<p>  </p> <p>Actinobacteria are a large and diverse group of bacteria that are known to produce a wide range of secondary metabolites, many of which have important biological activities, including antibiotics, anti-cancer agents, and immunosuppressants. The biosynthesis of these compounds is often highly regulated, with many natural products being produced at very low levels in laboratory settings. Environmental factors, such as small molecule elicitors, can induce the production of secondary metabolites. These elicitors can be natural products, including antibiotics or hormones, or synthetic compounds. The use of small molecule elicitors to induce the production of secondary metabolites has several advantages. First, addition of elicitors to fermentation media can result in increased titers of known natural products. Second, elicitors can enable the discovery of novel natural products typically produced at undetectable levels. In recent years, there has been a growing interest in the use of small molecule elicitors to induce the production of secondary metabolites from actinobacteria, especially for the discovery of “silent” natural products. In this work, we sought to expand on the method of chemical induction by utilizing oxytetracycline at a sub-MIC concentration to induce secondary metabolite production in Streptomyces. We have shown that translation-inhibiting antibiotics, specifically oxytetracycline, have a profound effect on the production of coeliomycin P1, actinorhodin, and calcium-dependent antibiotics (CDAs) in S. coelicolor and S. lividans. The expression of actinorhodin in S. lividans under these conditions is unique, unlike its counterpart, S. coelicolor, which can produce actinorhodin under standard conditions. In addition to the increased production of known secondary metabolites, we have also demonstrated the induction of BGCs in several other strains of Streptomyces, which were observed via LC-MS. </p> <p>In addition to exploring antibiotics as elicitors we have explored the traditional approach of natural product discovery by taking an bioactivity guided approach. Several strain that we isolated from soil collect of Hawaii were screened for activity against several pathogenic strains primarily looking for which strain will inhibit the growth of a. baumannii, which is an intriguing target because the rate of resistance to common antibacterial medication is rising and it’s membrane composition is vastly different compared to other gram negative bacterium like E.coli. From this preliminary screening 1 strain (Streptomyces sp. CS62) out of the 8 that tested exhibited the desired biological activity. The supernatant of Streptomyces sp .CS62 was processed and screen by LC-MS to gain insight on the type of molecules that Streptomyces CS62 could produce. Upon our initial screening process none of the masses observed in the mass spec were matched to knowns. However, after 2D NMR analysis and genomic analysis it was unveiled that Streptomyces sp. CS62 produces factumycin a known antibacterial agent that targets A.baumannii .This unfourtunate turn of events illustrates the issues with natural product discovery and the need to improve natural product databases.</p> <p>In conjunction to discovering a novel producer of factumycin we are also investigating the production of antifungal compounds from Staphylococcus lugdunensis  a commensal strain that modulates the microbiome by producing lugdunin. The supernatant collected of Staphylococcus lugdunensis  is exclusively being test against Candida auris due to the immense health risk it possess to society because of its innate resistance to many antifungal drugs and its ability to rapidly gain resistance to other classes of antifungals.</p> <p>In addition to exploring the influence of antibiotics on secondary metabolite production and using bioactivity as a guide to discovering antibiotics. We are evaluating the soils collected from unique environments as potential sources for novel natural products. Specifically, we are evaluating the biosynthetic potential of bacteria from ore-forming environments, specifically fluorspar and topaz mines. Soils from ore-forming environments tend have low pH, high saline content, low water holding capacity, and poor nutrient availability. Therefore, ore-forming environments pose a hostile environment for life. To date, no one has explored the natural product potential, or the bacterial diversity, exhibited in these harsh environments. To assess the bacterial diversity, bacteria were isolated from various ore-forming environments using a procedure that is selective for actinobacteria. Following bacterial isolation, genomic DNA was isolated and 16s rRNA gene sequencing was performed to gauge the type of bacteria that were isolated. To stimulate secondary metabolite production, bacteria were then subjected to 7 different media conditions. The supernatant collected from these media conditions were tested against ESKAPE pathogens utilizing the CTSI broth microdilution assay. LC-MS MS analysis was performed for samples exhibiting biological activity. GNPS molecular networking was then utilized to determine potential molecules present in each sample.  Through this process we were able to identify one strain, which we named Streptomyces sp. S1A that exhibited a board range of biological activity (anticancer and antibacterial) and possess a wide array of biosynthetic gene clusters ranging complex macrolides (PKS and NRPS) to terpenes. </p> <p>In summary this multifaced approach to natural product discovery may lead to the discovery of novel antibiotics, enable us to increase production of known or unknown antibiotics through chemical induction, and the characterization of metabolites from Streptomyces sp. S1A will shed insight on the biochemical potential of organisms that inhabit ore-forming environments </p>

<p dir="ltr">Phosphatidylinositol phosphates (PIPs) and soluble inositol phosphates (IPs) serve as critical secondary messenger molecules that regulate cellular processes. The INPP5 family of phosphatases play an essential role in regulating levels of PIP-5’ and IP-5’ molecules. Src homology 2-containing-inositol phosphatases (SHIP), are a subgroup of the INPP5 family that consists of two members, SHIP1 and SHIP2. Both SHIP proteins have been identified to hydrolyze PI(3,4,5)P3 into PI(3,4)P2. Interestingly, the dysregulation of PI(3,4,5)P3 and SHIP proteins have been observed in multiple diseases, such as cancer, diabetes, and neurodegenerative disease. Recently, SHIP1 was identified as a potential risk factor for the development of Late-onset Alzheimer’s Disease (LOAD). Furthermore, knockdown and inhibition of SHIP1 using small-molecule inhibitors were shown to reduce phenotypes associated with LOAD. Taking these studies together suggests SHIP1 to be a potential therapeutic target for the treatment of LOAD.</p><p dir="ltr"><br></p><p dir="ltr">Despite SHIP1’s therapeutic potential, the development of specific small-molecule inhibitors that target SHIP1 has been challenging. One explanation for this challenge is that very little is known about the overall structure and function of SHIP1. In this thesis I will discuss in detail how we generated multiple SHIP1 constructs to improve our understanding of SHIP1’s overall structure and function in an <i>in vitro </i>setting.</p><p><br></p><p dir="ltr">Efficient protein production is essential for studying enzyme structure and function. The choice of expression system can impact protein yield and stability. The E. coli (BL21) and Baculovirus expression systems are two commonly used systems for protein production. While E. coli is cost-effective and can yield a large amount of protein, the Baculovirus system offers advantages in terms of protein folding and post-translational modifications. Using both systems to generate SHIP1 protein, we demonstrate that the Baculovirus system significantly enhances SHIP1 solubility for all generated constructs, making it the preferable choice for investigating the structure and function of SHIP1.