Thesis (Doctoral)--Ludwig-Maximilians-Universität zu München, 1935.

Made available in DSpace on 2014-06-11T19:23:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-19Bitstream added on 2014-06-13T19:09:15Z : No. of bitstreams: 1 gargel_ca_me_sjrp.pdf: 289963 bytes, checksum: bef29e3ebeeee94612ad1bdd03849b34 (MD5) / Enzimas são de grande importância para a indústria de alimentos, a fim de facilitar e acelerar os processamentos. Pectinases são enzimas amplamente utilizadas no processamento de vinhos, facilitando o processo de maceração, clarificação e a filtração do mosto. Outra enzima de grande importância na fabricação de vinhos, é a tanase. A coloração do vinho se deve à presença de taninos; a oxidação destes compostos em contato com o ar pode causar uma turbidez indesejável e consequentemente perda da qualidade do produto final. Essa turbidez pode ser evitada com o emprego de tanases, que impedem a reação de oxidação. Como a maioria dos formulados comerciais destas enzimas são provenientes de fungos filamentosos nao pertencentes ao ambiente vinicola, estes formulados contêm também outras enzimas não adequadas para aplicação no vinho, produzindo alguns efeitos indesejáveis. Assim, a busca por leveduras autóctonas, isto é, do próprio ecossistema vínico, produtoras de tais enzimas faz-se necessária. Outra enzima de grande importância no processamento de vinhos é a invertase, que hidrolisa a sacarose liberando frutose e glicose. Ao lado da vinificação, a frutose é considerada 40% mais doce que a sacarose, sendo assim, de grande importância ao processamento de vinho. No Brasil, a região de Jales, no Noroeste Paulista, vem despontando como um importante centro de produção de uvas e recentemente alguns produtores começaram a processar vinho de maneira artesanal. Assim, o presente trabalho teve como objetivo a triagem de leveduras autóctonas isoladas de uma vinícola da região de Jales para a produção de poligalaturonases, tanases e invertases visando a aplicação no processamento e no melhoramento de vinhos. Foram rastreadas diferentes linhagens... / Enzymes are important for the food industry in order to facilitate and accelerate the processing of food. Pectinases are enzymes widely used in wine processing, facilitating the process of maceration, clarification and filtration of the must. Another enzyme of great importance in wine production is the Tanase. The color of the wine is due to the presence of tannins, the oxidation of these compounds in contact with air can cause an undesirable turbidity and consequently loss of product quality. This turbidity can be avoided by employing Tanase, which prevent the oxidation reaction. As most of these commercial enzymes are made from filamentous fungi, which do not belong to the winery environment, this formula also contains other enzymes that are not suitable for application in the wine, producing some side effects. Thus, the quest for autochthonous yeasts, ie, from the own wine ecosystem, producing such enzymes is necessary. Another enzyme of great importance in the processing of wine is invertase, which hydrolyzes sucrose releasing fructose and glucose. Besides the importance for winemaking, fructose is considered 40% sweeter than sucrose, therefore, of great importance to the food industry. In Brazil, the region of Jales, Sao Paulo in the Northwest, has emerged as an important center of production of grapes, and recently some farmer began to produce artisanal wine. Thus, this study aimed at screening of autochthonous yeasts isolated from a winery from Jales region, which are able to produce poligalaturonases, Tanase and invertase in order to apply in the processing of wines. From, thirteen different strains belonging to different species of yeast, there were no significant activities to Tanase and pectinases. However, one strain of Candida stellata produced invertase activity, reaching... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:23:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-09-13Bitstream added on 2014-06-13T18:50:18Z : No. of bitstreams: 1 coradello_lfc_me_araiq.pdf: 1578511 bytes, checksum: 9788032334af99e723890212a8a7529a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As destilarias brasileiras produzem etanol utilizando o processo fermentativo contínuo ou por batelada alimentada. Nas fermentações sucessivas, as células são recicladas e usadas continuamente durante toda a safra. No final de cada ciclo de fermentação as células são concentradas e tratadas com ácido para redução dos contaminantes antes da utilização como starter da fermentação seguinte. Alterações no estado fisiológico das células podem ocorrer dependendo das condições que ocorrem antes e durante o processo e também durante o tratamento ácido, bem como a concentração de subprodutos formados. Tais alterações podem afetar a produtividade e a eficiência do processo fermentativo. As variações no estado fisiológico podem ser avaliadas através da medida de alguns indicadores, tais como níveis de trealose e seu grau de mobilização, proteínas e glicerol. Além disso, são necessários estudos para selecionar e utilizar os parâmetros fisiológicos como ferramentas de diagnóstico do desempenho do processo. O objetivo do presente trabalho foi o de expor as células a tratamentos adequados entre os ciclos de fermentação, a fim de restaurar ou melhorar o seu estado fisiológico, evitando perdas na viabilidade, na atividade de crescimento e de fermentação. Dois métodos para a propagação da linhagem IQAr/45-2 foram estabelecidos e ambos mostraram ser eficientes no acúmulo de células. No entanto, maiores aumentos foram obtidos quando o lisado celular de levedura (LC) foi adicionado ao meio, o qual levou a um crescimento semelhante a linhagem PE-2. A obtenção LC consiste em um método rápido, fácil e de menor custo do que o uso do extrato de levedura comercial. Dois tipos de meio (meios MR1 e MR2) foram utilizados para manter as células entre os ciclos de fermentação. O meio de MR1 continha sais de zinco, amônio e magnésio e o meio MR2 era constituído... / Brazilian distilleries produce ethanol using continuous or fed-batch fermentations. In repeated fed-batch fermentations, cells are sequentially transferred from one fermentation cycle to the next. At the end of each fermentation, the yeast cells are harvested and acid-treated in order to kill contaminants prior to its use as starter of the next fermentation. Depending on stresses conditions occurring before and during the fermentation process, as well as the concentration of the by-products and acid treatment, changes in the physiological state of the cells can occur. Such changes can affect the productivity and fermentation efficiency of the process. The variations in physiological state can be evaluated by measuring indicators such as levels of trehalose, proteins and glycerol and their degree of mobilization. In addition, studies are required for selection and use of the physiological parameters as diagnostic tools of the process performance. The aim of present work was to improve and/or avoid losses in viability, growth and fermentation activities by exposing the cells to appropriate treatments between fermentation cycles, particularly in order to restore or improve their properties. Two methods for the propagation of the strain IQAr/45-2 were stablished and both showed efficiency on cell acumulation. However, greater biomass and viability were obtained for IQAr/45-2 strain when yeast cell lysate (LC) was added to the medium, having a similar growth when compared to the PE-2 strain. The procedure to LC obtention is a faster, easier and lower coast method than the commercial yeast extract. Two kinds of medium (MR1 and MR2 media) were used to storage the cells between the fermentation cycles. The MR1 medium was supplemented with ammonium, zinc and magnesium salts and MR2 medium was supplemented with LC... (Complete abstract click electronic access below)

Orientador: Márcia Justino Rossini Mutton / Banca: Marco Antonio de Castro e Souza / Banca: Clóvis Parazzi / Resumo: A cana-de-açúcar e uma cultura importante para a economia do país, devido à sua alta eficiência na produção e acúmulo de sacarose. Entretanto, com a utilização crescente de cana crua tem se observado aumento de impurezas vegetais nos carregamentos, tais como folhas e pontas, ricas em amido, que interferem negativamente na produção de açúcar e álcool. Este trabalho teve por objetivo avaliar o desempenho de leveduras com atividade amilolítica na produção do álcool. Os testes foram feitos com três linhagens de leveduras amilolíticas, J07, J32 e Saccharomyces diastaticus, em combinação com uma levedura alcoogênica (Saccharomyces cerevisiae - CAT-01) em mosto preparado de caldo de cana da variedade SP81-3250, que apresentava elevado teor de amido. No vinho foram realizadas análises de açúcares redutores totais, teor alcoólico, glicerol, acidez sulfúrica e viabilidade celular. Os resultados