Neste trabalho são apresentados alguns resultados relacionados a estudos sobre o comportamento eletroquímico do Cu(II) em meio contendo diferentes peptídeos (tri, tetra e pentaglicina) em função da acidez utilizando-se voltametria cíclica. Os experimentos foram realizados em uma faixa de pH entre 7,0 e 10,0. Experimentos espectrofotométricos e coulométricos também foram feitos, ambos para se caracterizar as espécies de Cu(II) e Cu(III) em solução e provar a existência da degradação dos complexos de Cu(III). Além disso, experimentos para esses sistemas também foram realizados em diferentes temperaturas utilizando eletrodo rotativo na configuração disco-anel. Estudos eletroanalíticos demonstraram a potencialidade de se empregar o sistema Cu(II)/Cu(III) no desenvolvimento de metodologia eletroanalítica para a determinação de sulfito. Neste contexto, um método amperométrico indireto para a determinação de sulfito em fluxo é proposto neste trabalho. O método baseia-se na medida amperométrica (OV vs Ag/AgCI (sat. NaCI)) do Cu(III) gerado quimicamente no processo de auto¬-oxidação do S(IV) em soluções de Cu(II)/tetraglicina na presença de traços de Ni(II) utilizando uma célula em fluxo com sistema adequado para separação de gases. A oxidação espontânea do complexo pelo oxigênio dissolvido e S(IV) é muito lenta ao contrário do que ocorre quando há traços de Ni(II). A faixa de trabalho foi de 20-100 µmoIL-1 com limite de detecção de 2µmoIL-1 (S/N=3). A repetibilidade para a determinação (100µmoIL-1) levou a um desvio de 4,9% (n = 38), valor superior aos encontrados na literatura para determinações de sulfito em vinho por FIA com detecção amperométrica indireta (1-4,6%), e a freqüência analítica foi de 40h-1, valor coerente com os dos outros métodos amperométricos. Ambas as análises de sulfito em amostras de vinhos e de sucos tiveram seus resultados comparados com aqueles obtidos utilizando-se o método padrão. / The electrochemistry of Cu(II) in aqueous solutions containing triglycine, tetraglycine and pentaglycine had been investigated at glassy carbon surfaces. Experiments were carried out in the pH range between 7.0 and 10.0. A correlation between voltammetric waves and the nature of the Cu(II) species involved in the electrochemical steps was performed. Spectrophotometric and coulometric studies were also done to characterise Cu(II) and Cu(III) species in solution and prove the existence of a degradation step related to Cu(III) complexes. Rotating ring-disc voltammetry was employed to follow this chemical step coupled to the electrode reaction, at three different temperatures. Data show that Cu(III)/triglycine species degrade, with faster kinetics, when the results are compared with those obtained for tetraglycine and pentaglycine. An indirect amperometric method for the determination of sulfite in a flow injection configuration is described. The method is based on the amperometric measurement (OV vs Ag/AgCI (sat. NaCI)) of Cu(III) chemically generated by the sulfite induced autoxidation of Cu(II)/tetraglycine complex in the presence of traces of Ni(II) with a manifold that incorporates flow extraction of sulfite as SO2 through a PTFE membrane. The operational range was 2µmoIL-1 with a detection limit of 2µmolL-1 (S/N = 3). The repeatability for the determination (100 2µmolL-1) was evaluated to be 4.9% (n = 20) and the analytical frequency was 40h-1. Results for 3 wines and 2 juices samples showed excellent agreement with those obtained by using a recommended procedure for sulfite analysis.

