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Ana Camila DOS SANTOS DIAS

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Ana Camila DOS SANTOS DIAS Ana Camila DOS SANTOS DIAS Mémoire présenté en vue de l’obtention du grade de Docteur de l'Université de Nantes sous le sceau de l’Université Bretagne Loire École doctorale : VENAM Discipline : Science de la Vie et de la Santé - Sciences Pharmaceutique (CNU 85-86-87) Spécialité : Substances Naturelles Bioactives d'Origine Marine Unité de recherche : EA 2160 - MMS - UFR Sciences Biologiques et Pharmaceutiques Soutenue le 28 avril 2017 Champignons marins : Lipidomique et lipides d'intérêt en santé et nutrition JURY Rapporteurs : Arlette BAILLET-GUFFROY , Professeur, Université Paris Sud François-Hugues POREE , Maitre de conférences, HDR, Université Paris Descartes Examinateurs : Joel BOUSTIE , Professeur, Université de Rennes I Joel FLEURENCE , Professeur, Université de Nantes Invité(s) : Samuel BERTRAND , Maitre de conférences, Université de Nantes Nicolas RUIZ , Maitre de conférences, Université de Nantes Directeur de Thèse : Gaetane WIELGOSZ-COLLIN , Maitre de conférences, HDR, Université de Nantes Co -directeur de Thèse : Aurélie COUZINET-MOSSION , Maitre de conférences, Université de Nantes Nicolas RUIZ , Maitre de conférences, Université de Nantes Remerciements Eu gostaria de primeiramente agradecer a minha familia, meu suporte, minha benção, sempre esteve ao meu lado, independente da distancia, das brigas e desse oceano que nos separa nunca deixou de me apoiar e de me amar. Ao meu pai, que tanto se esforçou para nos dar uma boa educação. A minha mãe, mulher de garra e força! Aos meus irmãos e irmã, meu muito obrigada. Essa tese também é de vocês. Essa é a nêga do pai! Je voudrais remercier ma directrice de thèse, Gaëtane Wielgosz-Collin, pour sa confiance et son encadrement tout aux long de ces années de thèse, et surtout pour sa patience à l'égard d'une "non-chimiste". Merci de m'avoir épaulée dans les moments difficiles d'énervement et de stress aux détours des petits thés et de rires partagés. Merci à mes deux encadrants officiels, Aurélie Couzinet-Mossion et Nicolas Ruiz, qui me suivent depuis mon M2. C'est grâce à vous que j'ai découvert, de façon passionnante, la Recherche dans le monde "des champignons". Merci, Aurélie, pour tes conseils avisés, pour ton partage d'expérience, notamment lors des cours de TPs, et pour ta compagnie ; toujours agréable. Merci, Nico, pour tout! Je n’ai pas de mot pour exprimer mon admiration et mon respect. Même si de temps en temps, nous ne partagions pas forcément les mêmes avis, particulièrement sur le comté et le café. Merci à mon encadrant officieux, Samuel Bertrand, et pour tout le temps que tu m'as consacré, malgré l'encadrement non officiel. Merci pour tout ce que tu m'as apporté avec ton regard sur l'approche lipidomique. Je souhaiterais remercier les rapporteurs que sont Mme Arlette Baillet-Guffroy et M. François- Hugues Porée ainsi que les membres du jury Joel Boustie et Joel Fleurence, qui ont gentiment accepté d'évaluer mon travail. Je remercie également la bourse d'études de la " Fundação CAPES - Ministério da Educação ", du gouvernement brésilien, qui a financé cette thèse et sans laquelle ce travail n'aurait pas eu lieu. Obrigada Lula, obrigada Dilma! Je voudrais remercier notre directeur de laboratoire, Yves François Pouchus, pour m'avoir accueillie au sein de son équipe depuis toutes ces années. Je voudrais remercier également les membres de mon comité de suivi de thèse, Olivier Goncalves et Edouard Kraffe, pour les discussions importantes qui ont permis de faire avancer ma recherche ainsi que pour leur regard extérieur qui m'a énormément aidé. Merci à l'équipe de chimie de la Faculté de Pharmacie de Nantes qui m'a donné la possibilité d'enseigner et de beaucoup