Made available in DSpace on 2014-08-20T14:37:52Z (GMT). No. of bitstreams: 1 tese_ana_albano.pdf: 1506402 bytes, checksum: 95dde41b6fa23dd1ab15dcbaaea21132 (MD5) Previous issue date: 2012-12-12 / The paracoccidioidomycosis (PCM) is one of the most important systemic mycoses in Latin America, especially in Brazil. Although the Rio Grande do Sul is considered endemic area of the disease, there are few studies on the ecology of Paracoccidioides brasiliensis in the state. In this sense, the study aimed to detect infection by P. brasiliensis in animals using them as sentinels for detection of the agent in the region. Serology was performed by the techniques of double radial immunodiffusion in agar gel (AGID) and ELISA for detection of anti-gp43 of P. brasiliensis in wild animals served by Núcleo de Reabilitação da Fauna Silvestre UFPel, and horses from different stables in the region of Bagé. Serology was performed in 128 wild animals, and about 20% of these were positive on ELISA. The presence of anti-gp43 was detected in animals from the mesoregion of Southeast Riograndense and Metropolitan of Porto Alegre. There was no significant difference in seropositivity with respect to gender, age, order and habits of animals (p> 0.05), however seropositivity differed between the conjugates used (p <0.001). The study regarding horses included 200 animals from different places, which 12% had antigp43 antibodies in ELISA, with rates ranging from 0 to 30% according to the place of origin (p <0.001). Neither the wild animals nor equines were positive by immunodiffusion. The results indicate the presence of the fungus P. brasiliensis in different mesoregions of Rio Grande do Sul. / A paracoccidioidomicose (PCM) é uma das micoses sistêmicas mais importantes da América Latina com destaque no Brasil. Embora o Rio Grande do Sul seja considerado como área endêmica da doença, existem poucos estudos a respeito da ecologia do Paracoccidioides brasiliensis no estado. Neste sentido, o estudo objetivou detectar a infecção por P. brasiliensis em animais, utilizando-os como sentinelas da presença do agente na região. Para tal foi realizada sorologia a partir das técnicas de imunodifusão radial dupla em gel de Ágar (IDGA) e ELISA indireto utilizando distintos conjugados para detecção de anticorpos anti-gp43 do P. brasiliensis em animais silvestres atendidos pelo Núcleo de Reabilitação da Fauna Silvestre da UFPel, e em equinos de distintos haras da região de Bagé. A sorologia foi realizada em 128 animais silvestres, e cerca de 20% destes foram soropositivos no ELISA. A presença de anticorpos anti-gp43 foi detectada em animais oriundos das mesorregiões do sudeste rio-grandense e metropolitana de Porto Alegre. Não foi encontrada diferença significativa de soropositividade considerando gênero, idade, ordem e hábito dos animais (p>0,05), no entanto a soropositividade diferiu entre os conjugados utilizados (p<0,001). O estudo referente aos equinos incluiu 200 animais provenientes de distintos haras, dos quais 12% apresentavam anticorpos anti-gp43 no teste de ELISA, com taxas variando de 0 a 30% de acordo com o local de origem (p<0,001). Tanto no estudo com animais silvestres quanto no estudo com equinos não foi encontrado nenhum animal positivo pela técnica de imunodifusão. Os resultados indicam a presença do fungo P. brasiliensis em distintas mesorregiões do Rio Grande do Sul.

Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: This study describes: a) the optimization of the current industrial-scale extraction and purification of Horseradish peroxidase from horseradish at BBI Enzymes, focussing on: a. Raw material quality, b. Extraction, c. Ultra-filtration, d. Salt fractionation, e. Diafiltration, f. Ion Exchange Chromatography, b) developing an new in-process microtitre plate calorimetric assay, c) characterization of main groups of HRP relevant to BBI Enzymes by SDS-PAGE- and HPLC analysis. / AFRIKAANSE OPSOMMING: Hierdie studie beskryf: a) die optimisering van die huidige industriële-skaal ekstraksie en suiwering van peperwortelperoksidase vanuit peperwortel by BBI Enzymes, deur te focus op: a. Rou material kwaliteit, b. Ekstraksie, c. Ultra-filtrasie, d. Sout fraksionering, e. Diafiltrasie, f. Ioon-uitruilchromatografie b) Ontwikkeling van ‘n nuwe in-proses mikro-titer gebaseerde kalorimetriese toetsmetode c) die karakterisering van die hoof groepe peperwortel-peroksidase belangrik vir BBI Enzymes.

