Binderless fiberboard production from Cynara cardunculus and Vitis viniferaTwo lignocellulosic materials, Cynara cardunculus and Vitis vinifera, were pretreated and used to produce fiberboards without synthetic adhesives. The lignocellulosic materials were steam exploded through a thermo-mechanical vapor process in a batch reactor. After pretreatment the materials were dried, ground and pressed to produce the boards. The effects of pretreatment factors and pressing conditions on the chemical and physicomechanical properties of the fiberboards were evaluated and the conditions that optimize these properties were found. Response surface methodology based on a central composite design and multiple response optimization were used. The variables studied were: pretreatment temperature, pretreatment time, pressing temperature, pressing pressure, and pressing time. Binderless fiberboards produced from Cynara cardunculus stalks at the optimum conditions found fulfilled the European standards for boards of internal use. Nevertheless, binderless fiberboards produced from Vitis vinifera prunings at the optimum conditions found for this material did not completely met the European standards; modulus of rupture and internal bond values for these boards were lower than required minimums.Simultaneously, commercial Kraft lignin was reacted in an alkaline medium to enhance its adhesive properties. Chemical changes in reacted Kraft lignins that include ash content, Klason lignin, acid-soluble lignin and sugars were determined, as well as, structural characteristics of these lignins in terms of phenolic hydroxyl, aliphatic hydroxyl, methoxyl, carbonyl, Mw, Mn and polydispersity. The effects of reaction temperature and reaction time on lignin properties were studied using response surface methodology, and optimal reaction conditions were found.Two different types of Kraft lignin were used, alkali treated Kraft lignin and crude acid-washed Kraft lignin, as additives to enhance the physicomechanical properties of binderless fiberboards produced from Vitis vinifera to reach and overcome the European standards completely. At the end fiberboards produced with 20% of Vitis vinifera fibers replaced by crude acid-washed Kraft lignin were able to meet the European standards completely.This research work was an effort to reduce our dependency upon petroleum derivates, to diminish deforestation and to increase the use of renewable and biodegradable materials with the intention of preserving the environment and to encourage a sustainable development of our society. / Producción de Tableros de Fibras a partir de Cynara cardunculus y Vitis viniferaEn el presente estudio trozos Cynara cardunculus y Vitis vinifera fueron pretratados, y usados para producir tableros de fibras sin adhesivos sintéticos. Estos materiales lignocelulósicos se explotaron con vapor a través de un proceso termomecánico de vapor en un reactor por lotes. Después del pretratamiento el material fue secado, molido y prensado en caliente para producir los tableros. Se evaluaron los efectos de los factores del pretratamiento (temperatura de reacción y tiempo de reacción) y las condiciones de prensado (presión de prensado, temperatura y tiempo) sobre las propiedades químicas y físico-mecánicas de los tableros de fibras y se establecieron las condiciones que optimizan dichas propiedades. Las propiedades físico-mecánicas de los tableros de fibras que fueron estudiadas son: densidad, módulo de elasticidad (MOE), módulo de ruptura (MOR), enlace interno (IB), absorción de agua (WA) y hinchazón en hinchazón (TS) y las propiedades químicas estudiadas de la materia prima y el material pretratado fueron las siguientes: Cenizas, contenido de lignina Klason, contenido de celulosa y contenido de hemicelulosas. Se uso una metodología de superficie de respuesta basada en un diseño de experimentos del tipo central compuesto y una metodología de optimización de respuesta múltiple.Los tableros de fibras sin adhesivos sintéticos producidos a partir de tallos de Cynara cardunculus a las condiciones óptimas encontradas cumplieron con las normas europeas para los tableros de uso interno. Sin embargo, los tableros de fibras sin adhesivos sintéticos producidos a partir de podas de Vitis vinifera a las condiciones óptimas encontradas para este material no cumplieron totalmente las normas europeas; los valores del módulo de ruptura y del enlace interno para estos tableros fueron inferiores a los mínimos requeridos. Una lignina Kraft comercial fue sometida a reacción en un medio alcalino para mejorar sus propiedades adhesivas. Se determinaron los cambios químicos en las ligninas Kraft tratadas, las propiedades medidas fueron: contenido en cenizas, lignina Klason, lignina soluble en ácido y azúcares, también se determinaron las características estructurales de estas ligninas en términos de hidroxilos fenólicos, hidroxilos alifáticos, metóxilos, carbonilos, Mw, Mn y polidispersidad. Se estudiaron los efectos de la temperatura de reacción y el tiempo de reacción sobre las propiedades de la lignina con una metodología de superficie de respuesta, y se encontraron la condiciones óptimas de reacción.Se usaron dos tipos diferentes de lignina Kraft, lignina Kraft tratada en medio alcalino y lignina Kraft cruda lavada con ácido, como aditivos para mejorar las propiedades físico-mecánicas de los tableros de fibras sin adhesivos sintéticos producidos a partir de Vitis vinifera, para alcanzar y superar las normas europeas completamente. Al final los tableros de fibras producidos con una substitución del 20% de fibras de Vitis vinifera por lignina Kraft cruda lavada con ácido fueron capaces de satisfacer las normas europeas por completo.Este trabajo de investigación fue un esfuerzo para reducir nuestra dependencia de los derivados del petróleo, para disminuir la deforestación y para aumentar el uso de materiales renovables y biodegradables con la intención de preservar el medio ambiente y fomentar un desarrollo sostenible de nuestra sociedad.