</p><p><br></p><p dir="ltr">SHIP1, a 133 kDa protein, which comprises five established domains: an N-terminal Src Homolgy 2 (SH2) domain, 2.) a pleckstrin homology-related (PH) domain, 3.) an inositol phosphatase catalytic (Ptase) domain, 4.) a C2 domain, and 5.) a C-terminal domain containing proline-rich regions (PXXP) and tyrosine phosphorylated (NPXY) motifs. Despite their regulatory roles in phosphatase activity, protein-protein interactions, and membrane association, limited information is available about their structures and how they contribute SHIP1’s biochemical functions. In this study, we utilized baculovirus-expressed SHIP1 constructs to investigate the impact of each domain on macromolecular structure. Interestingly, a previously unrecognized domain within SHIP1 that directly impacts the enzyme's oligomeric state was identified. This work highlights that SHIP1's individual domains can significantly impact its overall structure and function, providing valuable insights for the development of potential therapeutics in the treatment of LOAD.</p><p><br></p><p dir="ltr">Accurate determination of phosphatase kinetics is vital for understanding the enzymatic activity and its potential involvement in disease. Using our baculovirus generated SHIP1 constructs, we employed in-vitro assays, including the malachite green (MG) and the 2-amino-6-mercapto-7-methylpurine riboside (MESG) coupled enzyme assays, to gain insight into SHIP1 kinetics. Results from the MG assay shows that SHIP1 can hydrolyze the PI(3,4,5)P3 diC8 substrate more efficiently than I(1,3,4,5)P4. Additionally, SHIP1’s PH domain was observed to increase the turnover of PI(3,4,5)P3 diC8. Furthermore, dimerization of SHIP1 was not observed to alter SHIP1 kinetics in any way. Lastly, no major differences in I(1,3,4,5)P4 kinetics were observed with the addition of SHIP1’s N-terminus. These results offer the first comprehensive biochemical characterization of SHIP1 across its substrates and N-terminal domains.</p><p><br></p><p dir="ltr">The development of potent and specific small-molecule inhibitors that target SHIP1 remains challenging. One potential cause for this challenge is that no structures of SHIP1 have been solved in complex with active compounds, making structure-based drug design impossible. In this study, we developed a covalent compound, <b>TAD-58547</b>, from a previously published fragment-based screen that was conducted on SHIP1’s Ptase and C2 domain. <b>TAD-58547 </b>was shown to effectively inhibited SHIP1's Ptase and C2 domains at modest potency. Using X-ray crystallography, this compound was observed to form a covalent interaction with a cysteine residue near the Phosphatase-C2 domain interface. Intriguingly, the inhibitor's potency was observed to be reduced in the presence of the SH2 domain. In addition to testing <b>TAD-58547</b> against our SHIP1 constructs, we investigated the effect of SHIP1’s N-terminus on the potency of a literature compound, <b>TAD-58616</b>. This compound was shown to inhibit all our tested constructs at low µM concentrations. Furthermore, using x-ray crystallography <b>TAD-58616 </b>was solved in complex with SHIP1’s Ptase and C2 domain. Intriguingly, density for <b>TAD-58616 </b>was shown to interact with a site previously identified from the fragment-based screen. While we initially determined this site to be a result of crystal packing, fragments bound to this site may have the potential to inhibit SHIP1. The work presented in this study reinforced the importance of testing inhibitors against physiological relevant forms of SHIP1, when developing potential therapeutics.</p><p><br></p><p dir="ltr">Lastly, new evidence has suggested that the binding of phosphorylated immunoreceptor tyrosine-based activation motifs (p-ITAM) and immunoreceptor tyrosine-based inhibitory motifs (p-ITIM) to SHIP1’s N-terminal SH2 domain is essential for its “Anchorage and Activation” at the plasma membrane (PM). With this model it is believed that SHIP1’s SH2 domain, places the phosphatase into an auto-inhibited state. Upon binding to immune receptor proteins and adaptor proteins that contain ITAM/ITIM sequences, SHIP1 becomes un-auto-inhibited, allowing it to efficiently hydrolyze PI(3,4,5)P3 embedded in the PM. While this model does support the notion that SHIP1 activity is mediated by its PM localization, our biophysical and biochemical characterization add another level of complexity to this regulatory event. Taking all these results together, we propose a novel model for SHIP1 called “Anchorage and Assist” and suggest innovative therapeutic strategies for targeting SHIP1.</p><p><br></p><p dir="ltr">In conclusion, this thesis highlights the importance of choosing suitable expression systems for efficient protein production. Additionally, it offers insight into SHIP1's regulatory mechanisms through the discovery of a novel domain impacting its oligomeric state. Furthermore, the accurate determination of SHIP1 kinetics enhances our understanding of this phosphatase and its potential implications in disease. Also, the identification and crystallization of a novel and previously determined inhibitor scaffolds in complex with SHIP1 increases our ongoing efforts to develop a small-molecule inhibitor that specifically targets SHIP1. Lastly, using recently published data, detailing SHIP1 PM localization and activation, we proposed a new model for SHIP1 activity and suggest novel therapeutic strategies for targeting SHIP1.</p>

El polen apícola es uno de los productos de la colmena más conocido. Las abejas melíferas (Apis mellifera L.) lo recolectan como una de sus principales fuentes de alimento. Si bien ha sido utilizado por las civilizaciones egipcia y griega atribuyéndole propiedades terapéuticas y “rejuvenecedoras”, no fue hasta después de la Segunda Guerra Mundial que comenzó a aumentar la producción y su consumo. Actualmente sus características composicionales lo constituyen en un excelente suplemento nutricional y debido a sus propiedades bioactivas es considerado un alimento funcional. Por otra parte, el polen es uno de los productos de la colmena que presenta mayor variabilidad composicional atribuido principalmente a las diferentes floraciones visitadas por las abejas. En este sentido, el análisis entomopalinológico es apropiado para clasificarlo como multi o monofloral siendo este último más estable composicionalmente, aunque también puede sufrir algunas variaciones según la zona de producción. Por esta razón, es imprescindible caracterizar al polen apícola fisicoquímicamente para poder asegurar su calidad endógena y brindar información completa al potencial consumidor. Así, el principal objetivo de este trabajo de tesis es desarrollar metodologías analíticas que permitan determinar la composición del polen de abeja recolectado en la zona centro de Argentina. Inicialmente la clasificación entomopalinológica de las muestras se realizó mediante la acetólisis de los granos de polen evitando la separación por colores de las cargas corbiculares, que es la etapa del proceso que demanda más tiempo. Desde el punto de vista físico-químico se desarrolló una metodología rápida y de bajo costo para la determinación de humedad y proteínas totales en un pool de cargas corbiculares empleando espectroscopía en el infrarrojo cercano (NIR) y técnicas quimiométricas tales como SPA-MLR e iSPA-PLS. Además, la utilización del modelo iSPA-PLS-DA permitió la clasificación de las muestras en relación a un contenido proteico de 20%. Por otra parte, se pusieron a punto técnicas de microanálisis para evaluar el contenido de polifenoles, flavonoides y proteínas en cargas corbiculares individuales estableciendo los respectivos coeficientes de variación. De esta manera se confirmó la elevada variabilidad que presentan los mencionados parámetros en la matriz apícola probablemente relacionados al origen