La connaissance du métabolisme demeure incomplète mais sa compréhension reste un but majeur tant pour la recherche fondamentale qu’appliquée. Environ 40% des gènes des organismes procaryotes n’ont pas de fonction précise proposée. En conséquence, de nombreuses voies métaboliques restent incomplètes, rendant très difficile la prédiction du métabolisme d’un organisme. Les recherches et les avancées dans la compréhension de la cascade des « omiques » incluant la génomique, la transcriptomique, la protéomique et la métabolomique jouent aujourd’hui un rôle central dans la connaissance du métabolisme. En tant qu’intermédiaires des réactions biochimiques, les métabolites connectent les différentes voies métaboliques d’un organisme. La quantité et la nature de ces molécules étant la résultante de l’expression des gènes, la composition en métabolites de la cellule est donc liée de très près à son phénotype. La métabolomique, qui étudie l’ensemble de ces petites molécules présentes dans un échantillon biologique, constitue une source d’informations importante nécessaire à la compréhension du métabolisme. En ce sens, le travail présenté dans ce manuscrit décrit la mise en place d’une approche métabolomique au sein du Genoscope pour l’étude du métabolisme de la bactérie modèle Acinetobacter baylyi (ADP1) en utilisant la spectrométrie de masse à haute résolution (LTQ-Orbitrap) couplée à la chromatographie liquide (LC/MS).Nous avons suivi la réorientation du métabolisme d’ADP1 à la suite d’une perturbation environnementale, induite par un changement de source de carbone. Nous avons comparé les métabolomes d’ADP1 cultivée sur succinate (source de carbone de référence) et sur quinate (source de carbone alternative). Environ 450 métabolites potentiels ont été détectés et plus d’une centaine ont pu être identifiés. Plusieurs phénomènes ont pu être observés. Premièrement, l’utilisation du quinate comme source de carbone engendre une réponse spécifique attendue, liée à sa dégradation (intermédiaires cataboliques détectés et gènes associés surexprimés). Toutefois, alors que le métabolisme central est peu affecté par ce changement biotique, la concentration d’environ la moitié des métabolites détectés est significativement modifiée. Ce résultat inattendu est en accord avec les expériences de transcriptomique qui indiquent, qu’avec ce changement de source de carbone, 12% des gènes sont différentiellement exprimés. Nos résultats montrent que la perturbation du métabolisme dans ces conditions s’étend bien au-delà de la voie de dégradation du quinate ; elle engendre un bouleversement global du métabolisme (Stuani, Lechaplais et al. 2014).Cette étude a également permis de détecter de nouveaux métabolites, produits dans des cellules utilisant du quinate. L’élucidation structurale de l’un d’entre eux est décrite dans ce manuscrit. Il s’agit d’une tyrosine substituée en position benzylique par un groupement méthylamine (BMAT). Son identification aura nécessité des expériences de CID-MSn séquentielles avec détection à haute résolution et d’échanges H/D. Des études structurales effectuées sur deux autres molécules suggèrent des similarités structurales entre elles et avec le BMAT. Nous pourrions donc être en présence d’une famille de nouveaux métabolites secondaires, impliqués dans une même voie métabolique. C’est donc une approche métabolomique qui a permis à un simple changement de source de carbone de nous orienter vers la découverte de nouvelles voies métaboliques et d’appréhender une part cachée du métabolisme d’ADP1. / While incomplete, the knowledge of metabolism remains a priority for both basic and applied research. With approximately 40 % of genes of prokaryotic genomes with no accurate function, a number of metabolic pathways remain incomplete, which impedes the accurate prediction of the metabolism of a cell.Recent progress and breakthroughs in the understanding of the Omics Cascade that includes genomics, transcriptomics, proteomics and metabolomics play an important role in the knowledge of metabolism. As intermediaries of biochemical reactions, metabolites connect the different metabolic pathways of the cell. The amount and nature of these molecules being the result of gene expression, the metabolite content of a cell is thus closely related to its phenotype. Metabolomics, which aims at studying the whole set of metabolites present in a biological sample, is an invaluable source of information for the understanding of metabolism.This is why the work presented in this thesis reports the setup of a metabolomic approach at Genoscope for the study of metabolism of the model bacterium Acinetobacter baylyi ADP1 (ADP1) through high-resolution mass spectrometry (LTQ-Orbitrap) coupled to liquid chromatography (LC/MS).We investigated the metabolism of ADP1 after an environmental perturbation induced by a change of carbon source. We compared the metabolomes of ADP1 grown on succinate (reference carbon source) and on quinate (alternative source). About 450 potential metabolites could be detected and more than a hundred were identified. Many observations were made. First, quinate as a carbon source triggered an expected specific metabolic response in relation with its dissimilation (quinate catabolites were detected and the corresponding genes were up-regulated). However, while