Cette thèse de doctorat porte sur la valorisation du déchet solide issu de la préparation de la mélasse de caroube libanaise pour la production de bioénergie et de molécules à valeur ajoutée. L’analyse de la composition de ce déchet a montré qu’il contient 45 % (g/g) de sucres, substrats exploitables pour la fermentation alcoolique ou lactique, la production de biohydrogène, ou comme source de carbone pour la croissance d’une algue dans un procédé de phycoremédiation (traitement des eaux par les algues) pour l’élimination de micropolluants pharmaceutiques. Les résultats obtenus ont montré que la fermentation alcoolique en phase liquide à partir d’extraits de déchet de caroube peut conduire à des rendements élevés en bioéthanol sous réserve d’enrichir le milieu de culture par les éléments nécessaires pour la croissance de la levure Saccharomyces cerevisiae (en particulier l’azote), tandis qu’il est possible de produire directement le bioéthanol sur le déchet par fermentation en milieu solide sous réserve de contrôler précisément l’humidité. Nous avons également démontré que la fermentation lactique par immobilisation de Lactobacillus rhamnosus sur des billes d’alginate constitue une alternative possible au bioéthanol pour les sucres extraits du déchet. Cependant, un enrichissement du milieu de culture, ainsi que l’utilisation d’une invertase en prétraitement sont nécessaires afin de maximiser le rendement et la productivité de l’acide lactique. L’immobilisation des microorganismes a permis de réutiliser les mêmes billes d’alginate au cours de cinq cycles successifs de production. Une autre alternative potentielle aux voies précédentes est la fermentation sombre pour la production de biohydrogène directement à partir du déchet. Si des rendements intéressants ont été atteints, il faut noter que comme précédemment dans le cas de la fermentation solide du déchet, une forte dépendance aux conditions initiales de broyage a été mise en évidence ; de plus, à la carence en azote qui obligeait à supplémenter les milieux en fermentation en phase liquide s’ajoutent des besoins en fer. Enfin, l’utilisation du déchet comme substrat carboné dans un procédé de phycoremédiation avec l’algue Ankistrodesmus braunii a montré que l’élimination de 90% du diclofénac initial pouvait être atteinte en conditions de mixotrophie, même si seulement un tiers du diclofénac éliminé est effectivement métabolisé par l’algue. / This PhD thesis deals with the valorization of the solid waste generated by the Lebanese carob molasse process with the aim to produce bioenergy or high added-value platform molecules. The chemical analysis of this waste has highlighted that it contains 45% (w/w) sugars, which can be used as a substrate for alcoholic or lactic fermentation, the production of biohydrogen by dark fermentation, or as a carbon source for the growth of microalgae in a phycoremediation process (water treatment process based on algae/microalgae) for the removal of pharmaceutical micropollutants. Experimental results have shown that high ethanol yield and productivity could be achieved through alcoholic fermentation in the liquid phase using extracts from carob waste as the substrate, provided the culture medium was enriched by complementary nutrients (especially a nitro-gen source) to enhance the growth of the yeast Saccharomyces cerevisiae; in parallel, the direct production of ethanol from the solid waste based on solid-state fermentation was also proved to be efficient, provided humidity was accurately controlled. In addition, lactic fermentation per-formed with immobilized Lactobacillus rhamnosus on alginate beads was shown to constitute a potential alternative to bioethanol for the extracts from carob waste. An enriched culture medium was, however, necessary, together with the use of an invertase enzyme as a pretreatment so as to maximize the yield and the productivity of lactic acid. Using microorganism immobilization lead to the opportunity to recycle at least five times alginate beads in successive culture cycles. Another potential alternative to the above-mentioned valorization pathways was proved to lie in the dark fermentation process for biohydrogen production, directly using the solid waste. While attractive yields were reached, a strong dependence on the particle size from grinding pretreatment was emphasized, as in solid-state fermentation; moreover, iron supplementation in the culture medium was compulsory, in addition to nitrogen supplementation already described when fermentation was carried out in the liquid phase. Finally, when the carob waste was used as a substrate for the growth of the algae Ankistrodesmus braunii in a phycoremediation process, a removal yield of the initial diclofenac content about 90% could be reached under mixotrophic conditions, even though only one third of the diclofenac removed was effectively metabolized.