Thesis (Ph. D.)--University of Texas at Austin, 2006. / Vita. Includes bibliographical references.

A Serra Gaúcha apresenta uma excelente aptidão enológica para produzir vinhos espumantes de qualidade. Na produção de espumantes, o vinho base passa por uma segunda fermentação para tomada de espuma, durante a qual ocorre a produção de gás carbônico e etanol, assim como diversas transformações biológicas e químicas que influenciam as características organolépticas do produto final. Após a segunda fermentação, os espumantes maturam em contato com as borras (sur lie) por períodos variados durante o qual ocorre autofagia/autólise das leveduras. Neste trabalho foram avaliadas as características físico-químicas e compostos voláteis ao longo da segunda fermentação e maturação de espumantes, assim como a dinâmica da população de leveduras e expressão de genes relacionados ao processo autofágico/autolítico. Para tanto foram conduzidas segundas fermentações pelo método tradicional com vinho base elaborado com as variedades Pinot Noir, Chardonnay e Riesling Itálico e levedura S. cerevisiae EC1118. As amostras foram coletadas nos tempos 0, 7, 15, 30, 60, 90, 135, 180, 270 e 360 dias de fermentação, e avaliadas quanto as características físicoquímicas básicas, compostos voláteis (27 compostos), população de leveduras, concentração de compostos fenólicos e proteínas/peptídeos. A expressão de genes relacionados com autofagia foi determinada por qRT-PCR durante a segunda fermentação e início de maturação. Os resultados mostraram que a segunda fermentação de espumantes apresenta uma fase inicial de adaptação de 7 dias seguida por importante incremento do teor alcoólico até os 30 dias. Durante a fermentação e maturação ocorre redução da acidez total e leve aumento da acidez volátil que refletem no aumento do pH no produto final. Da mesma forma, foi observada variação significativa na concentração de distintos compostos aromáticos ao longo da segunda fermentação e maturação de espumantes, com redução de ésteres especialmente a partir dos 90 dias. Análise multivariada mostrou que o espumante apresenta mudanças importantes durante a segunda fermentação. Após 270 dias de maturação observou-se redução no conteúdo de propanol 1, 2-metil-1-butanol, 3-metil-1-butanol, octanoato de etila, ácido decanóico e ácido dodecanóico, e aumento de dietil succinato, dodecanoato de etila e feniletanol. Aumento da concentração de compostos fenólicos durante a maturação foi constatado, podendo interferir na cor do produto final. A população de leveduras (UFC/ml) exibiu importante aumento durante o período fermentativo com redução abrupta (~80%) nos 30 dias seguintes. Por outro lado, o número de células se mantem constante dos 30 aos 90 dias, com redução posterior indicativa de autólise. A redução no número de células integras foi acompanhada por aumento na concentração de proteínas/peptídeos no vinho, com estabilização a partir dos 270 dias. A avaliação da expressão de um conjunto de genes relacionados com a autofagia indica que tanto a micro quanto a macroautofagia são induzidas ainda na fase fermentativa com aumento da macroautofagia sobre o final da segunda fermentação, acompanhando a redução de viabilidade. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2015-02-24T17:38:07Z No. of bitstreams: 1 Dissertacao Andrelise Verzeletti.pdf: 2037402 bytes, checksum: eec916eb8804cd342d32a52df7d363bd (MD5) / Made available in DSpace on 2015-02-24T17:38:07Z (GMT). No. of bitstreams: 1 Dissertacao Andrelise Verzeletti.pdf: 2037402 bytes, checksum: eec916eb8804cd342d32a52df7d363bd (MD5) / The Serra Gaucha region shows an excellent oenological aptitude for the production of high quality wines and sparkling wines. Sparkling wines process involves a second fermentation with gas and ethanol production, and several biological and chemical transformations that influence the organoleptic properties of the final product. After the second fermentation sparkling wines mature in contact with the lees (sur lie) for long periods during which yeasts autophagy/autolysis occurs. In this work we evaluate the physic-chemical characteristics and volatile compounds during the second fermentation and maturation of sparkling wines, as well as the dynamics of yeast population and the expression of autophagic/autolytic related genes. For this purpose second fermentations were conducted by the traditional method using a base wine elaborated with Pinot Noir, Chardonnay and Riesling Italic and the S. cerevisiae EC1118 commercial strain. Samples were collected at 0, 7, 15, 30, 60, 90, 135, 180, 270 and 360 days of fermentation, and evaluated with respect to the basic physic-chemical characteristics, volatile compounds (27 compounds), yeast population, and the concentration of phenolic compounds and protein/peptides. The expression of autophagy/autolysis related genes during the second