central metabolism is not impacted by this biotic change, about half of the detected metabolites had their concentration changed. This unexpected result is consistent with transcriptomic experiments that revealed that with the change of carbon source, 12% of the total number of genes are differentially expressed. Our results show that in these conditions, metabolic alteration goes far beyond the quinate degradation pathway and causes a global overhaul of the metabolism of ADP1 (Stuani, Lechaplais et al. 2014).This study also led to the detection of novel metabolites that were exclusively produced in quinate-grown cells. The structural elucidation of one of them is reported in this manuscript. It is a tyrosine substituted in the benzylic position by an aminomethyl group (BMAT). Its identification required sequential CID-MSn experiments with high-resolution detection and isotopic H/D exchange experiments. Structural studies conducted on two other molecules suggest structural similarities with BMAT. We thus could be in the presence of a family of novel secondary metabolites, involved in the same metabolic pathway.In conclusion, a metabolomic approach allowed a mere change of carbon source to guide us toward the discovery of novel metabolic pathways and gain insight into the hidden part of ADP1 metabolism.

No / This article reports the structural elucidation of the Alangium alkaloid, tubulosine (1) on the basis of systematic 2D-NMR analyses (DEPT, COSY, TOCSY, NOESY, ROESY, HMQC and HMBC). The data obtained allowed the unambiguous assignment of all proton and carbon signals in 1 for the first time.

This thesis firstly describes a synthesis of the natural product taurospongin A, a potent DNA polymerase beta inhibitor. Sharpless asymmetric dihydroxylation on olefin <b>E-1.60</b> followed by selective deoxygenation at C(2) via Barton‒McCombie reaction delivers the desired C(1)–C(10) carboxylic acid core. Subsequent esterification of the C(1)–C(10) fragment with C(1′)–C(25′) fatty acid furnishes the natural product in 13.5% yield. The structure of an unnamed natural product <b>2.14</b> isolated in 1974 is proven to be misassigned by previous studies within the Robertson group. As described in this thesis, two proposed structures A and B are obtained via total synthesis in order to reveal the identity of the natural product. A synthesis of key intermediate spirocycles <b>2.148</b> and <b>2.158</b> with desired trans- diol moiety is described by a dihydroxylation reaction on an electron deficient gamma-keto unsaturated acid with subsequent spirocyclisation reaction. Finally, methodology for generating high-value synthetic intermediates by an asymmetric, one-pot enzymatic di/polycarbonyl reduction is described. The concept of such methodology is first proven by the synthesis of (3R)-hydroxybutyl (3R)-hydroxybutanoate <b>3.20</b>. This thesis then describes substrate scope studies and corresponding stereochemical proof to provide more information about this methodology.

A presença de resíduos de medicamentos antibacterianos em alimentos é um importante problema de saúde pública. Estas substâncias podem estar presentes nos alimentos em níveis inaceitáveis como resultados de práticas produtivas inadequadas. Devido a estas preocupações, são estabelecidos limites máximos de resíduos para estas substâncias (LMRs). No caso das sulfonamidas, este valor de LMR refere-se à soma do princípio ativo e de todos seus metabólitos. Neste trabalho, identificam-se e caracterizam-se metabólitos de sulfaquinoxalina (SQX) em diversas espécies animais. Dentro do processo investigativo, foram realizados estudos comparativos de métodos de extração, processos de validação e determinação de efeito de matriz. Foi elaborado e proposto um modelo para a priorização de fármacos baseado em análise de risco e discutiu-se o panorama atual da presença de resíduos de sulfonamidas em amostras ambientais. A investigação da formação de metabólitos de SQX in vitro e in vivo levaram à identificação de três compostos, dois deles ainda não descritos na literatura: N4-acetil-SQX, SQX-OH e N4-acetil-SQX-OH. O perfil de formação destes compostos em diversas espécies animais foi analisado e discutido. / The presence of antibacterial drugs residues in food is an important public health issue. These substances can be present in food at unacceptable levels due to inappropriate veterinary practices. Because of that, maximum residue levels (MRL) are established for these compounds. In the sulfonamide drugs case, this value corresponds to the sum of parent drug and their metabolites. In the present work, sulfaquinoxaline (SQX) metabolites were identified and characterized in several animal species. Inside that investigation process, several studies were developed about extraction methods, validation processes and matrix effects determination. A model for drugs residues prioritization based on risk analysis was proposed. Also, the state-of-art of sulfonamides residues analysis in environmental samples was discussed. The in vivo and in vitro investigation of SQX metabolites formation lead us to the identification of 3 compounds, 2 of them previously unreported: N4-acetyl-SQX, SQX-OH and N4-acetyl-SQX-OH. The formation profile of these compounds in several animal species was analyzed and discussed.

