Atualmente, a bioprodução de etanol a partir de materiais lignocelulósicos como o bagaço de cana é de grande interesse nacional e cientifico. Para utilização destes materiais em processos biotecnológicos procedeu-se sua hidrólise em função de sua constituição complexa em celulose, hemicelulose e lignina. Após hidrólise ácida branda obteve-se um hidrolisado hemicelulósico rico em xilose e outros açúcares juntamente com compostos tóxicos a atividade microbiana como ácido acético, furfural, hidroximetilfurfural e compostos fenólicos. Este trabalho avaliou a necessidade da suplementação nutricional bem como a concentração inicial de xilose e células no hidrolisado hemicelulósico de bagaço de cana-de-açúcar sobre os parâmetros fermentativos de sua bioconversão a etanol pela levedura Pichia stipitis UFMG-IMH 43.2. Os ensaios foram realizados de acordo com um planejamento fatorial completo 23 com face centrada tendo como resposta o fator de conversão de açúcares em etanol (YP/S). Na suplementação do hidrolisado avaliou-se a adição dos seguintes nutrientes: MgSO4 (0 a 1,0 g/L), extrato de levedura (0 a 5,0 g/L) e uréia (0 a 5,0 g/L). Neste estudo observou-se que o extrato de levedura (5,0 g/L) exerceu maior influência sobre o processo fermentativo alcançando máximos valores de YP/S (0,13 g/L) e produtividade volumétrica em etanol (0,07 g/L.h) não necessitando da adição dos demais nutrientes. Ao se avaliar a concentração inicial de xilose (30; 52,5; 75 g/L) bem como a concentração celular inicial (0,5; 1,0; 2,0 g/L) no hidrolisado observou-se uma melhoria significativa no valor de YP/S (0,19 g/g) utilizando máxima concentração celular inicial (2 g/L) associada com menor concentração de xilose inicial (30 g/L). Estes resultados indicaram que a levedura Pichia stipitis UFMG-IMH 43.2 favoreceu a conversão em etanol YP/S (0,19 g/g), mostrando que a levedura teve melhor comportamento a baixas concentrações de xilose. Este estudo demonstrou que a levedura P. stipitis UFMG-IMH 43.2 mostrou-se como uma alternativa viável para a produção de bioetanol em hidrolisado hemicelulósico de bagaço de cana-de-açúcar. / Currently, ethanol bioproduction from lignocellulosic materials such as sugarcane bagasse is of great national and scientific interest. For utilization of these materials in biotechnological processes according mild acid hydrolysis is required to complex constitution of cellulose, hemicellulose and lignin. After mild acid hydrolysis, a hemicellulosic hydrolyzate rich in xylose and other sugars together with toxic compounds to microbial activity as acetic acid, furfural, hydroxymethylfurfural and phenolics was obtained. The studies on the hydrolysate supplementation evaluated the nutrients addition: MgSO4 (0 to 1.0 g/L), yeast extract (0 to 5.0 g/L) and urea (0 to 5.0 g/L) and effects of initial xylose and cell concentration for ethanol production by Pichia stipitis UFMG-IMH 43.2 yeast were evaluated. The experiments were performed according to factorial design central composite face centered 23 using YP/S as response factor. This study showed that only the addition of 5.0 g/L yeast extract was enough to obtained YP/S equal to 0.13 g/L and productivity of 0.07 g/L-h. The evaluation of initial xylose concentration (30; 52,5; 75 g/L) and initial cell concentration (0,5; 1,0; 2;0 g/L) showed a significant influence on fermentation parameter YP/S. The best conditions were achieved using initial cell concentration of 2.0 g/L and initial xylose concentration of 30 g/L with YP/S equal to 0.19 g/g, showing that this yeast strain had better performance at low xylose concentrations. This study demonstrated that P. stipitis UFMG-IMH 43.2 yeast was a promising alternative for bioethanol production from sugarcane bagasse hemicellulosic hydrolysate.

