Le but de cette étude était de déterminer l'influence des différentes techniques de vinification sur la composition phénolique et les activités biologiques des moûts et des vins issus de raisins de Syrah et de Cabernet sauvignon appartenant à deux régions libanaises distinctes (vallée de la Bekaa et la région de Chouf) et à deux millésimes consécutifs (2014 et 2015). Parmi ces procédés, les effets de la macération pré-fermentaire à froid et à chaud, du traitement enzymatique, de deux souches de levures commerciales et les agents de collage ont été discutés dans notre étude. L'analyse des composés phénoliques par spectrophotométrie et HPLC a montré que la macération pré-fermentaire à chaud entraine une meilleure extraction des composés phénoliques que la macération pré-fermentaire à froid. L‟extraction des tanins et des polyphénols totaux sont favorisés par la température et le prolongement de la macération. L‟extraction des anthocyanes est aussi favorisée par la température mais à courte durée puisque le prolongement de la macération entraine une dégradation de ces composés. Les moûts et les vins issus de l‟addition d‟enzymes pectolytiques au début de la phase de macération montrent des activités antioxydantes et des concentrations en polyphénols totaux plus élevées comparés à celles réalisées sans ajout d‟enzymes. La fermentation alcoolique provoque une diminution de la concentration des polyphénols totaux ce qui révèle des différences significatives entre les vins fermentés par les deux souches de levures X et Y. Après fermentation alcoolique, la quasi-totalité des échantillons de vin ont présenté une augmentation de leur pourcentage d'inhibition avec l'apparition de nouveaux types d'activités biologiques qui n'existait pas au niveau des moûts. A la fin, les résultats montrent l‟importance de bien choisir le type de colle selon le type de vin ainsi que de minimiser la dose de collage appliquée afin de conserver la teneur en composés phénoliques du vin. / The aim of this study was to determine the influence of different winemaking techniques on phenolic composition and biological activities of musts and wines from grapes of Syrah and Cabernet sauvignon from two distinct Lebanese regions (Bekaa valley and Chouf district) and two consecutive vintages (2014 and 2015). Among these processes the impacts of pre-fermentative cold and heating maceration, enzymatic treatment, two different commercial yeast strains and fining agents were discussed in our study. Spectrophotometric and HPLC analysis of phenolic compounds showed that the pre-fermentative heating maceration leads to a better extraction of phenolic compounds than the pre-fermentative cold maceration. Tannins and total polyphenols extraction are favored by the temperature and the prolongation of maceration. Extraction of anthocyanins is also favored by the temperature with short duration since the extension of the maceration leads to a degradation of these compounds. Maceration enzymes addition at early stage of maceration, promoted higher concentration of total polyphenol and antioxidant activity compared to those macerated without added enzymes. Alcoholic fermentation results in a decrease of total polyphenols content which revealed differences between wines derived from X and Y strains. After alcoholic fermentation, almost all of the wine samples presented an increase of their percentage of inhibition with the occurrence of new types of biological activities which doesn‟t existed at must level. At the end, the results showed the importance of selecting a fining agent according to the type of wine and to minimize the dose of fining applied in order to conserve the content of phenolic compounds in wine.