Introdução: Os crustáceos ocupam o quarto lugar em volume de captura no mundo e o seu elevado valor comercial, os tornam relevantes em relação ao montante financeiro gerado pela atividade pesqueira. Atualmente, a garantia da qualidade é requisito para a permanência no mercado, sendo exigida dentro de especificações previamente estabelecidas, tanto pelas autoridades sanitárias brasileiras como dos países para os quais o pescado é exportado. Objetivo: Caracterizar a qualidade dos camarões sete-barbas desembarcados na região Metropolitana da Baixada Santista, visando a segurança alimentar e a sustentabilidade da cadeia produtiva e, a partir do método do índice de qualidade (QIM), desenvolver um protocolo de avaliação do frescor para o camarão marinho Xiphopenaeu kroyeri inteiro armazenado em gelo. Métodos: Após o levantamento dos principais pontos de desembarque do camarão sete-barbas na região Metropolitana da Baixada Santista, SP., considerando a produção e frota pesqueira; realizou-se a caracterização da qualidade do camarão desembarcado na região pela determinação da biometria e dos parâmetros físicos, químicos e microbiológicos regulados (pH, bases nitrogenadas voláteis totais (BNVT), trimetilamina (TMA), residual de dióxido de enxofre, mercúrio e pesquisa de Staphylococcus aureus e Salmonella spp). Foi realizada a determinação instrumental de cor e textura; a quantificação das substâncias reativas ao ácido tiobarbitúrico (TBARS), as contagens de coliformes totais e termotolerantes, de clostrídios sulfito redutores e testes sensoriais de aceitação. Para otimizar o protocolo de avaliação do frescor do camarão X. kroyeri armazenado em gelo, com e sem o uso de sulfito, foram conduzidos ensaios de vida útil empregando-se as análises física e químicas (pH, BNVT, TMA, TBARS e aminas biogênicas), microbiológicas (S.aureus, Salmonella spp, coliformes totais, bolores e leveduras, contagens padrão de bactérias heterotróficas aeróbias psicrotróficas e mesófilas) e testes sensoriais, onde se utilizou o protocolo elaborado. Resultados: A análise global dos parâmetros de qualidade de 20 desembarques mostra que os camarões apresentavam-se em desacordo com os limites propostos pelo Regulamento de Inspeção Industrial e Sanitária de Produtos de Origem Animal, embora atendessem aos padrões microbiológicos do Ministério da Saúde e, em alguns casos, foram qualificados sensorialmente como muito bons. Indicando a necessidade de adequação dos limites estipulados para os parâmetros físico e químicos nacionais vigentes para o consumo do camarão sete-barbas. Os padrões de coloração para a espécie parecem bem promissores, mas devem ser mais estudados, frente a gama de fatores que afetam a mesma. A rejeição sensorial do produto sem sulfito ocorreu primeiro, pelo escurecimento do cefalotórax do camarão (melanose) no 2° dia de armazenamento em gelo, tanto para o produto cru como para o cozido. A eficiência do uso de sulfitos, em solução a 1,25 por cento , na conservação do camarão foi indicada pela rejeição sensorial que ocorreu com 3 ou 4 dias e foi determinada pela presença de odores não característicos (amoniacal, ácido e/ou sulfídrico) no camarão cozido, os quais apresentaram correlação com o período de armazenamento em gelo (r= 0,70; p < 0,05). O limite residual de dióxido de enxofre em camarões provenientes de pescas longas (3 a 10 dias) apresentou-se sempre acima do limite permitido pela Agência Nacional da Vigilância Sanitária, confirmando que o uso do metabissulfito de sódio para retardar o aparecimento da melanose, deve ser cauteloso, com vistas a não causar prejuízos à saúde do consumidor. A regressão linear dos parâmetros sensoriais propostos no protocolo QIM (cor, odor, melanose, aderência do cefalotórax ao corpo e aderência da carapaça) mostrou a correlação dos mesmos com o período de estocagem em gelo, tanto para os camarões do controle, como para os tratados com sulfito. Conclusão: Com vistas à classificação de um lote de camarão sete-barbas como aceitável para o consumo são sugeridos os seguintes limites: pH 7,7; BNVT 35 mg.100 g -1 de músculo; firmeza muscular (entre os 2 primeiros segmentos da cauda) > 6 N, sendo desejável valores superiores para o camarão recém desembarcado. Há necessidade de melhorias nas práticas a bordo das embarcações pesqueiras. O protocolo QIM para o camarão X.kroyeri inteiro e conservado em gelo, consiste de 5 parâmetros e seus descritores, os quais somam 11 pontos de demérito e pode ser utilizado em camarões tratados ou não com sulfitos / Introduction: Crustaceans occupy the fourth place in world capture volume and their high economic value makes them relevant fishery activity in the market share. Nowadays, quality assurance is a mandatory requirement to be kept in market, due specific established requirements from national sanitary authorities or from imported countries. Objectives: Characterize the quality of the seabob-shrimp landed in Santos, SP (BR), to food security and sustainability of supply chain and, using the quality index method (QIM) to develop a protocol for freshness evaluation of Raw whole Xiphopenaeus kroyeri shrimp stored on ice. Methods: After the assessment