Experimentos industriais são realizados pelas empresas no intuito de melhorar as características de qualidade dos produtos e processos de fabricação. Nesse sentido, esta dissertação tem como objetivo estudar e aplicar as técnicas de planejamento e análise de experimentos na melhoria da qualidade industrial. Como parte do objetivo, desenvolve-se uma aplicação com as técnicas de planejamento fatorial fracionado '2POT. K-P', metodologia de superfície de resposta e análise de variância, em um processo de moldagem por injeção, utilizado por uma indústria que fabrica e comercializa componentes plásticos usados na construção civil. Com essa pesquisa experimental identificam-se os parâmetros mais importantes da injeção plástica: temperatura da máquina e pressão de injeção. Ao mesmo tempo, determinam-se os níveis ótimos de regulagem desses parâmetros. Assim, com esse estudo, avalia-se o procedimento de implantação das técnicas de experimentação e as dificuldades práticas encontradas, bem como busca-se contribuir na integração entre universidade e empresa. / Industrial experiments are made by companies in order to improve the quality characteristics of products and production processes. In this sense, the objective of this dissertation is to study and apply the design of experiments in the industrial quality improvement. In addition, as a part of the objective, an application of the techniques of design Fractional Factorial '2POT. K-P', Analysis of Variance and Response Surface Methodology is done. It is focused in an injection molding process applied by a company, that makes and trades plastic products for the civil construction. Using this experimental study, the most important parameters of plastic injection are identified: melt temperature and injection pressure. At the same time, the optimal levels of adjustment of these parameters are determined. From this study, it is evaluated both the implantation procedures of the designs of experiments as well as the difficulties faced. Also, this study tries to contribute to the university-company relationship.

O objetivo, no capítulo I deste trabalho, foi desenvolver e validar um novo método espectrofotométrico in vitro para a avaliação da eficácia de protetores solares, o qual pode ser adotado na rotina industrial como uma maneira de identificar desvios de qualidade e garantir a eficácia de protetores solares a cada novo lote produzido. O método proposto é baseado na absorção espectrofotométrica na região do UV (280-400 nm) de uma camada fina de protetor solar aplicada na face externa de uma cubeta de quartzo preenchida com uma solução padrão (SP) que absorve em toda a região do UV, a fim de obter uma medida de área sob a curva (ASC), empregada como valor de referência para cada FPS testado. Para o desenvolvimento do método, foram utilizados protetores solares da mesma marca, com valores de FPS de 15; 20; 30; 40; 50 e 60, o qual foi aplicado sobre a face externa da cubeta a uma concentração de 0,2 mg/cm2. A SP consistiu de uma solução de quercetina (10 ?g/mL) à qual foram adicionadas 6 gotas de solução de AlCL3 6%. A partir do espectro de absorvância de cada amostra na região de UV foi calculado o valor da ASC. O método demonstrou ser reprodutível, robusto e eficaz para controle de qualidade de protetores solares. Outro importante objetivo desse estudo, descrito no capítulo II, foi determinar as condições ótimas para a extração de compostos fenólicos do cranberry em pó utilizando metodologia de superfície de resposta, empregando quatro variáveis independentes: (i) tempo de extração; (ii) proporção planta:solvente; (iii) temperatura e (iv) velocidade de agitação, segundo desenho experimental do tipo Box-Behnken, com análise de três respostas: teor de polifenois totais (TPT); atividade antioxidante (CI50; DPPH) e teor de sólidos totais (TST). As condições ideais obtidas foram: tempo de extração 5 horas; proporção planta:solvente 1:2; velocidade de agitação 80 RPM e temperatura 45°C, correspondendo aos valores preditos de 433,1 mg/g; 0,100 mg/mL e 19,9% para o TPT; CI50 DPPH e TST, respectivamente. Em seguida, o extrato otimizado de cranberry foi avaliado com relação à sua atividade fotoprotetora/fotoquimioprotetora. Os resultados apresentados neste trabalho demonstraram que o extrato otimizado de cranberry apresenta elevado potencial antioxidante ao inibir a quimioluminescência (CI50 de 0,202 ± 0,32 ?g/mL) e reduzir a peroxidação lipídica (CI50 de 32,6 ± 5,8 ?g/mL). No ensaio de DPPH, obteve-se uma CI50 de 136,7 ±6,8 ?g/mL. Reduziu em 41% a peroxidação lipídica em células L929. Além disso, o extrato incorporado a uma formulação gel-creme, apresentou elevado potencial fotoprotetor in vitro. / The