Le bois est un matériau nanocomposite complexe, fortement anisotrope et hiérarchiquement organisé. La micro-structure à l'échelle nanométrique est caractérisée par des nanofibres rigides de cellulose cristalline parallèles les unes aux autres et noyées dans un matrice plus molle et moins anisotrope. Cette matrice est composée par hémicelluloses et lignine, avec une absorption contrôlée par l'entropie et les liaisons hydrogène entre hémicelluloses, lignine, et la surface de micro-cristaux de cellulose. La matrice est hygroscopique et se gonfle avec les changements de l'humidité relative, en fournissant une source de stress interne, tandis que les fibrilles de cellulose s'enroulent en spirale autour de la lumière centrale à un angle précis, qui est appellé AMF, angle des micro-fibrilles. Les micro-fibrilles se réorganisent passivement selon les contraintes osmotique appliquées. Selon l'AMF, le positionnement et les propriétés mécaniques des fibres de bois varient considérablement: le matériau rigide se rétrécit ou il s'allonge dans le sens de la longueur lors du gonflement, en générant de cette façon une grande déformation anisotrope.Une première forme d'équation d'état comprenant entropie et termes chimiques, colloïdaux (comme la force d'hydratation) et termes mécaniques macroscopiques a été établie et permet de prédire sans paramètres l'absorption d'eau en fonction de l'humidité relative du composite de bois pas traité.Le but de la thèse est d'étendre cette équation d'état au bois traité par absorption d'électrolytes en conditions hydrotermales, en intégrant l'équation avec des termes chimiques lié à la présence d'électrolytes. Les résultats du modèle sont comparés avec les données expérimentales de sorption d'électrolytes formulés en fonction de la balance entre chaotropes et cosmotropes de l'échelle de Hofmeister. Les données expérimentales sont produit à partir des isothermes de sorption et des mécanismes moteurs pour tester la prédiction de la théorie en explorant l'effet du soluté (introduit dans le bois par mise en équilibre avec une solution utilisée comme une réservoir osmotique) sur la structure et les dimensions du matériau. / The so-called “wood material” is a complex, highly anisotropic and hierarchically organized nanocomposite. At the nanometric scale, it is characterised by stiff crystalline cellulose nanofibres parallel to each others embedded in a matrix of a much softer, less anisotropic, gel of hemicelluloses, lignin and water. This matrix is hygroscopic and the solvent uptake is controlled by molecular forces like entropy, H-bonding of polysaccharides to cellulose nano-crystals and hydration force. The swelling provides a source of internal stress for the cellulose fibrils that, winding with a spiral angle (the microfibril angle, MFA) around the central lumen, passively reorient following the osmotic stress applied to them. Depending on the MFA, wood fibres exhibit a wide range of behaviors and mechanical properties, being able to act as stiff material to bear load, or shrink or expand in the longitudinal direction upon swelling, generating in this way either large tensile or compressive stresses or large strains.For the first time, the equation of state including entropic, chemical, colloidal terms (as the hydration force) as well as the mechanical, macroscopic, term has been established and allows to predict with a parameterless analytical expression, the water absorption of untreated softwoods as a function of relative humidity changes.The aim of the thesis is to extend this first equation of state to describe wood treatments by electrolytes adsorption in hydrothermal conditions. The experimental part is performed using specific sorption devices as well as X-Ray scattering techniques. The theoretical part is developed extending the established master equation linking molecular level chemical forces, colloidal interactions and mechanics. The results of the model are tested against experimental data of electrolytes adsorption and compared with their chaotropic/cosmotropic character according to the Hofmeister series.Nowadays, several wood treatments have been developed to reduce the water uptake by wood, in order to confer higher resistance to moist to low-quality wood materials. The final aim of the project is to test the prediction of the model against these known treatments and to develop a general model to describe the behaviour of the wood under different chemical and physical environments. This way, new treatments can be conceptually designed and pre-existing treatments (as for instance the patented "woodprotect" one) improved.

