Ce travail s’inscrit dans la convention CIFRE n°2012/0379 avec la startup Adionics pour le développement d’un procédé de dessalement par extraction liquide-liquide. La régénération de la phase organique est réalisée par désextraction du sel à chaud. Les isothermes de partage du NaCl, Na2SO4 et MgCl2 à 20 40 60 et 80°C ont été obtenues. Une stœchiométrie expliquant leur forme a été proposée et les constantes d’équilibre associées ont été déterminées. Les enthalpies d’extraction ont été déduites des variations des constantes d’équilibre avec la température.La prise en compte de l’effet de milieu en phase aqueuse a été réalisée à l’aide du modèle de Pitzer avec l’hypothèse des interactions spécifiques. Une correction pour les effets de milieu en phase organique a également été ajustée sur NaCl -Na2SO4 à 20 40 et 60°C.Les propriétés physico-chimiques du solvant (viscosité, masse volumique, chaleur spécifique, conductivité thermique, ...) ont été déterminées sur une plage de 20°C à 80°C. La tension interfaciale du solvant a également été obtenue à 20°C, avec de l'eau eau pure et salée. Une étude approchée de la solubilité du diluant en phase aqueuse a également été entreprise.Un prototype avec deux colonnes à garnissage de 4 mètres de haut a été élaboré pour tester la faisabilité du procédé en continu. L’extracteur a été modélisé par une cascade d’étages parfaitement agités avec prise en compte du mélange en retour. Les HUT (Hauteur d'une Unité de Transfert) expérimentales ont été comparées à celles calculées à l'aide de différentes corrélations des coefficients de transfert disponibles dans la littérature. Une adéquation satisfaisante a été obtenue entre modèle et expérience. Une réflexion sur le type d'extracteur doit être entreprise pour une amélioration de la performance de dessalement. / This work is a contribution to the development of an innovative desalination process by solvent extraction. The regeneration of the organic phase is achieved by desextraction of the salt at high temperature. Isotherms of distribution of NaCl, Na2SO4 and MgCl2 at 20 40 60 and 80°C were obtained. A stoichiometry able to account for their shapes was suggested and the associated equilibrium constants were determined. The extraction enthalpies were deduced from the variation of the constants with temperature.Non ideality in the aqueous phase was modeled with the Pitzer equations, simplified with the specific interaction principle. A thermodynamic correction to account for mixed salts effect in the organic phase was adjusted on binary data (NaCl, Na2SO4) at 20 40 and 60°C.Mechanical (viscosity, density) and thermal (specific heat, thermal conductivity) properties of the organic phase were characterised from 20 to 80°C. The interfacial tension was obtained at 20°C with pure water and filtrated natural seawater. The overall solubility of the solvant in water was measured. A prototype of the process with two extraction columns with a packed bed of 4 meters was tested. The HTU obtained were compared with a numerical model based on the classical stagewise backflow model expanded by a thermal balance.

Esse estudo teve por objetivo avaliar a viabilidade e qualidade in vitro do concentrado de hemácias felino (CHF), colhido e armazenado em bolsas e produtos nacionais, remanufaturadas, pelo Laboratório de Hemoterapia do Serviço de Anestesia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, de acordo com normas definidas por órgãos regulamentadores como a ANVISA. Foram analisados parâmetros bioquímicos e hematológicos de 24 unidades de CHF nos dias 0, 14 e 21 de armazenamento. Utilizou-se sistema fechado para colheita e armazenamento do sangue com solução de anticoagulante-preservativa CPDA-1. A avaliação consistiu na mensuração de porcentagem de hematócrito, hemoglobina total, hemoglobina extracelular, porcentagem de hemólise, concentrações de potássio, lactato, glicose, ATP, pH, bicarbonato, pressão de CO2 e O2, inspeção visual da bolsa e cultura microbiológica aeróbia e anaeróbia. Os resultados obtidos foram avaliados estatisticamente por meio de testes paramétricos, sendo que as determinações de hematócrito, hemoglobina total não apresentaram variação significante nos 21 dias de preservação, enquanto os de potássio, lactato, pO2 aumentaram gradativamente de forma significante. Os níveis de ATP, glicose, pH, bicarbonato e pCO2 reduziram de forma significante com o decorrer do tempo. Não houve alteração à inspeção visual das bolsas de sangue, nem crescimento de microorganismos nas culturas realizadas. Por meio destas avaliações constatouse que o sistema remanufaturado com produtos nacionais pode ser empregado com segurança para obtenção de sangue felino tendo-se em vista que se manteve estéril, com eficiente conservação do concentrado de hemácias felino em CPDA-1 até o 21º dia de armazenamento. / This study aimed to evaluate the feasibility and quality of in vitro feline packed red blood cells (CHF), harvested and stored in bags and domestic products, manufactured, Hematology Laboratory at the Department of Anesthesia, Faculty of Veterinary Medicine, University of São Paulo, according to standards set by regulatory bodies such as ANVISA. Were analyzed biochemical and hematological parameters 24 units of CHF on days 0, 14 and 21 of storage. We used a closed system for collection and storage of blood with preservative-anticoagulant solution CPDA-1. The evaluation consisted in measuring percentage of hematocrit, total hemoglobin, extracellular hemoglobin, percentage of hemolysis, potassium concentrations, lactate, glucose, ATP, pH, bicarbonate, CO2 and O2 pressure, visual inspection of the bag and aerobic and anaerobic microbiological culture . The results were statistically analyzed using parametric tests, and determinations of hematocrit, total hemoglobin showed no significant variation within 21 days of preservation, while potassium lactate, pO2 gradually increased significantly. ATP levels, glucose, pH, pCO2 and bicarbonate decreased significantly with time. There was no change to the visual inspection of blood bags, or growth of microorganisms in the cultures performed. Through these evaluations it was found that the system refilled with domestic products can be used safely for obtaining blood feline bearing in mind that remained sterile, efficient storage of red blood cells feline in CPDA-1 until the 21 th days of storage.

