Made available in DSpace on 2014-06-11T19:27:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-10-16Bitstream added on 2014-06-13T19:35:27Z : No. of bitstreams: 1 kiiper_pg_me_rcla.pdf: 773936 bytes, checksum: 1b480acb49acd45ba8d3f3f9a6e99ca8 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A utilização de solvente Organosolv em condições de acidez e temperaturas elevadas para a realização da deslignificação de bagaço de cana e hidrólise da fração de hemicelulose tem sido muito estudada e existem muitos parâmetros experimentais já descritos na literatura. Tem sido obtidos bons resultados de deslignificação e hidrólise parcial do bagaço a partir do prétratamento ácido com solvente Organosolv utilizando misturas de etanol e água, normalmente em proporções iguais de volume de cada. Este estudo descreve um método de deslignificação e hidrólise da hemicelulose do bagaço da cana combinando a utilização da mistura dos solventes orgânicos, acetona e álcool, com água e solução ácida diluída. A mistura do solvente organosolv (acetona+álcool/água – 1:1 v/v), concentração ácida de 0,5 - 2,0% de H2SO4, e tempo de reação de 30 a 120 minutos e 5,0% de bagaço (p/v) foi realizado para o pré-tratamento. O efeito do ácido, solvente e temperatura no rendimento e extensão da deslignificação e hidrólise da oram estudados. Os resultados obtidos indicam importantes diferenças no pré-tratamento ácido com solvente Organosolv, devido ao uso da acetona e concentração ácida empregados nos experimentos deste estudo. Os melhores resultados foram obtidos com 1,0% de H2SO4 e 120 °C. Sob estas condições o rendimento de açúcares obtidos da hidrólise ácida da porção de hemicelulose do bagaço de cana foi de 7.53%. Estes resultados demonstram um rendimento de açúcares fermentescíveis que podem ser utilizados como fonte de carbono para a fermentação alcoólica por Pachysolen tannophilus. A conversão de Xilose em etanol por Pachysolen tannophilus é relativamente ineficiente nas condições de fermentação estudada. Esta ineficiência é atribuída em partes, ao consumo de etanol pela levedura concorrente à sua produção, baixa eficiência... / The utilization of Organosolv solvents under acid and high temperature conditions in the delignification and hemicelulose and cellulose hydrolysis of wood and bagasse has already been reported in the literature for several experimental parameters. Organosolv treatment of sugarcane bagasse has also been reported and has shown good results with ethanol–water mixtures, normally at volume ratios close to unit. This study describes a method of the delignification hemicelulose hydrolysis of sugarcane bagasse combining the use of ethanol+acetone/water mixtures and diluted acid solutions. Ethanol+acetone/water mixture (1:1 v/v), acid concentration of 0,5 - 2,0% of H2SO4, reaction times from 30 to 120 min, and 5,0% of sugarcane bagasse (w/v) was carried out for bagasse pre-treatment. The effect of acid, solvent and temperature on the yield and extent of delignification and hemicelulose hydrolysis was studied. The obtained results indicate important differences from the Organosolv process, which may be due to the presence of acetone and acid concentration employed in this work. The best results were obtained at 1,0% of H2SO4 and 120 °C. Under these conditions the sugar yield obtained from hemicelulose hydrolysis from sugarcane bagasse 7.53%. These results shows that the sugar yield obtained can be used as carbon source for Pachysolen tannophilus fermentation to produce ethanol. The conversion of D-xylose to ethanol by the yeast Pachysolen tannophilus is relatively inefficient in batch culture. The inefficiency has been attributed in part to concurrent utilization of ethanol, slow fermentation metabolism of the yeast and to the formation of xylitol and other by-products. The xylose concentration in the must was 2,0% during 120 hours of fermentation. The best ethanol yield was obtained in the first 24 hours, as the ethanol began to be consumed and the sugar concentration became low... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:24:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-27Bitstream added on 2014-06-13T19:31:25Z : No. of bitstreams: 1 morais_aps_me_botfca.pdf: 687506 bytes, checksum: baad0742c0a39bca3a4a2b5908c9cedf (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O bioetanol é um combustível produzido por meio da fermentação do açúcar do caldo da cana, que representa apenas um terço do carbono (energia), presente na planta. Atualmente, estão sendo feitos esforços para o aproveitamento do restante da biomassa, divididos entre as folhas e bagaço do colmo. Esta biomassa lignocelulósica poderia ser aproveitada para produção de etanol, desde que submetida a processos hidrolíticos químicos (ácidos e bases) e enzimáticos gerando açúcares fermentescíveis. Pela fermentação alcoólica é possível a produção de etanol a partir da mistura de açúcares liberados. Neste trabalho procurou-se a padronização de procedimentos para avaliar o pré-tratamento físico e químico do bagaço da cana-de-açúcar, aliado a diferentes tratamentos térmicos a partir de duas granulometrias de bagaço (1,1 e 2,5 mm). Para o delineamento experimental, utilizou-se tratamentos ajustados em fatorial 4 X 5, sendo que as variáveis foram a influência do tempo de pré-tratamento (0, 15, 30, 45 e 60 minutos) e concentração de ácido sulfúrico (H2SO4) a 7 e 9%. Estes fatores exerceram influência sobre o desempenho da pré-hidrólise, medido pela liberação açúcares redutores (AR) na solução e a % de espécies químicas no bagaço prétratado / Bioethanol fuel is produced through the fermentation of sugar cane juice, which representes only a third of the carbon (energy) present in the plant. Currently, efforts are being made to the use of the remaining biomass, divided among the leaves and seed stalk. This lignocellulosic biomass could be used for ethanol production, provided that undergo hydrolytic process chemicals (acids and bases) and enzymatic generating fermentable sugars. For fermentation is possible to produce ethanol from mixed sugars released. This research is the standardization of procedures to assess the pre-treatment physical and chemical properties of bagasse from sugar cane, coupled with different thermal treatments from two particle sizes of mulch (1,1 and 2,5 mm). For this experiment, we used adjusted treatments in a factorial 4 x 5, and the variables were the influence of time of pretreatment (0, 15, 30, 45 and 60 minutes) And concentration of sulfuric acid (H2SO4) 7 and 9%. These factors have exerted influence on the performance of pre-hydrolysis, measured by the release sugars (RS) in the solution, and% of chemical species in the pretreated bagasse