La réduction de la consommation des carburants fossiles est l’un des enjeux majeurs du XXIème siècle. Le bioéthanol représente une alternative durable à l’essence, mais sa production est limitée puisqu’il est produit à partir de ressources alimentaires. L’éthanol de seconde génération offre une alternative pour relancer son développement. Encore au stade expérimental, il serait produit à partir de biomasse lignocellulosique (bois, paille, plantes annuelles..) et donc ne concurrencerait pas l’alimentation humaine. Cette thèse s’inscrit dans le cadre de production d’éthanol de 2nde génération dans une usine de pâte kraft. Dans ces usines, les hémicelluloses, qui représentent 20 à 30% du bois, ne sont pas valorisées. Cette étude porte sur l’extraction de ces hémicelluloses en amont du procédé et de leur fermentation en éthanol. Elle consiste à la mise au point et l’optimisation d’un procédé global allant du copeau de bois à l’éthanol. / Decreasing the consumption of crude oil derivatives has become one of the main world issues of the 21st century. The only green substitute for gasoline potentially available in large quantities today is bioethanol, but its production has to be limited as it is produced from crops. It is therefore important to develop the production of second generation ethanol, which consists in using lignocellulosic biomass as raw material to avoid to compete with food resources. Second generation ethanol is still at experimental scale nowadays. This thesis deals with the specific case of the production of ethanol from hemicelluloses in a kraft pulp mill. In pulp mills, hemicelluloses, which make up 20 to 30% of the wood, are not value added. It is proposed to extract them from wood prior to the kraft treatment and to ferment them into ethanol. The study talks more specifically of the optimisation of the global process from the woodchip to the ethanol.

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / In this work six galactomannans (Adenanthera pavonina, Caesalpinia pulcherrima, Delonix regia, Dimorphera mollis, Prosopis glandulosa, Schizolobium parahyba) and three xyloglucans (Hymenaea courbaril, Mucuna sloanei e Tamarindus indica) were isolated from seed endosperm and cotyledon, respectively, by aqueous extraction followed by precipitation with ethanol. Yield of extraction, monosaccharide ratio, macromolecular parameters as well as molar mass distribution were determined and compared to guar gum (Cyamopsis tetragonoloba), LBG (Ceratonia siliqua) and xanthan (Xanthomonas campestris). Extraction yield in relation to seed mass ranged from 7.0 to 40.63%, with xyloglucan yields higher than the galactomannans ones. Schizolobium parahyba and Caesalpinia pulcherrima galactomannans presented the lowest protein content of 0.05% e 0.08%, respectively. The Mw values ranged from 0.09 â 3.37 x107 g mol-1 Flow curves of hemicelluloses solutions at 1% (w:v) were measured by varying the shear rate from 0.1 to 100 s-1. The resulting data were fit to the Power law e Herschel-Bulkley models. All the hemicelluloses presented shear-thinning behavior. Galactomannan and xyloglucans with different monosaccharide ratio showed similar consistency index; it may be influenced of the galactose distribution pattern on the chains and the interactions among the polysaccharides molecules. Rheological properties were compared and the results suggest new hemicelluloses sources which offer more profound applications in areas such as materials science, medicine e biology. / As hemiceluloses sÃo polissacarÃdeos presentes na parede celular de vegetais, onde funcionam como polissacarÃdeos de reserva e possuem uma cadeia principal composta por ligaÃÃes β-(1→4) em configuraÃÃo equatorial. Neste trabalho, hemiceluloses de sementes foram avaliadas quanto as suas propriedades reolÃgicas. Como resultado, galactomananas endospÃrmicas de Adenanthera pavonina, Caesalpinia pulcherrima, Delonix regia, Dimorphera mollis, Prosopis glandulosa e Schizolobium parahyba e xiloglucanas cotiledonÃrias de Hymenaea courbaril, Mucuna sloanei e Tamarindus indica foram isoladas por extraÃÃo aquosa, seguida por precipitaÃÃo em etanol. O rendimento das extraÃÃes, razÃo monossacarÃdica, parÃmetros macromoleculares, assim como distribuiÃÃo de massa molar foram determinados e comparados Ãs jà comercializadas goma guar (Cyamopsis tetragonoloba), LBG (Ceratonia siliqua) e goma xantana (Xanthomonas campestris). O rendimento das extraÃÃes em relaÃÃo à massa das sementes mostrou um intervalo de 7,0 a 40,63%, com os maiores valores para as xiloglucanas. Galactomananas de Schizolobium parahyba e Caesalpinia pulcherrima apresentaram os menores percentuais de proteÃnas, 0,05% e 0,08%, respectivamente. Valores de Mw variaram dentro de um intervalo entre 0,09 â 3,37 x107 g mol-1. Curvas de fluxo das soluÃÃes de hemiceluloses a 1% (m:v) foram obtidas pela variaÃÃo da taxa de cisalhamento entre 0,1 a 100 s-1. Os dados obtidos foram analisados pelos modelos de Lei das PotÃncias e Hershel-Bulkley demonstrando carÃter pseudoplÃstico para todas as hemiceluloses estudadas nessa concentraÃÃo. Galactomananas e xiloglucanas com diferentes razÃes monossacarÃdicas apresentaram Ãndice de consistÃncia similar, provavelmente influenciado pelo padrÃo de distribuiÃÃo de galactose nas cadeias e as interaÃÃes entre as molÃculas desses polissacarÃdeos. As propriedades reolÃgicas foram comparadas e os resultados sugerem novas fontes de hemiceluloses que permitem mais aplicaÃÃes em Ãreas como ciÃncia dos materiais, medicina e biologia.