La levure Saccharomyces cerevisiae joue un rôle essentiel dans la production de composés aromatiques, tels que les esters, les alcools supérieurs et les acides organiques, ainsi que dans la transformation de précurseurs d'arômes du raisin pendant la fermentation du vin. Afin d'identifier les bases génomiques et métaboliques de ces propriétés, un croisement a été réalisé entre deux souches de levures de vin, sélectionnées pour leurs besoins en azote différents lors de la fermentation. 130 ségrégants de génération F2 ont été génotypés par séquençage complet du génome et individuellement phénotypés pendant la fermentation en mesurant les métabolites extracellulaires par HPLC et GC-MS. Les flux métaboliques intracellulaires ont été estimés à l’aide d’un modèle à base de contraintes. Une analyse QTL (quantitative trait locus) a été utilisée pour identifier les allèles influençant les variations d'arômes et de flux métaboliques. Plus de 80 QTL expliquant la variation de 59 caractères quantitatifs ont été détectés. Ces caractères comprennent des paramètres fermentaires, de consommation de substrat, la production de principaux métabolites et d’arômes fermentaires, ainsi que le métabolisme de composés aromatiques du raisin. L’intérêt de la cartographie QTL pour identifier les déterminants génétiques de variations de flux intracellulaires (f-QTLs) a par ailleurs été démontrée. Les QTL détectés ont été disséqués et des gènes dont les allèles contribuent spécifiquement aux variations phénotypiques ont été identifiés. Ces résultats soulignent la complexité génomique et métabolique de la synthèse et de la transformation d'arômes par la levure. L'identification de ces déterminants génétiques permet de mieux comprendre les liens entre variation génétique des levures et traits technologiques et fournit une base précieuse pour le développement de souches optimisées par des stratégies génétiques de croisement assisté par marqueurs. / The yeast Saccharomyces cerevisiae plays a vital role in the production of aroma compounds, such as esters, higher alcohols and organic acids, and the conversion of grape-derived aroma precursors during wine fermentation. To identify the genomic and metabolic bases for these processes, a cross was performed between two wine yeast strains selected because of their different nitrogen requirement during fermentation. 130 F2-segregants were genotyped by whole genome sequencing and individually phenotyped during wine fermentation by measuring extracellular metabolites using HPLC and GC-MS. Intracellular metabolic fluxes were estimated by constraint-based modeling. Quantitative trait locus (QTL) mapping was used to identify allelic variants influencing variations in the aroma profile and metabolic fluxes. More than 80 QTLs explaining variation in 59 quantitative traits were detected. These traits consisted of general fermentation parameters, substrate consumption, the production of main metabolites and fermentative aromas and the metabolism of grape aroma compounds. The applicability of QTL mapping to detect regions influencing intracellular fluxes (f-QTLs) was furthermore demonstrated. Found QTLs were dissected and genes with allele specific contributions to the phenotype were identified. These results emphasize the genomic and metabolic complexity of yeast aroma formation. In addition, the identification of genetic determinants increases knowledge about the links between genetic variation and industrial traits and provides a valuable foundation for the development of optimized strains by marker-assisted selection breeding strategies.

Orientador: Hamilton Cabral / Banca: Rodrigo Simões Ribeiro Leite / Banca: Fabiana Fonseca Zanoelo / Resumo: A produção de proteases de origem microbiana depende das condições de cultivo e da diversidade bioquímica de cada espécie. Estudos comparativos entre fermentação em estado sólido (FES) e fermentação submersa (FSm) usando farelo de trigo e meio sintético, respectivamente, foram realizados para a determinação dos parâmetros de produção de proteases pelo fungo Aspergillus fumigatus Fresenius. A melhor produção de protease foi em FES no período de 96 horas utilizando farelo de trigo, temperatura de 30 ºC e 1x106 esporos/5g de substrato com 1.517 U/mL. Em FSm o pico de produção foi em pH 6,0, a 30ºC, 5x105 esporos/mL de meio no período de 72 e 96 horas em meio contendo 0,5 e 0,25% de caseína, respectivamente, ambos com 40 U/mL. Conforme a produtividade dos processos fermentativos, o extrato enzimático da FES foi utilizado para estudos de purificação e caracterização bioquímica. Neste estudo, a protease purificada apresentou atividade ótima em pH 7,5 e 50ºC, sendo inibida por Fenil-metil-sulfonil-fluoreto (PMSF) e mais intensamente por antipaína (1,6 µM). Sobre efeito de íons, foi observado modulação da atividade proteolítica, principalmente com inibição por AlCl3, cuja atividade proteolítica residual foi de 18% após incubação com este íon. Na presença de Ditiotreitol (DTT) e uréia houve diminuição da atividade proteolítica, apresentando atividades residuais de 63% em 200 mM de DTT e 10% com 5 M de uréia. Comparativamente, na concentração de 0,1% de cada surfactante estudado, notou-se redução da atividade proteolítica, sendo 97% em presença de Brometo de cetil-trimetil amônio (CTAB), 79% para 4 - (1,1,3,3 - Tetrametilbutil) fenil- polietileno glicol (Triton X-100), 55% com Polyoxyethylenesorbitan monolaurato (Tween-20) e completa redução da atividade (0%) em... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The microbial protease production depends on growing conditions and the biochemical diversity of each species. Comparative studies between solid-state fermentation (SSF) and submerged fermentation (SmF) using wheat bran and synthetic medium, respectively, were performed to determine the optimum parameters for protease production by the fungus Aspergillus fumigatus Fresenius. The best protease production was in SSF within 96 hours using wheat bran, temperature 30°C and 1x106 spores/5g of substrate, with 1,517 U/mL. In SmF peak production was at pH 6.0 at 30°C, 5x105 spores/mL of media within 72 and 96 hours in medium containing 0.5 and 0.25% casein, respectively, with 40 U/mL. According to the productivity of the fermentative processes, enzymatic extract was used from SSF to study purification and biochemical characterization. In this study, purified protease showed optimum activity at pH 7.5 and 50°C, and inhibited by Phenylmethylsulfonyl fluoride (PMSF) and more intensely for antipain (1,6 µM). Concerning to the effect of ions, we observed modulation of the proteolytic activity, especially with inhibition by AlCl3, which residual activity was of 18 % after incubation with this ion. In the presence of Dithiothreitol (DTT) and ureia, we observed progressive decrease in proteolytic activity, presenting residual activities of 63% with 200 mM DTT, and 10% with 5 M ureia. Comparatively, in the concentration of 0.1% of each surfactant studied, there was a reduction in the proteolytic activity in 97% in presence of Cetyl trimethylammonium bromide (CTAB), 79% with 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), 55% with Polyoxyethylenesorbitan monolaurate (Tween-20) and a complete inactivation in the presence of Sodium dodecyl sulfate... (Complete abstract click electronic access below) / Mestre

Les souches œnologiques présentent une importante diversité dans leur besoin en azote, qui se traduit par des différences de capacité fermentaire. A l'heure actuelle, les mécanismes impliqués dans la variabilité des profils fermentaires, suite à un épuisement en azote dans le milieu, ne sont pas connus. L'identification de ces mécanismes serait un atout dans la compréhension des phénomènes conduisant aux fermentations problématiques et dans les reprises de fermentation. Afin d'identifier ces mécanismes, nous avons couplé une approche de physiologie et génomique classique, à une approche de génétique impliquant la recherche de QTL basée sur l'efficacité fermentaire en condition de carence en azote. Nous avons ainsi caractérisé cette différence de besoin en azote entre souches comme étant une variabilité dans la capacité à percevoir la carence en azote et à développer un programme de quiescence réduisant le flux d'énergie et augmentant un état de stress. Ces remaniements d'énergie se traduisant alors par des différences de capacités fermentaires. L'approche QTL a quant à elle permis de détecter 23 régions du génome potentiellement impliquées dans le maintien de la capacité fermentaire. Après analyse nous avons identifié 4 gènes dont les variations alléliques sont responsables des variations phénotypiques entre souches. L'utilisation de ces gènes pourrait permettre la conception de marqueurs génétiques, exploités pour la sélection de souches ayant de bonnes capacités fermentaires. Les données issues de cette approche QTL suggèrent une étroite corrélation entre les différences de réponse au stress par les souches et l'implication des mécanismes de perception et de signalisation de l'azote. Enfin, l'ensemble de nos travaux offre une nouvelle hypothèse, en désignant la voie TOR comme le mécanisme responsable de la variation des capacités fermentaires entre souches. / Oenological strains present an important diversity in nitrogen requirement, which result by difference in the fermentative performances. Nowadays, mechanisms involved in variability of fermentation profiles, result in nitrogen depletion in the medium, are not known. The identification of these mechanisms would be an advantage in the comprehension of phenomena leading to problematic fermentation and the fermentation restart. To identify these mechanisms, we have coupled a physiological and classical genomic approach with a genetic approach involving the QTL mapping based on the fermentation capacity in conditions of deficiency. We have characterized this difference in nitrogen requirement between strains as variability in the ability to sense nitrogen starvation and develop a quiescent program that reduces the flow of energy and increases its adaptation to stress. These energy rearrangements result in differences of fermentative performances. QTL approach allowed to detect 23 genome regions potentially involved in maintaining of the fermentative capacity. After analysis we have identified 4 genes for which allelic variations are responsible for the phenotypic variation between strains. The use of these genes may allow the design of genetic markers, exploited for the selection of strains with good fermentation capacity. The data from this QTL approach suggest a correlation between differences in stress response and the involving of mechanisms sensing and nitrogen signaling. Finally, all our work supplies a new hypothesis, pointing to the TOR pathway as the mechanism responsible of variation in fermentation capacity between strains.