fermentation and the beginning of maturation was determined by qRT-PCR. The results showed that the second fermentation involved an initial adaptation period of 7 days followed by an important increment in the alcoholic concentration during 30 days. During the fermentation and maturation it was observed a reduction in total acidity and a small increase of volatile acidity that led to a pH increase in the final product. A significant variation in several volatile compounds was detected during second fermentation and maturation of sparkling wines, with a reduction in esters after 90 days. Multivariate analysis showed that sparkling wines suffer important modifications during second fermentation. After 270 days of maturation sparkling wines exhibited a reduction in the concentration of 1-propanol, 2-methyl-1-buthanol, 3- methyl-1-buthanol, ethyl octanoate, decanoic acid, and dodecanoic acid, and an increase in the concentration of diethyl succinate, ethyl dodecanoate and phenylethanol. Increase in the concentration of phenolic compounds during maturation, can affect wine color. Yeast population (UFC/mL) exhibited an important increase during the fermentation period followed by a drastic reduction (~80%) in the next 30 days. However, the total number of cells remained constant between 30 and 90 days, a rapidly decrease after this period indicating yeast autolysis. The reduction of yeast cells was accompanied by an increase in the concentration of proteins/peptides in wine, which stabilized at 270 days. The evaluation of expression of a group of genes related with the autophagic process indicated that both micro and macroautophagy are induced during the second fermentation with an increase of macroautophagy at the end of the fermentation period, accompanying the decrease in yeast viability.

Capes; CNPq / O aroma de café é constituído por uma mistura de componentes voláteis, os quais podem ser encontrados no óleo obtido na torração do café. O café solúvel, em função das condições de processamento, perde parte do seu aroma que, a princípio, poderia ser reincorporado ao produto pela adição do óleo de café diretamente sobre o produto final. Contudo, alguns trabalhos apontam para dificuldades de se obter bons resultados quando se trabalha com o óleo in natura devido à sua volatilidade e à degradação dos aromas. A incorporação de nanocápsulas contendo o óleo de café pode ser uma alternativa para minimizar estas perdas, pois o polímero encapsulante pode agir como barreira de proteção e promover sua liberação de forma controlada. A proposta do trabalho é a obtenção de nanocápsulas de poli(L-ácido lático)(PLLA), contendo o óleo de café torrado pela técnica de miniemulsificação/evaporação do solvente e validar o método para quantificação do óleo de café por espectrofotometria UV-Vis. Os estudos de validação possibilitaram garantir a qualidade dos resultados obtidos. Formulações com diferentes concentrações de PLLA, óleo e tipos de surfactantes foram valiadas quanto ao tamanho e índice de polidispersão frente à recuperação de óleo. Todas as formulações testadas apresentaram valores de recuperação de óleo acima de 80% mostrando que a técnica é adequada e pode ser considerada uma alternativa promissora para produzir nanocápsulas contendo óleo de café. As análises da composição do aroma das nanocápsulas, que indica a qualidade da porção de óleo encapsulada, mostraram que as nanocápsulas contêm frações de todos os compostos encontrados no óleo. / Coffee flavor is a mixture of volatile components found in the oil obtained from the roasted coffee. Soluble coffee loses part of its aroma due to processing conditions which could be reintroduced to the product by the direct addition of the coffee oil. However, some studies point out difficulties in obtaining good results when working with in natura oil because of the high volatility and the flavor degradation. The incorporation of nanocapsules containing coffee oil could be an alternative to minimize these losses, because the encapsulating polymer could act as a protecting barrier and promote its controlled release. The aim of this work is obtain PLLA nanocapsules containing roasted coffee oil by the miniemulsification/solvent evaporation technique. The spectrophometric UV- Vis method used to quantify the coffee oil was validated. Formulations with different PLLA and oil concentrations and kind of surfactant were evaluated in respect to the average nanocapsules size and polydispersion index and oil recovery. All formulations demonstrated oil recovery values above 80 % demonstrating that the technique is adequate and can be consider a promising alternative to coffee oil nanocapsules production. Flavor analysis nanocapsules composition, which indicates the quality of the encapsulated oil, showed that the nanocapsules contain fractions of all the substances found on the in natura oil.