Throughout the study of polymer adsorption at the air/water and solid/water interfaces, surface tension measurements and surface plasmon resonance (SPR) spectroscopy have been identified as key methods for the acquisition of structural and thermodynamic information. These techniques were used to determine air/water and cellulose/water interfacial properties of pullulan (P) and pullulan cinnamates (PCs), 2-hydroxypropyltrimethylammonium xylans (HPMAXs), and hydroxypropyl xylans (HPXs). Hydrophobic modification of pullulan with cinnamate groups promoted adsorption onto model surfaces of regenerated cellulose. In order to understand the relative contributions of hydrophilic and hydrophobic interactions towards PC adsorption, PC adsorption onto self-assembled monolayers (SAMs) with different functional groups was also studied. As the degree of cinnamate substitution increased, greater adsorption onto cellulose, methyl-terminated SAMs (SAM-CH3), and hydroxyl-terminated SAMs (SAM-OH) was observed. This study showed that hydrogen bonding alone could not provide a complete explanation for PC adsorption onto cellulose. The adsorption of cationic 2-hydroxypropyltrimethylammonium (HPMA) xylans with different degrees of substitution (DS) onto SAMs and regenerated cellulose was studied by SPR. Surface concentration (Р) exhibited a maximum (Рmax) for HPMAX adsorption onto carboxylic acid-terminated SAMs (SAM-COOH) at an intermediate HPMA DS of 0.10. This observation was indicative of a relatively flat conformation for adsorbed HPMAXs with higher HPMA DS because of higher linear charge densities along the polymer backbone. Рobserved for HPMAX adsorption onto regenerated cellulose and SAM-OH surfaces was relatively low compared to HPMAX adsorption onto SAM-COOH surfaces. Surface tension measurements for aqueous solutions of HPX by the Wilhelmy plate technique showed that surface tension changes ("γ = γwater " γHPX(aq)) increased and critical aggregation concentrations generally decreased with increasing hydroxypropyl (HP) DS. Hence, even though HP substitution was necessary to induce aqueous solubility, excessive hydroxypropylation promoted aggregation in water. SPR studies indicated that HPXs did not adsorb significantly onto regenerated cellulose or SAM-OH surfaces (submonolayer coverage). In contrast, HPX did adsorb (~monolayer coverage) onto SAM-CH3 surfaces. Collectively, these studies showed natural polymers could be chemically modified to produce surface modifying agents with sufficient chemical control, whereby the surface properties of the resulting systems could be explained in terms of chemical structure and intermolecular interactions. / Ph. D.

The fractionation of two biomass resources, red oak (Quercus rubra) chips and oil palm (Elaeis guineensis) trunk solids, into constitutive chemical components, cellulose, hemicelluloses (called "other carbohydrates") and non-carbohydrates (includes lignin, tannins, etc.), was studied quantitatively in terms of relative cleanness. Red oak chips were steam exploded using a batch reactor at five different treatment severities, Ro 5,000, 10,000, 15,000, 20,000 and 35,000. Steam exploded fibers (SEF) of each severity were extracted with water and alkali. Mass fractionation and summative analysis data of all solid biomass fractions were determined. These data were interpreted in term of a unifying clean fractionation concept designed to evaluate the effectiveness of the fractionation processes. Within a series of severities applied to a single biomass resource, the quantitative clean fractionation can be used to choose an optimum severity for the isolation of any particular component fraction. The red oak results revealed that 25 % (on average) of biomass solids were lost during steam explosion. Cellulose remained almost unaffected (retained in fibers form) by water and alkali extraction. About 35-55 % of the hemicelluloses can be recovered in the water extracted liquor fraction (WEL). The remaining non-cellulosic carbohydrates were lost during steam explosion, especially at high severity. At Ro 10,000 and above, alkali extracted fibers (AEF) consists almost entirely of cellulose and non-carbohydrates. The majority of the non-carbohydrates component (> 50 %) can be isolated by alkali extraction. The non-carbohydrate component harvested increased with severity to 67% at Ro 35,000. / Master of Science

L'objectif de ces travaux vise à évaluer les performances techniques et environnementales d'un procédé de fractionnement et de purification d'hémicelluloses de pailles et sons de blé. La démarche employée consiste dans un premier temps à évaluer la faisabilité du procédé à une échelle pilote. Celui-ci implique notamment une opération de fractionnement par extrusion bi-vis et des étapes de concentration-purification par une combinaison d'ultrafiltration et de chromatographie sur résine. Il est comparé à un procédé plus classique où l'extrait extrudé subit une évaporation et une précipitation éthanolique. L'étude des compositions chimiques des extraits et des poudres produites à chaque étape du procédé permet d'observer que l'extrusion bi-vis donne des rendements plus faibles que d'autres techniques d'extraction, mais présente des avantages en termes d'échelle, de continuité du procédé, et de faibles consommations d'eau. Par ailleurs, l'ultrafiltration est une technique intéressante car elle permet une purification et une concentration des extraits simultanées et la chromatographie sur résine s'est avérée être une solution attractive pour la décoloration des extraits. Dans un second temps, la méthodologie d'analyse du cycle de vie (ACV) a été appliquée au procédé. Celle-ci a mis en avant le fort impact environnemental généré par certaines opérations unitaires et a donné lieu à l'identification de priorités d'actions pour l'amélioration du procédé. Ainsi, le scénario mettant en jeu une combinaison de l'ultrafiltration avec une chromatographie sur résine semble plus avantageux que celui mettant en jeu une évaporation sous vide combinée à une précipitation éthanolique. La démarche employée apporte également des réponses méthodologiques sur la