Orientador: Pedro de Oliva Neto / Banca: Dejanira de Franceschi de Angelis / Banca: Sandra Regina Ceccato-Antonini / Resumo: A utilização de solvente Organosolv em condições de acidez e temperaturas elevadas para a realização da deslignificação de bagaço de cana e hidrólise da fração de hemicelulose tem sido muito estudada e existem muitos parâmetros experimentais já descritos na literatura. Tem sido obtidos bons resultados de deslignificação e hidrólise parcial do bagaço a partir do prétratamento ácido com solvente Organosolv utilizando misturas de etanol e água, normalmente em proporções iguais de volume de cada. Este estudo descreve um método de deslignificação e hidrólise da hemicelulose do bagaço da cana combinando a utilização da mistura dos solventes orgânicos, acetona e álcool, com água e solução ácida diluída. A mistura do solvente organosolv (acetona+álcool/água - 1:1 v/v), concentração ácida de 0,5 - 2,0% de H2SO4, e tempo de reação de 30 a 120 minutos e 5,0% de bagaço (p/v) foi realizado para o pré-tratamento. O efeito do ácido, solvente e temperatura no rendimento e extensão da deslignificação e hidrólise da oram estudados. Os resultados obtidos indicam importantes diferenças no pré-tratamento ácido com solvente Organosolv, devido ao uso da acetona e concentração ácida empregados nos experimentos deste estudo. Os melhores resultados foram obtidos com 1,0% de H2SO4 e 120 °C. Sob estas condições o rendimento de açúcares obtidos da hidrólise ácida da porção de hemicelulose do bagaço de cana foi de 7.53%. Estes resultados demonstram um rendimento de açúcares fermentescíveis que podem ser utilizados como fonte de carbono para a fermentação alcoólica por Pachysolen tannophilus. A conversão de Xilose em etanol por Pachysolen tannophilus é relativamente ineficiente nas condições de fermentação estudada. Esta ineficiência é atribuída em partes, ao consumo de etanol pela levedura concorrente à sua produção, baixa eficiência... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The utilization of Organosolv solvents under acid and high temperature conditions in the delignification and hemicelulose and cellulose hydrolysis of wood and bagasse has already been reported in the literature for several experimental parameters. Organosolv treatment of sugarcane bagasse has also been reported and has shown good results with ethanol-water mixtures, normally at volume ratios close to unit. This study describes a method of the delignification hemicelulose hydrolysis of sugarcane bagasse combining the use of ethanol+acetone/water mixtures and diluted acid solutions. Ethanol+acetone/water mixture (1:1 v/v), acid concentration of 0,5 - 2,0% of H2SO4, reaction times from 30 to 120 min, and 5,0% of sugarcane bagasse (w/v) was carried out for bagasse pre-treatment. The effect of acid, solvent and temperature on the yield and extent of delignification and hemicelulose hydrolysis was studied. The obtained results indicate important differences from the Organosolv process, which may be due to the presence of acetone and acid concentration employed in this work. The best results were obtained at 1,0% of H2SO4 and 120 °C. Under these conditions the sugar yield obtained from hemicelulose hydrolysis from sugarcane bagasse 7.53%. These results shows that the sugar yield obtained can be used as carbon source for Pachysolen tannophilus fermentation to produce ethanol. The conversion of D-xylose to ethanol by the yeast Pachysolen tannophilus is relatively inefficient in batch culture. The inefficiency has been attributed in part to concurrent utilization of ethanol, slow fermentation metabolism of the yeast and to the formation of xylitol and other by-products. The xylose concentration in the must was 2,0% during 120 hours of fermentation. The best ethanol yield was obtained in the first 24 hours, as the ethanol began to be consumed and the sugar concentration became low... (Complete abstract click electronic access below) / Mestre

Orientador: Jorge Luís Akasaki / Banca: Haroldo de Mayo Bernardes / Banca: Malik Cheriaf / Resumo: O setor sucro-alcooleiro vem se expandindo nos últimos tempos, alavancado principalmente pelo advento da bioenergia. No Brasil, a produção de cana-de-açúcar vem avançando em áreas destinadas à pecuária extensiva e a outras culturas importantes como a soja, o milho e a laranja e concerne à geração de energia elétrica pela queima do bagaço (cogeração) e ao uso de etanol como combustível de veículos automotores. Contudo, a produção de açúcar, álcool e energia a partir da cana-de-açúcar pode causar problemas ambientais em decorrência da geração de resíduos como a cinza do bagaço de cana-de-açúcar. Diante do panorama mundial de crescimento do setor sucro-alcooleiro aliado ao desenvolvimento sustentável, é de grande importância a realização de estudos que visem o aproveitamento destes resíduos. Deste modo, este trabalho mostra a viabilidade técnica da utilização da cinza resultante do processo de queima do bagaço para a geração de energia como adição mineral, em argamassas. Foram realizados ensaios de caracterização química, difração de raios-X, microscopia eletrônica de varredura e atividade pozolânica da cinza do bagaço de cana-deaçúcar. Além disto, um estudo da moagem do material foi realizado, juntamente com a composição de argamassas contendo a cinza em diversos teores de substituição ao agregado miúdo. Através da resistência à compressão e à tração por compressão diametral e módulo de elasticidade, foram analisadas suas propriedades mecânicas. Também foram verificadas algumas propriedades relacionadas com a durabilidade do material, como a retração por secagem, absorção por capilaridade e reatividade acelerada pelo método acelerado de expansão em barras de argamassa / Abstract: The sugar-alcohol sector has expanding in the last years, mainly due to advent of the