No description available.

No description available.

No description available.

Pseudo-lignin, which can be broadly defined as aromatic material that yields a positive acid-insoluble (Klason) lignin value, has been reported to generate from biomass polysaccharides during dilute acid pretreatment (DAP). To investigate the fundamental chemistry of pseudo-lignin, a series of state-to-art analytical techniques including GPC, FT-IR and ¹³C NMR were applied to characterize pseudo-lignin extracted from poplar α-cellulose and holocellulose after DAP. The results showed that pseudo-lignin is polymeric (Mn ~ 1000 g/mol; Mw ~ 5000 g/mol) and consists of carbonyl, carboxylic, aromatic, methoxy and aliphatic structures, which can be produced from both dilute acid-treated cellulose and hemicellulose. During DAP, the hydrolysis of polysaccharides, which leads to some release of monosaccharides, and their subsequent dehydration reactions to form furfural and 5-hydromethylfurfural (HMF) takes place. Further rearrangements of furfural and/or HMF can produce aromatic compounds, which undergo further polymerization and/or polycondensation reactions to form pseudo-lignin. More importantly, pseudo-lignin was revealed to bind with cellulase enzymes unproductively and significantly retard enzymatic conversion of cellulose. As compared to native lignin after DAP, the inhibition effect arise from pseudo-lignin is much stronger, which clearly indicates pseudo-lignin formation should be avoided during DAP. Process optimization study indicated that addition of dimethyl sulfoxide (DMSO) to the DAP reaction medium can effectively increase sugar recovery and reduce pseudo-lignin formation, even under high-severity pretreatment conditions. The pseudo-lignin suppression property of DMSO has been attributed to the preferential arrangement of DMSO in the vicinity of the C1 carbon of the HMF molecule, thereby protecting HMF from further reactions to form pseudo-lignin.

L’identification de marqueurs susceptibles de mettre en évidence et quantifier l’altération du bois par différents agents de dégradation peut présenter un grand intérêt dans la mise au point de méthodes de contrôle du matériau. Dans la première partie de ce travail, la caractérisation et le dosage des composés extractibles de deux essences non durables, le hêtre et le chêne soyeux, exposées à différents champignons de pourritures blanches ou brunes préalablement traitées ou non avec du propiconazole a été effectué. L’évolution de la quantité et de la nature des extractibles dépend directement de la présence du biocide et permet de différencier clairement les éprouvettes en fonction du taux de dégradation de ces dernières. C’est le cas notamment de la catéchine présente dans les extraits de hêtre, qui disparaît rapidement lorsque le bois est exposé à un agent de pourriture et qui peut à ce titre constituer un marqueur de l’attaque du bois. Dans la seconde partie de ce travail, l’évolution de la composition chimique du bois traité par pyrolyse douce à 240°C sous azote a été étudiée. Bien que les résultats obtenus confirment en grande partie ceux décrits précédemment dans la littérature, le dosage des différentes fractions du bois (lignine et holocellulose) et l’analyse des monosaccharides constitutifs de la fraction holocellulose du bois ont permis de mettre en évidence un important phénomène de carbonisation rapporté jusqu’à présent pour des températures plus élevées. Ce phénomène confirmé par RMN13C peut être à l’origine de certaines des nouvelles propriétés du matériau. / The identification of markers able to highlight and quantify wood deterioration by different degradation agents can be of great interest in the development of material control methods. In the first part of this work, the characterization and the quantification of the two non durable wood species extractives, beech and silky oak, treated or not with propiconazole and exposed to various white rot or brown rot fungi were carried out. The evolution of the quantity and the nature of extractives depend directly on the presence of biocide and allowed to clearly differentiate degraded and non-degraded samples. This is particularly true in the case of catechin identified in the extracts of beech, which is rapidly degraded by rotting fungi constituting therefore a valuable marker of wood degradation. In the second part of this work, the evolution of the chemical composition of wood treated by mild pyrolysis at 240°C under nitrogen was investigated. Although the results mainly confirm those described previously in the literature, isolation of lignin and holocellulose fractions and analysis of monosaccharides constitutive of holocellulose fraction highlighted an important carbonization of wood reported up to now for higher temperatures. This phenomenon is confirmed by 13C NMR and could be at the origin of some of the new properties of the material.

No description available.

No description available.