The effect of different blanching treatments and packaging materials on the enzymatic (lipoxygenase and peroxidase) activity and fatty acid stability of two different varieties of sweet corn on the cob (Jubilee and GH 2684) was evaluated during nine months of frozen storage at -23.3°C. The initial moisture content in the kernels of the two sweet corn varieties averaged 72.5%. After nine months of frozen storage the moisture content in the kernels of corn depended greatly on the packaging material used. The ears stored in Cryovac B and E bags showed the best moisture retention (72.2% final moisture content), followed by the polyethylene bags (71.4%) while the ears stored without packaging material showed severe dehydration (70.1%). The peroxidase and lipoxygenase activities were determined using spectrophotometric assays on a crude extract obtained from liquid nitrogen powdered corn. Both unblanched varieties of sweet corn showed similar initial peroxidase specific activity and general behavior during the nine months of frozen storage. The presence of lipoxygenase isozymes with different thermal stabilities in both varieties was suggested by the higher lipoxygenase specific activity found in Jubilee after freezing and nine months of frozen storage (0.135 units/mg protein) compared with the GH 2684 variety (0.115 units/mg protein). Complete inactivation of lipoxygenase was obtained after 9 minutes steam blanching at 100°C. Peroxidase was more heat resistant showing some remaining specific activity after 9 minutes steam blanching with a complete inactivation after 15 minutes steam blanching. No regeneration of either enzyme was observed during the nine months of frozen storage suggesting a permanent disruption of the active site of both enzymes. Relative fatty acid content was determined by gas chromatographic analysis of fatty acids methyl esters. The major fatty acids present in both varieties were palmitic (14.93%), stearic (2.79%), oleic (31.54%), linoleic (46.87%) and linolenic (1.89%) acids. Good stability of the polyunsaturated fatty acids was observed during the nine months storage at -23.3°C, with autoxidation as the main mechanism responsible for the decrease in the relative percent of polyunsaturated fatty acids. Some enzymatic oxidation also occurred, decreasing the linolenic acid content. The control of the degradation of polyunsaturated fatty acids depended mostly on the frozen storage temperature (-23.3°C) and not on the oxygen permeability of the different packaging materials. The results obtained in our study suggested that blanching of the ears of sweet corn had an important effect on reducing the enzyme activity but little effect on the polyunsaturated fatty acid degradation after 9 months of storage at -23.3°C. / Graduation date: 1994

Peroxidasen des Typs ClassII sowie die Chlorperoxidase waren für mehrere Jahrzehnte die einzigen bekannten sekretorischen Hämperoxidasen aus Pilzen. Im Jahr 2004 wurde eine neue, ungewöhnliche Hämperoxidase aus dem Speisepilz Agrocybe aegerita isoliert und biochemisch charakterisiert. Diese Peroxidase, heute offiziell Unspezifische Peroxygenase (UPO, EC 1.11.2.1) genannt, vereint in sich Eigenschaften sowohl intrazellulärer Cytochrom-P450-Enzyme als auch klassischer extrazellulärer, pilzlicher Peroxidasen des Typs ClassII. In dieser Arbeit wurden für den UPO-Modellorganismus A. aegerita und für andere UPO-produzierende Pilze erstmals Nukleotidsequenzen der Enzyme ermittelt und phylogenetisch