of the main landing points of the seabob-shrimp in the metropolitan area of Santos, SP (BR), considering the production and shipping fleet; the characterization of the shrimp quality landed in the region was carried out by biometry determination and physical, chemical and regulated microbiological parameters (pH, total volatile basic nitrogen (TVB-N), trimethylamine (TMA), sulfur dioxide residue, mercury, S. aureus and Salmonella spp,). Instrumental determination of the color and texture, substances thiobarbituric acid reactive quantification (TBARS), total and fecal coliforms counts, Clostridium sulfide reductors and sensory acceptance tests were also performed. To optimize the protocol for the evaluation of ice stored X. kroyeri shrimp freshness, treated or not with sulfite, trials of shelf life were conducted using physical and chemicals analyzes (pH, TVB-N, TMA, TBARS and biogenic amines), microbiological (S.aureus, Salmonella spp, total coliforms counts, molds and yeasts, Mesophilic and Psychrotrophic Bacteria) and sensory analysis, where established protocol was used. Results: Overall analysis of the quality parameters of 20 landings has shown that the shrimps were in disagreement with limits proposed by the Regulation of Industrial and Sanitary Inspection of Animal Products, although the microbiological standards requirements of the Ministry of Health were attended and, in some cases, were sensorially qualified as \"very good\". This implies the necessity of physical and chemical parameters adequacy for the current national seabob-shrimp consumption. The staining patterns for the species look very promising, but should be further studied, since the great range of factors that affect it. The sensory rejection of products without sulfite occurred first with a darkening of the shrimp carapace (melanosis) in the 2nd day of storage on ice, as much to the crude product as cooked one. The efficiency of the sulphites, in solution at 1.25%, in the seabob-shrimp conservation was indicated by the sensory rejection that occurred in 3 or 4 days and was determined by the presence of uncharacteristic odors (ammonia, acid and/ or hydrogen sulfide) in cooked shrimp, that showed correlation with ice storage period (r = 0.70, p < 0.05). Results of residual sulfur dioxide in shrimps from long fisheries (3 to 10 days) were always above the limit allowed by ANVISA, which confirms that the usage of sodium metabisulfite to delay the onset of melanosis should be cautious to avoid consumer health. The linear regression of the parameters proposed in the protocol QIM (color, odor, melanosis, adhesion of the cephalothorax and of the carapace to the body) showed a correlation between the period of storage on ice for both, control shrimps and treated with sulphite. Conclusion: In order to classify a batch of seabob-shrimp as acceptable for consumption the following limits are suggested: pH 7.7; TVB-N 35 mg.100 g-1 of muscle; muscle firmness (between the first and second segment of tail) > 6 N, with higher values for freshly landed shrimp. It is necessary to improve practices onboard of fishing vessels. The Quality Index protocol for shrimp X. kroyeri \"in natura\" consists of five parameters and their descriptors, with total 11 demerit points and can be used in shrimps treated or not with sulfite

The goal of this thesis was to adopt a reverse flow injection analysis (rFIA) technique to the fluorometric analysis of the reaction o-phthaldialdehyde (OPA) with ammonium, allowing accurate measurements of ammonium concentrations lower than the detection limit of the widely used indophenol blue (IPB) colorimetric method while accounting for the background fluorescence of seawater. Ammonium is considered an essential nutrient for primary productivity, especially in the nutrient depleted surface ocean where as the most reduced form of dissolved inorganic nitrogen, it is readily assimilated via metabolic pathways. Challenges in the quantification of ammonium require more sensitive analytical techniques for a greater understanding of the biogeochemical cycling of ammonium in the oligotrophic ocean. On-line and automated flow analysis techniques are capable of mitigating some of the challenges. Fluorescent-based methods out-perform colorimetric methods in terms of detection limits and sensitivity. Presented here is the development of an rFIA technique paired with an OPA-sulfite chemistry. For this method, a sulfite-formaldehyde reagent is mixed with the sample stream and then injected with the OPA reagent before being heated. Fluorescence is measured before and at the peak of the OPA injection, differentiating the background fluorescence from the analyte signal. Experiments to optimize reaction parameters and characterize the effects of salinity and potentially interfering species were conducted. The newly developed method offers a reasonable throughput (18 samples per hour), low limit of detection (1.1 nM) ammonium analysis technique with automatic background fluorescence correction suitable for oligotrophic seawater as a preferable alternative to the low sensitivity and high limit of detection IPB colorimetric method.