chapter I of this work aims to develop and validate a new in vitro spectrophotometric method for assessing the effectiveness of sunscreens, which can be adopted in industrial routine production as a way to identify quality deviations and ensure the effectiveness of sunscreen product. Briefly, the proposed method is based on the spectrophotometric absortion on the UV region (280-400 nm) of a thin layer of sunscreen sample applied on the outside of a quartz cuvette which is filled with a standard solution (SS). The area under the curve of absorption of SS and sunscreen sample is obtained. For the experiment development, it was used sunscreens products of the same brand with SPF values of 15; 20; 30; 40; 50 and 60 which was applied on the cuvette outside at a concentration of 0,2mg/cm2. The reference standard used consisted of a quercetin solution added of AlCl3 6%. From the absorbance of each sample spectrum in UV region was calculated the value of the area under the curve (AUC). The spectrophotometric method developed was effective for confirm SPF sunscreen products produced batch to batch. In the chapter II, were determined the optimal conditions for phenolic compounds extraction from cranberry powder using the surface response methodology with four independent variables: (i) extraction time (hours); (ii) ratio plant : solvent; (iii) temperature and (iv) speed agitation (RPM) , according to Box-Behnken, with analysis of three answers: total polyphenol content (TPC); antioxidant activity (IC50; DPPH) and total solids content (TSC). The ideal conditions obtained were: extraction time 5 hours; ratio plant:solvent 1:2; agitation speed 80 RPM and temperature 45°C, corresponding to the values predicted of 433.1 mg/g; 0,100 mg/ml and 19.9% for TPC; IC50 DPPH and TSC respectively. Then, the optimized cranberry extract was evaluated with respect to its photoprotective/photochemoprotective activity. Preliminary results presented in this paper demonstrate the ability of optimized cranberry extract to scavenge free radicals, with a high antioxidant potential to inhibit satisfactorily chemiluminescence generated in the system xanthine/XOD/luminol; presenting IC50 0.202 ± 0.32 ?g/ml; lipid peroxidation using yolk egg as lipid substrate, with an IC50 of 32.6 ± 5,8 ?g/ml. In DPPH test, the optimized extract showed an IC50 of 136.7 ± 6.8 mg/mL. The extract was also able to reducing UVB radiation-induced damage at the cellular level in fibroblasts mice of L929 line, with a 41% reduction in lipid peroxidation in cells compared to irradiated control.

Nessa pesquisa avaliou-se a produção fermentativa de hidrogênio e ácidos orgânicos a partir do bagaço de cana-de-açúcar (BCA) usado como substrato em reatores em batelada. Três condições de pré-tratamento (hidrotérmico, autoclave e hidrotérmico mais autoclave) do BCA e condição in natura foram avaliadas a fim de favorecer a produção de hidrogênio. Verificou-se produção molar de hidrogênio de 3,79 mmol/L, 3,47 mmol/L, 1,67 mmol/L e 1,01 mmol/L para BCA autoclavado, BCA in natura, BCA pré-tratado em sistema hidrotérmico e BCA pré-tratado em sistema hidrotérmico seguido de autoclave, respectivamente. A partir desses valores, optou-se por usar o BCA autoclavado como substrato para otimização da produção de hidrogênio e ácidos orgânicos a partir de metodologias de delineamento do composto central e superfície de resposta. Foram monitorados 10 reatores em batelada (R1 a R10), em triplicatas, com diferentes concentrações de substrato (0,8 a 9,2 g/L) e pH (de 4,6 a 7,4). A maior produção de hidrogênio (24,1 mmol/L) e 6,4 g/L de ácidos orgânicos foram obtidos em R4 (8,0 g BCA/L e pH 7,0). Os açúcares glicose, arabinose, xilose, manose e galactose foram observados ao longo do tempo de operação em todos os reatores, sendo arabinose observado em maior concentração nas condições dos reatores R3 (8,0 g BCA/L e pH 5,0) e R8 (5,0 g BCA/L e pH 7,4), respectivamente, 1.415,3 e 1.372,5 mg/L. A produção de hidrogênio foi concomitante à formação de ácidos orgânicos, principalmente butírico (de 14,6 a 33,8% em R1 e R6, respectivamente) e succínico (de 19,5 a 26,4% em R3 e R9, respectivamente). Os dois fatores analisados, concentração de substrato e pH, exerceram efeitos significativos na produção de hidrogênio, ácido butírico e succínico. A partir dos resultados obtidos com o planejamento fatorial, foi possível verificar que o valor máximo de produção de hidrogênio estimado pelo modelo foi de 23,10 mmol/L, para 7,0 g BCA/L e pH 7,2. O valor obtido no experimento de otimização (Rotm) foi de 19,84 mmol/L, com grau de precisão do modelo de 85,9% para produção de hidrogênio a partir de BCA autoclavado. Sequenciamento massivo via