This is an experimental study into the permeability and compressibility properties of bagasse pulp pads. Three experimental rigs were custom-built for this project. The experimental work is complemented by modelling work. Both the steady-state and dynamic behaviour of pulp pads are evaluated in the experimental and modelling components of this project. Bagasse, the fibrous residue that remains after sugar is extracted from sugarcane, is normally burnt in Australia to generate steam and electricity for the sugar factory. A study into bagasse pulp was motivated by the possibility of making highly value-added pulp products from bagasse for the financial benefit of sugarcane millers and growers. The bagasse pulp and paper industry is a multibillion dollar industry (1). Bagasse pulp could replace eucalypt pulp which is more widely used in the local production of paper products. An opportunity exists for replacing the large quantity of mainly generic paper products imported to Australia. This includes 949,000 tonnes of generic photocopier papers (2). The use of bagasse pulp for paper manufacture is the main application area of interest for this study. Bagasse contains a large quantity of short parenchyma cells called ‘pith’. Around 30% of the shortest fibres are removed from bagasse prior to pulping. Despite the ‘depithing’ operations in conventional bagasse pulp mills, a large amount of pith remains in the pulp. Amongst Australian paper producers there is a perception that the high quantity of short fibres in bagasse pulp leads to poor filtration behaviour at the wet-end of a paper machine. Bagasse pulp’s poor filtration behaviour reduces paper production rates and consequently revenue when compared to paper production using locally made eucalypt pulp. Pulp filtration can be characterised by two interacting factors; permeability and compressibility. Surprisingly, there has previously been very little rigorous investigation into neither bagasse pulp permeability nor compressibility. Only freeness testing of bagasse pulp has been published in the open literature. As a result, this study has focussed on a detailed investigation of the filtration properties of bagasse pulp pads. As part of this investigation, this study investigated three options for improving the permeability and compressibility properties of Australian bagasse pulp pads. Two options for further pre-treating depithed bagasse prior to pulping were considered. Firstly, bagasse was fractionated based on size. Two bagasse fractions were produced, ‘coarse’ and ‘medium’ bagasse fractions. Secondly, bagasse was collected after being processed on two types of juice extraction technology, i.e. from a sugar mill and from a sugar diffuser. Finally one method of post-treating the bagasse pulp was investigated. The effects of chemical additives, which are known to improve freeness, were also assessed for their effect on pulp pad permeability and compressibility. Pre-treated Australian bagasse pulp samples were compared with several benchmark pulp samples. A sample of commonly used kraft Eucalyptus globulus pulp was obtained. A sample of depithed Argentinean bagasse, which is used for commercial paper production, was also obtained. A sample of Australian bagasse which was depithed as per typical factory operations was also produced for benchmarking purposes. The steady-state pulp pad permeability and compressibility parameters were determined experimentally using two purpose-built experimental rigs. In reality, steady-state conditions do not exist on a paper machine. The permeability changes as the sheet compresses over time. Hence, a dynamic model was developed which uses the experimentally determined steady-state permeability and compressibility parameters as inputs. The filtration model was developed with a view to designing pulp processing equipment that is suitable specifically for bagasse pulp. The predicted results of the dynamic model were compared to experimental data. The effectiveness of a polymeric and microparticle chemical additives for improving the retention of short fibres and increasing the drainage rate of a bagasse pulp slurry was determined in a third purpose-built rig; a modified Dynamic Drainage Jar (DDJ). These chemical additives were then used in the making of a pulp pad, and their effect on the steady-state and dynamic permeability and compressibility of bagasse pulp pads was determined. The most important finding from this investigation was that Australian bagasse pulp was produced with higher permeability than eucalypt pulp, despite a higher overall content of short fibres. It is thought this research outcome could enable Australian paper producers to switch from eucalypt pulp to bagasse pulp without sacrificing paper machine productivity. It is thought that two factors contributed to the high permeability of the bagasse pulp pad. Firstly, thicker cell walls of the bagasse pulp fibres resulted in high fibre stiffness. Secondly, the bagasse pulp had a large proportion of fibres longer than 1.3 mm. These attributes helped to reinforce the pulp pad matrix. The steady-state permeability and compressibility parameters for the eucalypt pulp were consistent with those found by previous workers. It was also found that Australian pulp derived from the ‘coarse’ bagasse fraction had higher steady-state permeability than the ‘medium’ fraction. However, there was no difference between bagasse pulp originating from a diffuser or a mill. The bagasse pre-treatment options investigated in this study were not found to affect the steady-state compressibility parameters of a pulp pad. The dynamic filtration model was found to give predictions that were in good agreement with experimental data for pads made from samples of pretreated bagasse pulp, provided at least some pith was removed prior to pulping. Applying vacuum to a pulp slurry in the modified DDJ dramatically reduced the drainage time. At any level of vacuum, bagasse pulp benefitted from chemical additives as quantified by reduced drainage time and increased retention of short fibres. Using the modified DDJ, it was observed that under specific conditions, a benchmark depithed bagasse pulp drained more rapidly than the ‘coarse’ bagasse pulp. In steady-state permeability and compressibility experiments, the addition of chemical additives improved the pad permeability and compressibility of a benchmark bagasse pulp with a high quantity of short fibres. Importantly, this effect was not observed for the ‘coarse’ bagasse pulp. However, dynamic filtration experiments showed that there was also a small observable improvement in filtration for the ‘medium’ bagasse pulp. The mechanism of bagasse pulp pad consolidation appears to be by fibre realignment. Chemical additives assist to lubricate the consolidation process. This study was complemented by pulp physical and chemical property testing and a microscopy study. In addition to its high pulp pad permeability, ‘coarse’ bagasse pulp often (but not always) had superior physical properties than a benchmark depithed bagasse pulp.