A biomassa lignocelulósica é uma fonte abundante de açúcares simples passíveis de serem fermentados em uma variedade de bioprodutos de maior valor agregado, além do etanol de segunda geração. Tal diversidade é relevante ao desenvolvimento e aprimoramento do conceito de biorrefinarias e da bioeconomia, em um viés mais amplo. Todavia, a elevada recalcitrância dos lignocelulósicos dificulta a sua sacarificação enzimática, resultando em bioprocessos mais onerosos. Por isso, coquetéis com diferentes enzimas ativas em carboidrato (CAZymes) são desenvolvidos, em busca de uma maior eficiência e melhor relação custo/benefício, para processos em larga escala. Dentre as CAZymes estudadas, encontram-se as mono-oxigenases líticas de polissacarídeo (LPMOs), tendo em vista a sua atestada capacidade de otimizar a hidrólise da lignocelulose, quando em sinergismo com diversas hidrolases. Levando isto em conta, selecionou-se, ao atual estudo, o ascomiceto termofílico Thermothelomyces thermophila (anteriormente denominado Myceliophthora thermophila), pois este tem se mostrado capaz de expressar e secretar ampla gama de LPMOs ativas em diferentes substratos. Objetivando-se estudar duas LPMOs derivadas deste organismo, as mesmas foram expressas, heterologamente, por Aspergillus nidulans linhagem A773, utilizando-se o vetor de expressão pEXPYR construído para viabilizar a secreção de altas concentrações de proteínas recombinantes. As proteínas heterólogas aqui analisadas foram denominadas TtLPMO1A9 e TtLPMO2A9. Embora ambas tenham sido capazes de gerar peróxido de hidrogênio na presença de oxigênio molecular e de um doador de elétrons, apenas TtLPMO2A9 apresentou atividade contra substratos celulósicos e bagaço de cana pré-tratado hidrotermicamente, atuando, em associação com hidrolases homemade e o preparo enzimático comercial Celluclast 1.5L, a degradação de tais materiais. / The lignocellulosic biomass is an abundant source of simple sugars that can be fermented to various value-added bio-based products. This diversity is seen as relevant to the improvement of biorefinery and bioeconomy concept. Nevertheless, the significant recalcitrance of lignocellulose imposes dificulties to its enzymatic saccharification, resulting in onerous bioprocessing. This scenario stimulates studies based on the development of efficient and cost-effective customizable carbohydrate-active enzyme (CAZymes) cocktails for large-scale processes. Among the available CAZymes, there are the lytic polysaccharide monooxygenases (LPMOs), a set of oxidative proteins capable of optimizing the lignocellulose hydrolysis, when acting in synergism with various hydrolases. Based on this fact, in the current study, the thermophilic ascomycete Thermothelomyces thermophila (previously known as Myceliophthora thermophila) was adopted, because of its ability of expressing and secreting large amounts of LPMOs. Thus, two LPMOs derived from this fungus was heterologously produced by an expression system composed by Aspergillus nidulans strain A773 and the vector pEXPYR: an expression vector built to increase the secretion of recombinant proteins. The heterologous proteins herein analysed were termed as TtLPMO1A9 and TtLPMO2A9. Although both enzymes were able to produce hydrogen peroxide in the presence of molecular oxygen and an electron donor, only the second one was active in reactions with cellulosic substrates and hydrothermally pre-treated sugar cane bagasse. When tailor-made hydrolases and the commercial enzymatic mixture Celluclast 1.5L were supplemented with TtLPMO2A9, it was noticed na improvement of the deconstruction of the aforementioned substrates.