renewable energy sources and bioenergy. In Brazil, the sugarcane production has advancing towards areas of cattle creation and agriculture, such as soya bean, corn and orange and concerning to electrical energy production through the burning bagasse and alcohol used like vehicle fuel. However, sugar, alcohol and energy production from sugarcane could cause other environmental problems in result to the wastes production such as sugarcane bagasse ash. In the presence of worldwide growth of sugar and alcohol industry and the sustainable development, it is very important the achievement of studies that reuse these wastes. Thus, this work shows technical viability on using sugar-cane bagasse ash (SCBA) obtained from sugar-cane bagasse burnt, as mineral addition, in mortars. It was made some tests on the SCBA, such as the chemical characterization, X-ray diffraction, pozzolanic activity, and also, images obtained by scanning electron microscope. Moreover, grinding of SCBA was carried out, and mortars were prepared with some different amounts of SBCA as replacement of fine aggregate by mass. Through the compressive strength, splitting tensile strength and modulus of elasticity its mechanical properties were verified, and also some properties related to material durability as drying shrinkage, water absorption by capillarity, potential alkali reactivity of aggregates by mortar-bar method / Mestre

O objetivo deste trabalho foi caracterizar a farinha do bagaço da cana-de-açúcar (FBCA) e avaliar o efeito de sua aplicação, como fonte de fibra insolúvel, em bolo. A FBCA foi caracterizada com relação a sua composição química (teor de umidade, cinzas, proteínas, carboidratos, fibras solúveis, fibras em detergente ácido e neutro e lipídeos), atividade de água (Aw), composição de minerais pela técnica de Espectrometria de Fluorescência de Raios-X (TXRF), determinação do tamanho e distribuição das partículas, avaliação da cor instrumental e contagem de bolores e leveduras e de coliformes totais. A FBCA foi aplicada em bolo tipo cupcake nas proporções de 3 e 6%. Os bolos recém-preparados foram avaliados com relação à umidade, atividade de água (Aw), densidade, cor instrumental e pH. Os bolos foram estocados durante 42 dias e em 5 pontos deste período foram avaliados com relação a umidade, Aw, perfil de textura (parâmetros dureza, elasticidade e coesão), contagem de bolores e leveduras e aceitação sensorial (atributos aparência, sabor, aroma, textura e aceitação global). Todas as análises foram realizadas também em uma amostra controle, preparada sem a adição de FBCA, para efeito de comparação. A FBCA demonstrou ser excelente fonte de fibra insolúvel, com 81,92% entre celulose, lignina e hemicelulose, bem como de minerais como o ferro, manganês e cromo. Todavia é uma fonte pobre de fibra solúvel, proteína, carboidratos e lipídeos. Em torno de 60% das partículas da FBCA possuem tamanho inferior a 80µm. A FBCA apresentou contagem alta de bolores e leveduras e cor tendendo para o amarelo e vermelho. Os bolos apresentaram em torno de 15% de umidade e Aw na faixa intermediária. Bolos com FBCA foram ligeiramente mais úmidos, escuros e densos que o controle. Porém, foram menos elásticos e tão coesos e duros quanto. Os bolos adicionados de 3% de FBCA apresentaram a mesma aceitação que o produto tradicional, com a vantagem do maior valor nutricional em função da presença das fibras e dos minerais oriundos da FBCA. / The aim of this study was to characterize the flour from sugarcane bagasse (FSB) and to evaluate the effect of its application as a source of insoluble fiber in cake. The FSB was characterized in respect to the chemical composition (moisture content, ash, proteins, carbohydrates, soluble fiber, fiber in acid and neutral detergent, cellulose, hemicellulose, lignin and lipids), water activity (Aw), mineral composition by Fluorescence spectrometry technique X-ray (TXRF), size and distribution of particles, evaluation of the instrumental color and counts of molds and total coliforms. The FSB was applied in cupcakes, in proportions of 3% and 6%. The freshly prepared cakes were evaluated in respect to moisture content, water activity (Aw), density, pH and instrumental color. The cakes were stored for 42 days and during this period were evaluated for moisture content, Aw, instrumental texture (hardness, elasticity and cohesion), counts of molds and sensory acceptance (appearance, taste, flavor, texture and overall acceptance). All analysis were also carried out in a control sample, prepared without the addition of FSB, for comparison. The FSB demonstrated to be an excellent source of insoluble fiber, containing 81.92% between cellulose, lignin and hemicellulose, and is source of minerals such as iron, manganese and chromium. However, it is a poor source of soluble fiber, protein, carbohydrates and lipids. Around 60% of the particles of FSB are smaller than 80µm. The FSB presented high counts of molds and the color tends to yellow and red. The cakes showed around 15% of moisture content and Aw in the middle range. Cakes with FSB were slightly damper, darker and denser than traditional. However were less elastic, but so cohesive and hard as the traditional one. Cakes added with 3% of FSB presented the same acceptance that the traditional product, with the advantage of greater nutritional value due to the presence of fibers and minerals from FSB.