Eight strains of bacteria (L1-L8) were isolated from the mud of a decantation lake (A) from the Station of Effluent Treatment (SET) of the S. A. Usina Coruripe Açúcar e Álcool , in the harvest season of 2004/05. Three of these bacteria were considered facultative anaerobes (L3, L7 and L8). The isolates were identified as species of the genus Achromobacter (L1 and L5), Acinetobacter (L2 and L6), Proteus (L3), Flavobacterium (L4), and Serratia (L7 and L8), with different enzymatic activities that are of environmental interest. On the other hand, 19 microorganisms were isolated from the wastewater samples of the same lake and of the sugarcane bagasse stored in that company, it means,8 from the effluent and 11 from the bagasse, being 14 bacteria G+ and G- (A1, A2, A3, A4, A5, A6, A7, A8, BC1, BC2, BC3, BC6, BC7 and BC11), and 5 were fungi - 3 yeast (BC4, BC5 and BC9) and 2 moulds (BC8 and BC10). All the isolates were mesophylic. The bacteria from the studied effluent were identified as Klebsiella sp (A1), Corynebacterium sp (A2 and A7), Arthrobacter sp (A3), Streptomyces sp (A4); Staphylococcus sp (A5), Acinetobacter sp (A6); Serratia sp (A8). The isolates from the sugarcane bagasse were species from the genus Bacillus (BC1),Clostridium. (BC2), Acinetobacter (BC3 and BC6); Corynebacterium (BC7) and Lactobacillus (BC11). Because of their high potential in degrading polysaccharides, lipids, proteins, and phenolic compounds such as tannin, the mud bacteria L1 (Achromobacter sp) and L3 (Proteus sp) were screened according to their kinetic of growth and degradation of tannic acid. Although the concentration of tannic acid at 0,8 % still allows the growth of Proteus sp, the concentration of the same substrate at 1% in solid medium inhibited its growth, suggesting that the microorganism do not tolerate high concentrations of this compound. Furthermore, even if Achromobacter sp from the mud had not grown in solid medium containing 0.8-1.0% of tannic acid, it was able to developed very well in medium with gallic acid, and also hydrolysed ester linkages of the tannic acid molecule in liquid medium, releasing gallic acid. The species of Proteus and Achromobacter sp, isolated from the mud from the decantation lake of the studied sugar-alcohol industry (harvest season 2004/05) appear promising for studies of co-metabolism and bioaugmentation for treatment of this type of wastewater containing phenolic compounds and other substances. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Foram isoladas 8 linhagens bacterianas (L1-L8) de lodo da lagoa de decantação A da Estação de Tratamento de Efluentes (ETE) da S. A. Usina Coruripe Açúcar e Álcool, na safra de 2004/05, sendo 3 deles anaeróbios facultativos com metabolismo fermentativo (L3, L7 e L8). Os isolados foram identificados como espécies de Achromobacter (L1 e L5), Acinetobacter (L2 e L6), Proteus (L3), Flavobacterium (L4), e Serratia (L7 e L8), apresentando diversas atividades enzimáticas de interesse ambiental. Quanto às amostras do efluente da mesma lagoa de decantação da ETE estudada, e do bagaço de cana-de-açúcar acumulado na empresa, foram isolados 19 microrganismos (8 do efluente e 11 do bagaço), sendo que 14 são bactérias G+ e G- (A1, A2, A3, A4, A5, A6, A7, A8, BC1, BC2, BC3, BC6, BC7 e BC11) e 5 são fungos, isto é, 3 leveduriformes (BC4, BC5 e BC9) e 2 filamentosos (BC8 e BC10). Todos os microrganismos obtidos são mesofílicos. As bactérias dos efluentes da ETE estudada foram identificadas como pertencentes aos gêneros Klebsiella (A1), Corynebacterium (A2 e A7), Arthrobacter (A3), Streptomyces (A4); Staphylococcus (A5), Acinetobacter (A6); Serratia (A8). Os isolados de bagaço-de-cana pertencem ao gêneros: Bacillus sp (BC1), Clostridium. Sp (BC2), Acinetobacter (BC3 e BC6); Corynebacterium (BC7) e Lactobacillus (BC11). Por terem apresentado um alto potencial em degradar polissacarídeos, lipídeos, proteínas e compostos fenólicos (tanino), os isolados de lodo L1 (Achromobacter sp) e L3 (Proteus sp) foram selecionados para testes de cinética de crescimento/degradação de ácido tânico. Embora concentrações de 0,8 % de ácido tânico ainda favoreçam o crescimento de Proteus sp, a concentração de 1 % desse substrato em meio sólido inibiu seu crescimento, sugerindo que o microrganismo não tolera altas concentrações desse composto. Apesar de não ter sido observado o crescimento do isolado de lodo Achromobacter sp. em meio sólido contendo 0,8-1,0% de ácido tânico, o mesmo foi capaz de degradar fenóis pelo seu desenvolvimento em meio contendo ácido gálico, ou por ter hidrolisado as ligações éster da molécula de ácido tânico em meio líquido, liberando ácido gálico. As espécies de Proteus sp e Achromobacter sp, isoladas de lodo da lagoa de decantação da ETE da indústria sucroalcooleira estudada (safra 2004/05), são promissoras para estudos de cometabolismo de microrganismos e bioaumentação de águas residuárias contendo compostos fenólicos e outras substâncias.

