Energy generating processes like the citric acid cycle are a pivotal part of metabolism. Members of the 2-oxoacid dehydrogenase multienzyme complex (OADHC) superfamily feed into and act within the citric acid cycle. OADHCs are composed of three enzymes: 2-oxoacid decarboxylase (E1), dihydrolipoamide acyltransferase (E2) and dihydrolipoamide dehydrogenase (E3). Covalent attachment of lipoic acid (LA) to E2 is essential for overall OADHC activity. Although thought to be absent in Archaea, it has recently been found that Thermoplasma acidophilum has all the components for an active recombinant OADHC (Heath et al., 2007). Recent studies have further suggested that Tp. acidophilum may have an enzyme to covalently attach LA to E2 (Sun et al., 2007; McManus et al., 2006). This work describes the cloning and recombinant expression of the Thermoplasma lipoate protein ligase (Tp. LplA), its C-terminal domain and a fusion protein composed of the above two proteins. Both proteins are required for lipoylation of E2 in vitro. For the first time, in vivo lipoylation of E2 in Tp. acidophilum cell cultures is also being reported. The effect of lipoylation and temperature on the Thermoplasma OADHC assembly has also been studied. This study revealed the temperature dependence of the E2 core and the whole complex assembly. These findings are in line with the optimum growth temperature of Tp. acidophilum. Dynamic light scattering and analytical ultracentrifugation were used to determine the molecular mass of whole OADHC. The molecular mass was determined to be 5 MDa with an octahedral geometry of the E2 core. The results of this work strengthen the assumption that these enzyme systems may have had or potentially have a role in the Archaea. This may hold further clues to the evolutionary relationship between the three kingdoms of life and the role of OADHCs/lipoylation in the Archaea. The temperature dependent assembly of the complex and thermostability of these proteins may also provide a model to study thermostability and protein-protein interactions at high temperatures.

Acclimation of photosynthesis allows plants to adjust the composition of their photosynthetic apparatus to adapt to changes in the environment, and is important in maintaining fitness. Dynamic acclimation refers to acclimation of fully developed leaves, after developmental processes have ceased. Rates of photosynthesis fluctuate with environmental change, and this requires metabolic adjustments. It has previously been shown that acclimation requires the chloroplastic glucose 6-phosphate/ phosphate translocator GPT2. Using label-free proteomics we characterised the acclimation deficient gpt2 mutant. High light acclimation involves changes in the composition of the photosynthetic proteome and increases in many other metabolic enzymes, but in gpt2 plants, a reduced ability to alter protein composition, and enhanced stress responses were seen. Using a combined transcriptomics and proteomics approach we also analysed acclimation to low temperature. We show that photosynthetic acclimation requires the cytosolic fumarase, FUM2. In fum2 mutants, an enhanced transcriptional response to low temperature was seen, which was impaired at the level of the proteome, relative to the WT. We also identified a protein encoding a β-Amylase, BAM5, that strongly responded to high light acclimation. The role of this protein was further characterised, and we show a nonchloroplastic location. Furthermore, suppression of this gene resulted in plants that were unable to acclimate, and had a reduced sugar content. This research highlights novel and diverse roles for proteins in acclimation, and provides a comprehensive proteomic profiling of high light and low temperature acclimation that has previously been lacking.

The variation in properties of Polypropylene (PP) / Ethylene Vinyl Acetate (EVA) blends upon EVA content, temperature, and light stability were followed by using tensile testing, impact testing, and differential scanning calorimetry (DSC). Young&rsquo / s modulus of the blends decreased with increasing drawing temperature and EVA content. The stress at break values of the blends slightly increased with EVA whereas decreased with drawing temperature. The percent strain at break values of the blends were found to increase abruptly by increasing EVA content and drawing temperature. These changes in the mechanical properties are the indication of compatibility. The impact tests were performed only at 0&deg / C, 25&deg / C and the impact strength increased with the temperature and EVA content, but none of the samples were broken at higher testing temperatures. The effect of stabilizer was very obvious because stabilizer-free samples failed after 400 hours whereas, the samples with stabilizer resisted up to 750 hours. Elongation values of the samples decreased because of increasing brittleness by UV irradiation. We also observed chain stiffening effect by crosslinking in all samples upon UV irradiation. Thermal analysis of the blends of the drawn samples showed an increasing trend of crsytallinity with increasing drawing temperature. Increasing drawing temperature made polymer chains more flexible because of the increasing chain mobility. These flexible chains were then oriented in one direction during tensile testing and therefore uniaxial crystallization occurred. The morphology of impact and tensile tests samples were also analyzed by scanning electron microscope (SEM). The fibrillation of pure PP is higher than the fibrillation of the blends.