analysiert. Die Genome zweier Stämme von A. aegerita wurden sequenziert. Die Ergebnisse legen nahe, dass UPOs gemeinsam mit der Chlorperoxidase Vertreter einer phylogenetisch alten Protein-Superfamilie (Häm-Thiolat-Peroxidasen) darstellen, die in allen echten Pilzen (Eumycota) zu finden ist, aber in echten Hefen und einigen hefeartig wachsenden Basidiomyceten sekundär reduziert wurde. Die natürliche Funktion dieser extrazellulären Enzyme ist bislang unbekannt. Im Labor können UPOs ausschließlich unter Verwendung leguminosenhaltiger Kulturmedien induziert werden. Die induzierende Komponente aus Soja wurde in dieser Arbeit als proteinogen identifiziert. Eine zweite neue, extrazelluläre Hämperoxidase-Superfamilie ausserhalb der ClassII-Peroxidasen wurde im Totholz bewohnenden Pilz Bjerkandera adusta entdeckt. Die als DyP-type-Peroxidasen benannten Enzyme (DyP, EC 1.11.1.19) wurden später auch in anderen Pilzen gefunden, von denen in dieser Arbeit ebenso Nukleotidsequenzen erhalten wurden. Die phylogenetische Analyse ergab, dass pilzliche Sequenzen der DyP-type-Peroxidase-Superfamilie sehr ungleichmäßig sowie ausschließlich in den Genomen höherer Pilze (Dikarya) verteilt sind und vermutlich einen prokaryotischen Ursprung haben. Transkripte von UPO- und DyP-Genen wurden in nahezu allen untersuchten Laubstreuproben aus unterschiedlich stark genutzten Waldgebieten der Deutschen Biodiversitätsexploratorien nachgewiesen. Dies legt eine biologische Funktion dieser neuen Hämperoxidase-Superfamilien in entsprechenden Habitaten nahe. / For several decades ClassII peroxidases and chloroperoxidase have been the only known secretory heme peroxidases from fungi. In 2004, a novel heme peroxidase was isolated and characterized from the edible mushroom Agrocybe aegerita. This enzym, nowadays called unspecific peroxygenase (UPO, EC 1.11.2.1), combines characteristics of intracellular cytochrom P450 enzymes as well as extracellular classII fungal peroxidases. In the course of this PhD work, for the first time nuleotide sequences of UPO model organism A. aegerita as well as other UPO producing fungi were obtained and phylogenetically analyzed. The whole-genome sequences of two strains of A.aegerita were obtained. The results suggest that UPOs and the chloroperoxidase together represent a phylogenetically old protein superfamily (of heme-thiolate peroxidases) that is found in all true fungi (Eumycota), but is secondarily reduced in true yeasts and a few yeast-like growing basidiomycetes. The natural function of these enzymes remains unclear. In the laboratory, fungi secrete UPOs solely in legume-containing culture media. The eliciting component of the legume soybean has been determined as proteinogenic. A second novel heme peroxidase superfamily outside the classII peroxidases was discovered in deadwood inhabiting fungus Bjerkandera adusta. These so-called DyP-type peroxidases (DyP, EC 1.11.1.19) were later found expressed in several other fungi for which nucleotide sequences were obtained in this work, too. Phylogenetical analysis revealed that fungal DyP sequences are exclusively but very unequally distributed in genomes of higher fungi (Dikarya). Fungal DyP sequences are seemingly decended from prokaryotes. Transcripts encoding UPOs and DyPs were found in almost every sample of leave litter (derived from forests with different intensity of human usage) investigated. This suggests biological functions of these new heme peroxidase superfamilies in respective habitats.