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第23939号 / 農博第2488号 / 新制||農||1089(附属図書館) / 学位論文||R4||N5374(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 渡邊 隆司, 教授 梅澤 俊明, 教授 森 直樹 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Conventional, advanced and fractionation pretreatment technologies were employed to recover and/or enhance the efficacy of the main constituents present in lignocellulosic biomass. Bamboo and triticale bran are novel feedstocks and hence their response towards treatment is unknown. Thus, to assist with the characterisation of these feedstock, in terms of the amount of sugar released during acid and enzymatic hydrolysis, use were made of conventional pretreatment technologies. Pretreatment involved the use of either the conventional single-stage dilute-acid or the conventional acid-catalysed steam-explosion process at times, temperatures and acid concentration ranging between 5 to 40 min, 120 to 214°C, and 0.002 to 0.055 (H3O+) gmol/L, respectively. For additional comparison, results were also obtained from an established feedstock namely sugarcane bagasse, by subjecting it to the single-stage dilute-acid process, at similar pretreatment conditions employed during the treatment of the other feedstocks. Sugarcane bagasse and bamboo, upon pretreatment and enzymatic hydrolysis, both yielded a similar combined sugar recovery yield of 78.0% and 81.2%, respectively. Alternatively, only 55.3% (w/w) of the total sugar content in triticale bran could be recovered. Triticale bran consists predominantly of hemicellulose which, compared to cellulose, the main constituent in sugarcane bagasse and bamboo, is more susceptible to degradation. Thus, to enhance the combined sugar recovery yield, it is recommended that triticale bran be treated at less severe pretreatment conditions, in order to preserve the hemicellulose. To further enhance the amount of sugar (i.e. cellulose and hemicellulose) that can be recovered from sugarcane bagasse, the use of a two- rather than one-stage dilute acid process was proposed. The single-stage dilute acid process, despite being the subject of many research efforts, failed to recover more than 83% (w/w) of the total sugar content in sugarcane bagasse. Following an extensive literature study, it was concluded that sugarcane bagasse comprises a hemicellulose and cellulose fraction which dictates the use of different pretreatment conditions in order to ensure their effective recovery. The use of a more advance two-step dilute acid process was therefore proposed as it allows for multiple-sets of pretreatment conditions which accommodate the requirements set forth by each of the polysaccharides present in sugarcane bagasse. With the assistance of response surface methodology, a 4.8% (w/w) improvement over the single-stage method was calculated for the two-stage process, by assuming both pretreatment technologies operated at optimum pretreatment conditions. This improvement, which is similar to the 7.7% (w/w) obtained with substrates other than sugarcane bagasse, was accredited to the use of multiple-sets of pretreatment conditions. Both low and high severity pretreatment conditions were applied to maximise the recovery of hemicellulose and cellulose, respectively. In the case of bamboo however, little can be gained by using the two- in lieu of the one-stage method, as similar pretreatment conditions are required to maximise both hemicellulose and cellulose recovery. Alternatively, a three- rather than a two-stage process needs to be applied, when treating triticale bran, to accommodate for an additional set of pretreatment conditions required to recover arabinan, a third polysaccharide. In order to realise the full potential of sugarcane bagasse, use was made of a fractionation method, which in addition to the recovery of polysaccharide (i.e. cellulose and hemicellulose), also allowed for the recovery of the polyphenolic content (i.e. lignin) thereof. Limited by the complexity of the fractionation process, sugarcane bagasse was selected as the preferred substrate to be subjected to fractionation, mainly because of its availability in the South African (established sugar milling industry), and ease of treatment using both conventional (e.g. single-stage) and advanced (e.g. two-stage) pretreatment technologies. A novel fractionation technology, involving the use of the ionic liquid 1-butyl-3- mehylimidazolium methyl sulphate ([BMiM]MeSO4), was devised to separate the main constituents of sugarcane bagasse. Although other ionic liquid fractionation examples also exist in literature, processes used caused production of multi-component product streams with 34% (w/w) of the original lignin and hemicellulose being recovered in the same product stream. Tests conducted during the present study confirmed these results and further indicated that the production of multi-constituent product streams could not be avoided by using acetone, an acetone-water mixture or a sodium hydroxide solution as the solvent in a subsequent solvent extraction step. Hence, to avoid the production of multi-component product streams, a hemicellulose pre-extraction step, comprising the single-stage dilute acid process, was introduced to extract and recover 75% (w/w) of the hemicellulose content. The remaining solid was subjected to ionic liquid treatment whereafter it was effectively separated into cellulose and lignin enriched product streams through solvent extraction. Up to 73% (w/w) of the original lignin and cellulose content was recovered using optimum operating conditions (120 min; 125°C). Comparison of the novel ionic liquid ([BMiM]MeSO4) with 1-ethyl-3-methylimidazolium acetate, an established ionic liquid, identified the former to be the superior delignification solvent whereas the latter contributed more towards the digestibility of the residual solids. In conclusion, methods for the effective recovery of polysaccharides and polyphenol units from established (i.e. sugarcane bagasse) and novel (i.e. bamboo, triticale bran) lignocellulosic feedstocks have been demonstrated through the application of conventional (single-stage), advanced (two-stage) and fractionation pretreatment technologies. Economic improvement is promised through the application of these constituents as they may serve as precursor chemicals for the production of value-added products that may replace the fuel and chemicals currently derived from fossil carbon resources. / AFRIKAANSE OPSOMMING: Konvensionele, gevorderde en fraksionering voorafbehandeling tegnologieë is gebruik om die herwinning en gebruiksaamheid van die hoofbestanddele wat in lignosellulose biomassa aanwesig is, te verbeter. Bamboes en tritikale semels is nuwe rou materiale en dus is hul reaksie op behandeling onbekend. Ten einde die karaktereienskappe van hierdie rou materiale, in terme van die hoeveelheid suiker wat gedurende die suur- en suiker ensimatiese-hidrolise vrygestel word, te bepaal, is gebruik gemaak van konvensionele voorbehandeling tegnologieë. Voorbehandeling behels die gebruik van óf die konvensionele enkelstadium vedunde-suur óf die konvensionele suur-gekataliseerde stoomontploffingsproses; by