botánico. Asimismo, se construyó un modelo quimiométrico en base a espectroscopía NIR para estimar el contenido de polifenoles totales en cargas individuales de polen corbicular. Finalmente, en relación al tratamiento poscosecha, se llevó adelante un ensayo cuantitativo in vitro de actividad antimicrobiana utilizando un extracto etanólico de propóleos como preservante de polen fresco como una alternativa de conservación de bajo costo. Los resultados obtenidos en esta tesis indican que el polen de abeja recolectado en la zona centro de Argentina presenta muy buenas características nutricionales y bioactivas en base al contenido de proteínas, polifenoles y flavonoides definidos principalmente por los taxones Brassicaceae (mayormente Diplotaxis tenuifolia), Eucalyptus sp. y Lotus sp. La incorporación de las metodologías desarrolladas en este trabajo para realizar el control de calidad de este producto apícola permitiría el correcto rotulado del producto comercial o, incluso definir su destino para las diferentes industrias (alimenticia, farmacéutica, cosmética). / Bee pollen is one of the best-known beehive products. Honey bees (Apis mellifera L.) collect it as one of their main food sources. Although it has been used by the Egyptian and Greek civilizations ascribing therapeutic and "rejuvenating" properties to it, it was after the Second World War that the production and consumption increased. Currently, its compositional characteristics make it an excellent nutritional supplement, and due to its bioactive properties, it is considered a functional food. On the other hand, bee-pollen is the hive product that presents the greatest compositional variability, attributed mainly to the different blooms visited by bees. In this sense, the entomopalynological analysis is suitable to classify it as multi or monofloral, being the latter more stable in its composition, although it may also undergo some variations depending on the production area. For this reason, it is essential to perform a physicochemical characterization of the bee-pollen to ensure its endogenous quality and provide complete information to the potential consumer. Thus, the main objective of this thesis is to develop analytical methodologies that allow determining the composition of the bee-pollen collected in central Argentina. Initially the entomopalynological classification of the samples was carried out through the acetolysis of the pollen grains, avoiding the separation by colours of the corbicular pellets, which is the stage of the process that requires more time. From the physicochemical point of view, a rapid and low-cost methodology was developed for the determination of moisture and total proteins in a pool of bee-pollen pellets using near-infrared spectroscopy (NIR) and chemometric techniques such as SPA-MLR and iSPA-PLS. Moreover, the use of the iSPA-PLS-DA model allowed the classification of the samples concerning a protein content of 20%. On the other hand, microanalysis techniques were developed to evaluate the content of polyphenols, flavonoids and proteins in individual corbicular loads, establishing the respective variation coefficients. In this way, the high variability of the mentioned parameters presented in the apicultural matrix was confirmed, probably related to botanical origin. Likewise, a chemometric model based on NIR spectroscopy was constructed to estimate the total polyphenols content in individual loads of corbicular pollen. Finally, concerning postharvest treatment, a quantitative in vitro antimicrobial activity test was carried out using an ethanolic extract of propolis as a preservative for fresh pollen as a low-cost conservation alternative. The results obtained in this thesis indicate that the bee-pollen collected in central Argentina has very good nutritional and bioactive characteristics based on the content of proteins, polyphenols, and flavonoids defined mainly by the Brassicaceae taxa (mostly Diplotaxis tenuifolia) , Eucalyptus sp. and Lotus sp. The incorporation of the methodologies developed in this work to carry out the quality control of this beekeeping product would allow the correct labelling of the commercial product or even define its destination for the different industries (food industry, pharmaceuticals, cosmetics).

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / O presente trabalho teve como objetivo avaliar as caracterÃsticas fÃsicas, fÃsico-quÃmicas e quÃmicas e atividade antioxidante total de frutos de viuvinha (Myrcia splendens (Sw.) DC.), mapirunga (Eugenia tinctoria Gagnep.) e araticum-do-brejo (Annona glabra L.) oriundos da vegetaÃÃo litorÃnea do CearÃ, em diferentes estÃdios de maturaÃÃo. Os frutos foram caracterizados fisicamente quanto à massa fresca, dimensÃes, coloraÃÃo, rendimento e firmeza. Para a avaliaÃÃo das caracterÃsticas fÃsico-quÃmicas e quÃmicas foram feitas as seguintes anÃlises: sÃlidos solÃveis (SS), acidez total titulÃvel (ATT), pH, relaÃÃo SS/ATT, aÃÃcares solÃveis totais e redutores, pectina solÃvel, vitamina C, clorofila e carotenÃides totais, antocianinas e flavonÃides amarelos, polifenÃis extraÃveis totais (PET) e atividade antioxidante total pelo mÃtodo ABTS. Os resultados indicaram que os frutos de viuvinha e mapirunga apresentaram um aumento na massa fresca, comprimento, diÃmetro e rendimento em polpa no decorrer do amadurecimento. Nos frutos de araticum-do-brejo observou-se um declÃnio nestes parÃmetros no final da maturaÃÃo. Observou-se tambÃm um aumento, no teor de sÃlidos solÃveis totais, na relaÃÃo SS/ATT, aÃÃcares solÃveis totais e aÃÃcares redutores nos frutos analisados à medida em que ocorria a hidrÃlise do amido. Apenas nos frutos de viuvinha ocorreu diminuiÃÃo da acidez ao longo do amadurecimento. Houve solubilizaÃÃo da pectina ao longo do amadurecimento, exceto nos frutos de mapirunga. Dentre os compostos bioativos observou-se aumento na vitamina C no decorrer do amadurecimento das frutas analisadas. Constatou-se alto teor de antocianinas nos Ãltimos estÃdios de maturaÃÃo na polpa de mapirunga. Apenas nos frutos de viuvinha houve uma diminuiÃÃo de flavonÃides amarelos no decorrer do amadurecimento. Foram encontrados altos valores de polifÃnois extraÃveis totais e elevada atividade antioxidante nos frutos de viuvinha e mapirunga. Houve correlaÃÃo positiva e significativa entre a atividade antioxidante total e o teor de polifenÃis extraÃveis totais de todas as espÃcies analisadas. As polpas de viuvinha e mapirunga sÃo promissoras tendo em vista considerÃveis teores de compostos bioativos como, carotenÃides, antocianinas, flavonÃides amarelos e polifenÃis totais extraÃveis que as fazem produtos diferenciais no mercado de frutas nativas. / O presente trabalho teve como objetivo avaliar as caracterÃsticas fÃsicas, fÃsico-quÃmicas e quÃmicas e atividade antioxidante total de frutos de viuvinha (Myrcia splendens (Sw.) DC.), mapirunga (Eugenia tinctoria Gagnep.) e araticum-do-brejo (Annona glabra L.) oriundos da vegetaÃÃo litorÃnea do CearÃ, em diferentes estÃdios de maturaÃÃo. Os frutos foram caracterizados fisicamente quanto à massa fresca, dimensÃes, coloraÃÃo, rendimento e firmeza. Para a avaliaÃÃo das caracterÃsticas fÃsico-quÃmicas e quÃmicas foram feitas as seguintes anÃlises: sÃlidos solÃveis (SS), acidez total titulÃvel (ATT), pH, relaÃÃo SS/ATT, aÃÃcares solÃveis totais e redutores, pectina solÃvel, vitamina C, clorofila e carotenÃides totais, antocianinas e flavonÃides amarelos, polifenÃis extraÃveis totais (PET) e atividade antioxidante total pelo mÃtodo ABTS. Os resultados indicaram que os frutos de viuvinha e mapirunga apresentaram um aumento na massa fresca, comprimento, diÃmetro e rendimento em polpa no decorrer do amadurecimento. Nos frutos de araticum-do-brejo observou-se um declÃnio nestes parÃmetros no final da maturaÃÃo. Observou-se tambÃm um aumento, no teor de sÃlidos solÃveis totais, na relaÃÃo SS/ATT, aÃÃcares solÃveis totais e aÃÃcares redutores nos frutos analisados à medida em que ocorria