A presença de resíduos de medicamentos antibacterianos em alimentos é um importante problema de saúde pública. Estas substâncias podem estar presentes nos alimentos em níveis inaceitáveis como resultados de práticas produtivas inadequadas. Devido a estas preocupações, são estabelecidos limites máximos de resíduos para estas substâncias (LMRs). No caso das sulfonamidas, este valor de LMR refere-se à soma do princípio ativo e de todos seus metabólitos. Neste trabalho, identificam-se e caracterizam-se metabólitos de sulfaquinoxalina (SQX) em diversas espécies animais. Dentro do processo investigativo, foram realizados estudos comparativos de métodos de extração, processos de validação e determinação de efeito de matriz. Foi elaborado e proposto um modelo para a priorização de fármacos baseado em análise de risco e discutiu-se o panorama atual da presença de resíduos de sulfonamidas em amostras ambientais. A investigação da formação de metabólitos de SQX in vitro e in vivo levaram à identificação de três compostos, dois deles ainda não descritos na literatura: N4-acetil-SQX, SQX-OH e N4-acetil-SQX-OH. O perfil de formação destes compostos em diversas espécies animais foi analisado e discutido. / The presence of antibacterial drugs residues in food is an important public health issue. These substances can be present in food at unacceptable levels due to inappropriate veterinary practices. Because of that, maximum residue levels (MRL) are established for these compounds. In the sulfonamide drugs case, this value corresponds to the sum of parent drug and their metabolites. In the present work, sulfaquinoxaline (SQX) metabolites were identified and characterized in several animal species. Inside that investigation process, several studies were developed about extraction methods, validation processes and matrix effects determination. A model for drugs residues prioritization based on risk analysis was proposed. Also, the state-of-art of sulfonamides residues analysis in environmental samples was discussed. The in vivo and in vitro investigation of SQX metabolites formation lead us to the identification of 3 compounds, 2 of them previously unreported: N4-acetyl-SQX, SQX-OH and N4-acetyl-SQX-OH. The formation profile of these compounds in several animal species was analyzed and discussed.

A presença de resíduos de medicamentos antibacterianos em alimentos é um importante problema de saúde pública. Estas substâncias podem estar presentes nos alimentos em níveis inaceitáveis como resultados de práticas produtivas inadequadas. Devido a estas preocupações, são estabelecidos limites máximos de resíduos para estas substâncias (LMRs). No caso das sulfonamidas, este valor de LMR refere-se à soma do princípio ativo e de todos seus metabólitos. Neste trabalho, identificam-se e caracterizam-se metabólitos de sulfaquinoxalina (SQX) em diversas espécies animais. Dentro do processo investigativo, foram realizados estudos comparativos de métodos de extração, processos de validação e determinação de efeito de matriz. Foi elaborado e proposto um modelo para a priorização de fármacos baseado em análise de risco e discutiu-se o panorama atual da presença de resíduos de sulfonamidas em amostras ambientais. A investigação da formação de metabólitos de SQX in vitro e in vivo levaram à identificação de três compostos, dois deles ainda não descritos na literatura: N4-acetil-SQX, SQX-OH e N4-acetil-SQX-OH. O perfil de formação destes compostos em diversas espécies animais foi analisado e discutido. / The presence of antibacterial drugs residues in food is an important public health issue. These substances can be present in food at unacceptable levels due to inappropriate veterinary practices. Because of that, maximum residue levels (MRL) are established for these compounds. In the sulfonamide drugs case, this value corresponds to the sum of parent drug and their metabolites. In the present work, sulfaquinoxaline (SQX) metabolites were identified and characterized in several animal species. Inside that investigation process, several studies were developed about extraction methods, validation processes and matrix effects determination. A model for drugs residues prioritization based on risk analysis was proposed. Also, the state-of-art of sulfonamides residues analysis in environmental samples was discussed. The in vivo and in vitro investigation of SQX metabolites formation lead us to the identification of 3 compounds, 2 of them previously unreported: N4-acetyl-SQX, SQX-OH and N4-acetyl-SQX-OH. The formation profile of these compounds in several animal species was analyzed and discussed.