revelaram que a combinação da CAT-01 com a linhagem J07 incrementou a produção de álcool. A levedura selvagem J32 não se constituiu boa fermentadora, podendo ser utilizada em combinação com outras linhagens de leveduras fermentadoras. / Abstract: Sugarcane is a very important crop to the economy of Brazil, because of its high efficiency in producing and accumulating sucrose. However, the rising rate of green cane harvest has resulted in higher levels of plant impurities as leaves and tops, which are rich in starch and affect both sugar and ethanol production. This work was carried out to evaluate the performance of amylolytic yeasts for ethanol production. The amylolytic strains J07 and J32, both isolated from the fermentation process, and Saccharomyces diastaticus were used in combination with Saccharomyces cerevisiae in a high-starch must from the sugarcane variety SP81-3250. The levels of total reducing sugars, ethanol, glycerol, sulfuric acidity and cell viability were determined. Results showed that the combination of S. cerevisiae with J07 increases ethanol production. The native yeast J32 has shown to be not good for fermentation, and for this reason must be used in combination with high fermentation yield strains. / Mestre

Orientador: Cecilia Laluce / Banca: Sandra Regina Pombeiro Sponchiado / Banca: Sandra Regina Ceccato Antonini / Resumo: As destilarias brasileiras produzem etanol utilizando o processo fermentativo contínuo ou por batelada alimentada. Nas fermentações sucessivas, as células são recicladas e usadas continuamente durante toda a safra. No final de cada ciclo de fermentação as células são concentradas e tratadas com ácido para redução dos contaminantes antes da utilização como starter da fermentação seguinte. Alterações no estado fisiológico das células podem ocorrer dependendo das condições que ocorrem antes e durante o processo e também durante o tratamento ácido, bem como a concentração de subprodutos formados. Tais alterações podem afetar a produtividade e a eficiência do processo fermentativo. As variações no estado fisiológico podem ser avaliadas através da medida de alguns indicadores, tais como níveis de trealose e seu grau de mobilização, proteínas e glicerol. Além disso, são necessários estudos para selecionar e utilizar os parâmetros fisiológicos como ferramentas de diagnóstico do desempenho do processo. O objetivo do presente trabalho foi o de expor as células a tratamentos adequados entre os ciclos de fermentação, a fim de restaurar ou melhorar o seu estado fisiológico, evitando perdas na viabilidade, na atividade de crescimento e de fermentação. Dois métodos para a propagação da linhagem IQAr/45-2 foram estabelecidos e ambos mostraram ser eficientes no acúmulo de células. No entanto, maiores aumentos foram obtidos quando o lisado celular de levedura (LC) foi adicionado ao meio, o qual levou a um crescimento semelhante a linhagem PE-2. A obtenção LC consiste em um método rápido, fácil e de menor custo do que o uso do extrato de levedura comercial. Dois tipos de meio (meios MR1 e MR2) foram utilizados para manter as células entre os ciclos de fermentação. O meio de MR1 continha sais de zinco, amônio e magnésio e o meio MR2 era constituído... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Brazilian distilleries produce ethanol using continuous or fed-batch fermentations. In repeated fed-batch fermentations, cells are sequentially transferred from one fermentation cycle to the next. At the end of each fermentation, the yeast cells are harvested and acid-treated in order to kill contaminants prior to its use as starter of the next fermentation. Depending on stresses conditions occurring before and during the fermentation process, as well as the concentration of the by-products and acid treatment, changes in the physiological state of the cells can occur. Such changes can affect the productivity and fermentation efficiency of the process. The variations in physiological state can be evaluated by measuring indicators such as levels of trehalose, proteins and glycerol and their degree of mobilization. In addition, studies are required for selection and use of the physiological parameters as diagnostic tools of the process performance. The aim of present work was to improve and/or avoid losses in viability, growth and fermentation activities by exposing the cells to appropriate treatments between fermentation cycles, particularly in order to restore or improve their properties. Two