive to the so-called breathing effect, a flexible structural behaviour upon guest adsorption/desorption or upon thermal treatment. It is shown that the aluminum ions can be substituted by vanadium but the octahedral coordination environment changes slightly to a pseudo-octahedral or a square-pyramidal coordination. Based on the hyperfine interactions between the electron spin and the nuclear spins of the surrounding atoms, structural models can be derived from orientation-selective measurements. In such a way, structural information of materials like powder samples and adsorbate complexes can be obtained which are hardly or even not accessible by other methods.

Le dioxyde de chlore est l’agent de blanchiment le plus utilisé pour le blanchiment des pâtes chimiques lignocellulosiques. Son utilisation s’accompagne malheureusement de la formation d’ions chlorates, composés inefficaces pour le blanchiment, ainsi que d’importants rejets de DCO et d’organo-chlorés (AOX), nuisibles pour l’environnement. Cette étude a consisté à concevoir un nouveau stade de blanchiment au dioxyde de chlore (stade D) à pH non-conventionnel, économe en réactifs, et visant à réduire l’impact environnemental d’une séquence classique. Le travail a été effectué sur des pâtes kraft de résineux, obtenues après cuisson ou à différents stades de la séquence de blanchiment. Les résultats ont montré que l’efficacité du nouveau stade D était meilleure lorsqu’il était placé en fin de séquence. Après optimisation et incorporation de peroxyde d’hydrogène comme réactif complémentaire, les pâtes obtenues présentent des caractéristiques équivalentes à celles d’un blanchiment conventionnel, en termes de blancheur et de degré de polymérisation moyen viscosimétrique ; et ce, avec une réduction drastique de la pollution (70% des AOX et 20 % de DCO), et un gain économique substantiel en productivité de séquence (diminution de la température et du temps de réaction). L’étude chimique de la réaction du dioxyde de chlore effectuée par différentes techniques (spectroscopie RPE, RMN, FTIR, chromatographie HPAEC-PAD), a révélé la présence de radicaux hydroxyles en milieu alcalin, et mis en évidence les principales étapes du mécanisme à différents pH. / Chlorine dioxide is the most widely used bleaching agent for the production of bleached chemical pulps. However, its main drawbacks are the formation of chlorate ions which decrease delignification efficiency and the reject of toxic chloro-organic molecules (AOX) in mill effluents. This study focused on the development of a new bleaching stage using chlorine dioxide (D stage) at non-conventional pH, to reduce environmental impact and production costs. The work was carried out on several softwood kraft pulps after cooking, and at different stages of the bleaching sequence. The best results of the novel D stage were obtained for pulps at low kappa number, i.e. at the end of the bleaching sequence. The D stage was optimized and coupled with hydrogen peroxide addition. The same brightness and viscosimetric average degree of polymerization as after conventional D bleaching were obtained. Interestingly, a very important decrease of pollution load (70% of AOX and 20% of COD) was obtained, accompanied by a significant gain of productivity and energy saving (lower temperature and reaction time).Chemical investigations on the reaction mechanisms, carried out by different techniques (ESR, NMR and FTIR spectroscopies, HPAEC-PAD chromatography), revealed the formation of hydroxyl radicals at alkaline pH. The structural modification of the pulp residual lignin was also studied; differences of the reaction mechanism as function of pH were highlighted.