Capes; CNPq / O aroma de café é constituído por uma mistura de componentes voláteis, os quais podem ser encontrados no óleo obtido na torração do café. O café solúvel, em função das condições de processamento, perde parte do seu aroma que, a princípio, poderia ser reincorporado ao produto pela adição do óleo de café diretamente sobre o produto final. Contudo, alguns trabalhos apontam para dificuldades de se obter bons resultados quando se trabalha com o óleo in natura devido à sua volatilidade e à degradação dos aromas. A incorporação de nanocápsulas contendo o óleo de café pode ser uma alternativa para minimizar estas perdas, pois o polímero encapsulante pode agir como barreira de proteção e promover sua liberação de forma controlada. A proposta do trabalho é a obtenção de nanocápsulas de poli(L-ácido lático)(PLLA), contendo o óleo de café torrado pela técnica de miniemulsificação/evaporação do solvente e validar o método para quantificação do óleo de café por espectrofotometria UV-Vis. Os estudos de validação possibilitaram garantir a qualidade dos resultados obtidos. Formulações com diferentes concentrações de PLLA, óleo e tipos de surfactantes foram valiadas quanto ao tamanho e índice de polidispersão frente à recuperação de óleo. Todas as formulações testadas apresentaram valores de recuperação de óleo acima de 80% mostrando que a técnica é adequada e pode ser considerada uma alternativa promissora para produzir nanocápsulas contendo óleo de café. As análises da composição do aroma das nanocápsulas, que indica a qualidade da porção de óleo encapsulada, mostraram que as nanocápsulas contêm frações de todos os compostos encontrados no óleo. / Coffee flavor is a mixture of volatile components found in the oil obtained from the roasted coffee. Soluble coffee loses part of its aroma due to processing conditions which could be reintroduced to the product by the direct addition of the coffee oil. However, some studies point out difficulties in obtaining good results when working with in natura oil because of the high volatility and the flavor degradation. The incorporation of nanocapsules containing coffee oil could be an alternative to minimize these losses, because the encapsulating polymer could act as a protecting barrier and promote its controlled release. The aim of this work is obtain PLLA nanocapsules containing roasted coffee oil by the miniemulsification/solvent evaporation technique. The spectrophometric UV- Vis method used to quantify the coffee oil was validated. Formulations with different PLLA and oil concentrations and kind of surfactant were evaluated in respect to the average nanocapsules size and polydispersion index and oil recovery. All formulations demonstrated oil recovery values above 80 % demonstrating that the technique is adequate and can be consider a promising alternative to coffee oil nanocapsules production. Flavor analysis nanocapsules composition, which indicates the quality of the encapsulated oil, showed that the nanocapsules contain fractions of all the substances found on the in natura oil.