démarche d'ACV appliquée aux procédés : l'importance de prendre en compte différentes méthodes de caractérisation, de tester la sensibilité de l'étude à l'unité fonctionnelle et aux hypothèses initiales ont été mises en avant. Enfin, dans le but de démontrer sa forte potentialité dans le domaine des procédés, l'ACV a été appliquée à certaines opérations unitaires. L'intérêt d'étudier l'influence des conditions opératoires des procédés sur les impacts environnementaux a été démontré. Par ailleurs, la nécessité de développer des outils de modélisation des procédés qui pourront être couplés à ceux de l'ACV afin d'intégrer pleinement les critères environnementaux au dimensionnement des procédés a été mis en avant. / The objective of this work was to evaluate the technical and environmental performance of a fractionation and purification process of hemicelluloses from wheat straw and bran. The approach used was first to assess the feasibility of the process on a pilot scale. This implied setting up twin-screw extrusion, concentration steps by ultrafiltration and a purification step by ion exchange chromatography. It was compared to a more traditional process where the extruded extract was treated by evaporation and ethanol precipitation. Study of the chemical composition of extracts and powders produced at each stage of the process, showed that the twin-screw extrusion gave lower yields than other extraction techniques, but had the advantage in terms of scale, continuity of the process and low water consumption. Furthermore, ultrafiltration is an interesting technique because it allows simultaneous purification and concentration of the extracts, and ion exchange chromatography proved to be an attractive solution for the removal of coloured compounds. In a second step, the life cycle assessment (LCA) methodology was applied to the process. This highlighted the strong environmental impact generated by some unit operations, and resulted in the identification of priorities to improve the process. Thus, the scenario involving a combination of ultrafiltration with ion exchange chromatography appears to be more advantageous than that involving vacuum evaporation combined with ethanol precipitation. The approach used also provides methodological answers for LCA applied to processes: the importance of taking into account different characterization methods, testing the sensitivity of the study to the functional unit, and reconsidering the initial assumptions, has been stressed. Finally, in order to demonstrate its high potential in the field of processes, LCA has been applied to some unit operations. The interest of studying the influence of process operating conditions on environmental impacts has been demonstrated. In addition, the need to develop tools for modelling processes that can be coupled to those of LCA in order to fully integrate environmental criteria into process design, has been highlighted.

Les panneaux de bois MDF (Medium-Density Fiberboard), principalement utilisés dans le secteur de l’ameublement, génèrent une quantité annuelle mondiale de déchets estimée à 40 millions de m³. Actuellement,aucune méthode viable ne permet leur recyclage du fait de la présence de contaminants de type résine ou finitions (peinture, laminé, plastique). Ces déchets,constitués de bois à hauteur de 85-90 %, représentent cependant une source de matière lignocellulosique non négligeable. Afin de transformer cette ressource inexploitée, en matériaux à haute valeur ajoutée, un procédé de fractionnement combinant traitement alcalin et délignification a permis la séparation des hémicelluloses et lignines, amenant à l’isolement de la cellulose. En premier lieu, cette méthode a été utilisée pour l'extraction de nanocelluloses, à savoir des nanocristaux de cellulose par hydrolyse acide et des nanofibrilles de cellulose par délamination mécanique. Les nanocristaux de cellulose extraits des déchets ont montré des caractéristiques semblables à ceux obtenus avec des fibres de bois vierges. Ensuite, la fraction riche en hémicelluloses isolée par un traitement alcalin à partir des déchets MDF, a été purifiée puis précipitée à l’éthanol permettant l’isolation d’une fraction hémicellulosique, ultérieurement intégrée à la préparation d’hydrogels à base d’acide acrylique. Les hydrogels présentent des caractéristiques similaires qu'ils soient élaborés avec la fraction issue des déchets ou fabriqués à base de xylanes commerciaux de hêtre. L'évaluation de la capacité de ces déchets à être recyclés ouvrirait une voie de conversion en une nouvelle ressource de produits à forte valeur ajoutée. / Wood-based medium-density fiberboards (MDF) are principally used for furniture and fittings and areproduced in great volumes around the world. The annual quantity of MDF waste generated world-wide is estimated to be over 40 million m3. Currently, there is noviable way to transform MDF into useful products due to the presence of contaminants such as resins and finishes (paint, varnish, laminated or plastic). This type of waste, composed of approximately 85-90 % of wood, represents a significant source of lignocellulosic material. This thesis describes research on transforming this under-utilized resource into high valued materials via a fractionation process. An alkali treatment followed by a delignification step, allows the extraction of hemicelluloses and lignin respectively, leading to the isolation of pure cellulose. This cellulose was converted to nanocelluloses in the form of nanocrystals by acidhydrolysis and nanofibrils by mechanical fibrillation. The cellulose nanocrystals obtained from waste MDF is shown to have similar characteristics to those obtained from virgin wood fibers. The hemicelluloses-rich fraction obtained by an alkali treatment from MDF waste was purified, precipitated by ethanol and then further used in the preparation of acrylic acid based hydrogels. The hydrogels generated had similar characteristics to those made from commercial beech xylans.This research clearly demonstrates the technical feasibility to convert MDF waste into high value added products, opening a new conversion route to recycle them.