plataforma Illumina (Miseq) foi realizado para a identificação de bactérias do reator do ponto central, (R9, 5,0 g BCA/L e pH 6,0), do reator otimizado (Rotm, 7,0 g BCA/L e pH 7,2), de amostras do BCA autoclavado e inóculo. No inóculo foram identificadas principalmente bactérias semelhantes a Clostridium bifermentans (62,69% de abundância relativa), Bacillus coagulans (31,67%) e Enterobacter aerogenes (2,72%). No BCA foram identificadas bactérias semelhantes a C. bifermentans (31,91%), C. cellobioparum (32,29%), C. cellulolyticum (5,69%), C. sartagoforme (14,63%) e Paenibacillus spp. (11,67%). Estas bactérias não foram favorecidas sob as condições impostas em R9 (5,0 g BCA/L e pH 6,0) e Rotm (7,0 g BCA/L e pH 7,2), uma vez que a abundância relativa das bactérias nas amostras dos reatores foram completamente diferentes. Em R9, bactérias semelhantes a Lactobacillus paracasei e Escherichia hermannii foram as principais identificas com 37,50 e 34,32% de abundância relativa, respectivamente. Em Rotm, as principais bactérias identificadas foram semelhantes a Bacteroides sp. e Enterobacter aerogenes, com 37,35 e 27,72% de abundância relativa, respectivamente. Assim, as populações bacterianas, bem como a produção de metabólitos, foram alteradas em função das condições impostas; ou seja, concentração de BCA, pH em reatores em batelada com BCA autoclavado como substrato. / This study evaluated the hydrogen and organic acids fermentative productions from sugarcane bagasse (SCB) as substrate in batch reactors. Three pre-treatment conditions (hydrothermal, autoclave and hydrothermal plus autoclave) of BCA and the in natura condition were evaluated in order to favor the hydrogen production. Hydrogen molar productions of 3.79 mmol/L, 3.47 mmol/L, 1.67 mmol/L and 1.01 mmol/L was found for SCB pretreated in autoclave, BCA in natura, SCB pretreated in hydrothermal system and SCB pretreated in hydrothermal system followed by autoclaving, respectively. From these values, it was decided to use autoclaved BCA as a substrate for optimization of hydrogen and organic acids productions from the design methodologies of the central compound and response surface. Ten batch reactors (R1 to R10) were monitored in triplicates with different substrate concentrations (0.8 to 9.2 g/L) and pH (4.6 to 7.4). The highest production of hydrogen (24.06 mmol/L) and 6.42 g/L of organic acids were obtained in R4 (8.0 g BCA/L and pH 7.0). Glucose, arabinose, xylose, mannose and galactose were produced and consumed throughout the operating time of all reactors, and arabinose was observed at higher concentration, 1,415.26 and 1,372.45 mg/L in R3 (8.0 g BCA/L and pH 5.0) and R8 (5.0 g BCA/L and pH 7.4), respectively. The production of hydrogen was concomitant to the formation of organic acids, mainly butyric (from 14.6 to 33.8% in R1 and R6, respectively) and succinic (from 19.5 to 26.4% in R3 and R9, respectively). The two factors analyzed, substrate concentration and pH, had significant effects on the production of hydrogen, butyric acid and succinic acid. From the results obtained with the factorial design, it was possible to verify that the maximum value of hydrogen production estimated by the model was 23.10 mmol/L, to 7.0 g BCA L and pH 7.2. The value obtained in the optimization experiment (Rotm) was 19.84 mmol/L, with an accuracy of 85.9% for hydrogen production from autoclaved BCA. Sequencing by the Illumina platform (Miseq) was performed for the identification of bacteria from the central point reactor (R9, 5.0 g BCA/L and pH 6.0), optimized reactor (Rotm, 7.0 g BCA/L and pH 7.2), autoclaved BCA and inoculum samples. In the inoculum were identified mainly bacteria similar to Clostridium bifermentans (62,69% of relative abundance), Bacillus coagulans (31,67%) and Enterobacter aerogenes (2,72%). Bacteria similar to C. bifermentans (31.91%), C. cellobioparum (32.29%), C. cellulolyticum (5.69%), C. sartagoforme (14.63%) and Paenibacillus spp. (11.67%). These bacteria were not favored under the conditions imposed on R9 (5.0 g BCA/L and pH 6.0) and Rotm (7.0 g BCA/L and pH 7.2), since the relative abundance of the bacteria in the reactor samples were completely different. In R9, bacteria similar to Lactobacillus paracasei and Escherichia hermannii were the main identified with 37.50 and 34.32% of relative abundance, respectively. In Rotm, the main bacteria identified were similar to Bacteroides sp. and Enterobacter aerogenes, with 37.35 and 27.72% relative abundance, respectively. Thus, bacterial populations, as well as the production of metabolites, were altered as a function of the imposed conditions; ie, BCA concentration, pH in batch reactors with autoclaved BCA as substrate.