Orientador: Maurício Boscolo / Banca: Sandra Helena da Cruz / Banca: Roberto da Silva / Resumo: No presente trabalho foi avaliada a utilização de pré-tratamentos com micro-ondas e ultrassom, associados ou não a agentes químicos, sobre o bagaço de cana-de-açúcar para posterior sacarificação enzimática para produção de açúcares fermentescíveis. Os sobrenadantes dos pré-tratamentos foram analisados por HPLC-RI para quantificação de açúcares e HPLC-UV para determinação de compostos fenólicos, provenientes da degradação da lignina e inibidores da sacarificação. Os bagaços pré-tratados foram hidrolisados com dois complexos enzimáticos fornecidos pela Novozymes, o complexo NS50012 contendo diversas enzimas como celulases, xilanases e hemicelulases e o complexo NS50013 mais específico, contendo maior atividade de -glicosidase. Verificou-se que as amostras controle tratadas com micro-ondas e ultrassom sofreram perda de massa de 3% e 1,6% respectivamente, porém, variações consideráveis ocorreram com as amostras tratadas com ácidos minerais, chegando a 55,8 % de perda de massa com ácido clorídrico, seguido do ácido nítrico, com perda de 51,5%, ambas com liberação de açúcares, como glicose e xilose. Em meio alcalino, a maior perda foi com o tratamento de NaOH , com 17,3%. O uso de peróxido de hidrogênio seguido de hidróxido de sódio mostrou-se eficiente na deslignificação, com perdas de massa na ordem de 22 e 17%, com o uso de micro-ondas e ultrassom, respectivamente. O uso de CaO proporcionou perda de massa de 45,7 % e o tratamento realizado com sais cujos cátions são ácidos de Lewis como o CaCl2 e o FeCl2 resultaram em redução de 30-35% na massa do bagaço. No tratamento ultrassônico destacam-se valores entre 11-29% para os ácidos, mas, de modo geral, os tratamentos com uso de ultrassom demonstraram serem menos efetivos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In this work it was tested the utilization of microwave and ultrasound as pretreatments of the sugarcane bagasse associated or not with chemical agents, for later enzymatic saccharification for the production of fermentable sugars. The supernatants of the pretreatments was analyze by HPLC-RI for sugars quantification and HPLC-UV for determination of fenolic compounds, from the lignin degradation and fermentation inhibitors. The treated bagasse was hydrolyzed with two different enzymatic complexes provided by Novozymes, the NS50012 complex that content some different enzymes like cellulases, xylanases and hemicellulases, and the NS50013 complex that is more specific, with larger values of -glucosidase. It was checked that the control samples treated with microwave and ultrasound suffered weight loss of 3% and 1,6% respectively, and considerable variations occurred with the samples treated with mineral acids, reaching to 55,8% of weight loss with chloridric acid, followed of nitric acid, with loss of 51,5%, both with sugar liberation, as such as glucose and xylose. In alkaline medium, the larger weight loss was with the treatment with NaOH, com 17,3%. The utilization of hydrogen peroxide followed by NaOH showed efficiency in the delignification, with weight losses in the order of 22% and 17%, with microwave and ultrasound, respectively. The utilization of CaO provided weight loss of 45,7% and the treatment realized with salts that the cation are Lewis acids like CaCl2 and FeCl2 resulting in reductions of 30-35% in bagasse weight. In the ultrasound treatment stand out values between 29-11% for the acids, but, in overall, the ultrasound treatments showed be fewer effective in weight loss when compared with microwave utilization. Was detect liberation of sugars like glucose, xylose and cellobiose, after treatments... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2016-06-02T18:55:26Z (GMT). No. of bitstreams: 1 5512.pdf: 1675884 bytes, checksum: 47d8b8c6a94d1de848c43c964a5f3257 (MD5) Previous issue date: 2013-08-13 / Financiadora de Estudos e Projetos / Bioethanol is an alternative low cost fuel, which is conventionally obtained in Brazil from the fermentation of sugarcane juice, molasses, or a mixture of these. However, alternatively, ethanol could be obtained from lignocellulosic materials, wastes or agroindustrial byproducts composed of cellulose, hemicellulose and lignin, such as sugarcane bagasse. For each ton of cane processed, 250 kg of bagasse are obtained on average, which is usually burned for power generation industry. The microbial cultivation using these materials depend on the availability of substrates, accordingly, acid hydrolysis has been considered one of the most widely used process for the depolymerization of the hemicellulose fraction of lignocellulosic materials, due to their low cost and high efficiency. However, under conditions of high temperature and pressure, glucose and xylose released are degraded to 5- hydroxymethylfurfural (HMF) and furfural, respectively. These compounds are characterized by inhibiting the yeast used in the fermentation step, which influences the microbial metabolism and the conversion of hexoses and pentoses obtained in ethanol. In this context, the aim of this work was to evaluate the ability of a yeast strain, from the species Dekkera bruxellensis (CCA155) in producing ethanol from sugarcane bagasse hydrolysates, since it has shown