Orientadores: Telma Teixeira Franco, Saartje Hermalsteens / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-14T02:25:44Z (GMT). No. of bitstreams: 1 LopezGarzon_CamiloSixto.pdf: 3882684 bytes, checksum: 507e1d93e97fc4494726832b744c6cf8 (MD5) Previous issue date: 2009 / Resumo: Neste trabalho foram desenvolvidos estudos visando o estabelecimento de um processo de produção de lipídeos microbianos a partir de fontes renováveis, particularmente xilose, carboidrato derivado do processo de hidrólise de bagaço de cana-de-açucar e que em breve deverá estar disponível para fermentação. Inicialmente foi verificada a influência da fonte de nitrogênio e a relação inicial carbono-nitrogênio sobre o processo de acúmulo delipídeos utilizando duas linhagens características. A utilização de sulfato de amônio juntocom extrato de levedura em uma relação C/N de 50 g/g foi adequada para o processo de produção de lipídeos. A seguir, 25 leveduras oleaginosas foram testadas quanto a sua capacidade de assimilação de xilose e produção de lipídeos. A produtividade de produção de lipídeos e elevado rendimento energético da levedura Lipomyces starkeyi DSM 70296 permitiram sua escolha para as etapas posteriores. Estudos de inibição por substrato foram desenvolvidos e diferentes modelos matemáticos foram ajustados aos dados experimentais. O modelo de Mulchandani representou adequadamente a cinética de inibição. Um modelo do cultivo em batelada simples foi desenvolvido baseado nos dados cinéticos obtidos, o qual foi validado para cultivo com limitação de nitrogênio em biorreator de dois litros. A análise de composição dos lipídeos obtidos mostrou semelhança à de fontes vegetais. Finalmente, dois modos de operação em batelada alimentada foram testados, o modo de alimentação simples aumentou significativamente a produtividade do processo quando comparado ao processo em batelada simples convencional. A alimentação usando pH-amônio/auxostat seguida de alimentação de carbono dobrou a concentração celular final obtida nos modos anteriores, mantendo níveis de acúmulo aceitáveis. Assim, um processo de produção de lipídeos a partir de pentoses foi viabilizado, adicionalmente o lipídeo produzido atende as especificações necessárias do valor cetano para a produção de biodiesel. / Abstract: Studies attempting the establishment of a microbial lipid production process from renewable resources, mainly xylose, were developed. This pentose, obtained from sugar cane bagasse hydrolysis, is expected to be available in large quantities and is going to be used as a substrate in fermentation processes in the near future. Initially, the effects of nitrogen source and initial carbon to nitrogen ratio over the lipid accumulation process were tested using two typical oleaginous strains. The joint use of ammonium sulphate plus yeast extract at an initial C/N ratio of 50 g/g were determined as adequate conditions for induction of lipid production with these yeasts. Subsequently in this work, 25 oleaginous yeasts were tested in order to evaluate the ability to consume xylose and lipid production. As a result of this stage, Lipomyces starkeyi DSM 70296 was selected regarding its highest productivity and energetic yield over all the set of yeasts evaluated. Thus, this strain was used in the development of the later studies. Substrate inhibition studies were developed and several mathematical models were adjusted to the experimental data. The Mulchandani model fitted well the substrate inhibition kinetics. Based on the kinetics obtained in shake flask fermentation, a batch model that represents the behavior of the principal variables in the time was developed. The model proposed was validated successfully using a set of data from two liters bioreactor cultivation. The composition of the obtained lipids was very similar of those from vegetal sources. Finally, two fed-batch operation modes were tested intending to get higher lipid productivities. In this direction, simple carbon fed-batch operation showed better productivities than those obtained from simple batch. Nitrogen fed using the pHamonium/auxostat mode followed by carbon fed duplicated the final cellular concentration obtained in the other modes tested, maintaining an adequate lipid content. Therefore, a process for lipid production was developed, additionally; the lipids obtained attend the cetane value specifications required for biodiesel production. / Universidade Estadual de Campi / Desenvolvimento de Processos Químicos / Mestre em Engenharia Química