A associação de tecnologias como transgenia, genômica e proteômica aos programas de melhoramento convencional de árvores, pode acelerar o processo de obtenção de exemplares com características específicas da madeira para a indústria de papel e celulose. A parede celular vegetal é extremamente importante para este setor industrial, pois fornece polissacarídeos como celulose e hemiceluloses. A UDP-glucose pirofosforilase (UGPase, EC 2.7.7.9) é uma enzima chave na produção de UDP-glucose, precursor relevante na interconversão de nucleotídeoaçúcares que constituem os monômeros desses polissacarídeos. O objetivo deste trabalho foi clonar os cDNAs que codificam a UGPase, extraídos do tubérculo de batata (Solanum tuberosum L.) e da raiz de eucalipto (Eucalyptus grandis), e inseri-los por meio de transformação genética via Agrobacterium tumefaciens, respectivamente, no genoma de plantas de fumo (Nicotiana tabacum) e de eucalipto, para alterar o fluxo de carbono de modo a aumentar o conteúdo de pentosanas (xilose e arabinose). O maior teor de pentosanas é benéfico economicamente para a indústria de papel e celulose, pois diminui a energia consumida no refino da polpa além de benificiar a qualidade do papel que se torna mais resitente ao rasgo. O impacto da superexpressão do gene ugp nas plantas de tabaco e de eucalipto obtidas da transformação genética, foi avaliado por meio de análises bioquímicas e morfológicas. A análise de Southern blot indicou uma a quatro cópias do transgene no DNA genômico de dez linhagens T2 de tabaco, e uma cópia em uma linhagem T0 de eucalipto. A análise de qPCR mostrou que todas as linhagens T2 de tabaco apresentaram expressão do gene ugp exógeno, sendo que em quatro o nível de expressão foi maior que nas demais. As linhagens com menor expressão do gene ugp exógeno, com exceção de uma delas, também expressaram menos o gene ugp endógeno, sugerindo que nessas linhagens algum mecanismo de regulação atuou alterando a expressão desse gene, tanto endógeno, quanto exógeno. As linhagens transgênicas de tabaco foram morfologicamente semelhantes às plantas controle quando comparadas quanto à altura da planta, comprimento de internós, área foliar total, diâmetro do caule e massa seca da raiz. As alterações nos tabacos transgênicos, em relação aos controles, verificadas através das análises químicas, foram estatisticamente mais significativas na medula caulinar, onde houve aumento no conteúdo dos monossacarídeos e dos ácidos urônicos. As poucas diferenças encontradas no tecido xilemático sugerem que a planta, de alguma forma, tende a alterar menos a composição da parede celular secundária, do que da parede primária. Também sugerem que o metabolismo dos carboidratos pode afetar o conteúdo de lignina, embora seja sintetizada por outra rota metabólica. A parede celular primária, analizada utilizando a medula das plantas de tabaco e folhas de eucalipto, mostrou um aumento no conteúdo de pentosanas, o que não foi observado na análise de plântulas de tabaco. / Assossiating technologies like transgenesis, genomics and proteomics with conventional forestry breeding can accelerate the process of obtaining new trees with specific wood properties for the paper and pulp industry. Plant cell wall is extremely important for this industry, providing polysaccharides like cellulose and hemicelluloses. UDP-glucose pyrophosphorylase (UGPase, EC 2.7.7.9) is a key enzyme producing UDP-glucose, an important forerunner for nucleotide sugars that constitute the monomers of these polysaccharides. The goal in this work was to clone the cDNAs that encode UGPase, extracted from potato (Solanum tuberosum L.) tuber, and eucalypt (Eucalyptus grandis) root, and transfer through genetic transformation via Agrobacterium tumefaciens, respectively, in tobacco (Nicotiana tabacum) plants and eucalypts, in order to change the carbon flux, increasing the pentosans (arabinose and xylose) content. Higher pentosans content is economicaly important for the paper and pulp industry, as decreases energy consumption at pulp refining and also can benefit paper quality that becomes more resistant to rip. The impact of the ugp gene overexpression over tobacco and eucalypt plants obtained through genetic transformation were morphological and biochemicaly evaluated. Southern blot analysis showed one to four copies of the transgene in the genomic DNA of ten tobacco T2 lines, and one copy in one eucalypt T0 line. The qPCR