to be a yeast tolerant to adverse conditions found in industrial fermentation processes. The results indicated that D. bruxellensis was capable of producing ethanol in a synthetic medium containing xylose or arabinose, and xylose or glucose as sole carbon sources, and when grown in concentrations of 50 or 100% sugarcane bagasse hydrolyzate, maximum specific speeds growth was similar, about 0.009 h-1, with cell productivity of about 0.035 gL-1h-1. In the bioreactor tests, the yeast displayed low specific growth rate during the first five days, 0.003 h-1, caused by a slow consumption of glucose. But in a second phase of growth, with a lower rate 0.001 h-1, there was an increase in xylose consumption, a period in that an increase in ethanol concentration with maximum yield of approximately 3.25 mg/L.h was observed. Therefore even with lower growth when compared to yeast fermentation conventionally used in ethanol industries, these results suggest the feasibility of the cultivation of D. bruxellensis in hydrolysates of sugarcane bagasse. / O bioetanol é um combustível alternativo, de baixo custo o qual convencionalmente é obtido no Brasil a partir da fermentação de caldo de cana-de-açúcar, melaço ou a mistura destes. Entretanto, alternativamente, o etanol poderia ser obtido a partir de materiais lignocelulósicos, resíduos ou subprodutos agroindustriais compostos por celulose, hemicelulose e lignina, tais como o bagaço de cana-de-açúcar. Para cada tonelada de cana processada, são obtidos, em média, 250 kg de bagaço, o qual é geralmente queimado para geração de energia na indústria. Os cultivos microbianos a partir destas matérias-primas dependem da disponibilização dos substratos, nesse sentido, a hidrólise ácida tem sido referida como um dos processos mais utilizados para a despolimerização da fração hemicelulósica dos materiais lignocelulósicos, devido ao seu baixo custo e alta eficiência. No entanto, em condições de alta pressão e temperatura, glicose e xilose liberadas são degradadas a 5-hidroximetilfurfural (HMF) e furfural, respectivamente. Estes compostos são caracterizados por inibirem as leveduras utilizadas na etapa de fermentação, o que influencia o metabolismo microbiano e a conversão das hexoses e pentoses obtidas em etanol. Neste contexto, o trabalho consistiu em avaliar a capacidade de uma linhagem da levedura Dekkera bruxellensis em produzir etanol a partir de pré-hidrolisados de bagaço de cana-de-açúcar, já que a mesma tem mostrado ser uma levedura tolerante a condições adversas encontradas em processos fermentativos industriais. Os resultados indicaram que D. bruxellensis foi capaz de produzir etanol em meio sintético contendo xilose ou arabinose, ou ainda xilose e glicose como únicas fontes de carbono, e quando cultivadas em concentrações de 50 ou 100% de pré-hidrolisado de bagaço de cana-de-açúcar, as velocidades específicas de crescimento máximas foram semelhantes, em torno de 0,009 h-1, com produtividade celular de aproximadamente 0,035 g.L-1 h-1. Os ensaios em biorreator demonstraram baixas velocidades específicas de crescimento nos primeiros cinco dias, 0,003 h-1, ocasionado por um lento consumo de glicose. Porém em uma segunda fase de crescimento, com velocidade específica menor 0,001 h-1, ocorreu um aumento no consumo de xilose, período que também se observou aumento na concentração de etanol com produtividade máxima de aproximadamente 3,25 mg/L.h. Portanto apesar de crescimento inferior quando comparado às leveduras convencionalmente usadas na fermentação etanólica industrial, estes resultados sugerem a viabilidade de cultivo da D. bruxellensis CCA155 em pré-hidrolisados de bagaço de cana-de-açúcar.

Made available in DSpace on 2014-06-11T19:24:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-23Bitstream added on 2014-06-13T20:32:10Z : No. of bitstreams: 1 zanelato_ai_me_sjrp.pdf: 1477010 bytes, checksum: e2437c2c0ba00be5a6b7df73d8be90f5 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Este trabalho teve como objetivo produzir enzimas celulolíticas por fermentação em estado sólido (FES), em um bioreator de leito fixo, empregando-se o fungo termofílico Myceliophthora sp. e utilizando-se como substratos bagaço de cana-de-açúcar e farelo de trigo. Os testes foram realizados inicialmente em sacos de polipropileno apresentando uma boa produção de CMCase (550U/g) no período de 96 horas de fermentação. As variáveis controladas foram a temperatura de fermentação (40, 45 e 50ºC), umidade inicial (75, 80 e 85% b.u.) e proporção do substrato de bagaço de cana e farelo de trigo (1:1, 3:7 e 1:9 peso). Os estimadores estatísticos não indicaram diferença significativa da produção de celulase com as variações da proporção de bagaço de cana e de farelo de trigo, com a temperatura e com a umidade inicial do substrato, contudo, os melhores resultados foram obtidos para a proporção de 70% de bagaço de cana e 30% de farelo de trigo, temperatura de 50oC e umidade inicial do substrato de 80%. Nos testes em escala de bioreator, foi empregado um leito fixo de 7cm de diâmetro e 50cm de comprimento, encamisado, operado nas condições experimentais de melhor resultados obtidos na escala de sacos, sendo a temperatura da camisa e vazão de ar as variáveis de interesse. O fungo apresentou um bom desenvolvimento no reator, tendo uma produtividade