京都大学 / 新制・課程博士 / 博士(人間・環境学) / 甲第25019号 / 人博第1097号 / 新制||人||257(附属図書館) / 京都大学大学院人間・環境学研究科相関環境学専攻 / (主査)教授 小松 直樹, 教授 津江 広人, 教授 藤田 健一, 准教授 Zhao Li / 学位規則第4条第1項該当 / Doctor of Human and Environmental Studies / Kyoto University / DFAM

O ambiente marinho tem sido reconhecido como uma importante fonte de metabólitos secundários biologicamente ativos. Neste contexto, fungos endofíticos associados a algas ganharam importância nas últimas décadas, como alvos alternativos para a pesquisa de produtos naturais. O presente trabalho teve como o objetivo o estudo químico e biológico de duas linhagens de fungos endofíticos associados à alga vermelha Bostrychia radicans. Inicialmente foi realizada a triagem química e biológica (atividade antitumoral e antimicrobiana) dos extratos brutos das duas linhagens selecionadas, linhagens M20 (Hypocrea lixii) e M23 (Eutypella sp), obtidos a partir de cultivos em escala piloto, tanto variando-se os meios de cultivo e bem como períodos de crescimento. O extrato da linhagem M20 cultivada em arroz apresentou potencial citotóxico interessante quando submetido a ensaios utilizando células tumorais HCT-116. Ainda, após a análise química, esta linhagem foi selecionada para o cultivo em escala ampliada, visando o isolamento e elucidação estrutural dos metabólitos secundários presentes neste fungo. O estudo químico em escala ampliada da linhagem M20, espécie Hypocrea lixii, proporcionou o isolamento e identificação de quatro metabólitos: ácido 3-hidroxi-5-metóxi-6-metil-1,3-diidro-isobenzofurano-4- carboxílico (S1), 3,7-dimetóxi-6-metil-1-oxo-1,3-diidro-isobenzofurano-4-carbaldeído (S3), galactitol (S4), convolvulol (S5), além do isolamento de dois metabólitos que ainda não foram completamente elucidados, S2 e S6. Os metabólitos S1 e S3 são metabólitos inéditos como produtos naturais. Além disso, foi possível a identificação de 14 substâncias via cromatografia gasosa acoplada à espectrometria de massas (CG-EM), entre elas hidrocarbonetos, ácidos graxos, inclusive insaturados, aldeídos, aldeídos ?,?-insaturados e esteróide. As substâncias S1 e S4 foram submetidas à avaliação de atividade biológica (atividade antibacteriana, antifúngica, anticolinesterásica e antitumoral), porém nenhum resultado positivo foi constatado. Foi realizada avaliação da atividade tumoral das frações da linhagem M20, e as frações M20F e M20H apresentaram atividade citotóxica seletiva para linhagens de células tumorais. Em um segundo momento foi realizado o cultivo em escala ampliada da linhagem M23 (Eutypella sp) que proporcionou o isolamento da R-5-metilmeleína (S7). Dessa forma, o estudo químico de fungos endofíticos associados à alga Bostrychia radicans mostrou-se promissor na busca de novas estruturas químicas, visto que já foram isoladas e identificadas duas estruturas inéditas como produtos naturais. / The marine environment has been recognized as an important source of biologically active secondary metabolites. In this context, endophytic fungi associated with algae gained importance in recent decades, as alternative to natural products research targets. The present work had as goal the chemical and biological study of two strains of endophytic fungi associated with red algae Bostrychia radicans. The chemical and biological screening (antimicrobial and antitumor activity) of the crude extracts of two selected strains, M20 (Hypocrea lixii) and M23 (Eutypella sp), were obtained from pilot-scale cultivation, by means of culture media and growth period variation. The M20 strain extract, grown in rice, showed an interesting cytotoxic potential front HCT -116 tumor cells and after chemical analysis, this strain was selected for cultivation on a large scale, with the purpose of secondary metabolites isolation. Chemical studies of M20 species strain Hypocrea lixii, performed on an enlarged scale, afforded the