tye, temperature en suurladings wat wissel tusen 5 en 40 minute, 120 tot 214°C, en 0.002 tot 0.055 (H3O+)gmol/L, onderskeidelik. Vir verdere vergelykings is resultate vanaf ’n gevestigde rou materiaal, naamlik suikerriet reste, verkry, deur dit te onderwerp aan ’n enkelstadium verdunde suur proses met dieselfde voorbehandeling toestande wat gegeld het gedurende die behandeling van die ander roumateriale. Tydens voorbehandeling en ensimaties-hidrolise het suikerriet reste en bamboes ’n gelyke gekombineerde terugwinnings opbrengs van 78.0% en 81.2% suiker, onderskeidelik, gelewer. Daar kon egter net 55.3% (w/w) van die totale suikerinhoud van tritikale semels herwin word. Tritikale semels bestaan hoofsaaklik uit hemisellulose, wat meer vatbaar is vir degradasie ten opsigte van sellulose, die hoof bestanddeel van suikerriet reste en bamboes. Om dus die gekombineerde suikerherwinnings-opbrengs te verbeter, word daar voorgestel dat tritikale semels aan minder strawwe toestande onderwerp word, om sodoende die hemisellulose te bewaar. Om verder die hoeveelheid suiker (d.w.s glukose en xilose) wat vanuit suikerriet reste herwin kan word, te verbeter, is die gebruik van ’n twee- in plaas van ’n eenstadium verdunde suur proses, voorgestel. Die enkelstadium verdunde suur proses het, ten spyte daarvan dat dit al die onderwerp van verskillende navorsing projekte was, nie daarin geslaag om meer as 83% (w/w) van die totale suikerinhoud vanuit die suikerriet reste te herwin nie. Na ’n uitgebreide literatuurstudie is daar tot die slotsom gekom dat suikerriet reste bestaan uit ’n hemisellulose deel en ’n sellulose deel wat die gebruik van verskillende voorbehandeling toestande noodsaak, om sodoende effektiewe herwinning daarvan te verseker. Die aanwending van ’n meer gevorderde twee-stap verdunde suur proses is dus voorgestel, aangesien dit van veelvoudige-stelle voorbehandelings toestande, soos vereis deur die polisakkariedes in suikerriet reste, gebruik maak. Deur die toepassing van reaksie oppervlakte metodologie, is ’n 4.2% verbetering op die enkel-stadium metode vir die twee-stadium proses behaal deur aan te neem dat beide van hierdie prosesse, by optimum kondisies vir maksimale suiker herwinning, uitgevoer is. Hierdie verbetering is soortgelyk aan die 7.7% (w/w) verbetering wat voorheen met andersoortige substrate (uitgesonder suikerriet reste) verkry is. Sodanige verhoging word toegeskryf aan die gebruik van veelvoudige stelle van voorbehandeling toestande, wat beide meer en minder strawwe toestande insluit, om sodoende die herwinning van xilose en glukose onderskeidelik te maksimeer. In die geval van bamboes, is die twee-stadium metode nie superieur tot die een-stadium metode nie, aangesien soortgelyke voorbehandeling-toestande benodig word om beide die herwinning van xilose en glukose te maksimeer. Alternatiewelik moet ’n drie-stadium in plaas van ’n twee-stadium proses gebruik word wanneer tritikale semels behandel word aangesien ’n bykomende stel voorbehandeling toestande benodig word om arabinan, ’n derde polisakkaried, te herwin. Om sodoende die volle potensiaal van suikerriet reste te ontgin, is gebruik gemaak van ʼn fraksioneringstegnologie. Waar konvensionele voorbehandeling metodes slegs fokus op die herwinning van polisakkariedes (naamlik sellulose en hemisellulose), kan die fraksioneringsproses ook die poli-fenoliese inhoud (naamlik lignien) in lignosellulose, herwin. Weens die ingewikkeldheid van die fraksioneringsproses, is suikerriet reste gekies as die mees geskikte substraat vir fraksionering, hoofsaaklik omdat dit veral in die Suid- Afrikaanse konteks, maklik beskikbaar is en omdat dit ook maklik is om met beide die konvensionele (bv. die enkel-stadium) en die gevorderde (bv. die twee-stadium) voorbehandeling proses behandel te word. ʼn Nuut ontwikkelde fraksionering tegnologie wat gebruik maak van die ioniese vloeistof 1-butiel-3-metielimidazolium metielsulfaat ([BMiM]MeSO4) is ontwikkel, om die hoofbestanddele in suikerriet reste van mekaar te skei. Literatuur bevat verskeie voorbeelde waar ioniese vloeistowe gebruik word vir fraksionering. Die doeltreffendheid van hierdie prosesse is egter deur die produksie van multi-komponent produkstrome, waartydens tot en met ekwivalente hoeveelhede hemisellulose en lignien in dieselfde produkstrome herwin is, beperk. Hierdie resultate is tydens die studie deur middel van toetse bevestig. Sodanige toetse het ook aangedui dat die produksie van multi-komponent produkstrome nie deur die gebruikmaking van asetoon, ʼn asetoon-water mengsel of ʼn natrium hidroksied oplosmiddel, wat deel maak van ʼn opvolgende oplossing-ekstraksie-stap, vermy kon word nie. Om gevolglik die vervaardiging van multi-komponent produkstrome te vermy, is ʼn hemisellulose vooraf-ekstraksie stap, bestaande uit die enkel-stap verdunde suur proses, gebruik, om sodoende 75% (w/w) van die hemisellulose inhoud in suikerriet reste, te herwin. Die oorblywende vastestof was onderwerp aan ioniese vloeistof behandeling waarna dit effektiewelik geskei is in onderskeidelik, sellulose en lignien verrykte produkstrome, verkry deur gebruikmaking van ʼn oplossing-ekstraksie-stap. Tot en met 73% (w/w) van die oorspronklike lignien en sellulose inhoud was herwin deur gebruik te maak van optimale behandelingskondisies (120 min, 125°C). ʼn Vergelyking tussen die nuut-geïdentifiseerde ioniese vloeistof ([BMiM]MeSO4) en 1-etiel-3-metielimidazolium asetaat, ʼn gevestigde ioniese vloeistof, het aangetoon dat eersgenoemde ʼn superieure delignifikasie oplosmiddel is terwyl laasgenoemde ʼn groter bydra tot die verteerbarheid van die oorblywende reste lewer. Ten slotte het hierdie studie, metodes vir die effektiewe herwinning van polisakkariedes en poli-fenoliese eenhede, vanaf gevestigde (bv. Suikerriet reste) en nuwe (bv. bamboes en tritikale semels) lignosellulose rou materiale, gedemonstreer, deur telkens van konvensionele (enkel-stadium), gevorderde (twee-stadium) en fraksionerings voorbehandeling tegnologieë gebruik te maak. Ekonomiese verbetering kan dus verwag word indien hierdie bestanddele as voorloper chemikalieë vir die produksie van waarde toegevoegde produkte aangewend word, aangesien sodanige produkte die plek kan inneem van brandstof en chemikalieë wat tans vanuit fossielbronne vervaardig word.