enzimática semelhante à encontrada em escala de saco. Foi observado um gradiente de umidade no interior do leito, sendo este crescente do fundo para o topo do bioreator, comportamento inverso ao da atividade enzimática. As vazões variaram entre 80 e 120 L/h e não afetaram estatisticamente a fermentação, possuindo pouca influência sobre as distribuições longitudinais de umidade e de atividade enzimática. Já os ensaios para as temperaturas de 45 e 50oC mostraram-se distintos, sendo a maior produção de enzimas após 96h... / This work aimed to produce celullolytic enzymes through solid state fermentation in a fixed bed bioreactor, using the fungus Myceliophtora sp. and wheat bran and sugar cane bagasse as substrate. Initial tests were performed in polypropylene plastic bags, and satisfactory production of CMCase (550U/g.d.s.) was observed at 96h of fermentation. The controlled variables were the temperature (40, 45, and 50o C), the initial moisture content (75, 80, and 85%, w.b.), and wheat bran to sugar cane bagasse proportion (1:1, 3:7, and 1:9, weight). The experimental results did not differ statistically for any experimental condition adopted, even though the best results were obtained using the proportion 3:7 wheat bran/sugar cane bagasse 3:7, 50o C temperature, and 80% initial moisture content. A jacketed fixed bed bioreactor 7.62cm ID and 50cm long was used for scaling-up, and temperature, solid initial moisture content, and air flow rate were used as variables. The fungus adapted well to the bioreactor and to the experimental conditions, and the production of enzymes was similar to the tests performed in the plastic bags. It was observed a longitudinal gradient of moisture, increasing from the bottom to the top of the reactor, opposite effect observed for the enzyme production. The air flow rate was varied from 80 to 120L/h and did not affect neither the moisture nor the enzyme distributions. However, the experiments carried out at 45 and 50o C were statistically distinct, and the enzyme production at 50o C at 96h was larger than at 45o C. At 144h of fermentation, the results at 45o C were similar to those observed at 50o C and 96h. It was observed temperature peaks, the higher one at 18h of fermentation, which was 6o C above the temperature fixed through the jacket and the air at the entrance, for 45o C temperature and 80L/h air flow rate

Made available in DSpace on 2016-06-02T19:56:28Z (GMT). No. of bitstreams: 1 DissREM.pdf: 1870524 bytes, checksum: 11256ff68568004b18adba99fa4ba12b (MD5) Previous issue date: 2007-01-29 / Cellulases are enzymes that catalyze the hydrolysis of cellulose producing glucose. Immobilization and stabilization of these enzymes could improve enzymatic process making it more attractive by reducing costs during their use. In this work, Celluclast (Novozymes A/S, Denmark) and CG 200 (Genencor) were immobilized into chitosan and into a chitosan/alginate hybrid support, after activation of the support with glutaraldehyde and glycidol under different conditions of immobilization such as temperature and pH. Immobilization process was evaluated by calculating the immobilization yield, coupling yield and thermal stability at 65°C. Temperature and pH optimum of enzymatic activity were analyzed as well. The best derivatives were obtained with chitosan alginate activated with glycidol, after immobilization at 27°C, pH 7.0, for 16 hours. The biocatalyst presented 56.3% of immobilization yield, 40.6% of coupling yield and it 11.2 fold more stable than free enzyme at 65°C. The temperature and pH for maximum activity of the derivatives and soluble enzyme was 60°C and 50°C, pH 2.5-3.0 and 4.2, respectively. Finally, the performance of the best biocatalyst developed in this work in the hydrolysis of sugarcane bagasse was compared to one observed using soluble enzyme. Although a higher conversion has been obtained with soluble enzyme in the beginning of the reaction, the immobilized enzyme allowed to reach a higher final conversion, 70%, than the one obtained with free Celluclast, 38.9%. / Celulases catalisam hidrólise de celulose à glicose e a imobilização e estabilização da mesma podem viabilizar o processo enzimático, pois reduz custos em sua utilização. Neste trabalho, Celluclast 1.5L (Novozymes A/S, Denmark) e CG 200 (Genencor) foram imobilizadas em géis de quitosana e quitosana-alginato ativados com glutaraldeído e glicidol para posterior hidrólise do bagaço de cana-de-açúcar. O processo de imobilização foi acompanhado medindo-se proteína e atividade hidrolítica da enzima em papel de filtro. Avaliaram-se rendimento de imobilização, atividade recuperada, estabilidade térmica, temperatura e pH ótimo de atividade enzimática. Os melhores derivados foram obtidos com quitosana-alginato, ativados com glicidol com temperatura de imobilização de 27°C e pH 7,0 sendo 11,2 vezes mais estáveis que a enzima livre, com rendimento em torno de 40,6% e atividade recuperada em torno de 56,3%. A temperatura ótima para os derivados foi cerca de 10 graus maior que a da enzima livre (50°C), enquanto que o pH deslocou-se de 4,2 (enzima livre) para a região entre 2,5 e 3, para os derivados. Com o melhor derivado, realizou-se a hidrólise do bagaço de cana-de-açúcar, que apresentou conversão de 1,8 vezes (70,0%) superior à enzima livre (38,9%).