Single stage and two-stage sodium sulfite cooking were carried out on either spruce, pine or pure pine heartwood chips to investigate the influence of several process parameters on the initial phase of such a cook down to about 60 % pulp yield. The cooking experiments were carried out in the laboratory with either a lab-prepared or a mill-prepared cooking acid and the temperature and time were varied. The influences of dissolved organic and inorganic components in the cooking liquor on the final pulp composition and on the extent of side reactions were investigated. Kinetic equations were developed and the activation energies for delignification and carbohydrate dissolution were calculated using the Arrhenius equation. A better understanding of the delignification mechanisms during bisulfite and acid sulfite cooking was obtained by analyzing the lignin carbohydrate complexes (LCC) present in the pulp when different cooking conditions were used. It was found that using a mill-prepared cooking acid beneficial effect with respect to side reactions, extractives removal and higher stability in pH during the cook were observed compared to a lab-prepared cooking acid. However, no significant difference in degrees of delignification or carbohydrate degradation was seen.  The cellulose yield was not affected in the initial phase of the cook however; temperature had an influence on the rates of both delignification and hemicellulose removal. It was also found that the  corresponding activation energies increased in the order:  xylan, glucomannan, lignin and cellulose. The cooking temperature could thus be used to control the cook to a given carbohydrate composition in the final pulp. Lignin condensation reactions were observed during acid sulfite cooking, especially at higher temperatures. The LCC studies indicated the existence of covalent bonds between lignin and hemicellulose components with respect to xylan and glucomannan. LCC in native wood showed the presence of phenyl glycosides, ϒ-esters and α-ethers; whereas the α-ethers  were affected during sulfite pulping. The existence of covalent bonds between lignin and wood polysaccharides might be the rate-limiting factor in sulfite pulping. / The sulfite pulping process is today practised in only a small number of pulp mills around the globe and the number of sulfite mills that use sodium as the base (cation) is less than five. However, due to the increasing interest in the wood based biorefinery concept, the benefits of sulfite pulping and especially the sodium based variety, has recently gained a lot of interest. It was therefore considered to be of high importance to further study the sodium based sulfite process to investigate if its benefits could be better utilized in the future in the production of dissolving pulps. Of specific interest was to investigate how the pulping conditions in the initial part of the cook (≥ 60 % pulp yield) should be performed in the best way. Thus, this thesis is focused on the initial phase of sodium based single stage bisulfite, acid sulfite and two-stage sulfite cooking of either 100 % spruce, 100 % pine or 100 % pine heartwood chips. The cooking experiments were carried out with either a lab prepared or a mill prepared cooking acid and the temperature and cooking time were varied. Activation energies for different wood components were investigated as well as side reactions concerning the formation of thiosulfate. LCC (Lignin carbohydrates complexes) studies were carried out to investigate the influence of different cooking conditions on lignin carbohydrate linkages.

Master of Science / Department of Grain Science and Industry / Sajid Alavi / Texturized vegetable protein (TVP) based foods offer several advantages compared to animal protein, including lower costs and improved health benefits. Wheat gluten is often processed using extrusion to produce TVP. Processing aids, such as reducing agents (example, cysteine and sodium metabisulfite) and pH modifiers (example, tetra potassium phosphate) aid in texturization. Reduction of sulfhydryl groups, cleavage of disulfide bonds, and reformation of bonds between elongated protein molecules results in protein aggregation and texturization. This study focused on development of a fundamental understanding of these mechanisms for texturization using analytical tools such as the phase transition analyzer (PTA), in combination with lab- and pilot-scale extrusion. The abovementioned three chemicals were added to four varieties of gluten. The control treatment had no additives. PTA was used to understand the operative flow properties of gluten in an environment similar to an extrusion system. Addition of sulfite (0.18%) and cysteine (0.18%) lowered the thermal softening (Ts:36.6-44.1 °C) and thermal flow (Tf:79.6-105.6 °C) temperatures of all varieties of gluten as compared to the controls (Ts:38.8-48.2 °C; Tf:91.7-112.2 °C). Phosphate (3%) did not have the same lowering effect on Ts (40.2-47.0 °C) and Tf (96.2-108.2 °C), indicating a different mechanism. Extrusion studies were conducted to gain an understanding of the reformation of disulfide bonds and texturization. Two of the varieties of gluten, a “superior” one that texturizes well and an “inferior” gluten requiring texturizing aids, were processed on a lab-scale extruder. Pilot scale extrusion was used to process the other two glutens (“superior” varieties) to obtain commercial quality products, which were evaluated for degree of texturization (hydration rate, absorption index and integrity). During lab-scale extrusion, texturization was observed only in the case of phosphate and corresponded with an increase in specific mechanical energy (SME) as compared to the control, indicating disulfide bond reformation. Phosphate also led to significantly (p<0.05) better texturization during pilot-scale extrusion, although SME trends were different due to higher in-barrel moisture and a more ideal extrusion system. Fourier Transform Infrared Spectroscopy was used to examine protein structural changes and indicated a loss of α-helix structure in TVP with an increase in β-sheet formation.

O objetivo desse trabalho foi avaliar a produção de \'H IND.2\' a partir da degradação de papel sulfite com a utilização de consórcio microbiano obtido do fluido ruminal, na presença e ausência da celulase. Para obtenção de consórcio de bactérias essencialmente produtoras de \' H IND.2\', fluido de rúmen in natura, utilizado como inóculo, foi submetido a tratamento ácido (pH 3 por 24 h), e posteriormente enriquecido em meio de cultura Del Nery modificado em diluições seriais. Nos ensaios de produção de \'H IND.2\' foi utilizado 10% (v/v) desse inóculo em reatores com diferentes concentrações de papel e celulase, e em reatores controle, nos quais não houve adição de celulase. Reator anaeróbio em batelada, em triplicata, com papel sulfite e meio Del Nery modificado, foi mantido a 37 ºC, pH inicial 7,0, com headspace preenchido com \'N IND.2\' (100%) para os seguintes ensaios: (1) 0,5 g papel/L e 4 mL celulase/L; (2) 2,0 g papel/L e 15 mL celulase/ L; (3) 4,0 g papel/L e 30 mL celulase/L; (CT 1) 0,5 g papel/L; (CT 2) 2,0 g papel/L; (CT 3) 4,0 g papel/L. Os rendimentos de \'H IND.2\' foram 42, 26,6 e 24 mmol\' H IND.2\'/g papel para os ensaios 1, 2 e 3, respectivamente. Não houve produção de \'H IND.2\' nos reatores controle. O consumo de substrato, avaliado como glicose, foi de 56%, 56,6% e 65,4% para os ensaios 1, 2 e 3, respectivamente. Nos reatores controle não foi detectado consumo de papel. Em todos os reatores com celulase foi detectado principalmente ácido butírico, além de ácido acético e álcoois. Nos reatores controle foram detectados principalmente ácido acético e ácido iso-butírico. Para os ensaios 1, 2, 3, CT 1, CT 2 e CT 3, os valores de pH final foram 4,6; 3,7; 3,5; 6,8; 6,6 e 6,5, respectivamente. Bacilos Gram positivos e formadores de endósporos foram predominantes em todos os reatores. A partir da análise de PCR/DGGE foi possível observar alteração de 78% entre a comunidade original do fluido de rúmen e a comunidade microbiana presente ao final dos ensaios. Por meio da clonagem e sequenciamento do gene RNAr 16S, verificou-se similaridade de 95% a 99% com Clostridium no consórcio bacteriano do reator com papel e celulase. A produção biológica de \'H IND.2\' nos reatores com celulase foi provavelmente devido as bactérias que estiveram presentes nas condições estudadas, identificadas, principalmente, como Clostridium sp., reconhecidas como produtoras de \'H IND.2\', ácidos voláteis e álcoois. A adição de celulase foi essencial para a hidrólise do papel em carboidratos solúveis, permitindo a produção de H2 pelas bactérias presentes no consórcio microbiano. / The aim of this study was to evaluate the production of \'H IND.2\' from office paper degradation by using a microbial consortium obtained from rumen fluid, in the presence and absence of cellulase. In order to obtain a microbial consortium consisting mainly of hydrogen producing bacteria, rumen fluid, used as inoculum, was subjected to acid pretreatment (pH 3 for 24 h), and subsequently enriched in the modified Del Nery medium in serial dilutions. In hydrogen production assays there was used 10% (v/v) of the pretreated inoculum in reactors with different concentrations of paper and cellulase, and in control reactors, without cellulase. Anaerobic batch reactor (5 L), in triplicate, with office paper and modificated Del Nery medium, was kept in 37 ºC and initial pH 7,0, with headspace filled with \'N IND.2\' (100%), for the following assays: (1) 0,5 g paper/L e 4mL cellulase/L; (2) 2,0 g paper/L e 15 mL cellulase/ L; (3) 4,0 g paper/L e 30 mL cellulase/L; (CT 1) 0,5 g paper/L; (CT 2) 2,0 g paper/L; (CT 3 ) 4,0 g paper/L. Hydrogen yields were 42, 26,6 and 24 mmol \'H IND.2\'/g paper for the assays 1, 2 and 3, respectively. Hydrogen production was not detected in the control reactors. Substrate consumption percentage, measured as glucose, was 56%, 56,6% and 65,4%, respectively for the assays 1, 2 and 3. Paper consumption was not detected in the control reactors. Butyric acid, low concentration of acetic acid, and alcohols were detected in the reactors with cellulase. In the control reactor it was observed mainly acetic and iso-butyric acids. Final pH values were 4,6; 3,7; 3,5; 6,8; 6,6 and 6,5 for the assays 1, 2, 3, CT 1, CT 2 and CT 3, respectively. It was observed a predominance of Gram positive endospore-forming rods. From PCR/DGGE analysis it was possible to observe an alteration of 78% between the original microbial community of the ruminal fluid and the microbial community present at the end of the assays. From cloning and sequencing analysis of RNAr 16S gene, there was verified similarity of 95-99% with Clostridium in the microbial consortium. The biological hydrogen production in the assays with cellulase was probably due to bacteria present in the conditions studied, mainly identified as Clostridium sp., known as producers of hydrogen, organic acids and alcohols. The addition of cellulase was essential to promote paper breakdown into soluble carbohydrates, which allowed hydrogen production by bacteria present in the consortium.

O presente trabalho apresenta estudos de lesão em biomoléculas (DNA e 2\'-deoxiguanosina) induzida por Cu(III)/tetraglicina (Cu(III)/G4), radicais de óxidos de enxofre (SO3&middot;-, SO4&middot;-, SO5&middot;-), HO&middot; e HSO5-, espécies estas geradas durante a autoxidação de S(IV) na presença de Cu(II)/G4 ou Cu(II) (ausência de tetraglicina) e traços de um segundo íon metálico (Ni(II), Co(II) ou Mn(II)). A formação dos radicais SO3&middot;- e HO&middot; foi detectada pela técnica de ressonância paramagnética eletrônica (EPR). As técnicas de espectrofotometria e dicroísmo circular foram empregadas para avaliar a formação de Cu(III)/G4 em diferentes condições experimentais, na presença e ausência de S(IV), e a interação entre os complexos de cobre (II)/(III) e a molécula de DNA. A eficiência da formação de Cu(III) depende da acidez, concentração de S(IV) e dos tampões utilizados. A lesão no DNA plasmidial pUC19 foi verificada empregando-se a técnica de eletroforese em gel de agarose. A extensão da lesão no DNA depende da acidez, concentração de S(IV), tempo de incubação e da presença de um segundo íon metálico. Usando a técnica de cromatografia líquida de alta eficiência (HPLC) foi possível estudar a oxidação de 2\'-deoxiguanosina a 8-oxo-7,8-dihidro-2\'-deoxiguanosina na presença dos oxidantes fortes gerados durante a autoxidação de S(IV) catalisada por Cu(II)/G4. Um estudo comparativo do efeito de vários íons metálicos evidenciou o sinergismo de Cu(II) e traços de um segundo íon metálico (Ni(II), Co(II) ou Mn(II), complexados ou não com tetraglicina). / The present work presents studies related to biomolecules damage (DNA and 2\'-deoxyguanosine) induced by Cu(III)/tetraglycine (Cu(III)/G4), oxysulfur radicals (SO3&middot;-, SO4&middot;-, SO5&middot;-) and HSO5-, species generated during S(IV) autoxidation in the presence of Cu(II)/G4 or Cu(II) (absence of tetraglycine) and trace level of a second metal ion (Ni(II), Co(II) or Mn(II)). The formation of SO3&middot;- and HO&middot; radicals was detected by electronic paramagnetic resonance technique (EPR). Spectrophotometric and circular dichroism techniques were used to evaluate the Cu(III)/G4 formation in different experimental conditions, in the presence and the absence of S(IV), and the interaction of copper (II)/(III) complexes and DNA molecule. The effectiveness of Cu(III) formation depends on the acidity, S(IV) concentration, and buffers used. The damage on pUC 19 plasmid DNA was verified by agarose gel electrophoresis. The extent on the DNA damage was related to acidity, S(IV) concentration, incubation time and to the presence of a second metal ion. Using the high performance liquid chromatography technique (HPLC) it was possible to study the oxidation of 2\'-deoxyguanosine to 8-oxo-7,8-dihydro-2\'-deoxyguanosine in the presence of strong oxidants generated during the S(IV) autoxidation catalyzed by Cu(II)/G4. A comparative study of the effect of several metal ions showed the synergism of Cu(II) and traces of a second metal ion (Ni(II), Co(II) or Mn(II), as tetraglycine complexes or not).

O presente trabalho teve como objetivo avaliar a fermentabilidade do hidrolisado enzimático do bagaço de cana pré-tratado com sulfito alcalino, visando a produção de etanol por leveduras. Inicialmente, o bagaço de cana foi pré-tratado com sulfito alcalino para remover parcialmente a lignina e alterar a estrutura da celulose e hemicelulose. Os efeitos deste pré-tratamento foram avaliados em termos da composição química dos materiais obtidos: um material lavado (BPL) e outro não lavado (BPNL) após tratamento. A digestibilidade enzimática dos materiais obtidos foi testada empregando dois extratos enzimáticos comercias e 3 cargas de enzimas, de acordo com um planejamento experimental 22. A partir destes resultados foi selecionado o extrato Cellubrix suplementado com ?-glicosidase na carga de 18 FPU/g de biomassa, e nesta condição os rendimentos de conversão de celulose e xilana foram cerca de 70% e 65%, respectivamente. Os hidrolisados enzimáticos do BPL e BPNL porém suplementados com nutrientes (2,0 g/L de (NH4)2SO4, 1,0 g/L de KH2PO4, 0,3 g/L de MgSO4.7H2O, 5,0 g/L de extrato de levedura), e avaliados quanto a produção de etanol por K. marxianus Y-6860 a 40°C. Os resultados mostraram que a levedura produziu etanol apenas a partir de glicose, e os parâmetros fermentativos obtidos com o hidrolisado BPNL QP = 3,7 g/L.h e YP/S = 0,40 g/g foram 20 e 5%, respectivamente superior ao BPL. O hidrolisado enzimático do BPNL foi então selecionado, para avaliar o efeito da suplementação nutricional nos parâmetros fermentativos da levedura K. marxianus Y-6860, em três temperaturas diferentes (30, 35 e 40°C) de