As lipases (EC 3.1.1.3) hidrolisam ligações éster de triacilgriceróis numa interface água-óleo. Por possuírem diversas aplicações industriais, são as enzimas mais utilizadas em sínteses orgânicas e mais de 20% de biotransformações são realizadas com lipases. Os objetivos deste trabalho foram avaliar as condições nutricionais de meios de cultivo (fontes de óleos vegetais e animais, nitrogênio e carbono) para melhorar a produção de lipase por Candida viswanathii além de purificar a enzima em sistema aquoso bifásico e realizar a caracterização parcial da lipase. Entre os meios de cultivos analisados o meio de Vogel proporcionou a maior produção de lipase após 36 horas de cultivo (11,73 U/mL). Após analisar as fontes de carbono lipídicas e não-lipídicas e fontes de nitrogênio, um planejamento experimental de misturas foi aplicado para analisar o efeito de glicerol, lactose e sorbitol sobre a produção de lipase. Os resultados obtidos mostraram que a maior produção de lipase (20,41 U/mL) foi observada com 0,5% de lactose (p/v) e 0,5% (p/v) de sorbitol, sendo observado um aumento de 74% da produção de enzima. Nesse estudo observou-se que a lactose e sorbitol foram utilizados como adjuntos para a produção de lipase. A lipase produzida nas condições anteriores foi purificada em sistema aquoso bifásico (SAB) formado por polietilenoglicol (PEG) e fosfato de potássio. O maior coeficiente de partição (1,34) foi encontrado no ensaio com PEG 4000/fosfato a Temperatura de 40 °C e pH 7,0, bem como o maior balanço de atividade (50,73%). Na caracterização parcial, a lipase não sofreu influência da temperaturana faixa de 20 – 60 °C e obteve atividade máxima em pH 8,1. A enzima apresentou também elevada estabilidade em solventes orgânicos como metanol e etanol, sendo estas propriedades consideradas importantes para aplicações em processos biotecnológicos. / Lipases (EC 3.1.1.3) hydrolyze ester bonds of triacylglycerols at a water-oil interface. By they have several industrial applications, they are the enzymes most used in organic synthesis and more than 20% of biotransformations are performed with lipases. The objectives of this work were to evaluate the nutritional conditions of culture media (plant and animal oils, nitrogen and carbon) to improve the production of lipase by Candida viswanathii in addition to purifying the enzyme in a biphasic aqueous system and to realize the partial characterization of lipase. Among the culture media analyzed, the Vogel medium provided the highest lipase production after 36 hours of culture (11,73 U/mL). After analyzing the lipid and non-lipid carbon sources and nitrogen sources, an experimental design of blends was applied to analyze the effect of glycerol, lactose and sorbitol on lipase production. The results showed that the highest production of lipase (20,41 U/mL) was observed with 0.5% lactose (w/v) and 0.5% (w/v) sorbitol, with an increase of 74% of enzyme production. In this study it was observed that lactose and sorbitol were used as adjuncts for the production of lipase. The lipase produced under the above conditions was purified in a aqueous two-phase systems (ATPS) consisting of polyethylene glycol (PEG) and potassium phosphate. The highest partition coefficient (1,34) was found in the PEG 4000/phosphate assay at temperature of 40 °C and pH 7,0, as well as the highest activity balance (50,73%). In the partial characterization, the lipase was not influenced by the temperature range of 20 - 60 °C and obtained maximum activity at pH 8,1. The enzyme also showed high stability in organic solvents such as methanol and ethanol, and these properties are considered important for applications in biotechnological processes.

A oxidação lipídica é uma das principais reações de deterioração em alimentos, provocando perdas em qualidade e valor nutricional. Para evitar este problema a indústria utiliza antioxidantes, substâncias capazes de retardar essas reações. Os compostos fenólicos constituem-se na principal substância antioxidante utilizada e encontrada em alimentos, sendo que estes podem estar presentes em várias matérias primas vegetais e em resíduos agroindustriais, desprezados pela indústria. O objetivo do presente trabalho foi estudar as condições de extração de compostos fenólicos de frutos de acerola (Malpighia emarginata), borra de café (Coffea arabica) e casca de lichia (Litchi chinensis), bem como avaliar sua atividade antioxidante in vitro e sua aplicação em emulsões. Para extração dos compostos fenólicos foi aplicada a metodologia de superfícies resposta, visando a definição da melhor combinação de variáveis (concentração de etanol, temperatura e tempo) para a máxima recuperação de compostos fenólicos. A avaliação da atividade antioxidante se deu pela estabilização do radical DPPH e estudo da cinética da reação em comparação ao antioxidante sintético TBHQ. Além disso, foi avaliada a atividade antioxidante dos extratos adicionados a emulsões em diferentes concentrações (50 a 200 mg.kg-1), submetidas a teste acelerado de oxidação. O estudo das condições de extração evidenciou que os compostos fenólicos de cada matriz devem ser extraídos em condições distintas. Extratos de frutos de acerola apresentaram o maior teor de compostos fenólicos totais quando foi utilizada apenas água como solvente (175,87 mg