methods for the propagation of the strain IQAr/45-2 were stablished and both showed efficiency on cell acumulation. However, greater biomass and viability were obtained for IQAr/45-2 strain when yeast cell lysate (LC) was added to the medium, having a similar growth when compared to the PE-2 strain. The procedure to LC obtention is a faster, easier and lower coast method than the commercial yeast extract. Two kinds of medium (MR1 and MR2 media) were used to storage the cells between the fermentation cycles. The MR1 medium was supplemented with ammonium, zinc and magnesium salts and MR2 medium was supplemented with LC... (Complete abstract click electronic access below) / Mestre

Orientador: José Roberto Ernandes / Banca: Rubens Monti / Banca: Jonas Contiero / Banca: Sandra Helena da Cruz / Banca: João Atílio Jorge / Resumo: O presente trabalho teve como objetivo principal realizar estudos do efeito da complexidade estrutural de fontes de nitrogênio no fluxo metabólico do carbono em leveduras industriais utilizadas nas indústrias de panificação, de produção de vinhos e cervejas. Os resultados obtidos com os carboidratos trealose e glicogênio estão de acordo com o comportamento esperado para o acúmulo destes compostos, uma vez que foi observado que a quantidade destes carboidratos de reserva nas células sofrem acentuadas variações em resposta a diferentes alterações nutricionais experimentadas pelas leveduras durante o processo fermentativo, e isto ocorre como conseqüência do complexo sistema regulatório que controla a produção dos carboidratos. A maioria dos dados indica que a quantidade de trealose e glicogênio foram sempre maiores na suplementação com amônio e menor com peptona. A concentração de trealose e glicogênio produzidas pelas linhagens talvez reflita o processo de seleção a que foram submetidas as linhagens, o tipo a e concentração da fonte de carbono, o tempo de fermentação e também com a natureza estrutural da fonte de nitrogênio. / Abstract: The present work had as main objective to carry out studies of effects of the structural complexity of nitrogen sources in the metabolic flux of carbon in industrial yeasts used in the production of bread, wines, and beers. The results obtained with the carbohydrates trehalose and glycogen are in accordance with the expected behavior for the accumulation of these compounds, once it was observed that the amount of these reserve carbohydrates of in the cells suffers accented variations in response to the different nutritional alterations experienced by the yeasts during the fermentative process. This may occurs as consequence of the complex regulatory system that controls the production of the carbohydrates. The majority of the data indicates that the amount of trehalose and glycogen was always higher under ammonium and casamino acids supplementation than with peptone. Perhaps the concentration of trehalose and glycogen produced by the strains reflects the process of selection that the strains were submitted, the type and concentration of the carbon source, the time of fermentation and also with the structural nature of the nitrogen source. / Doutor

Orientadores: Jose Francisco Hofling, Marta Cristina Duarte Teixeira / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-09T04:57:27Z (GMT). No. of bitstreams: 1 Obando-Pereda_GustavoAlberto_M.pdf: 3782174 bytes, checksum: d7ee038a5b8df1578af994efc4637544 (MD5) Previous issue date: 2007 / Resumo: Tem-se demonstrado que espécies de Candida podem também ser encontradas no biofilme oral, co-agregadas ou aderidas a espécies bacterianas ali presentes, preferencialmente à espécies de Estreptococos, como também há adesão destas leveduras às células do epitélio e aos aparatos protéticos (neste caso é muito comum que pacientes portadores de próteses sejam alvos para as infecções fúngicas). Assim, a habilidade das espécies de Candida para formar biofilme em dispositivos médicos tem ampliado a capacidade de causar doenças assim como a capacidade de resistir a antifúngicos. Por outro lado nas ultimas décadas, tem se observado um crescente interesse nas medicinas alternativas e nas terapias naturais, justificando o aumento significativo de pesquisas nessa área ampliandose no presente. O objetivo desta pesquisa é, primeiramente, avaliar a interação das espécies de Candida albicans, C. tropicalis e C. glabrata em biofilmes préformados por espécies de Streptococcus oralis, S. sanguis, S. mitis, S. mutans e Staphylococcus