Made available in DSpace on 2016-06-02T19:56:44Z (GMT). No. of bitstreams: 1 3648.pdf: 4034707 bytes, checksum: d61c1d29df91b4f878b199e7de814791 (MD5) Previous issue date: 2011-03-28 / Universidade Federal de Sao Carlos / This work presents a kinetic study of the enzymatic hydrolysis of three cellulosic substrates: filter paper (FP), used as a de-lignified substrate model; sugarcane bagasse (SB) steam exploded; and acid treated SB, the last two treated with 4% NaOH. All SB were chemically characterized. Hydrolysis experiments to study the influence of agitation and substrate concentration were performed in shaker, using Accellerase® 1500, Genencor, at pH 4.8, in 50mM sodium citrate buffer. To verify the substrate concentration effect, cellulose load (weightsubstrate/weighttotal) was 0.5%-13% (for FP) and 0.99%-9.09% (for SB). For FP, the role of the external mass transport resistance was not significant when the agitation speed was in the range 150-300 rpm. It was possible to fit a pseudo-homogeneous Michaelis-Menten model for substrate concentrations up to 13% (w/w). Preliminary tests fitting the model proposed by Chrastil (CHRASTIL J. Enzymic product formation curves with the normal or diffusion limited reaction mechanism and in the presence of substrate receptors, Int. J. Biochem., v. 20, No. 7, p. 683, 1988) indicated that the role of diffusion through the external film was a more relevant feature for steam exploded SB than for FP, at higher concentrations of substrate. It was possible to fit a pseudo-homogeneous model for steam exploded SB, within a range of cellulose concentrations from 0.99% to 3.85% (w/w). Product inhibition had to be considered by the model. For higher loads of steam exploded SB, a modified Michaelis- Menten model, appropriate for heterogeneous systems with high diffusion resistance, was fitted. Finally, for the highly recalcitrant acid treated SB, Chrastil models were fitted. As expected, the complexity of this system regarding to the substrate and to the pool of enzymes acting in synergy, makes difficult the use of one single lumped parameter model for all hydrolysis operation conditions. / Este trabalho apresenta um estudo cinético da hidrólise enzimática de três substratos celulósicos: papel de filtro (PF), usado como um modelo de substrato delignificado; bagaço de cana (BC) explodido a vapor e BC tratado com ácido, os dois últimos tratados com NaOH 4%. Todos os BC foram caracterizados. Experimentos de hidrólise para estudar a influência da agitação e da concentração do substrato foram realizados em shaker, usando Accellerase ® 1500, Genencor, em pH 4,8, em tampão citrato de sódio 50mM. Para verificar o efeito da concentração do substrato, a carga de celulose (m / m) foi de 0,5% -13% (para PF) e 0,99% - 9,09% (para o BC). Para o PF, o papel da resistência externa ao transporte de massa não foi significativo quando a velocidade de agitação foi na faixa de 150-300 rpm, em shaker. Foi possível ajustar um modelo pseudo-homogêneo de Michaelis-Menten para concentrações de substrato até 13% (m / m). Testes preliminares através do ajuste do modelo proposto por Chrastil (CHRASTIL J. Enzymic product formation curves with the normal or diffusion limited reaction mechanism and in the presence of substrate receptors, Int. J. Biochem., v. 20, No. 7, p. 683, 1988) indicaram que o papel da difusão através da película externa era uma característica mais relevante para o BC explodido a vapor que para o PF, em altas concentrações de substrato. Foi possível ajustar um modelo pseudo-homogêneo para o BC explodido a vapor, dentro de uma faixa de concentrações de celulose de 0,99% a 3,85% (m / m). Inibição pelo produto teve de ser considerada pelo modelo. Para cargas maiores de BC explodido a vapor, um modelo modificado de Michaelis-Menten, adequado para sistemas heterogêneos, com alta resistência à difusão, foi ajustado. Finalmente, para BC tratado com ácido (altamente recalcitrante), modelos de Chrastil foram ajustados. Como esperado, a complexidade deste sistema em relação ao substrato para o pool de enzimas agindo em sinergia, torna difícil o uso de um modelo único para todas as condições operacionais de hidrólise.

Submitted by Renata Cristina Julio da Silva (rcj.julio@hotmail.com) on 2018-04-11T17:50:35Z No. of bitstreams: 1 Dissertação.pdf: 2389871 bytes, checksum: 5ee2a84bc455331f9e1b35a12bed3205 (MD5) / Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2018-04-11T18:35:14Z (GMT) No. of bitstreams: 1 silva_rcj_me_sjrp.pdf: 2389871 bytes, checksum: 5ee2a84bc455331f9e1b35a12bed3205 (MD5) / Made available in DSpace on 2018-04-11T18:35:14Z (GMT). No. of bitstreams: 1 silva_rcj_me_sjrp.pdf: 2389871 bytes, checksum: 5ee2a84bc455331f9e1b35a12bed3205 (MD5) Previous issue date: 2018-03-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O processamento da cana-de-açúcar para a produção do etanol e do açúcar vem crescendo a cada ano no Brasil, gerando grandes quantidades de subprodutos de biomassa, a destacar a vinhaça e o bagaço de cana. O principal uso dessas biomassas é na geração de energia (bagaço de cana) e na fertirrigação (vinhaça). Porém a busca por novas alternativas de utilização do bagaço de cana e da vinhaça são necessárias na tentativa de agregar ainda mais valor a estes subprodutos. Sendo assim, estes subprodutos podem ser utilizados no processo de carbonização hidrotérmica (CHT), uma técnica capaz de converter termicamente a biomassa úmida em um material sólido rico em carbono, denominado carvão hidrotérmico. Devido às características físico-químicas do carvão hidrotérmico, diversas aplicações têm sido sugeridas, sendo umas delas o uso como possível fertilizante. No processo de CHT, também é gerada uma fração líquida denominada água de processo. Conhecer a composição orgânica do carvão hidrotérmico e da água de processo é fundamental para entender os efeitos do uso do carvão hidrotérmico como um fertilizante e para propor um destino adequado para a água de processo. Neste contexto, o presente trabalho teve como objetivo fazer um screening inicial dos principais compostos orgânicos presentes no carvão hidrotérmico e na água de processo, utilizando um espectrômetro de massas com ionização por electrospray (ESI-MS), seguido da identificação dos compostos orgânicos não voláteis, utilizando cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). A partir dos resultados obtidos com a análise por ESI-MS pode-se