a hidrÃlise do amido. Apenas nos frutos de viuvinha ocorreu diminuiÃÃo da acidez ao longo do amadurecimento. Houve solubilizaÃÃo da pectina ao longo do amadurecimento, exceto nos frutos de mapirunga. Dentre os compostos bioativos observou-se aumento na vitamina C no decorrer do amadurecimento das frutas analisadas. Constatou-se alto teor de antocianinas nos Ãltimos estÃdios de maturaÃÃo na polpa de mapirunga. Apenas nos frutos de viuvinha houve uma diminuiÃÃo de flavonÃides amarelos no decorrer do amadurecimento. Foram encontrados altos valores de polifÃnois extraÃveis totais e elevada atividade antioxidante nos frutos de viuvinha e mapirunga. Houve correlaÃÃo positiva e significativa entre a atividade antioxidante total e o teor de polifenÃis extraÃveis totais de todas as espÃcies analisadas. As polpas de viuvinha e mapirunga sÃo promissoras tendo em vista considerÃveis teores de compostos bioativos como, carotenÃides, antocianinas, flavonÃides amarelos e polifenÃis totais extraÃveis que as fazem produtos diferenciais no mercado de frutas nativas. / The objective of this work was to characterize physical, chemical and physico-chemical and evaluate the total antioxidant activity of fruits of viuvinha (Myrcia splendens (Sw.) DC.), mapirunga (Eugenia tinctoria Gagnep.) and araticum-do-brejo (Annona glabra L.) from the coastal vegetation of CearÃ, at different stages of maturation. Fruits were physically characterized by size, color, total weight, yield, and firmness. For the evaluation of the physico-chemical and chemical analyzes were made as follows: soluble solids (SS), acidity, pH, SS / TA ratio, total soluble sugars and reducing sugars, soluble pectin, vitamin C, chlorophyll and total carotenois, anthocyanins and yellow flavonoids , total extractable polyphenols (TEP) and total antioxidant activity (TAA) by ABTS method. The study found that the fruits of viuvinha and mapirunga showed an increase in total weight, length, diameter and yield of the pulp during ripening. In fruits of araticum observed a decrease in these parameters at the end of maturation. Were also observed an increase in soluble solids, SS / TA ratio, total soluble sugars, reducing sugars in the fruits analyzed as starch hydrolysis occurred. Only the fruits of viuvinha there was a decrease in acidity during the ripening. There solubilization of pectin during ripening, except in the fruits of mapirunga. Among the bioactive compounds reported an increase in vitamin C during ripening of fruits analyzed. There was a high content of anthocyanins in late-stage of viuvinha and mapirunga. Only in the fruits of viuvinha there were yellow flavonoids during ripening. Emphasis was placed high values of extractable polyphenols and total antioxidant activity in fruits of viuvinha and mapirunga. There was significant positive correlation between total antioxidant activity and total extractable polyphenol content of all species analyzed. The pulp of viuvinha and mapirunga are promising in view of considerable amounts of bioactive compounds such as carotenoids, anthocyanins, yellow flavonoids and total polyphenols total extractable differentials that make products on the market of native fruits. / The objective of this work was to characterize physical, chemical and physico-chemical and evaluate the total antioxidant activity of fruits of viuvinha (Myrcia splendens (Sw.) DC.), mapirunga (Eugenia tinctoria Gagnep.) and araticum-do-brejo (Annona glabra L.) from the coastal vegetation of CearÃ, at different stages of maturation. Fruits were physically characterized by size, color, total weight, yield, and firmness. For the evaluation of the physico-chemical and chemical analyzes were made as follows: soluble solids (SS), acidity, pH, SS / TA ratio, total soluble sugars and reducing sugars, soluble pectin, vitamin C, chlorophyll and total carotenois, anthocyanins and yellow flavonoids , total extractable polyphenols (TEP) and total antioxidant activity (TAA) by ABTS method. The study found that the fruits of viuvinha and mapirunga showed an increase in total weight, length, diameter and yield of the pulp during ripening. In fruits of araticum observed a decrease in these parameters at the end of maturation. Were also observed an increase in soluble solids, SS / TA ratio, total soluble sugars, reducing sugars in the fruits analyzed as starch hydrolysis occurred. Only the fruits of viuvinha there was a decrease in acidity during the ripening. There solubilization of pectin during ripening, except in the fruits of mapirunga. Among the bioactive compounds reported an increase in vitamin C during ripening of fruits analyzed. There was a high content of anthocyanins in late-stage of viuvinha and mapirunga. Only in the fruits of viuvinha there were yellow flavonoids during ripening. Emphasis was placed high values of extractable polyphenols and total antioxidant activity in fruits of viuvinha and mapirunga. There was significant positive correlation between total antioxidant activity and total extractable polyphenol content of all species analyzed. The pulp of viuvinha and mapirunga are promising in view of considerable amounts of bioactive compounds such as carotenoids, anthocyanins, yellow flavonoids and total polyphenols total extractable differentials that make products on the market of native fruits.

Cette thèse porte sur la mise en œuvre de procédés innovants d'extraction de composés naturels bioactifs de plantes. Nous avons considéré ici l'extraction d'un composé anti-oxydant, l'acide rosmarinique, à partir du végétal mélisse (Melissa officinalis L.), qui contient également d'autres composés d'intérêt (citral et caryophyllène). Différentes techniques d'extraction-purification, soit à haute pression (extraction au CO2 supercritique) ou à pression atmosphérique (extraction Soxhlet, extraction batch, nanofiltration etc.) ont été envisagées. L'objectif de ce travail a été d'étudier expérimentalement l'influence des paramètres opératoires (débit, composition et concentration du solvant, taille des particules, pression et température) sur la cinétique des processus afin de sélectionner les meilleures conditions pour chaque opération. Les résultats expérimentaux ont été ensuite comparés avec plusieurs modèles mathématiques décrivant les phénomènes de transfert de masse et l'écoulement au travers du milieu poreux constitué par la matière végétale broyée. Cette démarche, à partir de la détermination des paramètres physiques du modèle, a fourni les éléments pour une extrapolation potentielle à l'échelle industrielle. Du point de vue du procédé complet d'extraction-purification, l'originalité du travail a été de proposer plusieurs scénarii d'enchainement d'opérations, couplant en synergie des opérations conventionnelles à pression atmosphériques (macération, nano-filtration etc.) et des opérations de traitement au CO2 supercritique avec co-solvant. / This thesis deals with the extraction of natural bioactive compounds from plants (case study with Lemon Balm (Melissa officinalis L.)) by using different separation techniques at high (supercritical extraction) and atmospheric pressure (Soxhlet extraction, batch extraction, nanofiltration etc.). The influence of main operational parameters (solvent composition and flow rate, particle size of the raw material, pressure, temperature) on the process kinetics is studied experimentally with the aim to determine appropriate operational conditions for better extraction. The experimental results are confronted to a number of mathematical models in order to estimate the applicability of different theoretical concepts to the particular process and to select and apply appropriate models for determination of important parameters, characterizing the mass transfer process and necessary for scale-up and design purposes. Coupling between different separation methods is also considered and a number of integrated process schemes are proposed resulting in better yield of the targeted compounds.