isolation and identification of four metabolites: 3-hydroxy-5-methoxy-6- methyl-1,3-dihydro-isobenzofuran-4-carboxylic acid (S1), 3,7 dimethoxy-6-methyl-1-oxo- 1,3-dihydro-isobenzofuran-4-carbaldehyde (S3), galactitol (S4), convolvulol (S5), in addition the isolation of two metabolites which have not yet been fully elucidated, S2 and S6. The S1 and S3 metabolites are novel metabolites as natural products. Furthermore, it was possible to identify 14 compounds by gas chromatography coupled to mass spectrometry (GC-MS), including hydrocarbons, fatty acids, besides unsaturated ones, aldehydes, ?,?-unsaturated aldehydes and steroid. The S1 and S4 compounds were subjected to biological activity evaluation (antibacterial, antifungal, antitumor and acetylcholinesterase potential), but without any positive result. Assessment of tumor activity of fractions of the M20 strain was performed, and the M20F and M20H fractions showed selective cytotoxicity to tumor cell lines. In a second step, the M23 strain (Eutypella sp) was grown on a large scale, resulting in the R-5-metilmeleina (S7) isolation. Thus, the chemical study of endophytic fungi associated to Bostrychia radicans algae proved to be promising concerning the search for new chemical compounds discovery, since it yielded two new structures as natural products.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-29T12:15:24Z No. of bitstreams: 2 Tese - Luciane Dias Pereira - 2016.pdf: 4859479 bytes, checksum: e060870fade58076d056ab4d8121aa49 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-29T12:17:29Z (GMT) No. of bitstreams: 2 Tese - Luciane Dias Pereira - 2016.pdf: 4859479 bytes, checksum: e060870fade58076d056ab4d8121aa49 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-29T12:17:29Z (GMT). No. of bitstreams: 2 Tese - Luciane Dias Pereira - 2016.pdf: 4859479 bytes, checksum: e060870fade58076d056ab4d8121aa49 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-07-22 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Seeds and peels of jabuticaba (Myrciaria cauliflora (Mart) O. Berg) collected in the Jabuticabal Winery were separately extracted with aqueous acetone (50%). After partitioning between solvents, the fractions ethyl acetate and methanol were subjected to repeated column chromatography using Diaion HP-20 and Sephadex LH-20 as adsorbents. Pure substances and fractions were analyzed by 1H NMR and 13C, one and two dimensional, and ESI-TOF mass spectroscopy. In this work, fifteen compounds were isolated and identified: seven ellagitannins, one of them is novel, one galloyl ester, three phenolic acids, two glycosylated flavonoids, one depside and one organic acid. To optimize the extraction of phenolic compounds, a randomized experimental design was applied, using Box-Wilson central composite design for two factors. The independent variables were stirring time () and solvent volume (V), which consisted of ethanol/hydrochloric acid (9:1). Three independent experiments in duplicate were performed, as well as assays for quantitation of phenolic compounds total monomeric anthocyanins (TMA), tannins (T), total phenols (TP), and color parameters color index (CI) and tonality (Ton). According to the results, when the volume remained constant and the stirring time increased, all phenolic contents and color patterns intensified. The best conditions for greater extraction were total ultrasonic time of 75 min and solvent volume of 64 ml, yielding the concentrations of TP, T and TMA of 33.5; 7.9 and 5.57 mg g-1, and CI and Ton, 0.893 a.u. and 0.833, respectively. In the extraction of phenolic compounds from seeds, the following solvents were tested water, aqueous ethanol (50%), aqueous methanol (50%), and the latter mixture provided the highest levels of TP, T and flavonoids: 15.71; 12.65 and 64.14 mg g-1, respectively. The influence of soil on jabuticaba