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The energy crisis caused by the exhaustion of fossil fuels and environmental problems has caused concerns of researchers and, consequently, causing them to seek alternatives to replace fossil fuels with renewable sources. An interesting alternative is the use of solid-state fermentation for biofuels production using agricultural residues as a source of fermentable sugars. In Rio Grande do Sul, about 10 million tons of rice a year are produced in the state, producing around 3 million tons of rice husk and 1.5 million tons of rice bran. Such wastes have great potential for the production of bioethanol, but there are no proper techniques to date for the industrial production of ethanol by these wastes. The ethanol extraction technique from the solid state fermentation process is techniques should be improved so that the process becomes efficient and environmentally friendly. Thus, in this work was carried out a preliminary step to ethanol production process by solid state fermentation, evaluating an ethanol extraction method using distilled water as solvent. In this step, we evaluated parameters influencing the fermentation process (moisture content and initial concentration of ethanol) and the extraction process (temperature, agitation and solid-liquid ratio). Set the extraction conditions, the experimental design methodology was used in order to identify the significant variables in the process of solid state fermentation for ethanol production through the application of a design Plackett & Burmann experiments. The response surface methodology was used to perform process optimization, based on the evaluation of a central composite rotational design (CCRD). For end a scale-up of the simultaneous saccharification and solid state fermentation process was proposed, reaching a final ethanol concentration of 143.88 g EtOH / kg substrate. / A crise energética causada pela exaustão de combustíveis fósseis e problemas ambientais tem causado preocupações de pesquisadores e, consequentemente, fazendo com que os mesmos procurem alternativas para substituir os combustíveis fósseis por fontes renováveis. Uma alternativa interessante é o uso da fermentação em estado sólido para produção de biocombustíveis, utilizando resíduos agroindustriais como fonte de açúcares fermentáveis. No Rio Grande do Sul, cerca de 10 milhões de toneladas de arroz são produzidas por ano no Estado, produzindo em torno de 3 milhões de toneladas de casca de arroz e 1,5 milhão de toneladas de farelo de arroz. Tais resíduos apresentam grande potencial para a produção de bioetanol, porém não há técnicas adequadas até o momento para a produção industrial de etanol através destes resíduos. A técnica de extração de etanol proveniente do processo de fermentação em estado sólido é uma das técnicas que devem ser aprimoradas para que o processo se torne eficiente e ambientalmente correto. Com isso, neste trabalho foi realizada uma etapa preliminar ao processo de produção de etanol por fermentação em estado sólido, avaliando um método de extração de etanol utilizando água destilada como solvente. Nesta etapa, foram avaliados parâmetros que influenciam o processo fermentativo (teor de umidade, e concentração inicial de etanol) e o processo de extração (temperatura, agitação e razão sólido-líquido). Fixadas as condições de extração, a metodologia de planejamento de experimentos foi utilizada com objetivo de identificar as variáveis significativas no processo de fermentação em estado sólido para a produção de etanol, através da aplicação de um design de experimentos Plackett & Burmann. O método de superfície de resposta foi utilizado para realizar a otimização do processo, com base na avaliação de um delineamento composto central rotacional (DCCR). Por fim, foi proposto um aumento de escala do processo de sacarificação e fermentação simultânea em estado sólido, atingindo uma concentração de etanol final de 143,88 g EtOH/ kg substrato.

