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Etude De La Voie De Biosynthèse Des Phlorotannins Chez Les Algues

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Etude De La Voie De Biosynthèse Des Phlorotannins Chez Les Algues Université Pierre et Marie Curie Ecole Doctorale Complexité du Vivant ED515 Laboratoire de Biologie Intégrative des Modèles Marins Equipe Signalisation et défense chimique chez les macroalgues Spécialité Biologie Marine Etude de la voie de biosynthèse des phlorotannins chez les algues brunes, de la caractérisation biochimique d'enzymes recombinantes à l’étude des réponses écophysiologiques. Présentée par Emeline Creis Thèse de doctorat de Biologie Présentée et soutenue publiquement le 6 mars 2015 Devant le jury composé de : M. Alain Bouchereau Professeur IGEPP INRA (Rennes 1) Rapporteur M. Joël Boustie Professeur Faculté des Sciences (Rennes 1) Rapporteur Mme Geneviève Chiapusio Maître de Conférence Université de Franche- Examinatrice Comté (Montbéliard) M. Weinberger Florian Chercheur GEOMAR (Kiel) Examinateur M. Destombe Christophe Professeur Station Biologique CNRS (Roscoff) Examinateur M. Philippe Potin Directeur de recherche Station Biologique CNRS Directeur de thèse (Roscoff) M. Erwan Ar Gall Maître de Conférence LEMAR IUEM UBO(Brest) Co-Directeur de thèse Je dédie cette thèse à ma famille qui a toujours su me soutenir à tous moments, et en particulier à mon Grand-Père qui m’aura communiqué cette envie de découvrir la Nature et qui m’aura inculqué l’importance de toujours aller jusqu’au bout des choses entreprises. Je tiens tout d’abord à remercier la Station Biologique de Roscoff, son directeur Bernard Kloareg de m’avoir permis de réaliser cette thèse dans d’aussi bonnes conditions. Je tiens également à remercier la Région Bretagne pour le financement de cette thèse, le projet Phlorotan’ING soutenu par le GIS Europôle Mer ainsi que le projet Idealg. Merci également à l’UBO d’avoir financé la prolongation de cette thèse. Merci aux rapporteurs et examinateurs d’avoir accordé de leur temps pour juger ce travail et pour leur participation à ce jury. Merci au Gdr Biochimar, au Gdr Médiatec et à l’association des Jeunes Chercheurs en Ecologie Chimique de m’avoir fait découvrir une communauté de scientifiques aussi passionnés les uns que les autres. Merci à Catherine Boyen et Mirjam Czjzek directrices de l’unité 8227 pour votre travail et votre soutien qui nous permettent de travailler dans de très bonnes conditions. Un grand merci également à tous les membres de l’Equipe Signalisation et Défense Chimique chez les Macroalgues, Gaëlle, Céline, Léa, JB, merci Catherine pour ton soutien et ton dynamisme, merci Ludo mon pti chef sans toi cette thèse ne serait pas ce qu’elle est aujourd’hui, merci pour tout ce que tu as pu m’enseigner depuis maintenant plus de 4 ans, de ta disponibilité et de ton écoute. L’un n’allant pas sans l’autre, merci également à toi Laurence pour toute ton aide apportée pendant le master et le début de cette thèse ainsi qu’après ton départ. Philippe, comment te remercier à ta juste valeur, merci de m’avoir offert cette opportunité d’effectuer cette thèse avec toi, j’ai beaucoup de respect pour le scientifique passionné que tu es mais également pour l’homme qui sais rester juste, ouvert et à l’écoute des autres. Tu m’as permis de rencontrer Florian mais aussi Raj, des gens qui comme toi partagent ces mêmes valeurs morales et scientifiques. Erwan, notre histoire à commencé il y a un bout de temps maintenant (7 ans), c’est toi qui m’aura donné le virus « Algue » et je t’en remercie ! Merci pour ton écoute et ton soutien tout au long de cette thèse. Merci à toutes les personnes ayant séjournées plus ou moins longtemps au bureau 351, les actuelles Maria et Amandine, merci pour votre soutien lors des derniers moments de l’écriture. Plus largement et pour n’oublier personne, merci à tous les membres de l’UMR 8227 pour ces 4 années et ces bons moments passés en votre compagnie. Mon petit scarabé et oui je t’avais promis une place dans ces remerciements, alors merci pour ta bonne humeur ton accent brezhoneg qui en fait palir plus d’un, tu fais partie de ces rares personnes qui ont toujours la patate quel est donc ton secret ? Reste comme tu es, unique en ton genre ! Alexandra et Sophie, merci à toutes les deux pour tout ce que vous m’apportez pour votre bienvaillance, vos conseils, j’ai rencontré ici bien plus que des collègues merci pour tout ! Monique, un grand merci pour ta présence réconfortante toujours de bon conseil, calme, rationelle (enfin ça dépend ;)) merci d’être là. Comment vous dissocier tous les 4, Rémy, Kévin, Héloïse et Camille, les inséparables merci à tous les quatres, je pense que vous savez que les bretons ne donnent leur amitié qu’une seule fois, ça c’est fait, j’espère que l’avenir nous permettra de nous voir encore et encore dans les quatre coins du monde ! Ma petite Alexia, c’est ton tour, merci poulette pour tous ces bons moments passés avec toi j’ai hâte d’entendre le récit de tes aventures de l’autre côté de l’Atlantique ! Merci Adrien, Anaïs, Aurélie, Vanessa, Julien, Charlotte, Benoît, Perrine, Alice pour votre amitié. Je souhaite particulièrement remercier toute ma famille qui m’a toujours soutenue et qui est présente en toute circonstance, merci pour tous ces bonheurs que vous m’apportez. Le meilleur pour la fin, Steve, « merci » n’est pas un mot assez fort pour te remercier, tu auras réussi à me donner l’énergie nécessaire pour écrire cette thèse, ton enthousiasme, ton envie de toujours faire au mieux, me donnent chaque jour l’envie d’aller toujours plus loin Une thèse, bien plus qu’un diplôme c’est le temps privilégié pour passer de la vie d’étudiant à la vie d’adulte, dans un cadre comme Roscoff en passant par Kiel puis Truro, cette aventure aura certainement été pour moi la plus enrichissante autant professionnellement que personnellement, Merci à toutes et à tous ! Sommaire Introduction Générale ............................................................................................................................................ 1 I. De l’endosymbiose primaire, à l’émergence des macroalgues brunes ...................................................... 1 Phylogénie et classification des algues ................................................................................................................. 1 Les algues marines pluricellulaires: ...................................................................................................................... 3 Les macroalgues brunes dans l’Evolution: ............................................................................................................ 5 II. Les macroalgues brunes : une richesse de composés originaux ................................................................ 6 Des métabolismes originaux:................................................................................................................................ 6 Les polysaccharides :........................................................................................................................................ 6 Le mannitol ...................................................................................................................................................... 6 Le métabolisme halogéné : .............................................................................................................................. 7 Les phlorotannins : composés phénoliques des macroalgues brunes ................................................................. 7 Structures chimiques ................................................................................................................................ 7 Localisation cellulaire ............................................................................................................................... 9 Fonctions ................................................................................................................................................ 11 III. Connaissances acquises sur la synthèse des composés phénoliques: ...................................................... 17 Chez les plantes terrestres : Les flavonoïdes ...................................................................................................... 19 Chez les angiospermes et algues vertes : Les tannins hydrolysables ................................................................. 25 Chez les algues brunes : Les phlorotannins ........................................................................................................ 26 IV. Contexte et objectifs du projet de thèse ................................................................................................. 27 Partie I : Etude de la voie de biosynthèse des phlorotannins ................................................................. 28 Chapitre 1: Caractérisation de la Polyketide Synthase de type III d’Ectocarpus. .......................................... 29 Chapitre 2: Caractérisation biochimique et fonctionnelle de Chalcone Isomérases–like d’Ectocarpus. .............. 46 Abstract .............................................................................................................................................................. 48 Introduction ........................................................................................................................................................ 49 Materials and Methods: ..................................................................................................................................... 51 Results and discussion ........................................................................................................................................ 60
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