seed and peel chemical contents was evaluated. Data obtained from the analysis of centesimal composition, phenolic contents, color parameters and mineral nutrients of samples collected in five orchards were analyzed by canonical redundancy (RDA) and hierarchical clustering (HCA). These analyzes ordered samples in four groups: the peels in groups I and II were related to high levels of phenolics and fibers, respectively, while the tannins and flavonoids were main compounds present in the seeds of group III, the moisture content was higher in seeds of group IV. The separation of peels (Groups I and II) and seeds (Groups III and IV) in different groups was influenced by the levels of fruits nutrients, Mn2+ and Cu2+, whereas Mn2+ from soil distinguished samples of the clusters I and III from the others, II and IV, characterizing a strong influence of some soil and fruits nutrients on the chemical composition of the jabuticaba seeds and peels. / Sementes e cascas de jabuticaba (Myrciaria cauliflora (Mart.) O. Berg) coletadas na Vinícola Jabuticabal, foram extraídas separadamente com acetona:água 50%. Após partição entre solventes, as frações acetato de etila e metanólica foram submetidas a repetidas cromatografias em coluna utilizando-se Diaion HP-20 e Sephadex LH-20 como adsorventes. Substâncias puras e frações foram analisadas por RMN de 1H e 13C, uni e bidimensional e espectroscopia de massas ESI-TOF. Neste trabalho foram isolados e identificados quinze compostos sendo sete elagitaninos, um deles inédito, um éster galoílico, três ácidos fenólicos, dois flavonoides glucosilados, um depsídeo e um ácido orgânico. Para otimização da extração dos compostos fenólicos utilizou-se um delineamento experimental ao acaso, empregando-se a técnica de planejamento Box-Wilson composto central para dois fatores. Os fatores foram compostos pelo tempo de agitação () e volume de solvente (V) constituído por etanol/ácido clorídrico (9:1). Foram realizados três experimentos independentes em duplicata, além de ensaios para quantificação dos compostos fenólicos antocianinas totais monoméricas (ATM), taninos (T) e fenóis totais (FT), padrões de coloração índice de cor (IC) e tonalidade (Ton). De acordo com os resultados, quando o volume permaneceu constante e variou o tempo de agitação observou-se que todos os teores de fenóis e padrões de cor aumentaram. O tempo total de 75 min de ultrassom e volume de solvente de 64 mL apresentou maior poder de extração, obtendo-se teores de FT, T e ATM de 33,5; 7,91 e 5,57 mg g-1, com IC e Ton de 0,893 u.a. e 0,833, respectivamente. Na extração de compostos fenólicos nas sementes foram testados os solventes: água, etanol aquoso (50%) e metanol aquoso (50%), sendo que esta última mistura forneceu os maiores teores de FT, T e flavonoides: 15,71; 12,65 e 64,14 mg g-1, respectivamente. No estudo da influência do solo nas variações químicas de cascas e sementes de jabuticaba, os dados obtidos da análise de composição centesimal, compostos fenólicos, padrões de coloração e nutrientes minerais de amostras de cinco pomares foram analisados por redundância canônica (RDA) e de agrupamento hierárquico (HCA). Essas análises ordenaram as amostras em quatro grupos: as cascas do grupo I e II foram relacionadas aos altos níveis de fenóis totais e fibras, respectivamente, enquanto que os taninos e flavonoides foram os principais compostos presentes nas sementes do grupo III e o teor de umidade foi maior em sementes do grupo IV. A separação de cascas (grupos I e II) de sementes (grupos III e IV) em grupos distintos foi influenciada pelos teores dos nutrientes Mn2+ e Cu2+ dos frutos, enquanto Mn2+ do solo distinguiu amostras das classes I e III das outras, II e IV, caracterizando assim uma forte influência de alguns nutrientes dos solos e dos frutos na composição química das sementes e cascas de jabuticaba.