acordo com um planejamento experimental 22. Para efeito de comparação, foi também avaliada a fermentabilidade do hidrolisado BPNL pelas leveduras S. cerevisiae PE-2 e S. stipitis Y-7124. Os resultados mostraram que para as três leveduras estudadas, os valores dos parâmetros fermentativos, foram favorecidos a 30 °C na presença nutrientes. Nesta condição, as leveduras apresentaram valores de YP/S similares (~ 0,48 g/g), em quanto que o QP foi de 3,25; 2,17 e 0,95 g/L.h para K. marxianus, S. cerevisiae e S. stipitis, respectivamente. Ressalta-se ainda que nesta condição, a S. stipitis Y-7124 foi a única levedura, capaz de consumir completamente ambos os açúcares (glicose + xilose) e produzir uma concentração de etanol 46% maior comparado as outras leveduras. Foi avaliado também a fermentabilidade das três leveduras no hidrolisado enzimático BPNL com alta concentração de glicose (100 g/L), nas condições anteriormente selecionadas. Os resultados mostraram que, as três leveduras apresentaram potencial para produção de etanol a partir da glicose, com uma eficiência de conversão de etanol superior a 70%, no entanto deveriam se implementar estratégias para estimular o consumo de xilose. Com os resultados do presente trabalho se pode concluir que o hidrolisado enzimático do bagaço de cana pré-tratado com sulfito alcalino suplementado com nutrientes, é um meio com grande potencial para a produção de etanol com características favoráveis para ser eficientemente fermentado por diferentes espécies de leveduras. / The present work had the objective to evaluate the fermentability of the enzymatic hydrolyzate of sugar cane bagasse pre-treated with alkali sulfite process, aiming the ethanol production by yeasts. Initially, sugarcane bagasse was pretreated with alkaline sulfite process to remove lignin partially. The effects of this pre-treatment, were evaluated in the chemical composition of the materials: one material washed after treatment (WB) and another unwashed material (UWB). The enzymatic digestibility of the obtained materials was tested using two commercial enzymatic extracts and three enzyme loads according to an experimental design 22. From these results, the Cellubrix extract supplemented with ?-glucosidase, was selected in the load of 18 FPU/g of biomass, in this condition the cellulose and xylan conversion yields were about 70% and 65%, respectively. The enzymatic hydrolysates of WB and UWB supplemented with nutrients (2,0 g/L (NH4)2 SO4, 1,0 g/L KH2 PO4, 0,3 g/L MgSO4 .7H2O, 5,0 g/L yeast extract), were evaluated for ethanol production by K. marxianus Y-6860 at 40°C. The results showed that the yeast produced ethanol only from glucose, and the fermentative parameters obtained with the UWB hydrolyzate QP= 3,7 g/L.h and YP/S= 0,40 g/g, were 20 and 5% respectively higher to WB. The UWB hydrolyzate, was selected to evaluate the effect of nutritional supplementation, in the fermentative parameters of K. marxianus, at three different temperatures (30, 35 and 40°C) according to an experimental design 22. For comparison, were also evaluated fermentability of S. cerevisiae PE-2 and S. stipitis Y-7124 in the hydrolyzate. In the three yeasts, the results showed that the values of the fermentative parameters, are favored when the hydrolyzate is supplemented with nutrients to 30°C. In this condition , yeasts showed similar YP/S value (~0,48 g/g), whereas the QP was of 3,25; 2,17 and 0,95 g/L.h for K. marxianus, S. cerevisiae and S. stipitis, respectively. It is also worth noting that in this condition S. stipitis was the only yeast able to completely consume both sugars (glucose+xylose) and produce a concentration of ethanol of 46% higher than other yeasts. In the previously selected condition, the fermentability of the hydrolyzate with high glucose concentration (100 g/L) was also evaluated. The results showed that the three yeasts presented potential for the production of ethanol from glucose, with an ethanol efficiency of over 70%, however strategies should be implemented to stimulate the consumption of xylose. With the results of the present work it can be concluded that the enzymatic hydrolyzate of sugar cane bagasse pretreated with alkaline sulfite process supplemented with nutrients is a medium with great potential for the production of ethanol with characteristics favorable to be efficiently fermented by different species of yeasts.