Orientador: Maria Regina Wolf Maciel / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-24T00:04:45Z (GMT). No. of bitstreams: 1 Silva_BrunaTorresda_M.pdf: 1645900 bytes, checksum: cb35ae2b39e57ae64320dc4a3efe7b0b (MD5) Previous issue date: 2013 / Resumo: O presente trabalho descreve o estudo realizado para avaliar a viabilidade da produção de dois ácidos por uma metodologia nova e livre de sínteses químicas. Para a produção do ácido láctico, analisou-se o comportamento de três cepas bacterianas diferentes na fermentação do melaço de cana-de-açúcar: Lactobacillus delbrueckii, Leuconostoc mesenteroides e Lactobacillus plantarum. Foram realizados estudos em shaker, para simular o comportamento desses microrganismos frente a diferentes condições de temperatura (de acordo com cada bactéria), de concentrações de sacarose (15 g/L, 25 g/L e 35 g/L, aproximadamente) e de extrato de levedura (2 g/L, 4g/L e 6 g/L). Posteriormente, as melhores condições obtidas para cada bactéria foram reproduzidas em fermentador de grande volume, com controle de pH para avaliar a viabilidade do processo. A segunda etapa consistiu na aplicação do ácido láctico, obtido via fermentação, como substrato para a produção do ácido propiônico pela bactéria Propionibacterium acidipropionici. A manutenção da cepa e a preparação do pré-inóculo foram constituídos de lactato de sódio, em concentrações progressivas, até que a mesma estivesse pronta para ser inserida no meio de fermentação. Por ser um microrganismo anaeróbio, foi utilizada a alimentação de N2 ao reator, previamente ao início do processo, para manutenção da vida da cepa durante o processo, que contou com controle de pH, agitação e, consequentemente, taxa de formação de O2 durante o processo. Todas as amostras retiradas durante os processos foram quantificadas com relação aos açúcares e aos ácidos em cromatografia líquida de alta eficiência (HPLC). / Abstract: This paper describes the study conducted to evaluate the feasibility of production of two acids by a new methodology free of chemical syntheses. For the production of lactic acid, the behavior of three different bacterial strains in the fermentation of sugar cane molasses were analyzed: Lactobacillus delbrueckii, Lactobacillus plantarum and Leuconostoc mesenteroides. Shaker studies were performed to simulate the behavior of these micro-organisms to different temperature conditions (in accordance with each bacterium), and concentrations of sucrose (15 g/L 25 g/L and 35 g/L, approximately), and yeast extract (2 g/L, 4g/L and 6 g/L). Subsequently, the best conditions obtained for each bacterium were reproduced in large volume fermenter with pH control to assess the viability of the process. The second step consisted of the application of lactic acid obtained through fermentation, as a substrate for the production of propionic acid by the bacterium Propionibacterium acidipropionici. The maintenance of strains and the preparation of the pre-inoculum consisted of sodium lactate at progressive concentrations until it was ready to be inserted into the fermentation medium. Being an anaerobic microorganism, it was used to feed the reactor N2, prior to the start of the process, to maintain the life of the strain during the process, which had control of pH, agitation and the consequent rate of formation of O2 during the process. All samples taken during the process were quantified with respect to sugars and acids in high performance liquid chromatography (HPLC). / Mestrado / Desenvolvimento de Processos Químicos / Mestra em Engenharia Química