Submitted in the fulfilment of the requirement for the Degree of Doctor of Philosophy: Chemistry, Durban University of Technology, 2016. / The work conducted in this study comprised three aspects: syntheses, characterizations, and multi-component liquid-liquid extractions. The main objectives of the project were: (1) to evaluate the efficacy and efficiency of ionic liquids to extract aromatic components from catalytic reformates and paint solvent wastes, and (2) to validate the method(s) used in this project to qualitatively and quantitatively analyze the aromatic molecules (BTEX) in multi-component mixtures. Therefore, this research critically investigated the major effects of the chosen ionic liquids as extractive solvents for the recovery of BTEX components from model and industrial organic mixtures. The project was concerned with the nature of solvents currently used in most industries for the separation by extraction of aromatic hydrocarbons from non-aqueous or organic mixtures. Most solvents currently employed for this purpose are highly volatile; hence they contribute significantly towards environment pollution. In addition, the extraction efficiency of these conventional solvents is limited only to mixtures containing aromatic hydrocarbons of 20% or more. Furthermore, conventional solvents are organic compounds which are generally toxic, flammable, and expensive to recover or regenerate from extract phases due to methods which involve several steps. In addition, they demand high energy input for the distillation steps. used in the analysis of aromatic components were evaluated for validity. According to the literature no such work was carried out by previous researchers. The study targeted four ionic liquids, namely, 1-ethyl-3-methylimidazolium ethyl sulphate [EMIM][ESO4], 1-ethyl-3-methylpyridinium ethyl sulphate [EMpy][ESO4], 1- Butyl-1-methyl-2-pyrrolidonium bromide [BNMP][Br], and 1,1-Dimethyl-2- pyrrolidonium iodide [MNMP][I] in an attempt to address this concern. These ionic liquids were synthesized and characterized in our laboratories using previously accepted methods. After synthesis and purification, they were characterized by techniques including FTIR, 1H-NMR, and 13C-NMR. The densities and moisture content of both the synthesized and standard ionic liquids were also determined using density meters and Karl-Fischer apparatus, respectively. The extractions were carried out on both the model and industrial mixtures using ionic liquids. Each ionic liquid was mixed with a target mixture in a water-jacketed vessel and then stirred vigorously at constant temperature achieved by a thermostatically controlled water-bath. After a selected period of time the operation was stopped and the resulting mixture was left to stand overnight to allow phase equilibration to be reached. The two phases were then separated and analyzed for the content of individual aromatic components in each phase using GC-FID calibrated with external standards of the components present in the mixtures being investigated. According to the results obtained from the synthesis and characterization methods the percentages yield of ionic liquids were reasonably high (> 95%). In addition, spectral studies showed high purity with fewer traces of impurities based on the observed relative intensities. Results from GC-FID indicated a relatively lower concentration of aliphatic hydrocarbons in the extract phase. On the other hand, the concentrations of aromatic II components in the extract phase were relatively higher than those of aliphatic hydrocarbons. The results obtained from the three extraction stages showed the total recovery of greater than 50% for the aromatic components. This suggests that at least six extraction stages would be required in order to achieve a total recovery of 100% aromatic components which is an indication of good efficiency. Also noticeable was that the first extraction stages for all ionic liquids recovery values were much higher than those values obtained from successive stages which showed approximately the same extraction results. In most experiments, 1-ethyl-3-methylpyridinium ethyl sulphate gave higher recovery values than the other three ionic liquids. It was also noted that the recovery values obtained from the extractions performed on model mixtures of the entire concentration range (0.5 – 25%) of individual aromatic components did not show any significant difference. Proportional difference in recoveries occurred across the entire concentration range of model mixtures. The results also indicated that the solubility of aromatic hydrocarbons in the ionic liquids decreases in the order: benzene > toluene > ethyl benzene >xylenes. This phenomenon is attributed to a decrease in π-π, cation- π, cation- anion interactions occurring between the ionic liquid and each of the aromatic molecules in this order. The recovery values for BTEX ranged from 80 to 120 % by volume for the three extraction stages. This is in line with results previous research studies carried out on liquid-liquid extractions involving ternary systems containing only one aromatic component in each mixture. Therefore this study shows that ionic liquids are capable extraction solvents for simultaneous recovery of the aromatic components from any organic mixtures containing low to high BTEX concentrations. In addition, the outcomes of this project have proved that ionic liquids are economically viable as potential extraction solvents since they can be easily recycled and reusable many times without any noticeable degradation. The results of this study are envisaged to make significant contributions to the current research efforts aimed at achieving greener environments and minimization of global warming. The findings of this project are also geared to boost the economy of our country through job creation using economically viable methods. / D