Visando ampliar as rotas de produção de biodiesel, o objetivo deste trabalho foi utilizar uma tecnologia alternativa para a produção de biodiesel por via enzimática. Neste processo, células do fungo Mucor circinelloides URM 4182 foram cultivadas em meio apropriado, imobilizadas in situ em espuma de poliuretano e utilizadas diretamente como biocatalisador na síntese de ésteres de ácidos graxos em reações de transesterificação do óleo de coco usando etanol como agente acilante. O trabalho experimental foi desenvolvido em 4 etapas. Inicialmente, as propriedades bioquímicas do biocatalisador foram determinadas, incluindo valores ótimos de pH (7,3) e temperatura (40° C) e constantes cinéticas (Vmax= 610 U/g e Km= 263 mM). Na segunda etapa foram realizados testes utilizando reatores de tanque agitados operados em regime descontínuo confirmando a eficiência do M. circinelloides UMR 4182 para converter os ácidos graxos presentes no óleo de coco em ésteres correspondentes (rendimento de transesterificação da ordem de 96% em 144 horas). Com objetivo de aumentar a produtividade do sistema, na terceira etapa, estudos foram conduzidos em reator de leito fixo operando em fluxo contínuo empregando dois reatores (A e B) com diferentes relações geométricas altura/diâmetro (l/d=4,2 e l/d=14). Avaliou-se inicialmente no reator A o comportamento do sistema para substratos preparados em diferentes razões molares (1:6, 1:8 e 1:10 óleo/álcool). Comparativamente os rendimentos mais elevados (da ordem de 62%) foram obtidos para substratos preparados na razão molar de 1:8 (óleo/álcool), indicando a necessidade de investigar outros parâmetros do processo. A presença de caminhos preferenciais detectada visualmente no reator A pela injeção do corante lipossolúvel indicou limitações de transferência de massa, justificando a baixa conversão em ésteres de etila. Em seguida, mediante a avaliação da geometria dos reatores, foi selecionado o reator B (volume útil - 286 mL) para determinar a influencia do tempo espacial (60, 70 e 80 horas) no desempenho do processo utilizando a condição pré-estabelecida, etanol em excesso na razão molar 1:8 (óleo/etanol). O melhor desempenho do reator foi constatado para tempo espacial de 80h (vazão volumétrica = 0,06 mL.min-1). Nessa condição, 92,7 ??1,5% dos ácidos graxos presentes no óleo de coco foram convertidos nos ésteres de etila correspondentes, alcançando uma produtividade média de 51,0 ??1,4 mgéster.g-1meio.h-1 e fornecendo uma operação estável do sistema por 25 dias. As amostras purificadas apresentaram valores de viscosidade 5,14 mm2/s e teores residuais de monoglicerídeos (3,2 m/m%) e diglicerídeos (3,4 m/m%). Na quarta etapa foram realizados testes em reatores agitados operados em regime descontínuo na presença de solvente (terc-butanol e/ou líquido iônico), em diferentes proporções (10 - 50%). Os resultados foram comparados com a reação conduzida na ausência de solvente que apresentou baixa velocidade de reação e rendimento reduzido (44,5% em 6 dias de reação). Ambos os solventes aumentaram as conversões e velocidades de reação; no entanto, terc-butanol apresentou melhor desempenho em menor proporção (15%). Nessa condição, rendimentos da ordem de 96% foram alcançados em 96 horas, provavelmente devido à polaridade moderada e baixa viscosidade deste solvente que dilui o glicerol acumulado no suporte e melhora a transferência do substrato para o interior das células imobilizadas, impedindo a inibição da enzima por limitação da transferência de massa. De forma geral, os resultados foram promissores e demonstraram a potencialidade das células íntegras de M. circinelloides URM 4182 imobilizada em poliuretano para mediar a síntese de biodiesel em um reator de leito fixo em fluxo contínuo. / Aiming at expanding routes for biodiesel production, the goal of this work was to use an alternative technology for the production of biodiesel by enzymatic route. In this case, whole-cells from filamentous fungus Mucor circinelloides URM 4182 were growth in appropriate medium, simultaneously immobilized on low-cost support and used directly as a biocatalyst in the transesterification reactions of coconut oil using ethanol as the acylating agent. The experimental work was carried out in four steps. Initially, the biochemical properties of the immobilized biocatalyst were determined, including values for optimum pH (7.3) and temperature (40 °C) and kinetics parameters (Vmax 610 U/g and Km= 263 mM). Following this, tests were carried out in stirred batch reactors confirming the efficiency of M. circinelloides URM 4182 to produce biodiesel from coconut oil (96% in 144 hours). However, the low productivity attained motivated the replacement of the batch system for packed bed reactor operating under continuous flow using two reactor (A and B) having different geometric relations height/diameter (l/d = 4.2 and l/d = 14). Initially, the performance of the reactor A was assessed for substrates at different molar ratios (1:6, 1:8 and 1:10 oil/alcohol). Comparatively the best conversion (62%) was achieved for substrate at molar ratio of 1:8 (oil/alcohol); suggesting the need to investigate other process parameters to overcome such limitations. The presence of preferential paths visual detected for Reactor A by tracer assay