Spectroscopic characterization of ferric soybean peroxidase with peroxides were studied to determine the ligand coordination and to characterize the structure of the heme active site and its intermediates (ferryl species). The lifetime, chemical reactivity and distinctive colors of the ferryl species (FeIV) formed during the oxidation of peroxidase (FeIII) by peroxides enabled structure, dynamics and reaction mechanisms to be studied. Resonance Raman spectroscopy was used as a means of characterizing the structure of the soybean peroxidase and its intermediates. Excitation in the Soret absorption band at 406.7nm with 2-5mW laser power was used for this study. Resonance Raman spectra in the 200 to 1700 cm-1 region were obtained for the soybean peroxidase. However, the focus of this study was on the vibrational region of the resonance Raman spectra from 900 to 500cm-1 where the FeIV=O stretching frequencies for heme compound II intermediates are expected. Several pH and pD (deuterium substitution) samples of the soybean peroxidase were analyzed using resonance Raman spectroscopy. The vibrational stretching frequencies of the ferryl peroxidases varied with varying pH/pD were observed within the 773–787cm-1 range. From the deuterium experiment, accompanied with changes in the vibrational frequencies of the iron-ligand, a 3cm-1 upshift and intense resonant enhancement of the peaks, we observed the ferryl nature of compound II intermediate for soybean peroxidase. Badger’s rule was used to estimate the bond distances that existed within Fe-O which offers additional insight into the structure of the ferryl species. The estimated bond distance for the soybean peroxidase was significantly less than Fe-O bond distances proposed by X-ray crystallographers for other peroxidases in the same family. Comparing the vibrational frequencies of the ferryl intermediates in soybean peroxidase to that in heme proteins portrayed the effect the protein environment has on the vibrational frequencies.

Orientador: Ivan de Godoy Maia / Banca: Luiz Filipe Protasio Pereira / Banca: Jorge Mauríco Costa Mondego / Banca: Celso Luis Marino / Banca: Alessandra Ferreira Ribas / Resumo: A disponibilidade de promotores órgão/tecido-específicos responsáveis pela regulação de genes responsivos a estresses bióticos e abióticos constitui uma ferramenta fundamental para os programas de melhoramento genético do café (Coffea arabica) visando o incremento de resistência e tolerância. Relatamos aqui a caracterização de um promotor do gene que codifica uma provável peroxidase III em C. arabica (CaPrx), peroxidases da classe III (Prxs) são enzimas envolvidas numa variedade de processos fisiológicos relacionados com o estresse em plantas. A CaPrx é expressa em estágios iniciais da interação com o nematóide de galha (RKN). CaPrx mostrou expressão aumentada nas raízes de café inoculadas com RKN (Meloidogyne paranaensis) (12 h após a inoculação), mas nenhuma diferença significativa foi observada na expressão entre as plantas suscetíveis e resistentes. Ensaios com plantas de tabaco transgênicas portadoras do promotor CaPrx fusionado com o gene que codifica a β-glucuronidase (GUS), revelou que este promotor foi exclusivamente ativo nas galhas induzidas por RKN (Meloidogyne javanica). Em seções transversais de galhas, o repórter GUS foi detectado predominantemente em células gigantes. Um aumento na expressão do gene GUS em raízes de tabaco transgênicos foi detectado 16 horas pós-inoculação por RKN. Por outro lado, nenhuma alteração na expressão de GUS após tratamento com ácido jasmônico foi detectada. Um segundo estudo foi realizado a fim de desvendar o papel de um fator WRKY de café (CaWRKY1) na regulação do promotor do gene que codifica uma isoflavona redutase em café (CaIRL). Fatores de transcrição do tipo WRKY estão envolvidos na regulação da expressão de diversos genes de defesa em plantas. A região promotora do gene CaIRL possui diversos sítios W-boxes ao longo de sua sequência. Através de análises de deleção e ensaios de transativação ... / Abstract: Not available / Doutor