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: The world’s depleting fossil fuels and increasing greenhouse gas emissions have given rise to much research into renewable and cleaner energy. Biomass is unique in providing the only renewable source of fixed carbon. Agricultural residues such as Sugarcane Bagasse (SB) are feedstocks for ‘second generation fuels’ which means they do not compete with production of food crops. In South Africa approximately 6 million tons of raw SB is produced annually, most of which is combusted onsite for steam generation. In light of the current interest in bio-fuels and the poor utilization of SB as energy product in the sugar industry, alternative energy recovery processes should be investigated. This study looks into the thermochemical upgrading of SB by means of pyrolysis. Biomass pyrolysis is defined as the thermo-chemical decomposition of organic materials in the absence of oxygen or other reactants. Slow Pyrolysis (SP), Vacuum Pyrolysis (VP), and Fast Pyrolysis (FP) are studied in this thesis. Varying amounts of char and bio-oil are produced by the different processes, which both provide advantages to the sugar industry. Char can be combusted or gasified as an energy-dense fuel, used as bio-char fertilizer, or upgraded to activated carbon. High quality bio-oil can be combusted or gasified as a liquid energy-dense fuel, can be used as a chemical feedstock, and shows potential for upgrading to transport fuel quality. FP is the most modern of the pyrolysis technologies and is focused on oil production. In order to investigate this process a 1 kg/h FP unit was designed, constructed and commissioned. The new unit was tested and compared to two different FP processes at Forschungszentrum Karlsruhe (FZK) in Germany. As a means of investigating the devolatilization behaviour of SB a Thermogravimetric Analysis (TGA) study was conducted. To investigate the quality of products that can be obtained an experimental study was done on SP, VP, and FP. Three distinct mass loss stages were identified from TGA. The first stage, 25 to 110°C, is due to evaporation of moisture. Pyrolitic devolatilization was shown to start at 230°C. The final stage occurs at temperatures above 370°C and is associated with the cracking of heavier bonds and char formation. The optimal decomposition temperatures for hemicellulose and cellulose were identified as 290°C and 345°C, respectively. Lignin was found to decompose over the entire temperature range without a distinct peak. These results were confirmed by a previous study on TGA of bagasse. SP and VP of bagasse were studied in the same reactor to allow for accurate comparison. Both these processes were conducted at low heating rates (20°C/min) and were therefore focused on char production. Slow pyrolysis produced the highest char yield, and char calorific value. Vacuum pyrolysis produced the highest BET surface area chars (>300 m2/g) and bio-oil that contained significantly less water compared to SP bio-oil. The short vapour residence time in the VP process improved the quality of liquids. The mechanism for pore formation is improved at low pressure, thereby producing higher surface area chars. A trade-off exists between the yield of char and the quality thereof. FP at Stellenbosch University produced liquid yields up to 65 ± 3 wt% at the established optimal temperature of 500°C. The properties of the bio-oil from the newly designed unit compared well to bio-oil from the units at FZK. The char properties showed some variation for the different FP processes. At the optimal FP conditions 20 wt% extra bio-oil is produced compared to SP and VP. The FP bio-oil contained 20 wt% water and the calorific value was estimated at 18 ± 1 MJ/kg. The energy per volume of FP bio-oil was estimated to be at least 11 times more than dry SB. FP was found to be the most effective process for producing a single product with over 60% of the original biomass energy. The optimal productions of either high quality bio-oil or high surface area char were found to be application dependent. / AFRIKAANSE OPSOMMING: As gevolg van die uitputting van fossielbrandstofreserwes, en die toenemende vrystelling van kweekhuisgasse word daar tans wêreldwyd baie navorsing op hernubare en skoner energie gedoen. Biomassa is uniek as die enigste bron van hernubare vaste koolstof. Landbouafval soos Suikerriet Bagasse (SB) is grondstowwe vir ‘tweede generasie bio-brandstowwe’ wat nie die mark van voedselgewasse direk affekteer nie. In Suid Afrika word jaarliks ongeveer 6 miljoen ton SB geproduseer, waarvan die meeste by die suikermeulens verbrand word om stoom te genereer. Weens die huidige belangstelling in bio-brandstowwe en ondoeltreffende benutting van SB as energieproduk in die suikerindustrie moet alternatiewe energie-onginningsprosesse ondersoek word. Hierdie studie is op die termo-chemiese verwerking van SB deur middel van pirolise gefokus. Biomassa pirolise word gedefinieer as die termo-chemiese afbreking van organiese bio-materiaal in die afwesigheid van suurstof en ander reagense. Stadige Pirolise (SP), Vakuum Pirolise (VP), en Vinnige Pirolise word in hierdie tesis ondersoek. Die drie prosesse produseer veskillende hoeveelhede houtskool en bio-olie wat albei voordele bied vir die suikerindustrie. Houtskool kan as ‘n vaste energie-digte brandstof verbrand of vergas word, as bio-houtskoolkompos gebruik word, of kan verder tot geaktiveerde koolstof geprosesseer word. Hoë kwaliteit bio-olie kan verbrand of vergas word, kan as bron vir chemikalië gebruik word, en toon potensiaal om in die toekoms opgegradeer te kan word tot vervoerbrandstof kwaliteit. Vinnige pirolise is die mees moderne pirolise tegnologie en is op bio-olie produksie gefokus. Om die laasgenoemde proses te toets is ‘n 1 kg/h vinnige pirolise eenheid ontwerp, opgerig en in werking gestel. Die nuwe pirolise eenheid is getoets en vegelyk met twee verskillende vinnige pirolise eenhede by Forschungszentrum Karlsruhe (FZK) in Duitsland. Termo-Gravimetriese Analise (TGA) is gedoen om die ontvlugtigingskenmerke van SB te bestudeer. Eksperimentele werk is verrig om die kwaliteit van produkte van SP, VP, vinnige pirolise te vergelyk. Drie duidelike massaverlies fases van