Os sucos e bebidas de frutas são altamente consumidos pela população brasileira, feitos de modo caseiro ou industrializados, porém pouco se sabe sobre o conteúdo de compostos bioativos e capacidade antioxidante destes diferentes tipos de produtos. O objetivo deste trabalho foi a determinação e comparação de diferentes compostos bioativos e da capacidade antioxidante de sucos naturais, integrais, néctares, preparados líquidos e sólidos para refresco nos sabores de tangerina, goiaba, maracujá, laranja e uva. Os sucos naturais foram preparados descartando as cascas e sementes e para cada tipo de bebida industrializada foram selecionadas três marcas distintas adquiridas no comércio local em Ribeirão Preto/SP. Foram determinados os teores dos seguintes compostos bioativos: fenólicos totais, flavonoides totais e amarelos, antocianinas totais, ácido ascórbico, licopeno e beta caroteno nas bebidas, assim como, a capacidade antioxidante foi determinada pelos métodos ABTS, DPPH e FRAP, sendo os dados analisados estatísticamente atráves da diferença entre as médias, também foram feitas correlações entre os diferentes métodos de capacidade antioxidante. Entre os achados observa-se que os preparados sólidos para refresco de diferentes marcas e sabores apresentaram os menores teores de compostos bioativos e capacidade antioxidante em relação às demais bebidas. O conteúdo de ácido ascórbico, fenólicos, flavonoides totais e amarelos apresentaram uma variabilidade grande entre as bebidas. As diferentes marcas de mesmo tipo de bebida e sabor apresentaram diferença significativa no teor de compostos bioativos e capacidade antioxidante. As antocianinas totais apresentaram-se em maior quantidade nos preparados líquidos e sucos integrais de uva. Os maiores teores de licopeno foram encontrados nos sucos natural e integrais de goiaba. O beta caroteno foi detectado apenas em algumas amostras sendo que as bebidas de tangerina apresentaram os maiores valores deste carotenoide, no suco natural, alguns sucos integrais, néctares e preparados líquidos para refresco. Os três diferentes métodos de capacidade antioxidante apresentaram resultados divergentes entre si e dados de correlação variáveis em sabores de frutas, porém, em todos os métodos, os preparados sólidos para refresco de diferentes marcas e sabores apresentaram medidas mais baixas ou nulas de capacidade antioxidante, mostrando que estes produtos são os mais inadequados do ponto de vista nutricional. Os resultados deste trabalho também mostram a dificuldade na caracterização destes produtos, comparando diferentes formulalções assim como diferentes produtos industrializados, mas apontam para valorização dos produtos naturais e menos processados / Juices and fruit drinks produced at home or industrialized are highly consumed by the Brazilian population, but the content of bioactive compounds and antioxidant capacity of these different types of products is unknown.The objective of this work was the determination and comparison of bioactive compounds and antioxidant capacity of natural and whole juices, nectars, liquid and solid prepared for soft drinks of tangerine, guava, passion fruit, orange and grape. The natural juices were prepared with the fruit pulp, scorning the barks and seeds and for each type of industrialized beverages, three different brands were collected in the local market of Ribeirão Preto -SP.Total phenolics, total and yellow flavonoids, total anthocyanins, ascorbic acid, lycopene and beta carotene were determined and analyzed by the difference between the averages, the antioxidant capacity was evaluated by ABTS, DPPH and FRAP assays, also their correlations were determined. Prepared solids for soft drinks of different brands and flavors had the lowest levels of bioactive compounds and antioxidant capacity compared to other beverages. The content of ascorbic acid, phenolics, total and yellow flavonoids presented a great variability between the drinks. The different brands of the same type of beverage and flavor presented significant difference in the content of bioactive compounds and antioxidant capacity.The total anthocyanins were higher in liquid prepared for soft drinks and whole grapes juices. The highest levels of lycopene were found in natural and whole juices of guava. Beta carotene was not found in all beverages studied, tangerine beverages presented the highest values of this carotenoid in natural juice, some whole juices, nectars and liquid prepared for soft drinks. Three different methods of antioxidant capacity presented divergent results and variable correlation data on fruit flavors, however, in all methods, solid prepared for soft drinks of different brands and flavors presented the smallest and zero antioxidant capacity, showing that these products are the least nutritionally suitable. The results of this work show the difficulty in characterizing these products, comparing different formulations as well as different industrialized products, but stand out for the valorization of natural and less processed products

Flores comestíveis são cada vez mais utilizadas nas preparações culinárias, sendo também reconhecidas por seus potenciais efeitos benéficos na saúde humana, o que exige novas abordagens para melhorar a sua conservação e segurança. Estes produtos altamente perecíveis devem ser cultivados sem o uso de agrotóxicos. Tratamento de irradiação pode ser a resposta a estes problemas, garantindo a qualidade dos alimentos, aumentando seu prazo de validade e desinfestação. Tropaeolum majus L. (capuchinha) e Viola tricolor L. (amorperfeito) são flores amplamente utilizadas nas preparações culinárias, sendo também reconhecidas por suas propriedades antioxidantes e alto teor de compostos fenólicos. O objetivo deste estudo foi avaliar os efeitos dose-resposta da irradiação por gama e feixe de elétron (doses de 0, 0,5, 0,8 e 1 kGy) sobre a atividade antioxidante, compostos fenólicos, aspectos físicos e potencial antiproliferativo das flores comestíveis. O flavonoide Kaempferol-O-hexosídeo-Ohexosídeo foi o composto mais abundante em todas as amostras de flores de Tropaeolum majus, enquanto Pelargonidina-3-O-soporosídeo foi a principal antocianina. Em geral, as amostras irradiadas demonstraram maior atividade antioxidante. Nas amostras da Viola tricolor, os compostos fenólicos mais abundantes foram os flavonois, especialmente aqueles derivados da quercetina. Em geral, as amostras irradiadas com raios gama, independentemente da dose aplicada, apresentaram quantidades mais elevadas m compostos fenólicos, os quais também foram favorecidos pela dose de 1,0 kGy independente da fonte utilizada. A atividade antioxidante também foi maior entre as amostras irradiadas. As duas espécies de flores comestíveis não apresentaram as amostras não apresentaram potencial antiproliferativo e citotoxicidade. Assim, os tratamentos por irradiação aplicados, demonstraram ser uma tecnologia viável para preservar a qualidade de pétalas de flores comestíveis, considerando as exigências impostas para sua utilização. / Edible flowers are increasingly being used in culinary preparations, being also recognized for their potential valuable effects in human health, which require new approaches to improve their conservation and safety. These highly perishable products should be grown without using any pesticide. Irradiation treatment might be the answer to these problems, ensuring food quality, increasing shelf-life and disinfestation of foods. Irradiation treatment might be the answer to these problems, to ensure food quality, to increase shelf-life and disinfestation of foods. Tropaeolum majus L. (nasturtium) and Viola tricolor L. (johnny-jump-up) flowers are widely used in culinary preparations, being also acknowledged for their antioxidant properties and high content of phenolics. The purpose of this study was to evaluate the dose-dependent effects of gamma and electron beam irradiation (doses of 0, 0.5, 0.8 and 1 kGy) on the antioxidant activity, phenolic compounds, physical aspects and antiproliferative potential of edible flowers. Kaempferol-O-hexoside-O-hexoside was the most abundant compound in all samples of Tropaeolum majus flower while pelargonidin-3-O-sophoroside was the major anthocyanin. In general, irradiated samples gave higher antioxidant activity, probably due to their higher amounts of phenolic compounds, which were also favored by the 1.0 kGy dose, regardless of the source . The Viola tricolor samples displayed flavonols as the most abundant phenolic compounds, particularly those derived from quercetin. In general, gamma-irradiated samples, independently of the applied dose, showed higher amounts in phenolic compounds, which were also favored by the 1.0 kGy dose, regardless of the source. The antioxidant activity was also higher among irradiated samples. The two species of edible flowers have not provided the samples did not show potential antiproliferative and cytotoxicity. Accordingly, the applied irradiation treatments seemed to represent a feasible technology to preserve the quality of edible flower petals, considering the requirements imposed by their increasing uses.