GAE.g-1), seguido pelos extratos de borra de café e de casca de lichia extraídos com soluções etanólicas (26,37 e 25,87 mg GAE.g-1 respectivamente). O estudo da cinética de estabilização do radical DPPH demonstrou que o extrato de acerola foi o mais eficiente, seguido pelos extratos de casca de lichia e de borra de café (650,53; 43,09 e 38,74 de TEAC, respectivamente). O estudo de aplicação dos extratos em sistema modelo, com acompanhamento da oxidação por meio da determinação de hidroperóxidos formados (mmol.kg-1), indicou que somente o extrato de acerola, em todas as concentrações utilizadas, apresentou efeito comparável ao TBHQ. Os extratos de casca de lichia e de borra de café, em qualquer concentração utilizada, não apresentaram a mesma proteção à oxidação lipídica. Com os resultados obtidos pode-se concluir que cada matriz se comporta de uma forma na extração de compostos fenólicos, e daí vem a necessidade de se realizar estudo das condições de preparo dos extratos para cada amostra. O extrato de acerola se mostrou o mais eficiente, tanto na cinética de estabilização do radical DPPH quanto na inibição da oxidação no sistema lipídico estudado. No entanto mais trabalhos são necessários para avaliar diferentes sistemas de oxidação, bem como estudos toxicológicos para garantir níveis seguros de consumo. / The oxidation is one of major deterioration in food lipids, causing losses in quality and nutritional value. To avoid this, antioxidants are used and the phenolic compounds are the main component of antioxidant applied and presented in foods, and also may be present in agroindustrial residues, usually discarded by the industry. The objective of this study was to identify ideal conditions to extract phenolic compounds from acerola fruits (Malpighia emarginata), coffee grounds (Coffea arabica) and lychee skins (Litchi chinensis) by means of surface response methodology (RSM), evaluation of antioxidant activity of extracts in in vitro essays, application of selected extracts in emulsions systems during accelerated test. The response surface methodology was applied to each material in order to identify the effects of temperature, ethanol concentration and time in extraction processes. Antioxidant activity by DPPH stabilization and its kinectis evaluation were performed and compared to TBHQ. Furthermore, selected extracts (50 to 200 mg.kg-1) were added in emulsion system and subject to accelerated oxidation test. RSM have shown that phenolic compounds of each material are extractable in different conditions. Acerola fruits aqueous extracts present maximum phenolic content (175.87 mg GAE.g-1) and the higher DPPH antioxidant activity, followed by coffee grounds and lychee skins extracts (26.37 and 25.87 mg GAE.g-1). Acerola extracts also presented the best stabilization of DPPH in kinetic study, with results comparable to the TBHQ effect (650.53 TEAC). Acerola extract added to the emulsion showed a good antioxidant effect in all concentrations during accelerated test, resulting in final oxidation levels (mmol hydroperoxides kg-1) comparable to TBHQ in the same concentration. Coffee grounds and lychee skins extracts were not efficient in avoid hydroperoxides increasing. Results showed the importance of studying extractions conditions for each material. Acerola extract showed the most efficient antioxidant activity both in DPPH kinetics DPPH stabilization and inhibition of lipid oxidation in the system studied. However, further work is needed to evaluate different oxidation systems, as well as toxicological studies to ensure safe levels of consumption.

Made available in DSpace on 2017-07-21T18:53:04Z (GMT). No. of bitstreams: 1 Suelen Brazo.pdf: 3105120 bytes, checksum: d7c908a74b56c6c9c7dbcc45ef93c32a (MD5) Previous issue date: 2015-02-13 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Anthocyanins are natural colorants that provide attractive and bright colors to most plants, coloring them the bright red to violet. Due to its instability facing the food processing steps and their difficulty of extraction are still little used in the food industry. Flavonoids not anthocyanins, amino acids, nucleosides, organic acids and anthocyanins themselves may act as copigments enabling the anthocyanins as natural colorants. The objective was to evaluate the approximate composition, phenolic compounds, antioxidant activity of blackberries Brazo (Morus nigra L.) and the stability of anthocyanins. The effect of the addition of organic acids and polyphenolic extract of Camellia sinensis raw extract of blackberries anthocyanins as copigments. The copigmentation reaction buffer at pH 3.0, was confirmed by UV-Visible absorption and by high high-performance liquid chromatography analysis. The calculated half-life and the color retention percentage. The response surface methodology was effective to estimate the effect of three independent variables on anthocyanins extraction, the optimal values suggested for the extraction with citric acid were time of 60 min., ratio of 1 g / 30 mL and a temperature of 60 ºC and the extraction with hydrochloric acid have been suggested the same conditions of time and ratio, but at 20 ° C. The increase in relation copigmento / anthocyanins was evidenced by the bathochromic and hyperchromic effects. The interaction of anthocyanins