aureus em diferentes materiais protéticos como: titânio e resina acrílica, e, secundariamente, testar a ação de alguns extratos fitoterápicos como a Mentha piperita, Cymbopogum martinii e Cympopogum winterianus através da concentração mínima inibitória (CMI), na inibição da co-agregação das espécies de Candida spp. a estes microrganismos. Os dados obtidos nesta pesquisa demonstram que as espécies de Candida albicans desenvolvem biofilme sobre os biofilmes pré-formados das bactérias testadas independentemente do material avaliado. A presença de bactérias se demonstra determinante para o desenvolvimento de biofilme por espécies de Candida. A inibição de Candida spp. pelos extratos vegetais testados se mostrou parcial e semelhante quando comparadas, ao lado de se revelarem substâncias potencialmente antifúngicas / Abstract: Candida spp. have been demonstrated found in the oral biofilm, coaggregated or adhered to oral bacteria, especially Streptococcus spp., also too the adhesion of these yeast to mucosal cells and prosthetic devices (in this case is common that patients that carry a prosthetic device, could be targets to fungal infections). The ability of Candida spp. to form biofilm on medical devices has extended the capacity to cause diseases and to resist antifungal agents. The lasts decades, have been observed big interest concern to medicinal plants and natural therapies, justifying the significant increase of research on the area in the present. The aim of this research was firstly, evaluate the interaction of C. albicans, C. tropicalis and C. glabrata on S. oralis, S. sanguis, S. mitis, S. mutans and Staphylococcus aureus preformed biofilm in different prosthetics materials like titanium and acrylic resin. Secondly, was evaluated the action of some vegetal extracts like Mentha piperita, Cymbopogum martinii and Cympopogum winterianus, using the Minimal Inhibitory Concentration (MIC), against the Candida spp. coaggregation with these microorganisms. The data obtained in this research show that Candida spp. develop biofilm on preformed bacterial biofilm, independently of tested material. Bacterias are determinant to Candida spp. biofilm development. The Candida spp. inhibition for vegetal extract was partial and equal when compared, revealing itself like potentially antifungal substances / Mestrado / Microbiologia e Imunologia / Mestre em Biologia Buco-Dental

The crystal structure of yeast (Saccharomyces cerevisiae) iso-1- cytochrome c has been determined through molecular replacement techniques, and refined against X-ray diffraction data in the resolution range 6.0-1.23 Å to a crystallographic R-factor of 0.192. The yeast iso-1-cytochrome c molecule has the typical cytochrome c fold, with the polypeptide chain organized into five α-helices and a series of loops which serve to enclose almost completely the heme prosthetic group within a hydrophobic pocket Comparison of the structures of yeast iso-1-, tuna and rice cytochromes c shows that the polypeptide backbone fold, intramolecular hydrogen bonding, conformation of side chains and particularly packing within the heme crevice of protein groups against the heme moiety are very similar in the three proteins. Significant structural differences among the three cytochromes c can be explained by differences in amino acid sequence. X-ray crystallographic techniques have also been used to study the effect of single-site amino acid substitutions at Phe82 and at Arg38 in iso-1-cytochrome c. The structures of the various variant iso-1-cytochromes c have been determined at nominal resolutions in the range 2.8 to 1.76 Å. Conspicuous structural perturbations in the neighborhood of the substituted side chain are evident in all of the variant proteins. In wild-type iso-1-cytochrome c, the phenyl ring of Phe82 is positioned adjacent and approximately parallel to the heme group, and occupies a non-polar cavity within the heme crevice. In the Ser82 variant, a channel extending from the surface of the molecule down into the heme crevice is created. In the Gly82 variant, the polypeptide backbone has refolded into the space formerly occupied by the phenyl ring of Phe82. Steric conflicts prevent both the phenolic ring of Tyr82 and the side chain of Ile82 from being completely accommodated within the pocket normally occupied by a phenyl ring. Substitution of alanine at position 38 causes a slight reorganization of the hydrogen bonding network in which Arg38 normally participates, and