indicated severe mass transfer limitations, justifying the low conversion attained in ethyl esters. Following this, Reactor B was selected to run the continuous experiments using substrate at pre-established condition (molar ratio oil/ethanol 1: 8) under different space times (60, 70, and 80 hours). Better reactor performance was found for space time of 80 h (volumetric flow = 0.06 mL min-1). In this condition, 92.7±1.5% of the fatty acids present in the coconut oil have been converted into the corresponding ethyl esters, reaching an average volumetric productivity of 51.0 ??1.4 mgester.g-1medium.h-1 with no significant reduction in the efficiency during 25 days. The purified samples presented values of 5.14 viscosity mm2/s and residual levels of monoglycerides (3.2 wt %) and diglycerides (3.4 wt%). In the fourth step, transesterification reactions were performed in stirred tank reactors in the presence of solvent (tert-butanol and/or Ionic Liquid) at different proportions (10-50%) in relation to the total weight oil and ethanol. The results were compared with the reaction carried out in the absence of solvent which has low reaction rate and reduced yield (44.5% in 6 days of reaction). Both solvents enhanced the transesterification yields and reaction rates; however tert-butanol showed better performance at lower proportion (15%). Under this condition, yields in the order of 96% were attained in 96 hours, probably due to the moderate polarity and lower viscosity of this solvent which dilutes the glycerol accumulated in support and improved the substrate transfer into the interior of the immobilized cells by preventing the enzyme inhibition by limitation of the mass transfer. Overall, the results were promising and showed the potential of whole-cells from M. circinelloides URM 4182 immobilized in polyurethane to mediate the synthesis of biodiesel in a continuous packed bed reactor.

O presente trabalho teve como objetivo aprimorar a síntese de biodiesel por via enzimática empregando óleo de babaçu pela rota etílica. A motivação do uso do óleo de babaçu é referente ao seu forte potencial para a produção de combustível, tendo em vista que é de baixo custo e abundante em localidades carentes de fontes de energia, o que estimula a realização de pesquisas nesta área, procurando benefícios ambientais, energéticos e econômicos. Para atingir o objetivo proposto, teve-se como proposta a transesterificação do óleo de babaçu com etanol mediada pela enzima lipase em fluxo contínuo empregando reator de leito fixo. A enzima selecionada para condução dos experimentos foi a lipase microbiana de Burkholderia cepacia (lipase PS) imobilizada covalentemente em matriz híbrida sílica-PVA com comprovada eficiência na síntese de biodiesel a partir de diferentes matérias-primas lipídicas. Todos os experimentos foram efetuados na ausência de solventes a 50?C. A parte experimental foi iniciada pelos testes comparativos de desempenho da montagem experimental proposta utilizando a preparação de lipase previamente selecionada e a lipase Novozym® 435 disponível comercialmente na forma imobilizada, de comprovada eficiência para utilização em reatores de leito fixo. Os resultados indicaram que ambas as preparações de lipase são adequadas para catalisar a síntese de biodiesel em regime contínuo, entretanto a grande limitação de operação do reator está relacionada com a elevada composição em ácidos graxos saturados (? 83%) na matéria-prima lipídica, que acarreta entupimentos nas conexões e obstrução do leito. Para contornar as limitações identificadas foi proposto um sistema de aquecimento para manter constante a temperatura na faixa requerida (45-50?C) em todo o aparato experimental. O desempenho do reator (diâmetro interno X altura: 15 mm X 210 mm) foi avaliado para substratos constituídos de óleo de babaçu e etanol em duas razões molares (1:7 e 1:12), definindo os limites de operação em termos de vazão do substrato. Foi quantificado o desempenho do sistema para 4 diferentes vazões volumétricas correspondendo a tempos espaciais entre 7 a 13h e determinadas para cada condição a influência do tempo espacial na concentração de ésteres de etila formados, rendimentos de transesterificação e produtividade. A razão molar entre os materiais de partida interferiu de maneira marcante no alcance de elevadas conversões dos ácidos graxos presentes no óleo de babaçu em seus ésteres de etila correspondentes, sendo o sistema maximizado para substratos contendo maior excesso de etanol (razão molar óleo: etanol de 1:12). Nessas condições, o funcionamento do sistema foi comprovado quantitativamente para tempos espaciais no reator iguais ou superiores a 11h, fornecendo rendimentos de transesterificação médios de 95,00 ? 4,68 % e produtividade de 54,80 ? 