was significantly affected by temperature (5 ± 3 ° C and 25 ± 3 ° C) and the environment (light and dark). The stability of anthocyanins with added copigmento increased half-life and the color retention percentage of sample. Maximum stability was achieved in samples with the addition of caffeic acid at a temperature of 5 ° C ± 3 ° C in the dark. Anthocyanins added caffeic acid and malic showed half-life of 242.40 ± 25.40 and 200.48 ± 12.15 days, respectively, bringing prospects for application of this natural dye. / Antocianinas são corantes naturais que proporcionam cores atrativas e brilhantes à maioria das plantas, colorindo-as do vermelho vivo ao violeta. Devido à sua instabilidade frente às etapas de processamento de alimentos e a sua dificuldade de extração ainda são pouco utilizados na indústria alimentícia. Flavonóides não antociânicos, aminoácidos, nucleosídios, ácidos orgânicos e as próprias antocianinas podem agir como copigmentos viabilizando as antocianinas como corantes naturais. O objetivo do trabalho foi avaliar a composição aproximada, os compostos fenólicos totais, a atividade antioxidante de amoras Brazo (Morus nigra L.) e a estabilidade das suas antocianinas. Foi avaliado o efeito da adição dos ácidos orgânicos e extrato polifenólico de Camellia sinensis ao extrato bruto de antocianinas de amora como copigmentos. A reação de copigmentação em solução tampão pH 3,0, foi confirmada por espectrofotometria de absorção UV-Visível e por análise cromatográfica líquida de alta eficiência. Calculou-se o tempo de meia vida e a percentagem de retenção de cor. A metodologia de superfície de resposta foi eficaz para estimar o efeito de três variáveis independentes sobre a extração de antocianinas, os ótimos valores sugeridos para a extração com ácido cítrico foram tempo de 60 min., razão de 1g / 30 mL e uma temperatura de 60°C e para a extração com ácido clorídrico foram sugeridas as mesmas condições de tempo e razão, porém a uma temperatura de 20°C. O aumento na relação copigmento/antocianinas foi evidenciado pelos efeitos batocrômico e hipercrômico. A interação das antocianinas foi significativamente afetada pela temperatura (5±3ºC e 25±3ºC) e o ambiente (luz e escuro). A estabilidade das antocianinas com adição de copigmento aumentou o tempo de meia vida e a porcentagem de retenção de cor das amostras. A estabilidade máxima foi alcançada nas amostras com adição de ácido cafeico em temperatura de 5ºC±3°C e no escuro. As antocianinas adicionadas de ácido cafeico e málico apresentaram tempo de meia vida de 242,40 ± 25,40 e 200,48 ± 12,15 dias, respectivamente, trazendo perspectivas para aplicação deste corante natural.

O aumento nos rejeitos industriais e a contínua produção de resíduos causam muitas preocupações no âmbito ambiental. Neste contexto, o descarte de pneus usados tem se tornado um grande problema por conta da pequena atenção que se dá à sua destinação final. Assim sendo, essa pesquisa propõe a produção de uma mistura polimérica com polipropileno (PP), a borracha de etileno-propileno-dieno (EPDM) e o pó de pneu (SRT). A Metodologia de Superfície de Resposta (MSR), coleção de técnicas estatísticas e matemáticas úteis para desenvolver, melhorar e optimizar processos, foi aplicada à investigação das misturas ternárias. Após o processamento adequado em extrusora de dupla rosca e a moldagem por injeção, as propriedades mecânicas de resistência à tração e resistência ao impacto foram determinadas e utilizadas como variáveis resposta. Ao mesmo tempo, a microscopia eletrônica de varredura (MEV) foi usada para a investigação da morfologia das diferentes misturas e melhor interpretação dos resultados. Com as ferramentas estatísticas específicas e um número mínimo de experimentos foi possível o desenvolvimento de modelos de superfícies de resposta e a otimização das concentrações dos diferentes componentes da mistura em função do desempenho mecânico e além disso com a modificação da granulometria conseguimos um aumento ainda mais significativo deste desempenho mecânico. / The increase in industrial waste and solid waste production cause many concerns in the environment. In this context, the disposal of used tires has become a major problem because of the little attention given to their final destination. Therefore, this research proposes the production of a polymer blend of polypropylene (PP), rubber of ethylene-propylene-diene (EPDM) and tire dust (SRT). The Response Surface Methodology (RSM), a collection of mathematical and statistical techniques useful for developing, improving and optimizing processes, was applied to the investigation of ternary mixtures. After proper processing on twin screw extrusion and injection molding, the mechanical properties of tensile and impact strength were determined and used as response variables. At the same time, scanning electron microscopy (SEM) was used to investigate the morphology of different blends and better interpretation of results. With specific statistical tools and a minimum number of experiments it was possible to develop models of response surfaces and the optimization of the concentrations of different components of the mixture depending on the mechanical performance and even with the change in particle size could increase even more significant this mechanical performance.