Dans un contexte de transition énergétique, ce travail de recherche s’inscrit dans la dynamiqued’explorer la potentialité de nouvelles biomasses pour la production d’un vecteur énergétique propre,l’hydrogène (H2). Au cours de cette thèse, un procédé de production d’H2 par fermentation endogèneet sans dépense énergétique supplémentaire, a été élaboré à partir de biomasses viticoles, etl’influence de paramètres opératoires a été testée. Enfin, le type de biomasses influe sur laproduction d’H2, non seulement par sa composition biochimique, mais aussi par leur microfloreendogène qui oriente le métabolisme. La microflore endogène responsable de la production d’H2 àpartir de ‘bourbes’ (50 ± 6 L/Lbourbes avec un rendement de 2,0 ± 0,2 mol/mol) appartient à la familleClostridiaceae et est associée aux voies acétate/butyrate, alors que celle des ‘marcs’ (20 ± 4L/kgmarcs avec un rendement de 1,3 ± 0,3 mol/mol) appartient aux Enterobacteriaceae et estassociée aux voies acétate/éthanol. / In the context of energy transition, this work deals with the exploration of new potential biomasses forthe production of a clean energy vector, hydrogen (H2). During this thesis, a process of H2 productionby an endogenous fermentation from winery waste has been developed without any additionalenergetic consumption. The influence of operating parameters was studied. Finally, the type ofbiomass has an influence on the H2 production, not only because of initial biochemical composition,but also because of the endogenous microflora which orientates the metabolism. The endogenousmicroflora responsible of H2 production from ‘solid grape residues’ biomass (50 ± 6 L.L-1biomass with a2.0 ± 0.2 mol.mol-1 yield) through acetate/butyrate pathways belongs to Clostridiaceae family whilethe one responsible of H2 production from ‘grape pomace’ biomass (20 ± 4 L.kg-1biomass with a 1.3 ±0.3 mol.mol-1 yield) through acetate/ethanol pathways belongs to Enterobacteriaceae family.

Dans un contexte économique tendu pour les chocolatiers « premium », la maîtrise de la qualité organoleptique de la matière première est un impératif et un avantage compétitif. La qualité aromatique d'un chocolat est entre autre liée aux traitements post-récolte et particulièrement à l'étape de fermentation. Cette étape est essentielle au développement des caractéristiques sensorielles. Or, les outils de contrôle qualité du cacao sont basés principalement sur l'évaluation sensorielle d'échantillons. Cette méthode est onéreuse et chronophage. Le développement d'outils rapides et robustes d'analyse de la qualité devient nécessaire. Cette étude a pour objectif de caractériser le déroulement de la fermentation afin de sélectionner des marqueurs spécifiques de la qualité du cacao. L'étude s'est intéressée aux marqueurs mesurables chez le planteur (à travers l'évolution de la température et du pH), et à ceux issus des transformations biochimiques des fèves (de type azoté et polyphénolique). Ces marqueurs ont ensuite été confrontés aux données traditionnelles de qualité (cut-test et dosage de l'azote ammoniacal), et aux données sensorielles, afin de sélectionner les plus pertinents pour le pilotage de fermentation. L'analyse globale de l'ensemble des résultats a permis de faire ressortir huit bio-marqueurs analytiques et trois bio-marqueurs sensoriels, représentatifs de la qualité du cacao et en lien avec le développement de la fermentation. Par ailleurs, des recommandations de méthodes de mesure de ces bio-marqueurs à mettre en place et permettant de contrôler la qualité ont été proposés. / In a difficult economic context for "premium" chocolate makers, the control of the organoleptic quality of the raw material is an imperative and a competitive advantage.The aromatic quality of a chocolate is highly related to post-harvest treatment and particularly to the fermentation. This step is essential to the development of sensory characteristics.However, cocoa quality control tools are mainly based on sensory evaluation of samples. This method is expensive and time consuming.Fast and robust tools for quality analysis are needed. This study aims at characterizing the course of the fermentation in order to select specific markers of the quality of cocoa. The study focused on markers which can be measured on plantation (through the evolution of temperature and pH), and those from biochemical transformations of the beans (nitrogenous and polyphenol type). These markers have been then compared to data from traditional quality methods (cut-test and ammonia nitrogen quantification) and to sensory data in order to select the most relevant ones for the control of fermentation. The overall analysis of the database enabled to identify eight analytical bio-markers and three sensory bio-markers, representative of the quality of cocoa and in connection with the development of fermentation. In addition, recommendations for methods of measuring these bio-markers to implement and to monitor the quality have been presented.