Effects of temperature and light intensity on photofermentative hydrogen production by Rhodobacter capsulatus DSM1710 by use of acetic and lactic acids as substrates were studied. Experiments were conducted at 20, 30 and 38oC incubator temperatures under light intensities in the 1500 &ndash / 7000 lux range. pH of the medium and quantity of hydrogen forming together with quantity of biomass, and concentrations of acetic, lactic, formic, butyric and propionic acids in the medium were determined periodically. Growth took place and hydrogen was produced under all experimental conditions. Growth was found to increase with increase in temperature but to decrease with increase in light intensity. Total hydrogen produced increased with light intensity up to 6000 lux at 20oC, 5000 lux at 30oC and 3000 lux at 38oC and decreased beyond these values. Medium temperature of about 30oC was found to be optimum for cumulative hydrogen. pH was found to increase slightly and almost all of lactic acid and most of acetic acid was consumed while formic, butyric and propionic acids were first formed and then consumed in the experiments. Growth data fitted well to the logistic model and hydrogen production data fitted well to the Modified Gompertz Model. Lactic acid was found to be almost completely consumed by first order kinetics in early times. Consumption of acetic acid was found to follow zero order kinetics in the early times when lactic acid existed in the system but the order shifted to one later when most of lactic acid was consumed.

Le nouveau mode de combustion HCCI est adapté pour réduire les émissions d’oxydes d’azote et de particules fines issues de moteurs Diesel afin de respecter les futures normes d’émission Euro de plus en plus drastiques. Ce type de combustion se traduit par l’augmentation des émissions de monoxyde de carbone et des hydrocarbures et par une faible température des gaz d’échappement retardant ainsi leur conversion par le catalyseur d’oxydation Diesel (DOC). C’est dans ce contexte environnemental et économique que le couplage plasma-catalyseur apparait comme une solution intéressante afin d’améliorer l’efficacité du traitement des gaz d’échappement Diesel. Cette thèse est dédiée à l’étude du couplage d’un plasma non-thermique de type décharge à barrière diélectrique (DBD) et d’un catalyseur d’oxydation Diesel (Pt-Pd/Al2O3) pour le traitement de mélanges gazeux représentatifs d’un échappement de moteur Diesel HCCI (O2-NO-H2O-CO-CO2-CH4-C3H6- C7H8-C10H22-N2). Les expériences avec un réacteur plasma pilote ont été menées sur deux bancs expérimentaux : le premier à l’échelle laboratoire en vue de comprendre la physico-chimie impliquant le plasma et le catalyseur avec une attention particulière pour les sous-produits de réaction, et le second à l’échelle industriel afin de déterminer l’efficacité et la faisabilité d’un tel couplage dans les conditions de débit et de température les plus proches possibles de celles rencontrées en sortie moteur véhicule. L’étude menée en fonction de la puissance injectée dans le milieu, la VVH, la température des gaz, ainsi que la nature du cycle de roulage a permis de montrer l’efficacité du plasma pour abaisser de façon significative la température d’activation du DOC pour l’oxydation de CO et des hydrocarbures. Aussi, la présence du plasma en amont du DOC a permis, sur un cycle NEDC simulé, une réduction de 68% et 42% des masses de CO et des hydrocarbures émis en accord avec la norme Euro6 (2014). L’efficacité du plasma pour l’oxydation des hydrocarbures et de NO à basse température dans ces conditions de débits élevés (jusqu’à 900 Lmin−1 sur le cycle NEDC) a été confirmée et les principaux produits de réaction identifiés et quantifiés. / The new HCCI combustion mode is well adapted to improve nitrogen oxide and particulate matter reduction from Diesel engine in order to meet future emission regulations adopted in the Euro zone. However, HCCI engines emit relatively high amounts of unburned hydrocarbons and carbon monoxide due to lower engine exhaust temperature increasing the catalyst light-off time and decreasing the average efficiency of the Diesel oxidation catalyst (DOC). In this environmental and economic context, the combination of plasma with DOC has been considered especially for intermittent use during the cold start. The thesis presents the combination of nonthermal plasma upstream Diesel oxidation catalyst (Pt-Pd/Al2O3) applied to the treatment of simulating Diesel HCCI exhaust gas (O2-NO-H2O-CO-CO2-CH4-C3H6-C7H8-C10H22-N2). The studies were conducted at atmospheric pressure with a pilot-scale dielectric barrier discharge reactor (DBD) on two experimental devices. The first is a laboratory scale set-up (low flow rate : 20 Lmin−1) used to understand the physico-chemical involving the plasma and the catalyst by focusing on the by-products reactions. The second is an industrial scale (gas flow rate up to 260 Lmin−1) used to study the feasibility and the efficiency of the plasma-DOC system under conditions similar to those encountered in Diesel exhaust engine. The effects of the plasma, the DOC and the plasma-DOC systems on the exhaust gas have been investigated under various conditions. The main contribution of the plasma was to give a « thermal » and a chemical « push » to the DOC resulting in the decrease of light-off temperature for CO and HC oxidation. These improvements were shown to depend on the treatment conditions (injected energy i.e. energy density, space velocity, gas temperature and nature of the driving cycle). It is shown that for a simulated European Driving Cycle (NEDC), the combination of plasma upstream DOC reduces the cumulative mass of CO and hydrocarbons by about 68% and 42%, respectively, in accordance with the Euro 6 standard (2014). The efficiency of plasma for hydrocarbons and NO oxidation at low temperature in high flow conditions (up to 900 Lmin−1 on the NEDC) has been confirmed and the main reaction products identified and quantified.