2,70 mgéster.gmeio-1.h-1. A lipase PS imobilizada em SiO2-PVA foi estável quanto a suas características morfológicas e catalíticas revelando tempo médio de meia-vida (t1/2) de 40 dias, sendo ainda possível sua reutilização em um novo ensaio. As características globais do desempenho do sistema contínuo proposto tornam atrativo a continuidade dos estudos em escala ampliada de processo, entretanto recomenda-se a utilização de óleos vegetais com menor teor de ácidos graxos saturados. / The purpose of this work was to enhance the enzymatic synthesis of biodiesel from babassu oil by ethanolic route. Babassu oil was chosen as a feedstock due to its potential for fuel production, including low cost and availability in regions where energy sources are needed; stimulating the developing of research projects looking for environmental, energetic and economical benefits. To attain this purpose, the transesterification reaction of babassu oil with ethanol was mediated by the enzyme lipase on packed bed reactor running on continuous mode. The selected enzyme was the microbial lipase from Burkholderia cepacia (lipase PS) covalent immobilized on hybrid matrix silica-PVA (SiO2-PVA) which showed already satisfactory performance in the biodiesel synthesis from different lipidic feedstocks. The experiments were carried out in solvent free system at 50°C. The experimental work was started by performing comparative tests of the proposed experimental assembly, using the previously selected lipase preparation and a commercial available immobilized lipase Novozym® 435, which has consistent high performance to be used on fixed packed reactors. The results indicated that both lipase preparations are able to catalyze the biodiesel synthesis under continuous operation; however limitations were found to maintain stable operation. This was related to the high concentration of saturated fatty acids (83%) present in the feedstock, which blocked the tube connections and led to bed obstruction. To overcome these limitations, a warming system was proposed to ensure constant the temperature at the required range (45-50°C) in the entirely experimental apparatus. The performance of the reactor (inner diameter X height: 15 mm X 210 mm) was evaluated for substrates composed by babassu oil and ethanol in two molar ratios (1:7 e 1:12), determining its operation limits in terms of substrate flow rate. The system performance was quantified for four different flow rates corresponded to spatial times between 7 and 13 h. For each condition, the influence of spatial times in the ethyl esters formation, transesterification yields and productivities were determined. The raw materials molar ratio interfered, in a remarkable way, the conversion of babassu oil into the correspondent ethyl esters and the system was optimized for the substrates containing high excess of ethanol (oil-to-ethanol molar ratio of 1:12). In these conditions, the reactor operation was demonstrated for spatial times higher than 11h, attaining transesterification yields of 95,00 ? 4,60% and productivities of 54,80 ? 2,70 mgesters.gmedium-1.h-1The lipase PS immobilized on SiO2-PVA was found to be stable regarding its morphological and catalytic characteristics, showing half-life time (t1/2) of 40 days. The immobilized enzyme can be also reused in a new run and the presence of substrate containing high ethanol levels did not affected the lipase stability. The performance characteristics of the proposed continuous system made attractive to develop further studies aiming at scaling up the process, however it is recommended to use vegetable oils having lower saturated fatty acids levels.

A ciclodextrina glicosiltransferase (CGTase) é a única enzima capaz de catalisar a reação de ciclização a partir do amido e, assim, formar oligossacarídeos cíclicos conhecidos como ciclodextrinas (CDs). Através desta reação é produzida uma mistura de α-, β- e γ-CD que, respectivamente, contém 6, 7 e 8 resíduos de glicose. As CDs têm atraído enorme atenção devido ao seu grande potencial de aplicação em diversas áreas da indústria. Potencial este proporcionado por sua estrutura cônica, com interior hidrofóbico, capaz de encapsular sólidos, líquidos e gases, conferindo propriedades importantes e protegendo-os. Neste trabalho foi estudada a imobilização de uma CGTase em sílica mesoporosa de forma direcionada às cisteínas presentes em sua superfície, alterando a exposição do sítio ativo. A ligação via cisteínas nativas da proteína aumentou em quatro vezes a eficiência da imobilização, quando comparada a ligação via grupamento amino. Esta, no entanto, apresentou maior atividade enzimática em faixas mais amplas de temperatura e pH, além de maior estabilidade operacional, mantendo 100 % de sua atividade após 200 h de reação contínua a 60 °C e pH 4. Ainda que apresentando menor estabilidade da ligação, o derivado obtido por ligação dissulfeto manteve 40 % da atividade inicial durante 200 h e então, o suporte pôde ser recarregado e reutilizado por igual período. Os suportes desenvolvidos apresentaram estabilidade satisfatória, possibilitando o uso do derivado imobilizado em reator de leito fixo operado de forma contínua. Quando avaliado em relação a produção das três ciclodextrinas principais, o derivado cuja imobilização da enzima ocorreu via grupamento amino, evidenciou a possibilidade de modulação da produção apenas variando as condições de reação. α- e β-CD foram produzidas preferencialmente em pH 8,0 e 2 min (3,44 mg mL-1 e 3,51 mg mL-1, respectivamente), enquanto que pH mais ácido (4,0) e maior tempo de reação (141 min) favoreceram a formação de γ-CD (3,35 mg mL-1), com baixa formação α-CD (0,75 mg mL-1). Por fim, os resultados deste estudo evidenciam a importância da imobilização da CGTase para a estabilização de sua estrutura a