Metallocofactors are ubiquitous in nature, serving multiple purposes in proteins. These metallocofactors typically act as the site of catalysis or as an electron relay to move electrons within the protein, or within the cell, and are very energetically costly to manufacture. Yet, in nature it can appear that supernumerary, or ‘auxiliary’ cofactors are apparent, with no clear function. In this thesis, I address the question of what roles additional cofactors play, and why they are retained. The radical S-adenosylmethionine (AdoMet) enzyme superfamily has displayed great diversity in the cofactor requirements for its members. Some members of this family contain only the canonical [4Fe-4S] cluster, which reductively cleaves AdoMet to initiate chemistry, while others have additional [2Fe-2S] or [4Fe-4S] clusters. Even greater cofactor complexity is seen with the B12-dependent subclass, featuring a cobalamin-binding domain in addition to the canonical FeS cluster. The majority of this thesis has focused on using the technique of protein film electrochemistry (PFE) to study members of various subclasses of this superfamily: a dehydrogenase: BtrN, two methylthiotransferases: MiaB and RimO, as well as OxsB and TsrM, two B12-dependent enzymes. By evaluating the redox properties of members of different subclasses, we have been able to shed light on the redox properties of this superfamily, in general, and observed that the redox properties of auxiliary clusters can differ widely between subclasses (e.g. BtrN versus MiaB). PFE has also been used to evaluate five ferredoxins that are possible electron donors for MiaB from Thermotoga maritima. Additionally, bacterial cytochrome c peroxidases (bCCPs) are diheme enzymes catalyzing the detoxification of hydrogen peroxide; however, a novel subclass of bCCPs containing a third heme-binding motif has been identified in enteric pathogens. Protein film electrochemistry has been used to study the redox properties of Escherichia coli YhjA, a member of this subgroup. Further characterization of this novel bCCP was achieved with electron paramagnetic resonance, optical spectroscopy, and steady-state kinetics. Through characterizing YhjA and members of the AdoMet radical enzyme superfamily, we have shed light on the role these additional cofactors play in the mechanism and how these enzymes are tuned for their specific chemistries. / 2018-08-11T00:00:00Z

Thesis advisor: Clare O'Connor / Within cells, reactive oxygen species (ROS) are synthesized naturally and in response to environmental stimuli. However, ROS have deleterious effects on a wide range of cellular molecules. Oxidative stress, caused by the ROS generated by the partial reduction of oxygen, is a major cause of cell damage linked to the initiation and progression of numerous diseases. Thioredoxin peroxidase (Tpx1) plays important roles in cellular defense against ROS. Although homologous genes and their functions have been identified in other eukaryotes, the level of activity as well as the necessity of this protective enzyme in S. pombe exposed to oxidative stress has yet to be fully elucidated. To explore the role of the Tpx1 protein in oxidative stress resistance, novel strains were constructed in which the tpx1 gene was overexpressed. The polymerase chain reaction was used to amplify txp1, and the amplified sequence was cloned into the yeast overexpression plasmid, pNMT41, which allows overexpression under the control of the powerful promoter. DNA sequencing was used to determine that the sequences had been properly inserted into the vector. The plasmids were transformed into two leu- yeast strains: FWP6 and TP108-3C. Production of the Tpx1 protein was ensured using Western Blot techniques. Experimentation to test the responses of the tpx1 strain to oxidative stress will employ a variety of reactive oxygen generators, including hydrogen n peroxide, menadione, tert-butyl hydroperoxide, and paraquat. The results generally supported the proposed role of Tpx1 to confer additional resistance against the oxidative stress. In a complementary line of investigation, knockout strains are being constructed to reduce the levels of the Tpx1 in S. pombe. Gene deletion cassettes were constructed for tpx1. Currently, the strains are being analyzed for the successful replacement of the endogenous tpx1 gene by homologous recombination. If the absence of the protein results in decreased cell viability, the role of Tpx1 indicated by the overexpression experiments could be supported. / Thesis (BS) — Boston College, 2006. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: Biology. / Discipline: College Honors Program.