TGA is geïdentifiseer. Die eerste fase (25 – 110°C) is as gevolg van die verdamping van vog. Pirolitiese ontvlugtiging het begin by 230°C. Die finale fase (> 370°C) is met die kraking van swaar verbindings en die vorming van houtskool geassosieer. Die optimale afbrekingstemperatuur vir hemisellulose en sellulose is as 290°C en 345°C, respektiewelik, geïdentifiseer. Daar is gevind dat lignien stadig oor die twede en derde fases afgebreek word sonder ‘n duidelike optimale afbrekingstemperatuur. Die resultate is deur vorige navorsing op TGA van SB bevestig. SP en VP van bagasse is in dieselfde reaktor bestudeer, om ‘n akkurate vergelyking moontlik te maak. Beide prosesse was by lae verhittingstempo’s (20°C/min) ondersoek, wat gevolglik op houtskoolformasie gefokus is. SP het die hoogste houtskoolopbrengs, met die hoogste verbrandingsenergie, geproduseer. VP het hootskool met die hoogste BET oppervlakarea geproduseer, en die bio-olie was weens ‘n dramatiese afname in waterinhoud van beter gehalte. Die meganisme vir die vorming van ‘n poreuse struktuur word deur lae atmosferiese druk verbeter. Daar bestaan ‘n inverse verband tussen die kwantiteit en kwaliteit van die houtskool. Vinnige pirolise by die Universiteit van Stellenbosch het ‘n bio-olie opbrengs van 65 ± 3 massa% by ‘n vooraf vasgestelde optimale temperatuur van 500°C geproduseer. Die eienskappe van bio-olie wat deur die nuwe vinnige pirolise eenheid geproduseer is het goed ooreengestem met die bio-olie afkomstig van FZK se pirolise eenhede. Die houtskool eienskappe van die drie pirolise eenhede het enkele verskille getoon. By optimale toestande vir vinnige pirolise word daar 20 massa% meer bio-olie as by SP en VP geproduseer. Vinnige pirolise bio-olie het ‘n waterinhoud van 20 massa% en ‘n verbrandingswarmte van 18 ± 1 MJ/kg. Daar is gevind dat ten opsigte van droë SB die energie per enheidsvolume van bio-olie ongeveer 11 keer meer is. Vinnige pirolise is die mees doeltreffende proses vir die vervaardiging van ‘n produk wat meer as 60% van die oorspronklike biomassa energie bevat. Daar is gevind dat die optimale hoeveelhede van hoë kwaliteit bio-olie en hoë oppervlakarea houtskool doelafhanklik is.

Um dos grandes desafios nas indústrias alimentícia, farmacêutica e agropecuária é a busca por novos compostos para substituir os antibióticos. Como possíveis candidatos estão as bacteriocinas para serem utilizados paralelamente aos antibióticos ou até substituí-los. As bactérias ácido-láticas podem produzir substâncias inibitórias semelhantes às bacteriocinas (BLIS - bacteriocin-like inhibitory substances) que possuem efeito bacteriostático ou bactericida contra diferentes grupos de bactéria sendo largamente utilizadas como bioconservantes alimentares. Neste contexto, o objetivo deste trabalho foi o emprego de um resíduo agroindustrial, o hidrolisado de bagaço de cana-de-açúcar, como fonte de carbono em cultivos fermentativos para produção do BLIS pela cepa Pediococcus pentosaceus ET 34. Resultados revelaram que as células de ET34 foram capazes de crescer utilizando este resíduo agroindustrial como fonte de carbono e ensaios utilizando planejamento fatorial demonstraram que a agitação não influencia na produção de BLIS. Ao escalonar o cultivo em biorreatores, foi verificado que tanto o crescimento como a atividade antimicrobiana foram semelhantes aos obtidos em bancada com exceção da condição de 25% (v/v) de HBC (hidrolisado de bagaço de cana) que devido a maior viscosidade do meio, resultou em uma diminuição no crescimento e de produção de BLIS. O BLIS produzido por células de ET34 utilizando o HBC como fonte de carbono foi parcialmente purificado por sulfato de amônio e demonstrou atividade contra Listeria monocytogenes ATCC 934 e Salmonella enterica CECT 724. Desta maneira, pode-se concluir que Pediococcus pentosaceus ET34 foi capaz de crescer em meio contendo HBC como fonte de carbono produzindo BLIS em seu sobrenadante com ação frente a diferentes bactérias patogênicas. A possibilidade de utilizar uma fonte alternativa de carbono pode diminuir o custo de processo consideravelmente. Além disso, ensaios de planejamento fatorial, superfície de resposta e escalonamento em biorreator de bancada indicaram que concentrações baixas de HBC (5-15%, v/v) a 35 °C resultaram na maior produção de BLIS. / The great challenge in the food, pharmaceutical and agricultural industries is the search for new compounds to replace antibiotics. Bacteriocins are possible candidates that can be used in parallel with the antibiotics or even to replace them. Lactic-acid bacteria can produce bacteriocin inhibitory substances (BLIS - bacteriocin-like inhibitory substances) that have a bacteriostatic or bactericidal effect against different bacterial species and are widely used as food bioconservatives. In this context, the aim of this work was to use of an agroindustrial waste, hydrolyzed sugarcane bagasse, as a carbon source in fermentative cultures for the production of BLIS by Pediococcus pentosaceus ET 34 strain. Results revealed that ET34 cells were able to grow using this agroindustrial residue as a carbon source, and trials using factorial design showed that agitation did not influence on the production of BLIS. When it was perform cultivation scale up in bioreactors, it was verified that both the growth and the antimicrobial activity were similar to those obtained in the workbench with the exception of the condition of 25% (v/v) of HBC (sugarcane bagasse hydrolyzate) that due to its higher viscosity, resulted in a decrease in growth and BLIS production. BLIS produced by ET34 cells using HBC as a carbon source that was partially purified by ammonium sulfate showed activity against Listeria monocytogenes ATCC 934 and Salmonella enterica CECT 724. Thus, it can be concluded that Pediococcus pentosaceus ET34 was able to grow in medium containing HBC as carbon source producing BLIS in its supernatant with action against different pathogenic bacteria. The possibility of using an alternative carbon source can greatly reduce the process cost. In addition, factorial design, response surface and scale up trials in bench bioreactors indicated that low concentrations of HBC (5-15% v/v) at 35 ºC resulted in higher BLIS production.