also exposes to external solvent a normally buried propionic acid group of the heme. The altered functional properties of the position 82 variant proteins have been interpreted with respect to the observed structural perturbations. The drop in reduction potential, most notably for the Ser82 and Gly82 variants, can be explained by the elevated heme environment polarity arising from the increased access of solvent or polar protein groups to the heme pocket The reduced stability of the heme crevice, as indicated by lowered pKa's for alkaline isomerization, is likely due to the disruption of stabilizing packing forces formed by the Phe82 phenyl ring within its hydrophobic cavity. The lowered activity, in comparison to the wild-type protein and the Tyr82 variant, for electron transfer with Zn+-cytochrome c peroxidase is attributed to the loss of an aromatic group positioned adjacent to the heme group. The altered surface topography of the variant proteins (particularly the Gly82, Tyr82 and Ile82 variants) may further hinder productive complex formation between cytochrome c and its redox partners. These results suggest that the invariant Phe82 contributes in at least three ways to the proper functioning of cytochrome c. It has an important structural role in maintaining the integrity of the heme crevice and in establishing the appropriate heme environment The phenyl ring of Phe82 may also be required for efficient movement of an electron to and from the heme of cytochrome c. Finally, Phe82 may have a role in forming intermolecular interactions with enzymic redox partners of cytochrome c. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Protoplasts were prepared from two yeast strains P. rhodozyma (ATCC 24202) and K. fragilis (ATCC 8455). Protoplasts prepared from P. rhodozyma were facilitated by prior growth of the cells in a media containing S-(2-aminoethyl)-L-cysteine. Protoplasts from these two yeast genera were fused either by the use of electrofusion or polyethylene glycol treatment. Stable carotenoid producing cell lines were selected by growth at 30°C on yeast nitrogen base plus galactose. Selected single fusants display taxonomic characteristics common to both genera with a cellular morphology and a carotenoid composition similar to that of P. rhodozyma. / Land and Food Systems, Faculty of / Graduate

This thesis was focused on study of biotechnological utilization of waste substrates to produce lipids by yeast of the genus Metschnikowia. Waste materials and their subsequent transformation into high value-added products such as microbial lipids are currently considered as an alternative source for biofuel production. Therefore, the experimental part was aimed at investigating the influence of a carbon source to the controlled overproduction of lipids by yeast Metschnikowia. Total of 12 yeast strains of the genus Metschnikowia were selected. Yeast strains M. pulcherrima , M. pulcherrima 147, M. pulcherrima 149, M. andauensis 129 a M. fructicola 15 were purchased from Culture Collection of Yeasts (CCY, Bratislava, Slovakia). The growth characteristics of this yeast strains were also studied. It may serve to better understanding of the physiology of the yeast strains and also to help in further analysis of the produced metabolites. The other strains M. chrysoperlae 1158, M. pulcherrima 1232, M. fructicola 1235, M. andauensis 1241, M. sinensis 1244, M. zizyphicola 1247 a M. shanxiensis 1250 were purchased from CBS (Centraalbureau voor Schimmelcultures, Utrecht, the Netherlands).Yeast strains were cultivated on crude animal fat, glycerol and cheese whey under conditions of different C/N ratios. Because of higher lipid yields, cultivation was carried out at 14°C for 14 days. The accumulated lipid content was determined by gas chromatography and Raman spectroscopy. The glycerol-containing medium was evaluated as the most suitable for microbial lipids production. The total amount of lipids present in cells of M. pulcherrima 1232 was 36,31%. At the same time, quantitative screening of lipase enzymatic activity in Metschnikowia yeast was performed using spectrophotometric method with p-NPP. Controlled production of lipolytic enzymes has been monitored by using two types of media: crude animal fat and crude animal fat with addition of emulsifier (Tween 80). The conclusion of the work was supplemented by analysis of the karyotype of yeasts of the genus Metschnikowia using the technique of pulsed gel electrophoresis.