Bioremediation of aromatic pollutants using the ligninolytic enzymes of the white rot fungi has been thoroughly researched and has been shown to have considerable potential for industrial application. However, little success in scale-up and industrialisation of this technology has been attained due to problems associated with the continuous production of the pollutant-degrading enzymes using conventional bioreactor systems. The low productivities reported result from the incompatibility of conventional submerged culture reactor techniques with the physiological requirements of these fungi which have evolved on a solid-air interface, viz. wood. The enzymes are also produced only during the stationary phase of growth and can therefore be regarded as secondary metabolites. This study reports the conceptualisation, characterisation and evaluation of a novel bioreactor system as a solution to the continuous production of idiophasic pollutant degrading enzymes by the white rot fungus Phanerochaete chlysosporium. The reactor concept evolved from observation of these fungi in their native state, i. e. the metabolism of lignocellulosic material and involves the immobilisation of the organism onto a capillary ultrafiltration membrane. Nutrient gradients established across the biofilm, an inherent characteristic of fixed bed perfusion reactors, are exploited to provide both nutrient rich and nutrient poor zones across the biofilm. This allows growth or primary metabolism in the nutrient rich zone, pushing older biomass into the nutrient poor zone where secondary metabolism is induced by nutrient starvation. In effect, this represents a transformation of the events of a batch culture from a temporal to a spatial domain, allowing continuous production of secondary metabolites over time. Direct contact of the outer part of the biofilm with an air stream simulated the solid-air interface of the native state of the fungus. In order to facilitate the practical application of the membrane gradostat reactor (MGR) concept, conventional capillary membranes and membrane bioreactor modules were first evaluated. These were found to be unsuitable for application of the MGR concept. However, critical analysis of the shortcomings of the conventional systems resulted in the formulation of a set of design criteria for the development of a suitable membrane and module. These design criteria were satisfied by the development of a novel capillary membrane for membrane bioreactors, as well as a transverse flow membrane module, which is a novel approach in membrane bioreactor configuration. For the physiological characterisation of the MGR concept, a single fibre bioreactor unit was designed, which allowed destructive sampling of the biofilm for analysis. Using this system, it was shown that distinct morphological zones could be observed radially across the mature biofilm obtained through MGR operation. That these morphotypes do represent the temporal events of a typical batch culture in a spatial domain was confirmed by following the morphological changes occurring during batch culture of the immobilised fungus where the onset of primary and secondary metabolic conditions were manipulated through control of the nutrient supply. The different morphotypes were correlated to distinct growth phases by comparison of the morphology to the secretion of known enzymatic markers for secondary metabolism, viz. succinate dehydrogenase and cytochrome C oxidoreductase. Detailed structure-function analysis of the biofilm using transmission electron microscopy and adapted enzyme cytochemical staining techniques showed that the biofilm appeared to operate as a co-ordinated unit, with primary and secondary metabolism apparently linked in one thallus through nutrient translocation. This study provided new insights into the physiology of P. chrysosp,o rium and a detailed descriptive model was formulated which correlates well to existing models of wood degradation by the white rot fungi (WRF). Evaluation of the process on a laboratory scale using a novel transverse flow membrane bioreactor showed that a volumetric productivity of 1916 U.L.⁻¹day⁻¹ for manganese peroxidase, one of the pollutant degrading enzymes, could be attained, corresponding to a final concentration of 2 361 U.L.⁻¹ This may be compared to the best reported system (Moreira el at. 1997), where a volumetric productivity of 202 U.L.⁻¹day⁻¹was achieved with a final concentration of 250 U.L.⁻¹ However, MGR productivity is yet to be subjected to rigorous optimisation studies. The process could be operated continuously for 60 days. However, peak productivity could not be maintained for long periods. This was found to be due to physical phenomena relating to the fluid dynamics of the system which caused fluid flow maldistribution, which would have to be resolved through engineering analysis. In evaluation of the MGR concept for aromatic pollutant removal, in this case ρ- cresol, from growth medium, good performance was also achieved. The VmaxKm calculated by linear regression for the MGR was 0.8 (R² = 0.93), which compared favourably to that reported by Lewandowski et al. (1990), who obtained a Vmax/Km of 0.34 for a packed bed reactor treating chlorophenol. It was concluded that the MGR showed suitable potential to warrant further development, and that the descriptive characterisation of the biofilm physiology provided a sufficient basis for process analysis once engineering aspects ofthe system could be resolved.