A agroindústria da cana-de-açúcar no Brasil foi um dos setores que mais se desenvolveu nas últimas 4 décadas. Do caminho trilhado desde os canaviais até as bombas de combustível, cerca de 30% da massa de cana-de-açúcar utilizada para a produção de etanol é perdida na forma de palha e bagaço. O resíduo de natureza lignocelulósica, apresenta composição aproximada de 32-48% de celulose, 23-27% de hemicelulose e 19-32% de lignina. Em virtude da resistência física e mecânica oferecida por esse tipo de material, diversas metodologias como pré-tratamentos químicos ou enzimáticos têm sido propostos visando tornar mais acessível a fração polissacarídica do bagaço à quebra em açúcares fermentescíveis passíveis a conversão em etanol de segunda geração. Nesse contexto, o fungo Aspergillus fumigatus apesar de patogênico, apresenta enorme relevância no cenário da desconstrução de resíduos de origem lignocelulósica como o bagaço. Com a recém descoberta das chamadas mono-oxigenases líticas de polissacarídeos (LPMOs), metaloenzimas auxiliares à degradação de compostos lignocelulósicos que apresentam notoriedade por conta de seus mecanismos de ação oxidorredutivos, as mesmas já foram encontradas em diversos tipos de microorganismos, incluindo o próprio A. fumigatus. Nesse sentido, o projeto em questão realizou a clonagem e expressão heteróloga na cepa E. coli Rosetta(TM)(DE3)pLysS dos genes AFUA_1G12560 e AFUA_4G07850 responsáveis por codificar respectivamente as enzimas AfuLPMO9A e AfuLPMO9C de A. fumigatus Af293 identificadas em análises de secretoma e RNA-Seq previamente obtidos pelo nosso grupo de pesquisa. Através da análise desses dois genes, foi verificado que na presença de diferentes fontes de carbono, ambos foram induzidos, de modo que na presença de SEB, CMC e Avicel, o gene AFUA_4G07850 foi em até 3500X, 2000X e 1000X mais induzido, enquanto que o AFUA_1G12560 apresentou um aumento moderado de 16X, 13X e 7X, respectivamente. A avaliação da conformação tridimensional, bem como o alinhamento múltiplo com enzimas de maior grau de homologia revelaram algumas características estruturais importantes como a presença de núcleos ricos em estruturas ? em conformação de sanduíche e que importantes resíduos conservados como H20; H105; Y194 e H22; H107; Y196 encontravam-se coordenados ao centro metálico das enzimas AfuLPMO9A e AfuLPMO9C, respectivamente. Uma análise mais profunda baseada em DFT revelou que o sítio ativo de ambas assume uma geometria piramidal de base quadrada. Após purificação das LPMOs, ensaios de atividade revelaram que as enzimas AfuLPMO9A e AfuLPMO9C aumentaram respectivamente em até 1,26X e 1,20X a atividade do coquetel enzimático quando nas concentrações de 10 mg/g Avicel e presença de 0,02% NaN3. Análises mais profundas dessas enzimas auxiliares vão permitir uma maior compreensão sobre o papel desempenhado por elas na desconstrução do bagaço de cana-de-açúcar. / The sugarcane agroindustry in Brazil was one of the most developed sectors in the last 4 decades. In the path from cane fields to the gas stations, about 30% of the sugarcane mass used for ethanol production is lost in form of straw and bagasse. The lignocellulosic residue presents approximately 32-48% cellulose, 23-27% hemicellulose and 19-32% lignin. Due to the physical and mechanical resistance offered by this type of material, several methodologies such as chemical or enzymatic pre-treatments have been proposed to make the polysaccharide fraction of the bagasse accessible to the break in fermentable sugars that can be converted into second generation ethanol. In this context, the fungus Aspergillus fumigatus, despite being pathogenic, presents enormous relevance in the scenario of deconstruction of lignocellulosic residues such as bagasse. With the recently discovery of the so-called polysaccharide lytic monooxygenases (LPMOs), metalloenzymes that help the degradation of lignocellulosic compounds and are notorious for their oxidoreductive mechanisms of action, they have already been found in several types of microorganisms, including A. fumigatus. In this regard, this project performed the heterologous cloning and expression in E. coli Rosetta(TM)(DE3)pLysS of the AFUA_1G12560 and AFUA_4G07850 genes responsible for respectively encoding the A. fumigatus Af293 enzymes AfuLPMO9A and AfuLPMO9C identified in secretome and RNA-Seq results previously obtained by our research group. By analyzing these two genes, it was found that in the presence of different carbon sources, both were induced, so that in the presence of SEB, CMC and Avicel, the AFUA_4G07850 gene was up to 3500X, 2000X and 1000X more induced while AFUA_1G12560 presented a lower increase of 16X, 13X and 7X, respectively. The evaluation of the three-dimensional conformation as well as the multiple alignment with higher degree homology enzymes revealed some important structural features such as the presence of cores rich in ? structures in sandwich conformation and that important conserved residues as H20; H105; Y194 and H22; H107; Y196 were coordinated to the metal center of the enzymes AfuLPMO9A and AfuLPMO9C, respectively. Further analysis based on DFT revealed that the active site of both assumes a square-based pyramidal geometry. After purification of LPMOs, activity assays revealed that the enzymes AfuLPMO9A and AfuLPMO9C were able to increase cocktail activity up to approximately 1,26X and 1,20X at concentrations of 10 mg/g Avicel and presence of 0.02% NaN3. Further analysis of these auxiliary enzymes will allow a better understanding of their role in sugarcane bagasse deconstruction.