Les Composés Organiques Volatils d’origine Biogénique (COVB) émis par la végétation représentent 1PgC.an-1 à l’échelle globale. Ces COVB, une fois émis dans l’atmosphère, peuvent participer à la formation d’ozone troposphérique ainsi qu’à la formation d’aérosols organiques secondaires et donc à la pollution atmosphérique. C’est pourquoi, il est important de quantifier le plus précisément possible les taux d’émissions de COVB et de mieux comprendre quels sont les facteurs environnementaux qui contrôlent ces émissions. Il est bien connu que les émissions de COVB sont contrôlées par la lumière et/ou la température mais elles peuvent également être influencées par d’autres facteurs comme le stress hydrique, bien que son impact soit encore mal compris. En effet, il a été montré que le déficit hydrique pouvait augmenter ou diminuer les émissions de COVB selon son intensité, sa durée et l’espèce étudiée. Dans le cadre du changement climatique, une intensification de la sécheresse est attendue en région Méditerranéenne avec une augmentation de la température, une diminution des pluies ainsi qu’une prolongation de la période de sécheresse. Ce changement climatique pourrait donc modifier les émissions de COVB. De plus, les effets d’une sécheresse appliquée sur plusieurs années sont encore mal connus. Dans cette étude, nous nous sommes intéressés à la réponse des émissions de COVB du chêne pubescent (Quercus pubcescens Willd.)face au stress hydrique attendu en région méditerrannéenne avec le changement climatique. / Biogenic Volatile Organic Compounds (BVOC) emitted by vegetation represent 1PgC.yr-1 at the global scale. These BVOC, once emitted into the atmosphere, can participate in the troposheric ozone formation as well as secondary oragnic aerosols and, consequently, on the atmospheric pollution. That’s why, it is very important to quantify, as accurately as possible, the BVOC emissions and to improbe the knowledge about the environmental factors which drive these emissions. It is well known that BVOC emissions are controlled by the light and the temperature but they can be impacted by other factors such as water stress. Nevertheless, these mechanisms are not well understood yet, since it has been shown that water stress can increase or decrease BVOC emissions according to the intensity and the duration of stress. In a context of climate change, we can expected an intensification of summer drought in Mediteranean area with an incerase of temperature, a decrease of rainfall as well as an elongation of stress period. This climate change could modify BVOC emissions. Moreover, the effects of a water stress applied during several years are not known. In this study, we wanted to evaluate the impact of water stress, expected with climate change, on BVOC emitted by Downy oak (Quercus pubescens Willd.), main isoprene emitter of Mediterranean region.