Os neutrófilos representam a primeira linha de defesa do organismo contra microrganismos invasores, células infectadas com vírus e células tumorais. Em seres humanos saudáveis eles constituem a população de células brancas mais abundantes do sangue. O reconhecimento de componentes microbianos por neutrófilos desencadeia a fagocitose, geração de espécies reativas de oxigênio (EROs) e liberação de proteínas granulares, tais como mieloperoxidase (MPO) e elastase (NE). Essas EROs e enzimas agem como agentes microbicidas e contribuem para os processos inflamatórios. Em trabalho anterior constatamos um aumento da atividade do sistema NADPH oxidase fagocítico (NOX2) e da geração de EROs por neutrófilos de ratos diabéticos e não diabéticos tratados com aminoguanidina (AG), um conhecido inibidor da formação de produtos finais de glicação avançada (AGEs), que culminou num aumento da atividade microbicida destas células. Nosso objetivo é elucidar o mecanismo de ação pelo qual a AG está atuando no nosso modelo. Desta forma, neste trabalho, avaliamos a hipótese que a AG estaria estimulando a degranulação de neutrófilos, portanto, estudamos a modulação da MPO e da NE, principais enzimas presentes nos grânulos azurófilos de neutrófilos. Para isso, utilizamos neutrófilos humanos isolados a partir de sangue periférico colhido por punção venosa de voluntários sadios. Os neutrófilos foram incubados ou não com AG 0,5 mM por um período de 18 horas em estufa de cultivo celular. A atividade de MPO foi avaliada através da formação de HOCl e através de quimiluminescência amplificada por luminol. A atividade de NE foi avaliada através da formação do produto da reação p-nitroanilida, por espectrofotometria. Avaliamos também a expressão da MPO e NE por Western blotting e imunomarcação. Como controle das reações foi utlizado inibidor de MPO (azida) e de NE (inibidor de tripsina de glycine max (soybean) - (STI)). O isolamento de neutrófilos permitiu obter uma população celular com mais de 95% de neutrófilos, com viabilidade superior a 99%. Nossos resultados mostraram que a AG não foi capaz de influenciar a atividade de cloração e basal da MPO, porém, observamos um aumento de ERO pelo tratamento com a AG após estímulo. Vimos também que a atividade de NE permaneceu inalterada pelo tratamento com a AG. Além disso, a AG não interferiu na expressão e distruibuição da MPO e NE. Estes resultados em conjunto nos levam a sugerir que a AG não influencia diretamente no processo de degranulação de neutrófilos, uma vez que este processo é um mecanismo utilizado por fagócitos com o propósito de criar um ambiente totalmente hostil para patógenos invasores. Aparentemente, o aumento da atividade microbicida de neutrófilos tratados com AG, não é via NE ou MPO na atividade clássica desta enzima, ou seja, na produção de HOCl. Entretanto, vimos uma resposta importante na atividade de MPO, que nos permite propor várias atividades para esta enzima, ora como uma atividade oxidativa que pode contribuir para o killing, ora como uma atividade antioxidante e desta forma pode proteger outras proteínas granulares antimicrobianas dos danos oxidativos. / Neutrophils represent the organism´s first line of defense against invading microorganisms, infected cells with viruses and tumor cells. In healthy humans, they constitute the most abundant population of white blood cells. The recognition of microbial components by neutrophils initiate phagocytosis, generation of reactive oxygen species (ROS) and release of granular proteins, such as myeloperoxidase (MPO) and elastase (NE). These ROS and enzymes act as microbicidal agents and contribute to the inflammatory processes. In a previous study, we found an increase in phagocyte NADPH oxidase system activity (NOX2) and generation of ROS by neutrophils of diabetic rats and non-diabetics treated with aminoguanidine (AG), a known inhibitor of the formation of advanced glycation end products (AGEs), which resulted in an increased microbicidal activity of these cells. Our aim is to elucidate the mechanism of action by which AG is acting in our model. This way, this study evaluated the hypothesis that AG would be stimulating degranulation of neutrophils, therefore, we studied the modulation of MPO and NE, the main enzymes present in azurophilic granules of neutrophils. For this, we used human neutrophils isolated from peripheral blood collected by venipuncture from healthy volunteers. Neutrophils were incubated or not with AG 0,5 mM for a 18 hour period in cell culture incubator. MPO activity was evaluated through the formation of HOCl and luminol-amplified chemiluminescence. NE activity was evaluated through the formation of the product of p-nitroanilide reaction, by spectrophotometry. We also evaluated the expression of MPO and NE by western blotting and immunostaining. As control of reactions was used inibitor of MPO (azide) and of NE (trypsin inhibitor from Glycine max (soybean) - (STI)). The neutrophil isolation allowed the achievement of a cell population with over 95% of neutrophils, with viability greater than 99%. Our results