analysis showed that all tobacco T2 lines expressed exogenous ugp gene, being the expression level in four of them biger than the others. Lines that expressed less exogenous ugp gene, with exception of one of them, equally had less endogenous ugp gene expression level, suggesting that in these lines, some kind of regulation acted changing the gene expression. The transgenic tobacco lines were morphologicaly similar to the control plants when compared for total height, internode lenght, total leaf area, stem diameter and root dry mass. Alterations in transgenics tobacco plants, in relation to controls, checked by chemical analysis, were statisticaly more significants in the pith, where the monosaccharides and uronic acids contents increased. The few diferences founded in the xilematic tissue sugest that the plant, somehow, tends to change less the secondary cell wall composition than the primary wall. They also suggest that the carbohydrate metabolism can affect lignin content, thought it\'s sintetizaded in another metabolic pathway. The primary cell wall, evaluated using tobbaco plants pith and eucalypt leaves, showed an increase in pentosanas content, that wasn\'t observed analising tobacco seedlings.

Submitted by CÁROL CABRAL TERRONE null (36025505837) on 2018-01-09T15:20:42Z No. of bitstreams: 1 Tese - Cárol Terrone.pdf: 3140738 bytes, checksum: 563a37f8e4725bc76f326c128aae84bf (MD5) / Approved for entry into archive by Adriana Aparecida Puerta null (dripuerta@rc.unesp.br) on 2018-01-10T16:59:21Z (GMT) No. of bitstreams: 1 terrone_cc_dr_rcla.pdf: 3025804 bytes, checksum: 817c25d1a9eef55ee5d3fb7555bd7982 (MD5) / Made available in DSpace on 2018-01-10T16:59:21Z (GMT). No. of bitstreams: 1 terrone_cc_dr_rcla.pdf: 3025804 bytes, checksum: 817c25d1a9eef55ee5d3fb7555bd7982 (MD5) Previous issue date: 2017-11-14 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Biomassa lignocelulósica é a principal fonte de carbono renovável no planeta. Seus constituintes majoritários são celulose, hemiceluloses e lignina. Hemiceluloses são polissacarídeos ramificados sendo o principal tipo as xilanas, estruturas com cadeia principal de xilose. Dentre as diversas enzimas hidrolíticas, que atuam sobre a estrutura de xilanas, estão as α-L-arabinofuranosidases, que catalisam a hidrólise de ligações entre α-L-arabinofuranosídeos e a cadeia principal do polissacarídeo. Neste trabalho, uma linhagem de Aspergillus hortai CRM1919, isolada de solo próximo a lagoas salinas e alcalinas da região da Nhecolândia no Pantanal Matogrossense, foi utilizada para produzir α-L-arabinofuranosidases. Resíduos agroindustriais foram utilizados como substrato para o cultivo do fungo. O objetivo principal foi otimizar a produção de α-L-arabinofuranosidases e aumentar a produção a fim de se obter um filtrado enzimático rico nessa atividade, para posteriormente purificá-la e aplicá-la na degradação de biomassa. A linhagem foi cultivada em meio líquido de Vogel suplementado com diferentes substratos, dentre eles alguns subprodutos da agroindústria. As maiores produções de α-arabinofuranosidases ocorreram nos cultivos com polpa cítrica e casca de laranja a 1% (m/v) em cultivos estáticos por 4 dias, com pH inicial de 2,5 e temperatura de 30 ºC. Após todas as etapas de otimização, o extrato bruto apresentou atividade quinze vezes maior do que nos cultivos iniciais. A enzima foi purificada por uma estratégia de três etapas, sendo uma precipitação com sulfato de amônio, na concentração de 25 a 65%, uma cromatografia de troca iônica e outra de exclusão molecular, apresentando ao final uma recuperação de 10,1% com um fator de purificação de 58,7. As características da α-L-arabinofuranosidase purificada foram determinadas, sendo pH ótimo de 4,0 e temperatura ótima de 60 ºC, meia vida a 30 e 40 ºC de 265 e 230 minutos, respectivamente. A maior estabilidade da enzima ao pH ocorreu em pH 5,0, A atividade enzimática foi reduzida na presença de Zn+2, PMSF, SDS e etanol a 20%, mas foi ativada na presença de Mg+2, Na+, EDTA, Tween 20, Tween 80 e Triton X-100. A α-arabinofuranosidase apresentou tolerância a presença de 0,5 e 1,0 M de NaCl no meio reacional, e alta estabilidade nas concentrações de 0,5, 1,0, 2,0 e 4,0 M de NaCl. Os parâmetros cinéticos para o substrato ρ-nitrofenil-α-L-arabinofuranosídeo foram Km de 8,73 mM, Vmáx de 7,91 μmol/min.mg e Kcat de 0,59/min. A enzima purificada apresentou afinidade apenas sobre o substrato