apprendre. Qui m'a accompagné pendant la préparation des EDs et des TPs : Christine Herrenknecht, Isabelle Ourliac et Marjorie Albassier. Merci pour les fous rires Isabelle!! Tu me dois encore des kitkat au thé vert... Je voudrais remercier du fond du cœur ma petite Vony Rabesaotra, mon bras droit! A l'Isomer ou pendant les TPs en pharma, tu m'as toujours aidé, depuis le début, tu m'as énormément appris, tu as énormément donné de ta personne pour que cette thèse avance, je t'en serai éternellement reconnaissante. Et désolée pour les bourdes de temps en temps... Je souhaiterais remercier également Marie-Claude Boumard, Thibault Robiou Du Pont et Christophe Tomasoni (merci beaucoup Toto!), pour leur soutient technique qui m'a été indispensable depuis le début. Sois rassurée Marie-Claude, je ne piquerai plus ton petit chauffage (hahaha!). Merci à Chantal Simonato et à Nathalie Calvi pour m'avoir sauvé la vie plusieurs fois sur tout ce qui concerne papier au niveau administratif au secrétariat. Merci pour votre gentillesse et patience. Je sais que parfois je ne suis pas facile... Merci à Catherine Roullier, à Oliver Grovel et à Emanuel Gentil pour vos interventions, et l'apport de vos connaissances pour l'avancé de ma thèse. Catherine, ton rire est une bouffée d'air frais! Je l'adore! Merci également à Hassan Nazih et à son équipe pour leur disponibilité, leur gentillesse et leur aide avec les tests au sein de leur laboratoire. A ma chère Karina Petit, cette personne pétillante, toujours prête à m'aider et qui a beaucoup contribué à mon adaptation au laboratoire. Merci d'avoir apporté la joie de tes petits dans ma vie. Merci également de partager ton expérience et ton travail avec moi lors des séances de TP. I'm the bad cop! Merci à mes amis qui ont partagé le bureau des doctorants avec moi, aux anciens (Marie, Elodie, Mickael et Anne-Isaline) et aux nouvelles (Thuy et Lucie). Merci pour les apéros (hehe!), pour l'atelier nem, pour les fromages de chèvre, pour la bonne humeur, pour la joie de vivre tout au long de ce chemin et surtout et principalement pour supporter mon humeur exécrable!! Je remercie également tous les stagiaires que j'ai pu encadrer au long de cette thèse (y en a eu presque une dizaine!) qui m'ont également beaucoup appris et à qui j'ai pu transmettre un peu de mes connaissances. C'est toujours un plaisir de partager ces moments. C'est aussi grâce à eux que j'ai pu réaliser ce travail. Merci à ma famille en France : Celine Boché (Hector et Tess), Jailson et Frank, Rafaela et Yvonnick (et sa famille!), Larissa et Régis, Leonardo et Poliana, Carla, François et Matthieu Dupuis et Aurélie Pouchus. Mes amis, mes amours, qui m'accompagnent, pour la plus part, depuis bientôt 9 ans dans ce pays qui m'a adoptée, mais dans lequel je suis arrivée seule. C'est grâce à vous que j'ai tenu le coup... Parce que les amis, c'est la famille qu'on choisit! A mes amis du CrossFit, ce sport qui m'a montré que nous pouvons être des meilleurs athlètes, des meilleures personnes et que nous pouvons toujours évoluer. Ce sport qui m'a apporté tant de belles choses...et de magnifiques personnes... A mon Lucas Bryk, merci d'être mon support, mon rocher. Merci pour ton soutien quotidien, ton écoute, tes paroles pour me calmer. Merci de faire de moi une meilleure personne chaque jour, d'exercer ta patience avec moi et d'apporter un nouveau regard dans la relecture et corrections de mes publications. Merci également à ta famille pour son soutien. Mon rayon de soleil, Kocham cię kochanie . Sommaire Remerciements ........................................................................................................................... i Liste des travaux et publications ............................................................................................ iv Liste des abréviations .............................................................................................................. vi Liste des figures ..................................................................................................................... viii Liste des tableaux .................................................................................................................... xi Introduction générale ............................................................................................................... 1 Chapitre I. Étude bibliographique .......................................................................................... 3 1. LES CHAMPIGNONS : DÉFINITION ET CLASSIFICATION 4 2. L' IMPORTANCE DES FUNGI 7 2.1. Métabolites d'origine fongique 9 2.2. Nouvelles sources de molécules fongiques 11 3. LES LIPIDES DES CHAMPIGNONS MARINS 19 3.1. Les lipides : Définition générale 19 3.2. Les principales classes lipidiques étudiées 21 3.3. Métabolisme lipidique 29 4. ACTIVITÉS BIOLOGIQUES DES LIPIDES DES CHAMPIGNONS MARINS 30 5. VALORISATION DES LIPIDES DE CHAMPIGNONS 35 Chapitre II. Analyse lipidique à haut débit .......................................................................... 37 1. LIPIDOMIQUE 38 2. DÉRÉPLICATION 40 3. CRIBLAGE DES PROFILS LIPIDIQUES PAR APPROCHE OSMAC 41 4. CONCLUSION 44 Chapitre III. Matériel et méthodes ....................................................................................... 46 1. LES CULTURES FONGIQUES 47 i 1.1. Le matériel biologique 47 1.2. Les milieux de culture 49 1.3. Cultures fongiques pour l’étude OSMAC sur les lipides totaux 49 1.4. Culture s fongiques pour l’étude OSMAC sur les glycolipides 50 1.5. Cultures fongiques pour le criblage 51 2. ARRÊT DES CULTURES 51 3. EXTRACTION ET ANALYSE LIPIDIQUE 52 3.1. Extraction classique par macération 52 3.2. Séparation en classes lipidiques 53 3.3. Chromatographie sur couche mince (CCM) 53 4. ISOLEMENT ET PURIFICATION DES GLYCOLIPIDES 54 5. ISOLEMENT ET PURIFICATION DES STÉROLS 55 6. SAPONIFICATION 55 6.1. Acétylation des composés lipidiques
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    Aanen, D. K. & T. W. Kuyper (1999). Intercompatibility tests in the Hebeloma crustuliniforme complex in northwestern Europe. Mycologia 91: 783-795. Aanen, D. K., T. W. Kuyper, T. Boekhout & R. F. Hoekstra (2000). Phylogenetic relationships in the genus Hebeloma based on ITS1 and 2 sequences, with special emphasis on the Hebeloma crustuliniforme complex. Mycologia 92: 269-281. Aanen, D. K. & T. W. Kuyper (2004). A comparison of the application of a biological and phenetic species concept in the Hebeloma crustuliniforme complex within a phylogenetic framework. Persoonia 18: 285-316. Abbott, S. O. & Currah, R. S. (1997). The Helvellaceae: Systematic revision and occurrence in northern and northwestern North America. Mycotaxon 62: 1-125. Abesha, E., G. Caetano-Anollés & K. Høiland (2003). Population genetics and spatial structure of the fairy ring fungus Marasmius oreades in a Norwegian sand dune ecosystem. Mycologia 95: 1021-1031. Abraham, S. P. & A. R. Loeblich III (1995). Gymnopilus palmicola a lignicolous Basidiomycete, growing on the adventitious roots of the palm sabal palmetto in Texas. Principes 39: 84-88. Abrar, S., S. Swapna & M. Krishnappa (2012). Development and morphology of Lysurus cruciatus--an addition to the Indian mycobiota. Mycotaxon 122: 217-282. Accioly, T., R. H. S. F. Cruz, N. M. Assis, N. K. Ishikawa, K. Hosaka, M. P. Martín & I. G. Baseia (2018). Amazonian bird's nest fungi (Basidiomycota): Current knowledge and novelties on Cyathus species. Mycoscience 59: 331-342. Acharya, K., P. Pradhan, N. Chakraborty, A. K. Dutta, S. Saha, S. Sarkar & S. Giri (2010). Two species of Lysurus Fr.: addition to the macrofungi of West Bengal.
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