showed that AG was not capable of influencing the chlorination and basal activity of MPO, however, an increase of ROS by the treatment with AG after stimulation was noted. We also noted that NE activity remained unaltered by the treatment with AG. In addition, AG did not affect expression and distribution of MPO and NE. These results together lead us to suggest that AG does not influence directly the process of neutrophils degranulation, as this process is a mechanism used by phagocytes in order to create a fully hostile environment for invading pathogens. Apparently, the increase of microbicidal activity of neutrophils treated with AG is not via NE or MPO in the classical activity of this enzyme, in other words, in the production of HOCl. However, we observed a significant response in MPO activity, allowing us to propose various activities for this enzyme, either as a oxidative activity that can contribute to the killing or as an antioxidant activity, and therefore may protect other antimicrobial granular proteins from oxidative damage. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

A indução de resistência em plantas contra fitopatógenos é um método alternativo de controle de doenças, o qual envolve a ativação de mecanismos de resistência latentes da planta. Hoje no mercado existem poucos produtos que atuam segundo este princípio. Na busca de novas moléculas que possam ser utilizadas em campo, diversos compostos de origem microbiana, com capacidade de estimular uma ou mais respostas de defesa, já foram isolados e caracterizados. A Saccharomyces cerevisiae é uma levedura capaz de induzir resistência e elicitar respostas de defesa em algumas plantas. Com o objetivo de extrair da levedura um ou mais compostos capazes de induzir o acúmulo de fitoalexinas em cotilédones de soja e na proteção dessa leguminosa contra Microsphaera diffusa (agente causal do oídio da soja) e de pepino contra Colletotrichum lagenarium (agente causal da antracnose do pepino), células em suspensão foram autoclavadas. Os compostos assim extraídos foram inicialmente separados através de precipitação etanólica. Em seguida, foram realizadas cromatografias de troca iônica e de afinidade para separar as frações com maior poder elicitor das de baixo poder elicitor. A fração não adsorvida à resina DEAE-Celulose foi a que induziu maior acúmulo de fitoalexinas. No entanto, nenhum dos preparados testados foi capaz de conferir proteção a plantas de soja contra M. diffusa. Já no caso de pepino, plântulas tratadas com as frações resultantes da cromatografia de afinidade apresentaram reduções entre 50 e 70 % de área lesionada causada por C. lagenarium e aumento na atividade de peroxidases. Extratos incorporados à meio de cultivo não apresentaram efeito inibitório sobre o crescimento e esporulação de C. lagenarium. Com base nesses resultados, concluímos que existe na parede da levedura compostos capazes de induzir resistência local em pepino contra C. lagenarium, sendo que pelo menos um destes compostos é um carboidrato, contendo provavelmente manana e glucosamina. / The acquired resistance of plants to pathogens is an alternative method to control diseases which includes the activation of resistance mechanisms in the plants. A few products already commercially available have their action based upon this mechanism. In the search for novel molecules that can be used under field conditions, many compounds from microbes with the ability to stimulate one or more defense responses were already isolated and characterized. Saccharomyces cerevisiae is an yeast with the ability to induce defense responses and resistance in some plants. A suspension of cells from the yeast was autoclaved with the purpose of extracting one or more compounds with the ability to induce the accumulation of phytoalexins in soybean cotyledons and to protect soybean plants against Microsphaera diffusa (causal agent of powdery mildew) and plants of cucumber against Colletotrichum lagenarium (causal agent of anthracnose). The compounds extracted by this method were separated using ethanolic precipitation. After this step, the fractions of higher elicitation activity were separated from those of lower one by using ion exchange cromatography and affinity cromatography. The non-adsorbed fraction to DEAE-Cellulose was the one that induced the highest accumulation of phytoalexins. However, none of the fractions were able to protect soybean plants from M. diffusa. In the case of cucumber, seedlings treated with the fractions from affinity chromatography were able to reduce disease symptoms caused by C. lagenarium by 50 to 70 % and to increase the activity of peroxidases. Extracts that were incorporated into growing media did exhibit any inhibitory effect on in vitro growth and sporulation of C. lagenarium. According to these results, it is possible to conclude that there are compounds in the cell walls of the yeast that are able to induce local resistance to C. lagenarium in cucumber and that at least one of these compounds is a carbohydrate that likely contains mannan and glucosamine.