A biomassa lignocelulósica é uma fonte abundante de açúcares simples passíveis de serem fermentados em uma variedade de bioprodutos de maior valor agregado, além do etanol de segunda geração. Tal diversidade é relevante ao desenvolvimento e aprimoramento do conceito de biorrefinarias e da bioeconomia, em um viés mais amplo. Todavia, a elevada recalcitrância dos lignocelulósicos dificulta a sua sacarificação enzimática, resultando em bioprocessos mais onerosos. Por isso, coquetéis com diferentes enzimas ativas em carboidrato (CAZymes) são desenvolvidos, em busca de uma maior eficiência e melhor relação custo/benefício, para processos em larga escala. Dentre as CAZymes estudadas, encontram-se as mono-oxigenases líticas de polissacarídeo (LPMOs), tendo em vista a sua atestada capacidade de otimizar a hidrólise da lignocelulose, quando em sinergismo com diversas hidrolases. Levando isto em conta, selecionou-se, ao atual estudo, o ascomiceto termofílico Thermothelomyces thermophila (anteriormente denominado Myceliophthora thermophila), pois este tem se mostrado capaz de expressar e secretar ampla gama de LPMOs ativas em diferentes substratos. Objetivando-se estudar duas LPMOs derivadas deste organismo, as mesmas foram expressas, heterologamente, por Aspergillus nidulans linhagem A773, utilizando-se o vetor de expressão pEXPYR construído para viabilizar a secreção de altas concentrações de proteínas recombinantes. As proteínas heterólogas aqui analisadas foram denominadas TtLPMO1A9 e TtLPMO2A9. Embora ambas tenham sido capazes de gerar peróxido de hidrogênio na presença de oxigênio molecular e de um doador de elétrons, apenas TtLPMO2A9 apresentou atividade contra substratos celulósicos e bagaço de cana pré-tratado hidrotermicamente, atuando, em associação com hidrolases homemade e o preparo enzimático comercial Celluclast 1.5L, a degradação de tais materiais. / The lignocellulosic biomass is an abundant source of simple sugars that can be fermented to various value-added bio-based products. This diversity is seen as relevant to the improvement of biorefinery and bioeconomy concept. Nevertheless, the significant recalcitrance of lignocellulose imposes dificulties to its enzymatic saccharification, resulting in onerous bioprocessing. This scenario stimulates studies based on the development of efficient and cost-effective customizable carbohydrate-active enzyme (CAZymes) cocktails for large-scale processes. Among the available CAZymes, there are the lytic polysaccharide monooxygenases (LPMOs), a set of oxidative proteins capable of optimizing the lignocellulose hydrolysis, when acting in synergism with various hydrolases. Based on this fact, in the current study, the thermophilic ascomycete Thermothelomyces thermophila (previously known as Myceliophthora thermophila) was adopted, because of its ability of expressing and secreting large amounts of LPMOs. Thus, two LPMOs derived from this fungus was heterologously produced by an expression system composed by Aspergillus nidulans strain A773 and the vector pEXPYR: an expression vector built to increase the secretion of recombinant proteins. The heterologous proteins herein analysed were termed as TtLPMO1A9 and TtLPMO2A9. Although both enzymes were able to produce hydrogen peroxide in the presence of molecular oxygen and an electron donor, only the second one was active in reactions with cellulosic substrates and hydrothermally pre-treated sugar cane bagasse. When tailor-made hydrolases and the commercial enzymatic mixture Celluclast 1.5L were supplemented with TtLPMO2A9, it was noticed na improvement of the deconstruction of the aforementioned substrates.

Dynamic (or 2D) FT-IR spectroscopy in combination withpolarized IR irradiation has been used in this work to studywood polymer orientation and interactions on theultrastructural level in wood fibers in the native state aswell as the effects of different pulping processes. The woodpolymer interactions were studied under both dry and humidconditions. The matrix of lignin and hemicelluloses located between thewell-ordered cellulose fibrils in the wood cell wall of sprucewas here shown to be more highly ordered than has earlier beenrevealed. It was confirmed that glucomannan is orientedparallel to the cellulose fibrils and is highly coupled to it.The lignin was also shown to have a main orientation in thestructure although this is probably not as strong as that ofglucomannan. The orientation of the lignin may derive from thefact that the polysaccharides act as templates during thelignification of the cell wall. This organization implies thatnot only the cellulose but also the lignin and thehemicelluloses have different mechanical properties in thelongitudinal and cross-fiber directions. The ability to gain molecular information on the stresstransfer in polymers with dynamic FT-IR spectroscopy made itpossible to verify experimentally earlier molecularcalculations on the stress transfer within the cellulose chain.It was also possible to show, on the molecular level, thedominant importance of the cellulose fibrils for the stresstransfer in the longitudinal direction of pulp fibers,including lignin-rich mechanical pulp fibers. The glucomannanof softwood fibers was also shown to participate in the stresstransfer in the fiber direction indicating a close associationwith the cellulose, whereas the xylan showed no dynamicresponse. Already under dry conditions, the lignin was shown tohave a more viscoelastic response than the polysaccharidesduring the loading of pulp fibers and it was thus able to moveindependently of the cellulose. The enhanced spectral resolution obtained with dynamic FT-IRspectroscopy made it possible to study the crystalstructure/chain order of cellulose in pulp fibers. Thepossibility of following changes in the relative cellulose Iallomorph composition of pulp fibers was demonstrated for somechemical pulps. Dynamic FT-IR experiments under humid conditions and ofelevated temperatures made it possible to study the softeningof the biopolymers in their native environment. This was alsodemonstrated for some different pulps, and this may be apromising tool for obtaining viscoelastic information on themolecular level in composite systems such as wood fibers. <b>Keywords:</b>cellulose, cooperation, crystallinity, dynamictest, glucomannan, hardwood, holocellulose, humidity, infraredspectroscopy, kraft pulp, lignin, mechanical pulp, orientation,polarised light, softwood, strain, sulphite pulp,viscoelasticity, xylan