concluir que as condições de preparo do carvão hidrotérmico e da água de processo, como temperatura e acidez, influenciam na composição orgânica destas amostras. As principais classes de compostos observados apresentaram estruturas semelhantes a proteínas, lipídeos, compostos fenólicos e compostos nitrogenados. Na análise dos extratos de carvão hidrotérmico obtidos por GC-MS foram identificados principalmente compostos derivados de benzeno, compostos cíclicos, compostos fenólicos e ácidos graxos. Já nas amostras de água de processo os principais compostos identificados foram compostos fenólicos, ácidos carboxílicos e compostos nitrogenados. / The processing of sugarcane for ethanol and sugar production has been growing every year in Brazil, generating large quantities of biomass by-products, including vinasse and sugarcane bagasse. The main use of these biomasses is for energy generation (sugarcane bagasse) and fertigation (vinasse). However the search for new alternatives of sugarcane bagasse and vinasse uses are necessary in an attempt to add even more value to these byproducts. Thus, these by-products can be used in the hydrothermal carbonization (HTC) process, a technique capable to convert thermally the wet biomass into a solid carbon-rich material called hydrochar. Due to the physical-chemical characteristics of the hydrochar, several applications have been suggested, one of them being the use as a possible fertilizer. In the HTC process, a liquid fraction called process water is also generated. Knowing the organic composition of hydrochar and process water is fundamental to understand the effects of the use of hydrochar as a fertilizer and to propose a suitable destination for process water. In this context, the present work aimed to perform an initial screening of the main organic compounds present in hydrochar and process water using an electrospray ionization mass spectrometer (ESI-MS), followed by the identification of the non-volatile organic compounds, using gas chromatography coupled to mass spectrometry (GC-MS). From the results obtained by ESI-MS analysis, it can be concluded that the conditions of production for hydrochar and process water, such as temperature and acidity, influence the organic composition of these samples. The main classes of compounds observed showed similar structures proteins, lipids, phenolic compounds and nitrogen compounds. In the analysis of hydrochar extracts obtained by GC-MS, benzene derivatives, phenolic compounds and fatty acids were identified. In the process water samples, the main compounds identified were phenolic compounds, carboxylic acids, and nitrogen compounds.

Made available in DSpace on 2014-06-11T19:29:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-21Bitstream added on 2014-06-13T20:38:18Z : No. of bitstreams: 1 magalhaes_ts_me_sjrp.pdf: 702611 bytes, checksum: 13fc3a06f187539258af5a2b0088eb89 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Utilizando o bagaço de cana como matéria prima para a produção do etanol brasileiro pode-se aumentar a produção em 50% deste combustível sem o aumento de área cultivada com a cana de açúcar. A hidrólise química deste material demonstrou ser ineficiente à medida que gera um grande número de substâncias inibidoras do processo fermentativo alcoólico, como aldeídos furânicos e fenóis. A hidrólise enzimática vem se destacando como a forma mais viável de sacarificação do bagaço, em função de não gerar tais inibidores, mas a recalcitrância da fibra vegetal impede tal processo. A ação das enzimas hidrolíticas é muito dificultada pela forma como se encontra naturalmente a fibra vegetal: regiões de alta cristalinidade e a presença de lignina revestindo as fibras. O preparo do bagaço para a ação enzimática, normalmente denominado de pré-tratamento, utiliza condições extremas de temperatura e pressão que podem formar produtos tóxicos para a fermentação alcoólica e apresentam sérios riscos operacionais. O objetivo deste trabalho é avaliar o grau de desestruturação do bagaço após pré-tratamentos em soluções ácidas e alcalinas em glicerol e irradiadas com micro-ondas (MO) em dois modos: estático e rotativo. A melhor condição para a ação hidrolítica do complexo enzimático empregado foi obtida em meio não tamponado (água destilada) durante 24 horas. As maiores liberações de açúcares redutores totais (ART) após a hidrólise enzimática foram alcançadas quando o bagaço foi pré-tratado com irradiação de MO por dois minutos em modo rotativo em meio ácido (0,07 mol/L H2SO4) em 100% glicerol (617 ± 24 mg de ART/g de bagaço) e em meio alcalino (0,05 mol/L NaOH) em 100% glicerol... / The production of the Brazilian bioethanol can be increased in 50% without expanding the area cultivated with sugarcane by using sugarcane bagasse as raw material. Its recalcitrance is an obstacle to enzymatic hydrolysis, which can be overcome by physical-chemical treatments. Common pre-treatments using extreme conditions of temperature and pressure are dangerous, promote the generation of toxic compounds (phenols and aldehydes) besides being potentially inhibitory for yeast. The aim of this work is to assess the degree of disruption of the bagasse after pre-treatment in acidic and alkaline glycerol, irradiated with microwaves both in the static and rotational modes. The enzyme complex showed the best results with the reactions promoted in a non-buffered (distilled water) at 24 hours. The best results for the pre-treatment were found after inserting a rotation system in the microwave oven inasmuch as the samples were subjected to irradiation in a more homogeneous way. The best pre-treatment was obtained after enzymatic hydrolysis in sulfuric acid (0.07 mol/L), in pure glycerol, with the rotation of the balloon inside the microwave oven (617 ± 24 mg of ART/g bagasse). In alkaline medium, the best result was found in the absence of water in the reaction medium (0.05 mol/L NaOH) with the rotation of the balloon inside the microwave oven (601 ± 58 mg of ART/g of bagasse). Differential Scanning Calorimetry (DSC) allows to evaluate the degree of disruption of bagasse comparing the enthalpy change, which can be associated with disruption of bounds in pre-treated bagasse. Among the pre-treated bagasse, it is observed that the bagasse treated with NaOH 0.05 mol/L and 5% of water in glycerol had the lowest ΔH, suggesting greater breakdown of carbohydrates after pre-treatment