fim de aplicá-la em sistemas contínuos de produção de CDs onde é possível modular o perfil dos produtos gerados em função das condições de reação, aumentando assim a produtividade do biocatalisador. / Cyclodextrin glycosyltransferase (CGTase) is the only enzyme capable of catalyzing the cyclization reaction from the starch and thus forming cyclic oligosaccharides known as cyclodextrins (CDs). Through this reaction, is produced a mixture of α-, β- and γ-CD containing, 6, 7 and 8 glucose residues respectively. Cyclodextrins (CD) have been attracting considerable attention because of its great potential for application in various areas of industry. This potential is provided by its conical structure with hydrophobic interior, capable of encapsulating solids, liquids and gases, changing important features and protecting them. In this work, the immobilization of CGTase in mesoporous silica was studied in a way directed to cysteines present on its surface, altering the exposure of the active site. The connection via native cysteine of the protein increased by four times the efficiency of immobilization compared to amino groups connection. The binding of amino groups, however, showed greater enzymatic activity in wider ranges of temperature and pH, and higher operational stability, while maintaining 100 % of its activity after 200 h of continuous reaction at 60 °C and pH 4. Although showing less stable connection, the derivative obtained by disulfide bond retained 40 % of the initial activity for 200 h and then, the support could be reloaded and reused for the same period. Developed supports showed satisfactory stability, enabling the use of the derivative assets in a packed bed reactor and operated continuously. It was demonstrated the possibility of modulating the CDs production just varying the reaction conditions, using the derivative of which the enzyme immobilization occurred via amino group, to evaluate the production of three main cyclodextrins. α- and β-CD were produced preferentially at pH 8.0 and 2 min (3.44 mg mL-1 and 3.51 mg mL-1, respectively), whereas the more acid pH (4.0) and longer reaction (141 min) favored the formation of γ-CD (3.35 mg mL-1 and 0.75 mg mL-1 of α-CD). Finally, the results of this study show the importance of the immobilization of CGTase to the stabilization of its structure in order to apply it in continuous CD production systems, where it is possible to modulate the profile of the products generated as a function of the reaction conditions, thus increasing the productivity of the biocatalyst.

Visando ampliar as rotas de produção de biodiesel, o objetivo deste trabalho foi utilizar uma tecnologia alternativa para a produção de biodiesel por via enzimática. Neste processo, células do fungo Mucor circinelloides URM 4182 foram cultivadas em meio apropriado, imobilizadas in situ em espuma de poliuretano e utilizadas diretamente como biocatalisador na síntese de ésteres de ácidos graxos em reações de transesterificação do óleo de coco usando etanol como agente acilante. O trabalho experimental foi desenvolvido em 4 etapas. Inicialmente, as propriedades bioquímicas do biocatalisador foram determinadas, incluindo valores ótimos de pH (7,3) e temperatura (40° C) e constantes cinéticas (Vmax= 610 U/g e Km= 263 mM). Na segunda etapa foram realizados testes utilizando reatores de tanque agitados operados em regime descontínuo confirmando a eficiência do M. circinelloides UMR 4182 para converter os ácidos graxos presentes no óleo de coco em ésteres correspondentes (rendimento de transesterificação da ordem de 96% em 144 horas). Com objetivo de aumentar a produtividade do sistema, na terceira etapa, estudos foram conduzidos em reator de leito fixo operando em fluxo contínuo empregando dois reatores (A e B) com diferentes relações geométricas altura/diâmetro (l/d=4,2 e l/d=14). Avaliou-se inicialmente no reator A o comportamento do sistema para substratos preparados em diferentes razões molares (1:6, 1:8 e 1:10 óleo/álcool). Comparativamente os rendimentos mais elevados (da ordem de 62%) foram obtidos para substratos preparados na razão molar de 1:8 (óleo/álcool), indicando a necessidade de investigar outros parâmetros do processo. A presença de caminhos preferenciais detectada visualmente no reator A pela injeção do corante lipossolúvel indicou limitações de transferência de massa, justificando a baixa conversão em ésteres de etila. Em seguida, mediante a avaliação da geometria dos reatores, foi selecionado o reator B (volume útil - 286 mL) para determinar a influencia do tempo espacial (60, 70 e 80 horas) no desempenho do processo utilizando a condição pré-estabelecida, etanol em excesso na razão molar 1:8 (óleo/etanol). O melhor desempenho do reator foi constatado para tempo espacial de 80h (vazão volumétrica = 0,06 mL.min-1). Nessa condição, 92,7 ??1,5% dos ácidos graxos presentes no óleo de coco foram convertidos nos ésteres de etila correspondentes, alcançando uma produtividade média de 51,0 ??1,4 mgéster.g-1meio.h-1 e fornecendo uma operação estável do sistema por 25 dias. As amostras purificadas apresentaram valores de viscosidade 5,14 mm2/s e teores residuais de monoglicerídeos (3,2 m/m%) e diglicerídeos (3,4 m/m%). Na quarta etapa foram realizados testes em reatores agitados operados