L’électro-fermentation est un nouveau levier permettant le contrôle des procédés fermentaires à travers l'utilisation d'électrodes au potentiel contrôlé. Parmi de nombreux substrats fermentaires, le glycérol est une source de carbone largement utilisée issue de l’industrie du biodiesel, et permettant la production de molécules à valeur ajoutée comme le 1,3-propanediol. L'objectif de cette thèse est d'évaluer le potentiel de l’électro-fermentation du glycérol comme moyen de mieux maîtriser les spectres de produits fermentaires dans les procédés mettant en œuvre des cultures mixtes.La thèse étudie dans un premier temps la fermentation du glycérol en cultures mixtes afin de caractériser les principales voies métaboliques d'intérêt en réponse au paramètre environnemental le plus influent pour la fermentation du glycérol (pH). L'effet de l'introduction d'électrodes colonisées par des bactéries électro-actives, capables d'échanger des électrons avec l'électrode et d’autres microorganismes, est ensuite étudié. Ce travail est réalisé en cultures mixtes dans l'objectif d'améliorer le procédé de fermentation en termes de spécificité des métabolites formés et de leur rendement de production. Enfin, un système modèle composé d’une souche fermentaire et une souche électro-active a ensuite été conçu afin de mieux comprendre les mécanismes mis en jeu lors de l’électro-fermentation. Cette thèse ouvre de nouvelles possibilités quant à la régulation des balances redox lors de fermentation. L’électro-fermentation ainsi que l’utilisation de bactéries électro-actives ont le potentiel de devenir de puissants outils permettant d’améliorer les rendements et spécificité de production du 1,3-propanediol et d’autres molécules à valeur ajoutée. / Electro-fermentation is a novel tool allowing to control classic fermentation through the use of polarized electrodes. Among all possible fermentation substrates, glycerol is a widely used by-product from the biodiesel industry that can be converted in value-added chemicals such as 1,3-propanediol. This PhD thesis aims at evaluating the potential of glycerol electro-fermentation for the improvement of product specificity in mixed-culture fermentation.As a first step, classic fermentation of glycerol by mixed bacterial consortia was studied in order to characterize the main metabolic pathways according to the main influencing environmental parameter (pH). Then, the addition in fermentation broth of electrodes and electro-active bacteria, able to exchange electrons either with an electrode or other microorganisms has been investigated. This work was carried out in mixed-culture glycerol fermentation in order to optimize products selectivity and yields towards 1,3-propanediol. Finally, a model co-culture constituted of one fermentative and one electro-active species was used to elucidate part of the mechanisms underlying electro-fermentation. This thesis opens a whole new range of possibility regarding the regulation of redox balances in fermentation. Hence electro-fermentation and the use of electro-active bacteria could become efficient tools for improving specificity and yield of 1,3-propanediol and other value-added products in fermentation.