ρ-nitrofenil-α-L-arabinofuranosídeo. Na caracterização das biomassas in natura, o bagaço de cana-de-açúcar apresentou em sua composição 30% de celulose, 16% de hemiceluloses e 20% de ligninas totais, enquanto bagaço de malte apresentou 20% de celulose, 20% de hemiceluloses e 20% de ligninas totais. Na caracterização da hemicelulose extraída, a de bagaço de cana-de-açúcar apresentou composição em açúcares redutores de 7,8% de glicose, 23,7% de xilose e 2,6% de arabinose; a hemicelulose de bagaço de malte apresentou em sua composição 20,6% de glicose, 28,3% de xilose e 9,4% de arabinose. A hidrólise das hemiceluloses e de xilanas comerciais foi realizada na presença da α-arabinofuranosidase purificada e do filtrado bruto de cultivo contendo a α-arabinofuranosidase, associados com uma xilanase purificada, também produzida por Aspergillus hortai. Em ambos os casos foi possível verificar a cooperação entre enzimas do complexo xilanolítico e a atuação sequencial de cada uma na hidrólise das estruturas de xilano. / Lignocellulosic biomass is the main source of renewable carbon on the planet. Its major constituents are cellulose, hemicelluloses and lignin. Hemicelluloses are branched polysaccharides being the main type the xylan, xylose main chain structures. Among the various hydrolytic enzymes that act on the xylan structure there are the α-L-arabinofuranosidases, which catalyze the hydrolysis of linkages between α-L-arabinofuranosides and the polysaccharide main chain. In this work, a strain of Aspergillus hortai CRM1919, isolated from soil near saline and alkaline lagoons of the Nhecolândia region in Pantanal Matogrossense, was used to produce α-L-arabinofuranosidases. Agroindustrial wastes were used as substrate for the fungus cultivation. The main objective was to optimize the production of α-L-arabinofuranosidases and increase the yield in order to obtain a rich enzymatic filtrate in this activity, to later purify it and apply it in the hydrolysis of biomass. The strain was cultivated in Vogel liquid medium supplemented with different substrates, among them some by-products of agroindustry. The highest production of α-arabinofuranosidases occurred in cultures with citrus pulp and orange peel at 1% (w/v) in static cultures for 4 days, with an initial pH of 2.5 and temperature of 30 ºC. After all the optimization steps, the crude extract presented activity fifteen times higher than the initial cultures. The enzyme was purified by a three-step strategy, with ammonium sulfate precipitation at 25-65% concentration, ion exchange chromatography and molecular exclusion chromatography, with recovery of 10.1% and purification fold of 58.7. The characteristics of the purified α-L-arabinofuranosidase were determined, being optimum pH of 4.0 and optimum temperature of 60 °C, half-life at 30 and 40 °C of 265 and 230 minutes, respectively. The higher stability of the enzyme to pH occurred at pH 5.0. Enzymatic activity was reduced in the presence of Zn+ 2, PMSF, SDS and 20% ethanol and was activated in the presence of Mg+ 2, Na+, EDTA, Tween 20, Tween 80 and Triton X-100. The α-arabinofuranosidase presented tolerance to the presence of 0.5 and 1.0 M NaCl in the reaction medium, and high stability at the concentrations of 0.5, 1.0, 2.0 and 4.0 M NaCl. The kinetic parameters for the ρ-nitro phenyl-α-L-arabinofuranoside substrate were Km of 8.73 mM, Vmax of 7.91 μmol / min.mg and Kcat of 0.59 / min. The purified enzyme had affinity only on the ρ-nitro phenyl-α-L-arabinofuranoside substrate. In the characterization of in natura biomasses, sugarcane bagasse presented 30% cellulose, 16% hemicelluloses and 20% total lignins, while brewer’s spent grain presented 20% cellulose, 20% hemicelluloses and 20% total lignins. In the characterization of extracted hemicellulose, the sugarcane bagasse presented composition in reducing sugars of 7.8% of glucose, 23.7% of xylose and 2.6% of arabinose; the hemicellulose of brewer’s spent grain presented 20.6% glucose, 28.3% xylose and 9.4% arabinose. Hydrolysis of the hemicelluloses and commercial xylans was performed in the presence of the purified α-arabinofuranosidase and the crude culture filtrate containing α-arabinofuranosidase, associated with a purified xylanase, also produced by Aspergillus hortai. In both cases it was possible to verify the cooperation between xylanolytic complex enzymes and the sequential action of each one in the hydrolysis of the xylan structure.