em regime descontínuo na presença de solvente (terc-butanol e/ou líquido iônico), em diferentes proporções (10 - 50%). Os resultados foram comparados com a reação conduzida na ausência de solvente que apresentou baixa velocidade de reação e rendimento reduzido (44,5% em 6 dias de reação). Ambos os solventes aumentaram as conversões e velocidades de reação; no entanto, terc-butanol apresentou melhor desempenho em menor proporção (15%). Nessa condição, rendimentos da ordem de 96% foram alcançados em 96 horas, provavelmente devido à polaridade moderada e baixa viscosidade deste solvente que dilui o glicerol acumulado no suporte e melhora a transferência do substrato para o interior das células imobilizadas, impedindo a inibição da enzima por limitação da transferência de massa. De forma geral, os resultados foram promissores e demonstraram a potencialidade das células íntegras de M. circinelloides URM 4182 imobilizada em poliuretano para mediar a síntese de biodiesel em um reator de leito fixo em fluxo contínuo. / Aiming at expanding routes for biodiesel production, the goal of this work was to use an alternative technology for the production of biodiesel by enzymatic route. In this case, whole-cells from filamentous fungus Mucor circinelloides URM 4182 were growth in appropriate medium, simultaneously immobilized on low-cost support and used directly as a biocatalyst in the transesterification reactions of coconut oil using ethanol as the acylating agent. The experimental work was carried out in four steps. Initially, the biochemical properties of the immobilized biocatalyst were determined, including values for optimum pH (7.3) and temperature (40 °C) and kinetics parameters (Vmax 610 U/g and Km= 263 mM). Following this, tests were carried out in stirred batch reactors confirming the efficiency of M. circinelloides URM 4182 to produce biodiesel from coconut oil (96% in 144 hours). However, the low productivity attained motivated the replacement of the batch system for packed bed reactor operating under continuous flow using two reactor (A and B) having different geometric relations height/diameter (l/d = 4.2 and l/d = 14). Initially, the performance of the reactor A was assessed for substrates at different molar ratios (1:6, 1:8 and 1:10 oil/alcohol). Comparatively the best conversion (62%) was achieved for substrate at molar ratio of 1:8 (oil/alcohol); suggesting the need to investigate other process parameters to overcome such limitations. The presence of preferential paths visual detected for Reactor A by tracer assay indicated severe mass transfer limitations, justifying the low conversion attained in ethyl esters. Following this, Reactor B was selected to run the continuous experiments using substrate at pre-established condition (molar ratio oil/ethanol 1: 8) under different space times (60, 70, and 80 hours). Better reactor performance was found for space time of 80 h (volumetric flow = 0.06 mL min-1). In this condition, 92.7±1.5% of the fatty acids present in the coconut oil have been converted into the corresponding ethyl esters, reaching an average volumetric productivity of 51.0 ??1.4 mgester.g-1medium.h-1 with no significant reduction in the efficiency during 25 days. The purified samples presented values of 5.14 viscosity mm2/s and residual levels of monoglycerides (3.2 wt %) and diglycerides (3.4 wt%). In the fourth step, transesterification reactions were performed in stirred tank reactors in the presence of solvent (tert-butanol and/or Ionic Liquid) at different proportions (10-50%) in relation to the total weight oil and ethanol. The results were compared with the reaction carried out in the absence of solvent which has low reaction rate and reduced yield (44.5% in 6 days of reaction). Both solvents enhanced the transesterification yields and reaction rates; however tert-butanol showed better performance at lower proportion (15%). Under this condition, yields in the order of 96% were attained in 96 hours, probably due to the moderate polarity and lower viscosity of this solvent which dilutes the glycerol accumulated in support and improved the substrate transfer into the interior of the immobilized cells by preventing the enzyme inhibition by limitation of the mass transfer. Overall, the results were promising and showed the potential of whole-cells from M. circinelloides URM 4182 immobilized in polyurethane to mediate the synthesis of biodiesel in a continuous packed bed reactor.

An External Loop Airlift Bioreactor with a small amount (99% porosity) of stainless steel mesh packing inserted in the riser section was used for bioremediation of a phenol polluted air stream. The packing enhanced VOC and oxygen mass transfer rates and provided a large surface area for cell immobilization. Using a pure strain of Pseudomonas putida, fed-batch and continuous runs at three different dilution rates were completed with phenol in the polluted air as the only source of growth substrate. 100% phenol removal was achieved at phenol loading rates up to 33120 mg/h.m3 using only one third of the column, superior to any previously reported biodegradation rates of phenol polluted air with 100% efficiency. A mathematical model has been developed and is shown to accurately predict the transient and steady state data.

Many industrial activities discharge sulphate- and metal-containing wastewaters, including the manufacture of pulp and paper, mining and mineral processing, and petrochemical industries. Acid mine drainage (AMD) is an example of such sulphate- and metal-containing waste streams. Formation of AMD is generally the result of uncontrolled oxidation of the sulphide minerals present in the terrain in which the drainage flows with concomitant leaching of the metals. Acid mine drainage (AMD) and other sulphate- and metal-containing waste streams are amenable to active biological treatment. Anaerobic reduction of sulphate, reaction of produced sulphide with metal ions present in the waste stream, and biooxidation of excess sulphide are three main sub-processes involved in the active biotreatment of AMD. Anaerobic reduction of sulphate can be achieved in continuous stirred tank bioreactors with freely suspended cells or in immobilized cell bioreactors. The application of freely suspended cells in a continuous system dictates a high residence time to prevent cell wash-out, unless a biomass recycle stream is used. In an immobilized cell system biomass residence time becomes uncoupled from the hydraulic residence time, thus operation of bioreactor at shorter residence times becomes possible. In the present work, kinetics of anaerobic sulphate reduction was studied in continuous immobilized cell packed-bed bioreactors. Effects of carrier matrix, concentration of sulphate in the feed and sulphate volumetric loading rate on the performance of the bioreactor were investigated. The bioreactor performance, in terms of sulphate reduction rate, was dependent on the nature of the carrier matrix, specifically the total surface area which was provided by the matrix for the establishment of biofilm. Among the three tested carrier matrices, sand displayed the superior performance and the maximum volumetric reduction rate of 1.7 g/L-h was achieved at the shortest residence time of 0.5 h. This volumetric reduction rate was 40 and 8 folds faster than the volumetric reduction rates obtained with glass beads (0.04 g/L-h; residence time: 28.6 h) and foam BSP (0.2 g/L-h; residence time: 5.3 h), respectively. Further kinetic studies with sand as a carrier matrix indicated that the extent of volumetric reduction rate was dependent on the feed sulphate concentration and volumetric loading rate. At a constant feed sulphate concentration, increases in volumetric loading rate caused the volumetric reduction rate to pass through a maximum, while increases in feed sulphate concentrations from 1.0 g/L to 5.0 g/L led to lower volumetric reduction rates. The maximum volumetric reduction rates achieved in the bioreactors fed with initial sulphate concentration of 1.0, 2.5 and 5.0 g/L were 1.71, 0.82 and 0.68 g/L-h, respectively. The coupling of lactate utilization to sulphate reduction was observed in all experimental runs and the rates calculated based on the experimental data were in close agreement with calculated theoretical rates, using the stoichiometry of the reactions involved. The maximum volumetric reduction rates achieved in the immobilized cell bioreactors were significantly faster than those reported for freely suspended cells employed in the stirred tank bioreactors.

Podophyllotoxin is a natural medicine possessing an outstanding anti-tumour activity. It can be extracted from the rhizome of Podophillum peltatum (American Podophyllum). Volumetric heating of a packed bed of particles including solvent during the extraction can eliminate the solvent pre-heating time and provide uniform and quick heating of the bed. RF-assisted extraction has a potential to be a promising extraction alternative over conventional methods. The characterization and assessment of RF-assisted extraction of podophyllotoxin is crucial. Thermal properties including specific heat capacity, thermal conductivity, and thermal diffusivity of a packed bed of P. peltatum with and without ethanol solutions were determined and the associated multiples regression equations were obtained for the purpose of thermal analysis of RF-assisted packed bed extraction process and related modeling investigations.<p> The dielectric properties of the packed bed of rhizome particles were measured from 10 to 30 MHz using a precision LCR meter and a liquid test fixture. The effects of temperature, particle moisture content, volumetric concentration of ethanol and bed porosity on the dielectric constant, dielectric loss factor and power penetration depth were investigated. The dielectric loss factor significantly increased with the particle moisture content for the beds with 100% and 70% ethanol but not with 30% ethanol. The dielectric loss factor was proportional to temperature directly and to frequency inversely. With 30% ethanol (and therefore 70% water), the dielectric loss factor of the bed dramatically increased compared to 70% and 100% ethanol. Porosity had a significant effect on the dielectric constant but not on the dielectric loss factor. The power penetration depth of a packed bed with 100% ethanol was significantly larger than those of the packed bed with 30% and 70% ethanol. Empirical regression equations were developed for simulation and design of an RF-assisted packed bed extraction of podophyllotoxin.<p> A RF-transparent batch reactor was made of glass filled Teflon and the extraction kinetics of podophyllotoxin was characterized. The effects of temperature, ethanol volumetric concentration, solid/liquid ratio, RF heating and particle moisture content on the extraction rate and yield of podophyllotoxin were investigated at different extraction conditions. A generalized diffusion mathematical model taking into account three major particle geometries was developed and coupled with genetic algorithm for determination of effective diffusivity and partition coefficient through an inverse simulation approach. The approach was first verified by reported experimental data of andrographolide extraction followed by determining the effective diffusivity and partition coefficient of podophyllotoxin for different conditions. The optimum batch extraction condition was achieved with 30% ethanol-water solution at 53¢XC. A prototype was developed for RF-assisted extraction of podophyllotoxin using two optical and RF-transparent reactors with horizontal and vertical orientations. Applying the optimum conditions obtained from batch experiments, the potential of RF heating for providing a uniform temperature in the packed bed was evaluated. The effect of solvent dielectric loss factor on uniform RF heating was investigated and the chemical effect of NaCl used for increasing dielectric loss factor of the solvent on podophyllotoxin was assessed. The horizontal packed bed demonstrated a large temperature gradient across the thickness of the bed during RF heating; however, a uniform RF heating was achieved when the vertical packed bed reactor was used for RF-assisted extraction of podophyllotoxin. The concentration of 2.5 g NaCl/L of the solvent at the temperature controller set point of 40aC provided a relatively good uniform temperature of 50aC within the bed. Evaluating three flow rates of 130, 160 and 200 ml/min for the solvent of 30% ethanol with 2.5 g NaCl/L indicated that the flow rate of 160 ml/min could provide better temperature overlap of four positions of the bed height.

El entendimiento de los fenómenos de transferencia de calor y de masa en medios porosos implica el estudio de modelos de transporte de fluidos en la fracción vacía del medio; este hecho es de fundamental importancia en muchos sistemas de Ingeniería Química, tal como en procesos de extracción o en reactores catalíticos. Los estudios de flujo realizados hasta ahora (teóricos y experimentales) usualmente tratan al medio poroso como un medio efectivo y homogéneo, y toman como válidas las propiedades medias del fluido. Este tipo de aproximación no tiene en cuenta la complejidad del flujo a través del espacio vacío del medio poroso, reduciendo la descripción del problema a promedios macroscópicos y propiedades efectivas. Sin embargo, estos detalles de los procesos locales de flujo pueden llegar a ser factores importantes que influencien el comportamiento de un proceso físico determinado que ocurre dentro del sistema, y son cruciales para entender el mecanismo detallado de, por ejemplo, fenómenos como la dispersión de calor, la dispersión de masa o el transporte entre interfaces.La Dinámica de Fluidos Computacional (CFD) como herramienta de modelado numérico permite obtener una visión mas aproximada y realista de los fenómenos de flujo de fluidos y los mecanismos de transferencia de calor y masa en lechos empacados, a través de la resolución de las ecuaciones de Navier - Stokes acopladas con los balances de materia y energía y con un modelo de turbulencia si es necesario. De esta forma, esta herramienta permite obtener los valores medios y/o fluctuantes de variables como la velocidad del fluido, la temperatura o la concentración de una especie en cualquier punto de la geometría del lecho empacado.El objetivo de este proyecto es el de utilizar programas comerciales de simulación CFD para resolver el flujo de fluidos y la transferencia de calor y de masa en modelos bi/tri dimensionales de lechos empacados, desarrollando una estrategia de modelado aplicable al diseño de equipos para procesos de extracción o de reacción catalítica. Como referencia se tomaran procesos de tecnología supercrítica debido a la complejidad de los fenómenos de transporte involucrados en estas condiciones, así como a la disponibilidad de datos experimentales obtenidos previamente en nuestro grupo de investigación. Estos datos experimentales se utilizan como herramienta de validación de los modelos numéricos generados, y de las estrategias de simulación adoptadas y realizadas durante el desarrollo de este proyecto. / An understanding of the heat and mass transfer phenomena in a porous media implies the study of the fluid transport model within the void space; this fact is of fundamental importance to many chemical engineering systems such as packed bed extraction or catalytic reaction equipment. Experimental and theoretical studies of flow through such systems often treat the porous medium as an effectively homogeneous system and concentrate on the bulk properties of the flow. Such an approach neglects completely the complexities of the flow within the void space of the porous medium, reducing the description of the problem to macroscopic average or effective quantities. The details of this local flow process may, however, be the most important factor influencing the behavior of a given physical process occurring within the system, and are crucial to understanding the detailed mechanisms of, for example, heat and mass dispersion and interface transport.Computational Fluid Dynamics as a simulation tool allows obtaining a more approached view of the fluid flow and heat and mass transfer mechanisms in fixed bed equipment, through the resolution of 3D Reynolds averaged transport equations, together with a turbulence model when needed. In this way, this tool permit to obtain mean and fluctuating flow and temperature values in any point of the bed. The goal of this project is to use commercial available CFD codes for solving fluid flow and heat and mass transfer phenomena in two and three dimensional models of packed beds, developing a modeling strategy applicable to the design of packed bed chemical reaction and extraction equipment. Supercritical extraction and supercritical catalytic reaction processes will be taken as reference processes due to the complexity of the transport phenomena involved within this processes, and to the availability of experimental data in this field, obtained in the supercritical fluids research group of this university. The experimental data priory obtained by our research group will be used as validation data for the numerical models and strategies dopted and followed during the developing of the project.