Les unités de conditionnement des dattes génèrent des quantités importantes de déchets issues des écarts de triage. Cette biomasse, considérée jusqu'alors comme un déchet avec un impact sur l'environnement peut être transformée en produit à haute valeur ajouté. La valorisation des sous-produits de l'industrie des dattes en biocarburant s'inscrit dans une démarche économique et environnementale. Mais les fortes teneurs en sucres dans les sirops de dattes induisent une forte pression osmotique qui limite la capacité fermentaire des micro-organismes tel que Saccharomyces cerevisiae. Ceci nous a conduit à utiliser des souches osmotolérantes comme Zygosaccharomyces rouxii et Candida Pelliculosa. Différents essais ont été menés dans des milieux de culture à base de sirop de dattes, à différentes teneurs en sucres, en milieu discontinue et parfaitement agité. Les essais menés dans un milieu de culture à base de jus de dattes à 17,4 °Brix, conduisent à la production d'éthanol aux concentrations de 63 g L-1, 41 g L-1 et 33 g L-1 respectivement pour les levures Saccharomyces cerevisiae, Candida Pelliculosa et Zygosaccharomyces rouxii. Les essais menés dans le milieu à 35,8 °Brix (milieu de culture se rapprochant le plus des sirops de dattes brutes) montrent que la croissance des levures Saccharomyces cerevisiae et Candida Pelliculosa est inhibée par la pression osmotique élevée causée par la haute concentration en sucres. Seule la levure xérotolérante Zygosaccharomyces rouxii s'est adaptée au milieu en produisant 55 g L-1 de bioéthanol. La souche isolée des sous-produits de des dattes comme étant un Bacillus amyloliquifaciens est une souche allochtone capable de se developper dans un milieu hyper-osmotique et convertir les trois sucres (glucose, fructose et saccharose) en éthanol, contrairement à Z. rouxii qui est une souche fructophile. Le rendement en éthanol de B. amyloliquefaciens pour le milieu M-S175 est de 0,45 g.g-1, bien supérieur à ceux de S. cerevisiae, C. pelliculosa et Z. rouxii qui donnent respectivemnt 0,38, 0,29 et 0,34 g.g-1. En produisant 90 g.L-1 d'éthanol pour le milieu M-S350, B. amyloliquefaciens nous semble prometteuse pour valoriser un substrat très riche en sucres fermentescible. / Conditioning unit's dates generate large amounts of wastage after sorting. This biomass, previously regarded as waste product with a high impact on the environment can be transformed into a highly valued product. The use of by-products of date fruit into biofuel industry is a part of an environmentaly friended economic process. But high levels of sugars in the syrups of dates induce high osmotic pressure which limits the ability of fermentative microorganisms such as Saccharomyces cerevisiae. This led us to use osmotolerant strains, Zygosaccharomyces rouxii and Candida pelliculosa. Various tests were conducted in culture media containing date juice at different levels in sugars, and perfectly stirred in batch conditions. The tests performed in a culture medium based on date juice at 17.4 ° Brix, led to the production of 63 g L-1, 41 g L-1 and 33.1 g L-1 of ethanol for the yeasts Saccharomyces cerevisiae, Candida pelliculosa and Zygosaccharomyces rouxii, respectively. Tests conducted at 35.8 °Brix (culture medium close to syrup raw dates) show that the growth of yeast Saccharomyces cerevisiae and Candida pelliculosa is inhibited by high osmotic pressure caused by the high concentration of sugars. Only the xerotolerante yeast Zygosaccharomyces rouxii is adapted to the environment and produced 55 g L-1 of bioethanol. The strain isolated from by-products of dates, identified as bacillus amyloliquefaciens, is able to develop in a hyper-osmotic environment and convert the three sugars (glucose, fructose and sucrose) into ethanol, unlike Z. rouxii which is a fructophilic strain. The yield of ethanol for the M-S175 is 0.45 g.g-1, higher than those of S. cerevisiae, C. pelliculosa and Z. rouxii which give respectivemnt 0.38, 0.28 and 0.34 g.g-1. By producing 90 g.L-1 of ethanol for M-S350, B. amyloliquefaciens seems promising to valorize a very rish substrate in fermentable sugars.