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-15T14:00:25Z No. of bitstreams: 1 Nattany Tayany Gomes de Paula.pdf: 885461 bytes, checksum: 787ab63496a1306c113ee5c12e2dac92 (MD5) / Made available in DSpace on 2017-02-15T14:00:25Z (GMT). No. of bitstreams: 1 Nattany Tayany Gomes de Paula.pdf: 885461 bytes, checksum: 787ab63496a1306c113ee5c12e2dac92 (MD5) Previous issue date: 2015-03-12 / Sulfites are considered food additives that after chemical reaction releasing sulfur dioxide (SO2), sulfite ion (SO32-), or bisulfite ion (HSO3-). These additives act as antioxidants in food, in the control of bacterial growth and enzymatic and non-enzymatic browning reactions during processing and storage. Although widely used in the food industry, the use of sulfite as a preservative causes some problems, reducing the nutritional quality of foods processed and in some cases are caused adverse effects to human health. In this work it is proposed to determine sulfite in samples of processed juices and wines through an analytical flow system with biamperometric detection. The system was built employing a peristaltic pump, proportional injector, a Gas Diffusion Cell (GDC) and Reagent Generation Cell (RGC) and polyethylene tubes with an internal diameter of 0.8 mm for making the analytical path. The RGC was coupled to a potentiostat for applying current to the electrochemical generation of triiodide ions. For detection it was used a biamperometric detector, adapted to a coulometer for applying the constant potential. For the data acquisition and processing was performed using a software developed in Lab View® language. The analysis method was based on a redox reaction between the I3- ions SO32- and providing a decrease of the generated current by the pair and this decrease in the signal is proportional to the concentration of SO32- ions. After the system optimization, the proposed procedure was operated with KI concentrations of 0.25 mol L-1, HCl 1.0 mol L-1, Na2SO4 and 0.1 mol L-1, with 0.25 mA current applied in RGC and the length of the sampling loop was of 20 cm. In the proposed procedure, a linear response was obtained between 1 and 12 mg L-1 of sulfite E = - (0,2371 ± 0,0028) C + (6,7097 ± 0,0182), R = -0,9997), LD = 0,26 mg L-1 and LQ = 0,86 mg L-1, relative standard deviation less than 2% (n = 10), with sampling frequency of 40 h−1. Potential interfering studies demonstrated the selectivity of the proposed method. Addition and recovery tests with samples of juices and wines were performed to give results of recovery between 91.63 and 110.34%, demonstrating the possibility of operation of the proposed procedure in real samples. / Os agentes sulfitantes ou sulfitos são considerados aditivos alimentares que, após reação química, liberam dióxido de enxofre (SO2), íon sulfito (SO32-) ou íon bissulfito (HSO3-). Estes aditivos são agentes multifuncionais e atuam como antioxidantes em alimentos, no controle do crescimento bacteriano e de reações (enzimáticas e não enzimáticas) de escurecimento durante o processamento e estocagem. Mesmo sendo muito utilizado na indústria de alimentos, o emprego de sulfitos como conservantes provoca alguns problemas, reduzindo a qualidade nutricional dos alimentos tratados e em alguns casos são provocados efeitos adversos à saúde humana. Neste trabalho propõe-se determinar sulfitos em amostras de sucos industrializados e vinhos empregando um sistema de análise em fluxo com detecção biamperométrica. O sistema foi construído empregando-se uma Bomba Peristáltica, um Injetor Proporcional, uma Câmara de Difusão Gasosa (CDG) e uma Câmara de Geração de Reagente (CGR). A CGR foi acoplada a um potenciostato para a aplicação de corrente necessária para a geração eletroquímica de íons triiodeto. Para detecção foi utilizado um detector biamperométrico. Para o tratamento e aquisição de dados, foi utilizado um software desenvolvido em linguagem Lab View®. A metodologia de análise foi baseada em uma reação de oxirredução entre os íons SO32- e I3-, que proporciona uma diminuição da corrente biamperométrica gerada pelo par redox I3-/I-, sendo essa diminuição de corrente proporcional a concentração de íons SO32-. Após a otimização do procedimento proposto obteve-se as melhores condições: 0,25 mol L-1 de KI, 1,0 mol L-1 de HCl, 0,1 mol L-1 de Na2SO4, corrente de 0,25 mA aplicada na CGR e volume de amostra de 100 μL. Nestas condições, obteve-se uma resposta linear entre 1 e 12 mg L-1 de sulfito (I = - (0,2371 ± 0,0028) C + (6,7097 ± 0,0182), R = -0,9997), LD = 0,26 mg L-1 e LQ = 0,86 mg L-1, desvio padrão relativo menor que 2% (n = 10), com frequência de amostragem de 40 determinações por hora. Estudos de potenciais interferentes demonstraram a seletividade do método proposto. Testes de adição e recuperação com amostras de sucos e vinhos foram realizados, obtendo-se resultados de recuperação entre 91,63 e 110,34%, demonstrando a possibilidade de operação do procedimento proposto em amostras reais.

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