O ambiente marinho tem sido reconhecido como uma importante fonte de metabólitos secundários biologicamente ativos. Neste contexto, fungos endofíticos associados a algas ganharam importância nas últimas décadas, como alvos alternativos para a pesquisa de produtos naturais. O presente trabalho teve como o objetivo o estudo químico e biológico de duas linhagens de fungos endofíticos associados à alga vermelha Bostrychia radicans. Inicialmente foi realizada a triagem química e biológica (atividade antitumoral e antimicrobiana) dos extratos brutos das duas linhagens selecionadas, linhagens M20 (Hypocrea lixii) e M23 (Eutypella sp), obtidos a partir de cultivos em escala piloto, tanto variando-se os meios de cultivo e bem como períodos de crescimento. O extrato da linhagem M20 cultivada em arroz apresentou potencial citotóxico interessante quando submetido a ensaios utilizando células tumorais HCT-116. Ainda, após a análise química, esta linhagem foi selecionada para o cultivo em escala ampliada, visando o isolamento e elucidação estrutural dos metabólitos secundários presentes neste fungo. O estudo químico em escala ampliada da linhagem M20, espécie Hypocrea lixii, proporcionou o isolamento e identificação de quatro metabólitos: ácido 3-hidroxi-5-metóxi-6-metil-1,3-diidro-isobenzofurano-4- carboxílico (S1), 3,7-dimetóxi-6-metil-1-oxo-1,3-diidro-isobenzofurano-4-carbaldeído (S3), galactitol (S4), convolvulol (S5), além do isolamento de dois metabólitos que ainda não foram completamente elucidados, S2 e S6. Os metabólitos S1 e S3 são metabólitos inéditos como produtos naturais. Além disso, foi possível a identificação de 14 substâncias via cromatografia gasosa acoplada à espectrometria de massas (CG-EM), entre elas hidrocarbonetos, ácidos graxos, inclusive insaturados, aldeídos, aldeídos ?,?-insaturados e esteróide. As substâncias S1 e S4 foram submetidas à avaliação de atividade biológica (atividade antibacteriana, antifúngica, anticolinesterásica e antitumoral), porém nenhum resultado positivo foi constatado. Foi realizada avaliação da atividade tumoral das frações da linhagem M20, e as frações M20F e M20H apresentaram atividade citotóxica seletiva para linhagens de células tumorais. Em um segundo momento foi realizado o cultivo em escala ampliada da linhagem M23 (Eutypella sp) que proporcionou o isolamento da R-5-metilmeleína (S7). Dessa forma, o estudo químico de fungos endofíticos associados à alga Bostrychia radicans mostrou-se promissor na busca de novas estruturas químicas, visto que já foram isoladas e identificadas duas estruturas inéditas como produtos naturais. / The marine environment has been recognized as an important source of biologically active secondary metabolites. In this context, endophytic fungi associated with algae gained importance in recent decades, as alternative to natural products research targets. The present work had as goal the chemical and biological study of two strains of endophytic fungi associated with red algae Bostrychia radicans. The chemical and biological screening (antimicrobial and antitumor activity) of the crude extracts of two selected strains, M20 (Hypocrea lixii) and M23 (Eutypella sp), were obtained from pilot-scale cultivation, by means of culture media and growth period variation. The M20 strain extract, grown in rice, showed an interesting cytotoxic potential front HCT -116 tumor cells and after chemical analysis, this strain was selected for cultivation on a large scale, with the purpose of secondary metabolites isolation. Chemical studies of M20 species strain Hypocrea lixii, performed on an enlarged scale, afforded the isolation and identification of four metabolites: 3-hydroxy-5-methoxy-6- methyl-1,3-dihydro-isobenzofuran-4-carboxylic acid (S1), 3,7 dimethoxy-6-methyl-1-oxo- 1,3-dihydro-isobenzofuran-4-carbaldehyde (S3), galactitol (S4), convolvulol (S5), in addition the isolation of two metabolites which have not yet been fully elucidated, S2 and S6. The S1 and S3 metabolites are novel metabolites as natural products. Furthermore, it was possible to identify 14 compounds by gas chromatography coupled to mass spectrometry (GC-MS), including hydrocarbons, fatty acids, besides unsaturated ones, aldehydes, ?,?-unsaturated aldehydes and steroid. The S1 and S4 compounds were subjected to biological activity evaluation (antibacterial, antifungal, antitumor and acetylcholinesterase potential), but without any positive result. Assessment of tumor activity of fractions of the M20 strain was performed, and the M20F and M20H fractions showed selective cytotoxicity to tumor cell lines. In a second step, the M23 strain (Eutypella sp) was grown on a large scale, resulting in the R-5-metilmeleina (S7) isolation. Thus, the chemical study of endophytic fungi associated to Bostrychia radicans algae proved to be promising concerning the search for new chemical compounds discovery, since it yielded two new structures as natural products.