O feijão tem destaque por ser base da alimentação brasileira juntamente com o arroz, fornecendo nutrientes essenciais ao desenvolvimento humano, como proteínas e minerais. Os benefícios do consumo de feijão vão além do fornecimento de nutrientes essenciais, já que esta leguminosa tem se destacado também por seu potencial benéfico à saúde, apresentando efeito antioxidante e antimutagênico. Estes efeitos têm sido associados à presença de compostos bioativos, em especial os compostos fenólicos. O processamento térmico é essencial para o consumo destes grãos, entretanto, pode causar modificações nos seus fitoquímicos, seja na composição ou na concentração. Além disso, é preciso ressaltar que nem todo conteúdo de polifenóis, ou de qualquer nutriente, é absorvido e aproveitado integralmente pelo organismo, é preciso compreender como o processo digestivo influencia a bioacessibilidade destes compostos. Sendo assim, este trabalho teve como objetivo avaliar a influência de diferentes formas de tratamento térmico sobre os principais compostos fenólicos (mais relevantes de acordo com a literatura), conteúdo de fenólicos totais e atividade antioxidante dos feijões carioca e preto (variedades mais consumidas no Brasil), além de verificar a influência do tratamento térmico e do processo digestivo simulado in vitro sobre a concentração e a bioacessibilidade dos compostos fenólicos encontrados nas amostras. Foram avaliados grãos crus; grãos macerados em água por 12 horas; grãos cozidos em panela aberta com ou sem a água de maceração; grãos cozidos sob pressão com ou sem a água de maceração. Foram realizadas análises de compostos fenólicos totais, atividade antioxidante pelos métodos de DPPH, ABTS e ORAC, além da análise cromatográfica dos principais compostos fenólicos dos feijões. As amostras cozidas foram submetidas à digestão in vitro e analisadas quanto ao seu conteúdo de compostos fenólicos. O tratamento térmico promoveu a liberação de compostos fenólicos ligados a matriz, aumentando assim, o conteúdo de polifenóis livres acompanhado do aumento do potencial antioxidante em relação ao grão cru. A utilização da água de maceração e de pressão durante a cocção preservou maior conteúdo de polifenóis em relação aos tratamentos onde a água foi descartada e aos tratamentos em panela aberta. A avaliação da bioacessibilidade permitiu verificar que o processo digestivo tem influência na concentração dos polifenóis, sendo que a etapa duodenal, em especial, foi marcada por redução na concentração dos compostos fenólicos analisados. As amostras em que a água de maceração foi utilizada na cocção tinham maior conteúdo de polifenóis livres, entretanto, este conteúdo não estava totalmente bioacessível. Sendo assim, o descarte da água de maceração e a utilização de pressão para cocção são os procedimentos mais indicados para preservação dos compostos fenólicos da fração bioacessível. Neste estudo foi evidenciado que o tratamento térmico, a matriz e o processo digestivo in vitro têm influência sobre os compostos fenólicos da fração bioacessível. / Beans are highlighted as being the basis of the Brazilian diet along with rice, providing essential nutrients for human development, such as proteins and minerals. The benefits of bean consumption go beyond the supply of essential nutrients, since this legume has also been highlighted by its beneficial potential to health, showing antioxidant and antimutagenic effect. These effects have been associated with the presence of bioactive compounds, especially the phenolic compounds. Thermal processing is essential for the consumption of these grains, however, it can cause changes in their phytochemicals, either in composition or in concentration. In addition, it should be noted that not all polyphenol content, or any nutrient, is fully absorbed and utilized by the body, so it is necessary to understand how the digestive process influences the bioaccessibility of these compounds. Therefore, the objective of this work was to evaluate the influence of different forms of thermal treatment on the main phenolic compounds (more relevant according to the literature), total phenolic content and antioxidant activity of the carioca and black beans (most consumed varieties in Brazil), in addition to verifying the influence of the heat treatment and simulated in vitro digestive process on the concentration and bioaccessibility of the phenolic compounds found in the samples. Raw grains; grains macerated in water for 12 hours; grains cooked in an open pan with or without soaking water; grains cooked under pressure with or without soaking water were evaluated. Analyzes of the total phenolic compounds, antioxidant activity by the DPPH, ABTS and ORAC methods, as well as the chromatographic analysis of the main phenolic compounds of the beans were performed. Cooked samples were submitted to in vitro digestion and analyzed for their content of phenolic compounds. The heat treatment promoted the release of phenolic compounds bound to the matrix, thus increasing the content of free polyphenols accompanied by the increase of the antioxidant potential in relation to the raw grain. The use of the soaking water and pressure during cooking preserved a higher content of polyphenols than the treatments in which the water was discarded and the treatments cooked in an open pan. The evaluation of the bioaccessibility allowed to verify that the digestive process has influence in the concentration of the polyphenols, and that the duodenal stage, in particular, was marked by reduction in the concentration of the phenolic compounds analyzed. The samples in which the soaking water was used in the cooking process had higher content of free polyphenols, however, this content was not totally bioaccessible. Therefore, the discard of the soaking water and the use of cooking pressure are the most suitable procedures for the preservation of the phenolic compounds of the bioaccessible fraction. In this study it was evidenced that the thermal treatment, the matrix and the in vitro digestive process have influence on the phenolic compounds of the bioaccessible fraction.

As espécies patogênicas do gênero Colletotrichum apresentam importância mundial, pois causam danos a várias culturas de interesse agronômico. Diversas medidas de controle são empregadas, mas estas nem sempre são eficazes devido à ocorrência de linhagens resistentes. Desta forma, se faz necessário a busca por novos compostos que possam ser utilizados no manejo integrado desta doença. Os produtos naturais isolados de micro-organismos podem ser uma alternativa para o desenvolvimento de novos defensivos agrícolas. Dentre os micro-organismos, as actinobactérias são conhecidas pela produção de inúmeros compostos antimicrobianos. Neste contexto, o presente estudo teve como objetivo o isolamento e a identificação de compostos antifúngicos produzidos por actinobactérias da rizosfera de guaranazeiros. Para isto, a seleção de actinobactérias foi baseada em dois ensaios. No primeiro, as 65 actinobactérias foram avaliadas em ensaio de cultivo pareado frente ao fitopatógeno Colletotrichum gloeosporioides. Destas, os isolados mais promissores foram AM1 (43,78 % de inibição do crescimento micelial), AM3 (43,98 %), AM18 (37,86 %), AM25 (43,17 %), AM30 (47,12 %), AM61 (40,12 %) e AM68 (47,94 %). No segundo ensaio, estes isolados foram cultivados em meio BD e, após o cultivo, o meio metabólico foi submetido a três métodos de extração: (a) partição líquido-líquido com n-butanol; (b) partição líquido-líquido com acetato de etila e (c) coluna sílica gel C18. As frações obtidas a partir das três metodologias foram avaliadas pelo método de difusão em disco de papel contra C. gloeosporioides. Neste ensaio de difusão em disco foram selecionadas as linhagens AM1(n-butanol), AM3 (acetato de etila) e AM25 (C18) para o estudo de bioprospecção. Estas foram identificadas por técnicas moleculares como pertencentes ao gênero Streptomyces. A partir do extrato bruto da Streptomyces sp. AM1 foi isolado um composto análogo do ácido proclavamínico, o qual apresentou atividade mínima inibitória (MIC) de 1,25 mg mL-1 contra o fitopatógeno C. gloeosporioides. Da linhagem Streptomyces sp. AM3 foi isolado o composto streptimidona que apresentou MIC de 1,25 mg mL-1. Já no estudo de Streptomyces sp. AM25 um composto não identificado apresentou MIC de 2,50 mg mL-1. Estes três compostos apresentaram atividade superior aos fungicidas Captan SC&reg; (Captana) e Dithane NT&reg; (Mancozeb), e inferior ao Score&reg; (Difenoconazol). A atividade antifúngica destes compostos ao C. gloeosporioides está sendo relatada pela primeira vez. / The pathogenic species of the genus Colletotrichum present importance worldwide because they cause damage to numerous crops of agronomic interest. Several control methods are employed, but they are not always effective due to the occurrence of resistant strains. Thus, it is necessary searching for new compounds that can be used in the integrated management of this disease. Natural products isolated from microorganisms can be an alternative for the development of new agricultural pesticides. Among microorganisms, actinobacteria are known to produce numerous antimicrobial compounds. In this context, the present study aimed to isolate and identify antifungal compounds produced by actinobacteria from guarana rhizosphere. For this, the selection of actinobacteria was based on two tests. In the first one, the 65 actinobacteria were evaluated in paired cultivation test against the plant pathogen Colletotrichum gloeosporioides. Among them, the most promising isolates were AM1 (43.78% inhibition), AM3 (43.98%), AM18 (37.86%), AM25 (43.17%), AM30 (47.12%), AM61 (40.12%) and AM68 (47.94%). In the second assay, these isolates were cultured in BD medium and, after culturing, the metabolic medium was subjected to three extraction methods: (a) liquid-liquid partition with n-butanol; (B) liquid-liquid partition with ethyl acetate and (c) silica gel column C18. The fractions obtained from the three methodologies were evaluated by paper disc diffusion method against C. gloeosporioides. In this disk diffusion assay, the strains AM1 (n-butanol), AM3 (ethyl acetate) and AM25 (C18) were selected for the bioprospecting study. These were identified by molecular techniques as belonging to the genus Streptomyces. From the crude extract of Streptomyces sp. AM1 the analogous compound proclavaminic acid was isolated, which presented minimal inhibitory activity (MIC) of 1.25 mg mL -1 against the plant pathogen C. gloeosporioides. From Streptomyces sp. AM3, the compound streptimidone was isolated, which presented MIC of 1.25 mg mL-1. In the study of Streptomyces sp. AM25 an unidentified compound had MIC of 2.50 mg mL-1. These three compounds presented superior activity to the fungicides Captan SC&reg; (Captan) and Dithane NT&reg; (Mancozeb), and inferior to the Score&reg; (Difenoconazole). The antifungal activity of these compounds to C. gloeosporioides is being reported here for the first time.

O oxigênio, em determinadas condições, pode promover efeitos prejudiciais à saúde, pois pode produzir moléculas reativas e radicais livres. Como mecanismo de proteção, os organismos desenvolveram sistemas defensivos que, quando em desequilíbrio, podem desencadear o estresse oxidativo. Estudos têm indicado que antioxidantes naturais presentes em alguns alimentos são capazes de auxiliar na contenção deste processo. Os compostos fenólicos, presentes em várias plantas, têm despertado a atenção dos pesquisadores devido à atividade antioxidante. O guaraná (Paullinia cupana), planta originária da Amazônia, contém altas concentrações de substâncias bioativas. No entanto, a literatura científica ainda é escassa em relação à semente e praticamente inexistente em relação à casca e polpa no que diz respeito à caracterização química do guaraná, bem como sua atividade antioxidante. Desta forma, o presente projeto pretendeu determinar as principais substâncias bioativas e a atividade antioxidante nestas três frações do guaraná e do guaraná processado, visando ampliar o conhecimento acerca deste alimento e verificar possível utilização de partes hoje descartadas. Inicialmente, foi otimizado um método de extração de compostos fenólicos do guaraná em pó empregando-se diferentes etapas, incluindo solventes e suas concentrações, método de homogeneização, tempo e número de extrações, e os resultados baseados no teor de compostos fenólicos totais. O teor de CF totais foi analisado utilizando o reagente Folin-Ciocalteu. Em seguida, foi desenvolvido um delineamento composto central rotacional (DCCR), utilizando 2 variáveis independentes (22 = 4 ensaios + 4 pontos axiais + 3 pontos centrais, totalizando 11 ensaios), sendo cada variável avaliada em 5 níveis diferentes. Neste caso, além do teor de CF totais, avaliou-se também o teor de proantocianidinas (PA) totais por hidrólise ácida (butanol-HCl). Após esta padronização, as amostras foram submetidas à análise de composição centesimal, quantificação dos compostos fenólicos e proantocianidinas totais (extraíveis e não extraíveis), carotenóides totais por método espectrofotométrico, avaliação do perfil dos principais fenólicos e metilxantinas por Cromatografia Líquida de Alta Eficiência e fitosteróis por Cromatografia Gasosa, e análise da atividade antioxidante in vitro pelos métodos de Teste da capacidade de absorbância de oxigênio radical (ORAC) e Ensaio radical 1,1-difenil-2-2picrilhidrazil (DPPH). O método de extração mais eficiente foi o que consistiu em: agitação magnética por 1 h com metanol 30%, centrifugação, re-extrações com metanol 30% seguidas de pelo menos uma etapa de extração com acetona 70%, perfazendo um total de 4 extrações da mesma amostra. O modelo de extração de CF totais apresentou regressão não significativa. O modelo de extração de PA totais apresentou regressão significativa e R2 de 96%, indicando que para atingir uma extração acima de 80% de PA totais a porcentagem de água deve ser inferior a 20% e a proporção dos solventes acetona/metanol não interferiu na extração desses compostos. As frações polpa, casca e semente do guaraná e o guaraná em pó comercial apresentaram, em média, 2, 14, 51 e 129 mg EAG/ g amostra e 0, 3, 2 e 4 mg EAG/ g de amostra de fenólicos extraíveis e não extraíveis, respectivamente. Os teores de proantocianidinas extraíveis e não extraíveis das frações foram 6, 15, 28 e 46 mg de PA em EqCI/ g de amostra e 0, 3, 2 e 4 mg de PA em EqCI/ g de amostra, respectivamente. Os compostos bioativos encontrados nas amostras foram catequina, epicatequina, PA B1, PA B2, cafeína e teobromina nas seguintes concentrações: 30,05; 20,23; 3,72; 3,37; 39,82 e 0,14 mg/g de pó comercial, respectivamente; 6,63; 0,26; 0,17; 0,06; 35,63 e 0,25 mg/g de semente, respectivamente; e 0,07; 0,01; 0,03; 0,03; 0,02 e 1,17 mg/g de casca, respectivamente. Não foram encontrados CF na polpa. A casca apresentou maior conteúdo de teobromina e nenhuma das amostras apresentou teofilina. Os carotenóides só foram encontrados na casca (34 &#181;g/g). Os fitosteróis encontrados nas amostras foram o brassicasterol, campesterol, stigmasterol e &#946;-sitosterol nas seguintes concentrações: 0; 8,01; 6,45 e 40,45 &#181;g/ g de polpa, respectivamente; 225,79; 1110,34; 126,41 e 830,49 &#181;g/g de casca, 4,19; 7,94; 9,86 e 98,59 &#181;g/ g de semente; e 5,85; 23,90; 11,92 e 151,33 &#181;g/g de pó comercial, respectivamente. A atividade antioxidante dos extratos de fenólicos extraíveis pelo método ORAC foi de 258, 1007, 948 e 2693 &#181;M ET/g de polpa, casca, semente e pó, respectivamente. Por DPPH foi possível avaliar a fração extraível e a não extraível das amostras, sendo necessários 119, 67 e 24 g de casca, semente e pó de guaraná para inibir 1g de DPPH, e 475, 347 e 249 g de casca, semente e pó de guaraná para inibir 1g de DPPH, respectivamente. Os resultados obtidos sugerem que o guaraná é uma boa fonte de compostos bioativos, oferendo possível atividade antioxidante in vitro. / The oxygen, in certain circumstances, can promote adverse health effects due to its capacity to produce reactive molecules and free radicals. As a protective mechanism, organisms have developed defensive systems that, when unbalanced, can initiate oxidative stress. Studies have shown that natural antioxidants present in some foods are capable of assisting in the contention of this process. Phenolic compounds present in various plants, have attracted researchers attention due to their antioxidant activity. Guarana (Paullinia cupana), an Amazon plant, contains high concentrations of bioactive substances. However, the scientific literature is scarce in relation to guarana seed and practically nonexistent in relation to peel and pulp regarding chemical characterization, as well as their antioxidant activity. Thus, this project aimed to determine the main bioactive compounds and antioxidant activity in these three fractions of guarana and guarana processed in order to enhance our understanding of this food and verify the possible use of the by-product generated from the guarana powder production. Initially, was optimized a method of phenolic compounds extraction of guarana powder using different steps, including solvents and their concentration, homogenization method, time and number of extractions, and the results based on content of total phenolic compounds. The total CF content was analyzed using Folin-Ciocalteu reagent. Then, a central composite rotational design (DCCR) was developed using two independent variables (22 = 4 + 4 trials axial points + 3 center points, totalizing 11 trials), being each variable evaluated at 5 different levels. In this case, besides the total of CF content, was also evaluated the content of total proanthocyanidins (PA) by acid hydrolysis (HCl-butanol). After this standardization, samples were submitted to analysis of chemical composition, quantification of total phenolics and proanthocyanidins (extractable and non extractable), total carotenoid content by spectrophotometric method, evaluation of major phenolic profile and methyxanthines by High Performance Liquid Chromatography and phytosterols by Gas Chromatography, and analysis of in vitro antioxidant activity by the methods of the ability of oxygen radical absorbance test (ORAC) and radical 1,1-diphenyl-2-2picrilhidrazil (DPPH). The most efficient extraction method was consisting of: magnetic stirring for 1 hour with 30% methanol, centrifugation, re-extracted with methanol 30% followed by at least one extraction step with acetone 70%, making a total of 4 extractions. The extraction model of total CF showed no significant regression. The extraction model of total PA showed significant regression and R2 of 96%, indicating that to achieve an extraction above 80% of total PA the percentage of water should be less than 20% and the proportion of acetone/methanol did not interfere in the extraction these compounds. The fractions pulp, peel and seeds of guarana and commercial guarana powder showed, on average, 2, 14, 51 and 129 mg EAG/g sample, 0, 3, 2 and 4 mg EAG/g sample extractable and non extractable phenolics, respectively. The fractions contents of extracted and non-extractable proanthocyanidins were 6, 15, 28 and 46 mg of PA in EqCI/g sample and 0, 3, 2 and 4 mg of PA in EqCI/g sample, respectively. The bioactive compounds found in the samples were catechin, epicatechin, PA B1, PA B2, caffeine and theobromine in the following concentrations: 30.05, 20.23, 3.72, 3.37, 39.82 and 0.14 mg/g commercial powder, respectively, 6.63, 0.26, 0.17, 0.06, 35.63 and 0.25 mg/g seed, respectively, and 0.07, 0.01, 0.03; 0.03, 0.02 and 1.17 mg/g of peel, respectively. These CF were not found in the pulp. The peel had a higher content of theobromine and none of the samples showed theophylline. Carotenoids were found only in the peel (34 g/g). The phytosterols found in the samples were brassicasterol, campesterol, stigmasterol and &#946;-sitosterol in the following concentrations: 0, 8.01, 6.45 and 40.45 mg/g pulp, respectively, 225.79, 1110.34, 126.41 and 830.49 mg/g of peel, 4.19, 7.94, 9.86 and 98.59 mg/g of seed, and 5.85, 23.90, 11.92 and 151.33 mg/g of commercial powder, respectively. The antioxidant activity of extractable samples by the ORAC method was 258, 1007, 2693 and 948 mM ET/ g pulp, peel, seed and powder, respectively. For DPPH was possible to evaluate the extractable and non extractable fraction of the samples, which required 119, 67 and 24 g of peel, seed and guarana powder to inhibit 1g of DPPH and 475, 347 and 249 g of peel, seed and powder 1g guarana to inhibit DPPH, respectively. The results suggest that guarana is a good source of bioactive compounds, offering possible in vitro antioxidant activity.