La banane plantain est un fruit couramment consommé au Congo-Brazzaville sous la forme cuite en tant qu’accompagnement. Le procédé de friture profonde de la banane plantain est une pratique très répandue, souvent mal maitrisé, compte tenu de l’utilisation prolongée des huiles de friture entrainant la formation de composés néfastes à la santé. En effet, depuis un certain nombre d’années, la consommation d’huiles locales (soja, palme) et leurs dégradations après chauffage constituent une des préoccupations principales pour la sécurité sanitaire du consommateur. Les objectifs de cette thèse consistaient à étudier l’influence du procédé de la friture profonde appliqué à la banane plantain Musa AAB, variété « Harton » au stade de maturité 7, consommée sous la forme de disque. Nous avons dans un premier temps appliqué la méthodologie des surfaces de réponse au procédé de friture profonde, afin de déterminer l’influence de trois paramètres (temps / température / quantité) sur différentes réponses (couleur, dureté, teneurs en huile et en eau des produits frits) en fonction de la nature de l’huile utilisée (soja, palme, huile Frial). Une fois les conditions optimales obtenues sur la banane plantain en condition domestique, l’étude s’est focalisée sur l’évolution de la stabilité des huiles de friture par le suivi des paramètres physico-chimiques de dégradation de ces dernières (composés polaires totaux, acides gras libres, composés primaires et secondaires d’oxydation). Des analyses de la couleur, du point de cristallisation et de la viscosité sont venues compléter l’étude et ont permis d’établir la durée d’utilisation optimale de ces huiles en fonction de la quantité de banane plantain à frire. Enfin, les mécanismes de transfert d’eau et d’absorption d’huile dans la matrice ont été étudiés et modélisés en fonction de la température, de l’épaisseur et du temps d’immersion. / Plantain is a fruit commonly consumed in Congo - Brazzaville in its cooked form, as an accompaniment. The process of deep frying plantain is a widespread practice, often poorly mastered, given the prolonged use of frying oils, leading to the formation of compounds harmful to health. Indeed, for a number of years, the consumption of local oils (soybean, palm) and their degradation after heating are cause of major concern for consumer safety. The objectives of this study were to investigate the influences of the deep frying process applied on the Musa AAB plantain of "harton" variety, maturity level 7, as consumed in sliced form. We initially applied response surface methodology (RSM) to the deep frying process to determine the influence of three parameters (time / temperature / quantity) on different responses (color, hardness, oil uptake and water content) according to the type of oil used (soybean, palm, Frial oils). Once optimal conditions were obtained from the plantain in domestic conditions, the study focused on the evolution of frying oil stability, by monitoring physicochemical parameters of the degradation of the aforementioned (total polar compounds, free fatty acids, primary and secondary oxidation products). Analyses of color, crystallization point and viscosity, were added to the study and helped establish the optimal duration of use for these oils depending on the quantity of plantains fried. Lastly, the mechanisms of water transfer and oil uptake in the matrix has been studied and modeled according to temperature, thickness and immersion time.

&alpha / -Galactosidase is an exo-glycosidase that hydrolyses non-reducing, &alpha / -1,6-linked &alpha / -galactose units from oligosaccharides, galactomannans, and galactolipids. &alpha / -Galactosidase activity has biotechnological, industrial, and medical importance. &alpha / -Galactosidase from A. fumigatus IMI 385708, in particular, can catalyse unique hydrolysis and transgalactosylation reactions on polymeric substrates. In this study, &alpha / -galactosidase of the human pathogen A. fumigatus IMI 385708 was first produced in a GRAS organism, Aspergillus sojae. For this aim, &alpha / -galactosidase gene (aglB) of A. fumigatus IMI 385708 was ligated onto pAN52-4 vector (Acc. No: Z32699) and transformed into Aspergillus sojae ATCC11906, under the control of the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (gpdA) of A. nidulans and the signal sequence of glucoamylase gene (glaA) of A. niger. This allowed high level of &alpha / -galactosidase production on glucose instead of locust bean gum (2.45 U/mL), corresponding to a 3-fold increase in volumetric production. Next, using response surface methodology, carbon and nitrogen sources and agitation speed were optimized (10.5% molasses (w/v) / 1.3% NH4NO3 (w/v) / 276 rpm). Compared to non-optimized cultivation, a further 4-fold increase in &alpha / -galactosidase production (10.4 U/mL) was achieved. Recombinant &alpha / -galactosidase was purified 18.7-fold using Anion Exchange and Hydrophobic Interaction Chromatography with an overall yield of 56% and 64.7 U/mg protein. The Vmax and Km values for the hydrolysis of p-nitrophenyl &alpha / -D-galactopyranoside were 78 U/mg protein and 0.45 mM, respectively. Optimum pH and temperature for &alpha / -galactosidase activity were between pH 4&ndash / 6 and 50&ndash / 60 &deg / C, respectively. Among the tested chemical agents, Ag+, Hg2+, and Fe2+ drastically decreased the activity, while biotin, I+1, Mn+2, Pb+2, Li+1, and Mg+2 enhanced between 12&ndash / 29%. To analyse the influence of osmotic stress as a means of further inducing &alpha / -galactosidase production, salt was added into the complete growth medium. In addition to enzyme production, fungal growth and morphology were analysed for both &lsquo / salt-adapted&rsquo / and &lsquo / salt non-adapted&rsquo / A. sojae Ta1 cells in the presence of KCl, MgCl2, MgSO4, NaCl, and Na2SO4 at 1 M and 2 M. Accordingly, 3-fold increase in &alpha / -galactosidase production was achieved by non-adapted cells in the presence of 1 M NaCl. Exposure of A. sojae Ta1 cells to salt resulted in predominantly mycelial form, rather than the pellet form observed under normal conditions. Finally, the transgalactosylation ability of &alpha / -galactosidase was studied. &alpha / -Galactosidase efficiently catalysed galactose transfer to different monosaccharides and disaccharides in the presence of pNP&alpha / Gal as monitored by TLC, ESI-MS, and HPLC.