Sasol endeavors to expand its current business through the beneficiation of commodity feedstreams having marginal value into high-value chemicals via cost-effective processes. In this regard Merisol, a division of Sasol has access to phenolic and cresylic feedstreams, which have the potential to be converted to fine chemicals. Targeted products include para-anisaldehyde, ortho- and para-hydroxybenzaldehyde which are important intermediates for the manufacture of chemicals used in the flavor and fragrance market and various other chemicals. The use of CO technology (HF/BF3) to produce these aldehydes in a two-step process from phenol as reagent is economically attractive due to the relative low cost and other benefits associated with syngas as reagent. The aim of the study was to evaluate and understand this relatively unexplored approach to the formylation of aromatic compounds. The reactivity of both phenol and anisole proved to be much lower than that of toluene. The main aldehyde isomer (para) produced in these HF/BF3/CO formylations as well as of other ortho-para directing mono-substituted benzenes tested in our laboratories were in accordance with published results. Efforts to increase substrate conversion resulted in substantial secondary product formation and mechanistic investigations showed this to be a consequence of the inherent high acidity of the reaction environment. The effect of different substituents on the relative formylation rates of benzene derivatives was investigated. These results showed that methyl groups are activating while halogens are deactivating relative to benzene as substrate. The decrease in reactivity from fluorobenzene> chlorobenzene> bromobenzene is in accordance with formylation trends observed in other acidic systems. Deuterium labeling experiments were applied to gain additional information on the formylation reaction mechanism. This study provided interesting but inconclusive results in support of the so-called intra-complex mechanism. All reported studies as well as our own work suggested that HF and BF3 in (at least) stoichiometric amounts are required for effective formylation with CO. Under these conditions this methodology for effecting aromatic formylation is not economically viable. Industrial application of formylation using CO will require the development of new catalysts or methodology to allow the use of HF/BF3 in a catalytic way. In this regard ionic liquids as a new and ecological-friendly field was explored. Chloro-aluminate ionic liquids promote the carbonylation of alkylated aromatic compounds, but fails in the case of oxygenated aromatics. Aldehyde yields of formylation in the acidified neutral ionic liquids were generally similar compared to reactions conducted in HF as solvent/catalyst. Formylation of anisole and toluene, but not of phenol in the neutral ionic liquids resulted in increased secondary product formation in comparison with hydrogen fluoride used as solvent/catalyst. This difference in behavior is not understood at present, but suggests that phenol is a good substrate for formylation in this medium, particularly with the development of a system catalytic with respect to HF/BF3 in mind. / Prof. C.W. Holzapfel

A Serra Gaúcha apresenta uma excelente aptidão enológica para produzir vinhos espumantes de qualidade. Na produção de espumantes, o vinho base passa por uma segunda fermentação para tomada de espuma, durante a qual ocorre a produção de gás carbônico e etanol, assim como diversas transformações biológicas e químicas que influenciam as características organolépticas do produto final. Após a segunda fermentação, os espumantes maturam em contato com as borras (sur lie) por períodos variados durante o qual ocorre autofagia/autólise das leveduras. Neste trabalho foram avaliadas as características físico-químicas e compostos voláteis ao longo da segunda fermentação e maturação de espumantes, assim como a dinâmica da população de leveduras e expressão de genes relacionados ao processo autofágico/autolítico. Para tanto foram conduzidas segundas fermentações pelo método tradicional com vinho base elaborado com as variedades Pinot Noir, Chardonnay e Riesling Itálico e levedura S. cerevisiae EC1118. As amostras foram coletadas nos tempos 0, 7, 15, 30, 60, 90, 135, 180, 270 e 360 dias de fermentação, e avaliadas quanto as características físicoquímicas básicas, compostos voláteis (27 compostos), população de leveduras, concentração de compostos fenólicos e proteínas/peptídeos. A expressão de genes relacionados com autofagia foi determinada por qRT-PCR durante a segunda fermentação e início de maturação. Os resultados mostraram que a segunda fermentação de espumantes apresenta uma fase inicial de adaptação de 7 dias seguida por importante incremento do teor alcoólico até os 30 dias. Durante a fermentação e maturação ocorre redução da acidez total e leve aumento da acidez volátil que refletem no aumento do pH no produto final. Da mesma forma, foi observada variação significativa na concentração de distintos compostos aromáticos ao longo da segunda fermentação e maturação de espumantes, com redução de ésteres especialmente a partir dos 90 dias. Análise multivariada