Establishing a commercial, lignocellulose-based, second-generation ethanol process has received several decades of attention by both researchers and industry. However, a fully economically viable process still remains a long-term goal. The main bottleneck to this achievement is the recalcitrance of lignocellulosic feedstocks, although there are several other factors, such as the huge investment required for second-generation ethanol facilities. An intelligent alternative solution discussed in this thesis is an integrated approach using firstgeneration ethanol plants for second-generation processes. Wheat is the major feedstock for first-generation ethanol in Europe; therefore, wheat-based lignocellulose waste, such as wheat straw, bran, and whole stillage fiber (a waste stream from first-generation wheat-based ethanol plants) was the primary focus of the integration model in this thesis. Since the major share of first-generation ethanol plant economics focuses on the animal feed DDGS (Distillers’ dried gains with solubles), the integration of lignocellulose should be designed in order to maintain DDGS quality. An ethanol-producing edible filamentous fungus, Neurospora intermedia, a potential protein source in DDGS, was considered for use as the fermenting microbe. The morphological and physiological aspects of this fungus were studied in the thesis, leading to the first report of fungal pellet development. An alternative approach of using dilute phosphoric acid to pretreat lignocellulose, as it does not negatively affect fungal growth or DDGS quality, was demonstrated in both the laboratory and on a 1m3 pilot scale. Furthermore, the process of hydrolysis of pretreated lignocelluloses and subsequent N. intermedia fermentation on lignocellulose hydrolysate was also optimized in the laboratory and scaled up to 1 m3 using an in-house pilot-scale airlift bioreactor. Fungal fermentation on acid-pretreated and enzyme-hydrolyzed wheat bran, straw and whole stillage fiber resulted in a final ethanol yield of 95%, 94% and 91% of the theoretical maximum based on the glucan content of the substrate, respectively. Integrating the first- and second-generation processes using thin stillage (a waste stream from first-generation wheat-based ethanol plants) enhanced the fungal growth on straw hydrolysate, avoiding the need for supplementing with extra nutrients. Based on the results obtained from this thesis work, a new model for integrated first- and second-generation ethanol using edible filamentous fungi processes that also adds value to animal feed (DDGS) was developed.

Dans une démarche d'optimisation et de maîtrise d'un bioprocédé, une préoccupation importante concerne la mesure et le suivi de l'état des cellules. Au cours de cette thèse, des mesures in situ et/ou permettant de mettre en évidence une variabilité phénotypique au sein d'une population de levures ont été recherchées. La spectroscopie d'impédance, basée sur la capacité des cellules viables à se polariser sous l'effet d'un champ électrique, a été retenue pour estimer l'état des cellules en-ligne. Pour effectuer des mesures de morphologie et de viabilité à l'échelle de la cellule, un système complet de microscopie et d'analyse d'image automatisées a été développé, en parallèle de l'utilisation d'un compteur de cellules de type Coulter. Enfin, des suivis individuels de croissance sur milieu gélosé ont été réalisés afin de caractériser la population en termes de temps de latence et de vitesse de croissance. Le procédé modèle de cette étude est la production de bioéthanol, qui expose les levures utilisées (S. cerevisiae) à d'importantes contraintes physicochimiques (température, acétate, furfural,...) qui affectent leur état physiologique et limitent l'efficacité de l'étape de fermentation. La population, homogène sur le plan cinétique dans des conditions de culture non stressantes, devient hétérogène lorsqu'une perturbation est appliquée. La mesure de cette hétérogénéité peut être utilisée comme marqueur de la sévérité du stress subi. Au cours des phases de déclin, la mort s'accompagne d'une diminution de la taille des cellules et d'une modification de leur aspect en microscopie. Ces changements permettent d'estimer la proportion de cellules viables à partir des distributions de taille obtenues avec le compteur d'une part, et l'analyse des images de microscopie d'autre part. La spectroscopie d'impédance donne une estimation fiable de la fraction volumique de cellules viables et permet de mesurer la capacitance membranaire Cm, ainsi que la conductivité intracellulaire sin, des paramètres liés à l'état de la membrane et du cytoplasme. Cm, constante tant que les cellules sont viables, s'annule à leur mort, tandis que sin varie selon la phase de culture et en réponse aux stress. / For bioprocess control and optimization, biomass monitoring and physiological state evaluation is an important issue. During this work, in situ and at-line measurements have been used to evaluate cell state and detect a phenotypic variability within a yeast population. Dielectric spectroscopy, based on the polarization of viable cell membranes exposed to an electrical field, has been selected to infer cell state on-line. In parallel with the use of a Coulter-type cell counter, a dedicated system of automated microscopy and image analysis has been developed to measure cell morphology and viability. Single-cell growth on agar medium was monitored to characterize individual cells with regard to lag-time and initial growth rate. Bioethanol production with S. cerevisiae has been chosen as a model process since the yeast cells are exposed to strong physicochemical stresses (temperature, acetate, furfural,?) which affect their physiological state and impair fermentation efficiency. The cell population, kinetically homogeneous during stress-free fermentations, became heterogeneous when a perturbation was applied. The mean and the variance of lag-time distributions were related to the stress severity. During the decline phase, cell death went along with a decrease in cell size and changes of their microscopy aspect. These changes were significant enough to infer the proportion of viable cells directly from the size distributions obtained with the cell counter or from microscopy image analysis. Dielectric spectroscopy gave reliable estimates of the viable cell volume fraction and enabled the measurements of membrane capacitance Cm and intracellular conductivity sin, parameters related to membrane and cytoplasm states. The Cm value remained constant as long as the cells were viable and dropped to zero at cell death, while sin varied significantly depending on the growth phase and in response to stress.

L'adoption d'une méthodologie est cruciale pour la conception de procédés chimiques optimaux. L'optimisation multi-objective de modèles rigoureux en est un exemple, jouissant d'une application extensive dans la littérature. Cette méthode retourne un ensemble de solutions, dit de Pareto, présentant un compromis optimal entre les fonctions objectives. Ceci est suivi par une étape de sélection d'une solution d'intérêt répondant à des critères définis. Cette méthodologie s'appliquait, dans le cadre de cette thèse, à deux procédés. Le premier consistait en une distillerie d'éthanol à partir de la canne à sucre, combinée à un système de cogénération et de combustion à partir de la biomasse de canne à sucre. Le deuxième contenait en plus un système d'hydrolyse enzymatique de cette biomasse. Notre première contribution traitait de la construction d'une procédure pour la modélisation, simulation, intégration thermique et évaluation du coût des équipements. La deuxième contribution traitait de l'analyse des résultats réalisée à travers un suivi de variables mesurées, une fragmentation de la courbe de Pareto, une hiérarchisation des variables de décision et une comparaison avec la littérature. La dernière contribution traitait de l'étape de sélection qui s'est réalisée à travers une évaluation économique des solutions, sous des scénarii différents, avec la Valeur Nette Présente comme critère de sélection. En conclusion, cette thèse constitue une première application intégrale de la méthodologie proposée. Elle représente, de par ses contributions, un tremplin pour des applications futures à des procédés chimiques ou biochimiques, plus spécialement pour la canne à sucre / The use of a systematic methodology is crucial for the design of optimal chemical processes, namely bio-processes. Multi-objective optimization of rigorous process models is therein a prime example, with extensive use in literature. This method yields a Pareto set of optimal compromise solutions, from which one optimal solution is chosen based on specific criteria. This methodology was applied, in the course of this thesis, to two studied processes. The first consisted in a distillery converting sugarcane to ethanol, combined with a sugarcane biomass combustion and power cogeneration system. The second contained an additional biomass hydrolysis system. Our first contribution deals with the construction of an organized procedure for the modeling, simulation, heat integration and equipment and capital cost estimation of chemical processes. The second contribution deals with the analysis of the optimization results through a tracking of measured variables, the fragmentation of the Pareto curve, an ordering of optimization variables, and a comparisons with literature results. The final realization deals with the selection step realized through an economic evaluation of optimal solutions for various scenarios, with the Net Present Value as the selection criterion. In conclusion, this thesis constitutes a first integral application of the said methodology. It sets, through its contributions, a stepping stone for future application in the field of chemical and biochemical processes, namely for sugarcane processes

The purpose of diploma thesis is to provide an overview of biofuels market in Russian Federation, Belarus and Ukraine and estimate prospects of future development. First chapter characterizes biofuels in regard of the feedstock and further utilization. Next part of the thesis describes development of biofuel industry in the key selected countries and analyses current trends on the global market. Next three sections review Russian, Ukrainian and Belarusian sectors of biofuels including feedstock potential, supportive policies, strategies, international collaboration and issues in biofuel production, distribution and consumption in each country. Last section of the thesis investigates potential from exported raw materials and its contribution to achievement of established targets with the following outlook for the future development.

Thesis is concentrated on two biotechnologies: biofuels and genetically modified crops. Defines technical and economical characteristics, lists international agreements and czech and european legislation regarding both biotechnologies. Introduces opinions and analyses positions of selected political subjects as UNO, WTO, OECD, EU, catholic church and main czech political parties.

Orientadores: Maria Aparecida Silva, Adriano Viana Ensinas / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-22T07:39:18Z (GMT). No. of bitstreams: 1 Albarelli_JulianaQueiroz_D.pdf: 4345487 bytes, checksum: 3b35163c3127eaa2e83d7a92e58466c2 (MD5) Previous issue date: 2013 / Resumo: Nesta tese, utilizou-se o simulador comercial Aspen Plus® para simular o processo de produção conjunta de açúcar, etanol e eletricidade em uma usina convencional e a produção de etanol e eletricidade em uma destilaria autônoma. A integração dos processos e diferentes usos para o bagaço excedente foram estudados. Foram avaliadas modificações nos processos convencionais para aumentar a produção de eletricidade e a produção de etanol de segunda geração, utilizando como matéria prima o bagaço ou fração fina do bagaço composta por células de parênquima (fração-P). Os casos estudados também foram avaliados economicamente e quanto à captação de água, propondo alternativas para minimização desta. Os resultados indicaram que a integração energética dos processos convencionais minimizou a quantidade de vapor necessária, disponibilizando uma maior quantidade de bagaço para outros usos. O uso do bagaço excedente, em conjunto com a palha, como combustível em um sistema de cogeração com a instalação de uma turbina de condensação, permitiu o aumento da eletricidade disponível em 244%, quando utilizada a maior quantidade de palha admitida para o caso estudado para a destilaria autônoma. Os casos que estudaram a fração-P apresentaram maior produção de energia elétrica e produção de etanol/t material lignocelulósico do que os casos de segunda geração a partir do bagaço. Todos os casos apresentaram captação de água superior à permitida no Estado de São Paulo, 1 m3/t de cana de açúcar, sendo desta forma considerada a concentração da vinhaça e o aproveitamento, após tratamento, da água de lavagem do material pré-tratado, para os casos de segunda geração. A análise econômica mostrou que é possível aumentar a lucratividade dos processos convencionais, a longo prazo, com modificações no sistema de cogeração resultando em pouca alteração no tempo de retorno do investimento. O uso da fração-P para produção de etanol de segunda geração, tendo como pré-tratamento LHW e utilizando custo baixo para as enzimas, se apresentou igualmente atrativo, do ponto de vista econômico, ao melhor caso estudado de modificações no sistema de cogeração. Menores tempo de retorno, rentabilidade do investimento e lucratividade, a longo prazo, foram encontrados para os casos que consideraram a produção conjunta de açúcar e etanol. A viabilidade econômica dos processos de segunda geração apresentou-se altamente dependente do custo das enzimas / Abstract: In this thesis, the commercial simulator Aspen Plus® was used to simulate the conventional mill producing sugar, ethanol and electricity and the autonomous distillery producing ethanol and electricity. The integration of processes and different uses for the surplus bagasse were studied. It was evaluated changes in conventional processes to increase electricity production and the production of second generation ethanol using bagasse or bagasse fine fraction composed by parenchyma cells (fraction-P). The cases were economically evaluated and water uptake was analyzed, alternatives were proposed to its minimization. The results indicated that the energy integration of the conventional processes minimized the amount of steam, it provided a larger amount of bagasse for other uses. The use of bagasse, together with sugarcanetrash, as a fuel to the cogeneration system considering the installation of a condensation turbine led to an increase of 244% in the available electricity when using the higher amount of sugarcane-trash studied for the autonomous distillery. The cases that studied the P-fraction showed higher production of electricity and ethanol/t lignocellulosic material than the cases of second generation that used bagasse. All cases presented water uptake higher than allowed in the state of São Paulo, 1 m3/t de sugarcane, therefore, it was considered the concentration of vinasse and recovery of wash water from the pre-treated material, for the second generation cases. The economic analysis showed that it is possible to increase the profitability of the conventional processes, in the long term, with changes in the cogeneration system resulting in little change in the payback time of the investment. The use of P-fraction for production of second generation ethanol, with the LHW pretreatment and considered low cost of the enzymes, was equally attractive, from an economic point of view, to the best studied case of changes in the cogeneration system. Lower payback time, return on the investment and long-term profitability were found for cases that considered the joint production of sugar and ethanol. The economic viability of the second generation process presented itself highly dependent to the enzyme cost / Doutorado / Engenharia Química / Doutor em Engenharia Química

Diploma thesis deals with use of Zymomonas mobilis for the production of bioethanol from waste paper. There were used three kinds of substrate (cardboard, drawing and office paper) to optimize of bioethanol production. Individual papers were subjected to the same pre treatment, namely a milling, a combination of microwave irradiation and NaOH, a combination of microwave irradiation and H2SO4 and combination microwave irradiation, H2SO4 and NaOH. The substrates were decomposed by enzymatic hydrolysis after pre treatment to evaluate the best pre-treatment. Simultaneous saccharification and fermentation was carried out for each substrate (with two of the best pre-treatment). The samples were taken during the hydrolysis and the simultaneous saccharification and fermentation, and were determined by HPLC. Growth curves of Zymomonas mobilis were constructed, as the most appropriate for SSF was chosen temperature of 40 ° C in which the exponential phase took place at the time of 6 15 hours. During hydrolysis was monitored glucose concentration in the solution. The maximum concentration of glucose was in the cardboard (microwaves + H2SO4 + NaOH) 16.46 gdm-3, a drawing (microwaves + H2SO4 + NaOH) 31.78 gdm-3, and office paper (microwaves + H2SO4) 25.04 gdm-3. The concentration of ethanol for SSF was highest in the same cases as in the hydrolysis. The cardboard was the maximum concentration of bio ethanol 9.5 gdm-3, for the drawing 16.1 gdm-3 and for the office paper 12.13 gdm-3.

This diploma thesis deals with the optimization of the production of ethanol from waste paper using yeast. There were used 4 kinds of paper as a substrate - office paper, non-recycled workbook, recycled workbook and newspaper. All papers were pretreated with the following procedures: grinding, microwaves + NaOH, microwave + H2SO4 and microwave + H2SO4 + NaOH. The glucose concentration was determined in enzymatic hydrolysis by HPLC. Saccharomyces cerevisiae were chosen for ethanol production. The production of ethanol was carried out with all the pretreated papers in simultaneous saccharification and fermentation. During hydrolysis, the pretreated papers have reached the highest results in the combination with microwave + H2SO4 + NaOH. Non-recycled workbook was the only exception, where the highest concentration of glucose has been obtained by the pretreatment of microwaves + H2SO4. Following results have been acquired: office paper 24,69 gdm-3, non-recycled workbook 22,47 gdm-3, recycled workbook 16,94 gdm-3 and newspapers 15,36 gdm-3. SSF was carried out again with all the papers and their pretreatments. The highest concentration of ethanol has been achieved in microwave pretreatment + H2SO4 + NaOH. The highest overall concentration has been gained from the office paper, amounted to 16,98 gdm-3. The maximum concentration of ethanol for non-recycled workbook has been 15,25 gdm-3, for recycled workbook 12,2 gdm-3 and for newspapers 12,59 gdm-3.