The natural ability of filamentous fungi to synthesize, glycosylate, and secrete high levels of protein products has made them potentially attractive hosts for heterologous protein production. Advances in fungal genetics enabled the expression of several high value proteins in filamentous fungi. Particularly the genus, Aspergillus has proven to be potentially useful for the expression of eukaryotic gene products. This thesis pertains to the optimization of recombinant protein production by the fungal host, Aspergillus niger. The target recombinant protein of interest is hen egg white lysozyme (HEWL). This protein encoded in the genome resulting in relatively stable gene construct; however, it is subject to extracellular protease attack. The objective of the proposed research is the development and application of engineering methodology for the analysis and optimization of a fungal bioprocess for recombinant protein production. The underlying hypothesis is that a significant improvement of target protein productivity is achievable by using appropriate optimization techniques. To accomplish this, during the first phase of this study a statistically based experimental method was used to systematically elucidate the effect of medium components (starch, peptone, ammonium sulfate, yeast extract, and CaCl₂.2H₂O) on hen egg white lysozyme production by Aspergillus niger HEWL WT-13-16. A 2⁵⁻¹ fractional factorial design augmented with center points revealed that peptone, starch, and ammonium sulfate were the most significant factors, whereas the other medium components were not important within the levels tested. Then, the method of steepest ascent was employed to approach the proximity of optimum. This task was followed by a central composite design to develop a response surface for medium optimization. The optimum medium composition for lysozyme production was found to be: starch 34 g/L, peptone 34 g/L, ammonium sulfate 11.9 g/L, yeast extract 0.5 g/L, and CaCl₂.2H₂O 0.5 g/L. This medium was projected to produce theoretically 212 mg/L lysozyme. Using this optimized medium, an experimentally observed maximum lysozyme concentration of 209±18 mg/L verified the applied methodology. A second optimization approach was based on metabolic flux analysis (MFA). A comprehensive metabolic network comprising three intracellular compartments (cytoplasm, mitochondrion and peroxisome) was developed for Aspergillus niger. The metabolic flux network included carbohydrate and amino acid metabolism in both anabolic and catabolic reactions. According to experimental observations, the time course of fermentation was divided into five phases, each with unique physiological properties. The network was used to form a set of linear algebraic equations based on the stoichiometry of the reactions by assuming pseudo-steady state for intracellular metabolites. The metabolic flux model consists of 137 metabolites and 287 processes, of which 181 represent biochemical conversions and 106 represent transport processes between the different compartments and the extracellular environment. In addition, due to the physiological evidence some biochemical reactions considered to be active only in one direction. Linear programming was used for optimizing of the specific growth rate as the objective function in combination with 37 measured input and output fluxes of the key metabolites to evaluate corresponding intracellular flux distributions throughout the batch fermentations. The general applicability of the methodology was evaluated by establishing commonality to optimize recombinant HEWL production. The proposed model was able to predict correctly the specific growth rate, oxygen uptake rate, and carbon dioxide evolution rate with good precision. The results of the metabolic flux and sensitivity analysis were employed for medium design. Growth was biphasic; glucose was utilized initially as the carbon source and was followed by its oxidation product, gluconate, later. Logarithmic sensitivity analysis revealed that the addition of proline, alanine and glutamate benefited growth in defined media. The experimental observations and flux analysis showed that tyrosine was a potential candidate for biomass production improvement. The two amino acids, namely proline and tyrosine benefited biomass production during the initial growth phases. Glutamate and alanine were particularly important during the latter stages of the batch process. A series of growth studies were conducted with the identified amino acids added in the medium. In these preliminary nutritional experiments the contribution to growth enhancement was 46% for proline, 23% for glutamate, and 22% for tyrosine. Model predictions were further verified by conducting batch and fed-batch fermentations in a 7- liter bioreactor. The programmed addition of four amino acids (proline, glutamate, alanine, and tyrosine) according to a predetermined schedule resulted in a 44% improvement in biomass and 41% improvement in recombinant protein production. The experiments also confirmed the model prediction that extra amount of amino acids besides the identified ones would not significantly enhance biomass and the recombinant protein production. A computer-based control system was developed for the on-line monitoring and control of the major state variables (e.g., temperature, pH, and DO) during the time course of fermentation. The graphical programming environment, LabVIEW was used to acquire and integrate these variables in a supervisor computer. The temperature of the bioreactor during sterilization and fermentation was controlled using a cascade methodology. The controller parameters of the master and slave loops were determined experimentally to yield a smooth response with minimum overshoot of both the bioreactor and jacket temperatures. The program scheduled various required steps in an established order during the fermentation. This feature of the software guarantees that every necessary operation will be met. The graphical representation of the process is displayed on the screen and helps the user to follow the process and perform the required adjustments. Furthermore, different variables can be observed simultaneously and saved in text or spreadsheet files for further analysis.