mostrou que o espumante apresenta mudanças importantes durante a segunda fermentação. Após 270 dias de maturação observou-se redução no conteúdo de propanol 1, 2-metil-1-butanol, 3-metil-1-butanol, octanoato de etila, ácido decanóico e ácido dodecanóico, e aumento de dietil succinato, dodecanoato de etila e feniletanol. Aumento da concentração de compostos fenólicos durante a maturação foi constatado, podendo interferir na cor do produto final. A população de leveduras (UFC/ml) exibiu importante aumento durante o período fermentativo com redução abrupta (~80%) nos 30 dias seguintes. Por outro lado, o número de células se mantem constante dos 30 aos 90 dias, com redução posterior indicativa de autólise. A redução no número de células integras foi acompanhada por aumento na concentração de proteínas/peptídeos no vinho, com estabilização a partir dos 270 dias. A avaliação da expressão de um conjunto de genes relacionados com a autofagia indica que tanto a micro quanto a macroautofagia são induzidas ainda na fase fermentativa com aumento da macroautofagia sobre o final da segunda fermentação, acompanhando a redução de viabilidade. / The Serra Gaucha region shows an excellent oenological aptitude for the production of high quality wines and sparkling wines. Sparkling wines process involves a second fermentation with gas and ethanol production, and several biological and chemical transformations that influence the organoleptic properties of the final product. After the second fermentation sparkling wines mature in contact with the lees (sur lie) for long periods during which yeasts autophagy/autolysis occurs. In this work we evaluate the physic-chemical characteristics and volatile compounds during the second fermentation and maturation of sparkling wines, as well as the dynamics of yeast population and the expression of autophagic/autolytic related genes. For this purpose second fermentations were conducted by the traditional method using a base wine elaborated with Pinot Noir, Chardonnay and Riesling Italic and the S. cerevisiae EC1118 commercial strain. Samples were collected at 0, 7, 15, 30, 60, 90, 135, 180, 270 and 360 days of fermentation, and evaluated with respect to the basic physic-chemical characteristics, volatile compounds (27 compounds), yeast population, and the concentration of phenolic compounds and protein/peptides. The expression of autophagy/autolysis related genes during the second fermentation and the beginning of maturation was determined by qRT-PCR. The results showed that the second fermentation involved an initial adaptation period of 7 days followed by an important increment in the alcoholic concentration during 30 days. During the fermentation and maturation it was observed a reduction in total acidity and a small increase of volatile acidity that led to a pH increase in the final product. A significant variation in several volatile compounds was detected during second fermentation and maturation of sparkling wines, with a reduction in esters after 90 days. Multivariate analysis showed that sparkling wines suffer important modifications during second fermentation. After 270 days of maturation sparkling wines exhibited a reduction in the concentration of 1-propanol, 2-methyl-1-buthanol, 3- methyl-1-buthanol, ethyl octanoate, decanoic acid, and dodecanoic acid, and an increase in the concentration of diethyl succinate, ethyl dodecanoate and phenylethanol. Increase in the concentration of phenolic compounds during maturation, can affect wine color. Yeast population (UFC/mL) exhibited an important increase during the fermentation period followed by a drastic reduction (~80%) in the next 30 days. However, the total number of cells remained constant between 30 and 90 days, a rapidly decrease after this period indicating yeast autolysis. The reduction of yeast cells was accompanied by an increase in the concentration of proteins/peptides in wine, which stabilized at 270 days. The evaluation of expression of a group of genes related with the autophagic process indicated that both micro and macroautophagy are induced during the second fermentation with an increase of macroautophagy at the end of the fermentation period, accompanying the decrease in yeast viability.

The purpose of this investigation was to study the known methods of forming acetals and find one or several that could be used in further investigation. In connection with the reactions, the use of various catalysts was pertinent. Some of the catalysts used were hydrogen chloride, calcium chloride, ferric chloride, aromatic sulfonic acids, and other. There was no data available comparing the sulfonic acids with the inorganic salts and mineral acids. For that reason one of the subsidiary problems of this investigation was such a comparison. In order to investigate stability, several acetals were subjected to conditions intended to produce ring acetylation. These conditions are the same imposed by a Friedel Crafts reaction or a Fries rearrangement.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第20392号 / 工博第4329号 / 新制||工||1671(附属図書館) / 京都大学大学院工学研究科物質エネルギー化学専攻 / (主査)教授 村田 靖次郎, 教授 近藤 輝幸, 教授 小澤 文幸 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM