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Thèse Présentée Par : KITOUNI Mahmoud En Vue De L'obtention Du Diplôme De : DOCTORAT D'etat En : MICROBIOLOGIE APPLIQUEE

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Thèse Présentée Par : KITOUNI Mahmoud En Vue De L'obtention Du Diplôme De : DOCTORAT D'etat En : MICROBIOLOGIE APPLIQUEE République Algérienne Démocratique et Populaire Ministère de l’Enseignement Supérieur et de la Recherche Scientifique Université Mentouri-Constantine Faculté des Sciences de la Nature et de la Vie Département des Sciences de la Nature et de la Vie N ° d’ordreN° : 84 IT.E / 2007 SERIE : 05 ISN / 2007 Thèse présentée par : KITOUNI Mahmoud En vue de l’obtention du Diplôme de : DOCTORAT D’ETAT en : MICROBIOLOGIE APPLIQUEE Intitulée : Isolement de bactéries actinomycétales productrices d’antibiotiques à partir d’écosystèmes extrêmes. Identification moléculaire des souches actives et caractérisation préliminapréliminaireire des substances élaborées Membres du jury : Mr BENGUEDOUAR A. Professeur Président Univ. Mentouri-Constantine Mr BOULAHROUF A. Professeur Directeur de thèse Univ. Mentouri-Constantine Mr BOIRON P. Professeur Examinateur Univ. Lyon 1 Mr KARAM N. Professeur Examinateur Univ. Essania-Oran Mr BELLAHCENE M. Maître de Examinateur Univ. Mostaganem Conférences Mr HADDI M.L. Maître de Examinateur Univ. Mentouri-Constantine Conférences Session 2007 REMERCIEMENTS Ce travail a été réalisé au laboratoire de génie microbiologique et applications de l’Université Mentouri de Constantine. Que Monsieur Abderrahmane Boulahrouf (Professeur à l’Université Mentouri de Constantine) trouve ici l’expression de ma très vive reconnaissance pour avoir accepter la responsabilité de ce travail. Je remercie Monsieur Amar Benguedouar (Professeur à l’Université Mentouri de Constantine) de m’avoir fait l’honneur de présider mon jury de thèse. Mes remerciements vont également à Monsieur Patrick Boiron (Professeur à l’Université Claude Bernard Lyon 1) pour la confiance et l’accueil chaleureux qui ma réservé à Lyon, ses précieux conseils et d’avoir accepté de se déplacer à Constantine pour participer à ce jury. Je ne saurais oublié Messieurs Noureddine Karam (Professeur à l’Université Essania Oran) et Miloud Bellahcene (Maître de Conférences à l’Université de Mostaganem) qui sans hésitation ont accepté de se déplacer pour siéger dans ce jury. Mes remerciements s’adressent également à Monsieur Mohamed Laid Haddi (Maître de Conférences à l’Université Mentouri de Constantine) d’avoir accepter d’examiner ce travail. La partie identification moléculaire a été réalisée au laboratoire de Mycologie de l’Université Claude Bernard Lyon 1. Je suis très reconnaissant à Madame André Couble et Delphine Mouniee de m’avoir initié aux techniques de biologie moléculaire et qui ont veillé sur la réalisation de ces travaux et avec qui les discussions ont été toujours fructueuses. Je dois une profonde reconnaissance à ma chère épouse Lamia qui a suivi de très près ce travail avec une grande attention et une disponibilité remarquable. C’est avec gentillesse et patience qu’elle m’a fait toujours bénéficier de ses conseils. Mes remerciements vont également à Madame Hayet Goudjil ep. Benhizia qui a réalisé la prise des photos au microscope optique, ainsi que Monsieur Abdennour Zertal et Mademoiselle Hajira qui ont réalisé les spectres UV-Visibles et L’analyse des échantillons par HPLC. Que mes remerciements aillent aussi a mes amis pour l’amitié et la sympathie qu’ils m’ont témoigné durant ces années. DEDICACES A mon défunt papa, qui ma toujours aidé et guidé vers le chemin de la réussite. A ma très chère maman, pour m’avoir toujours écoutée, pour son soutien moral et ces précieux conseils. A ma tendre épouse Lamia qui ma encouragé, qui ma donné la force et la volonté de surmonter tous les obstacles et les difficultés. A mon poussin Moncef, ma joie, mon bonheur et ma raison de vivre Quoique que je fasse, je ne pourrais vous rendre ce que vous avez fait pour moi. Si je suis arrivé là, c’est bien grâce à vous. Que dieu vous bénisses, vous donne longue vie et vous protège pour moi. A ma famille, à mes amis, je dédie ce mémoire Said. SOMMAIRE REMERCIEMENTS DEDICACES Introduction générale ..................................................................................................................1 Revue bibliographique 1- Résistance aux antibactériens.................................................................................................4 1-1 L’adaptation des bactéries à l’agression des antibactériens .............................................4 1-1-1 Stratégie quantitative.................................................................................................4 1-1-2 Stratégie qualitative...................................................................................................4 1-2 Epidémiologie de la résistance aux antibactériens ...........................................................5 2- Actinomycètes ........................................................................................................................7 2-1 Définition et caractéristiques principales ........................................................................7 2-2 Ecologie..........................................................................................................................10 2-3 Biologie du développement............................................................................................14 2-3-1 Mycélium de substrat et mycélium aérien . ............................................................14 2-3-2 Structures particulières ...........................................................................................15 2-3-3 Formation des spores...............................................................................................16 - Les endospores ...............................................................................................................16 - Les exospores .................................................................................................................16 2-4 Taxonomie......................................................................................................................17 2-4-1 Taxonomie phénétique............................................................................................18 2-4-2 Taxonomie numérique ............................................................................................19 2-4-3 Chimiotaxonomie....................................................................................................19 - Composition en aminoacides..........................................................................................21 - Composition en sucres...................................................................................................21 - Composition en lipides...................................................................................................23 2-4-4 Taxonomie moléculaire...........................................................................................23 - Détermination du G + C %.............................................................................................23 - Les hybridations d'acides nucléiques .............................................................................25 - Etude de l’ARN 16S.......................................................................................................26 - Phylogénie des actinomycètes........................................................................................28 2-4-5 Taxonomie mixte et consensuelle (polyphasic taxonomy) .....................................28 2-5 Classification ..................................................................................................................29 3- Les antibiotiques produits par les actinomycètes .................................................................34 3-1 Généralités......................................................................................................................34 3-2 Antibiotiques inhibant la synthèse de la paroi bactérienne ............................................39 3-2-1 Paroi bactérienne et synthèse du peptidoglycane....................................................39 3-2-2 Structure et mode d’action des antibiotiques de type β-lactame.............................41 3-2-3 Structure et mode d’action des glycopeptides.........................................................43 3-2-4 Structure et mode d’action de la fosfomycine.........................................................45 3-3 Antibiotiques inhibant la synthèse des protéines............................................................47 3-3-1 Structure des Macrolides et apparentés (MLS)......................................................47 3-3-2 Structure et mode d’action des phénicolés..............................................................49 3-3-3 Structure et mode d’action des aminoglycosides....................................................52 3-3-4 Structure et mode d’action des Tétracyclines .........................................................54 3-4 Antibiotiques agissant au niveau de l’ARN polymérase (les Rifamycines)...................56 Partie expérimentale Optimisation d’un protocole d’isolement, étude de la diversité des actinomycètes dans les écosystèmes explorés et mise en évidence de l’activité antibiotique 1- Prélèvement des échantillons ...............................................................................................57 1-1 Sol...................................................................................................................................57 1-2 Eau..................................................................................................................................57 1-3 Ecorces d’arbres .............................................................................................................57
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