{"id":14885,"date":"2024-09-11T17:39:00","date_gmt":"2024-09-11T15:39:00","guid":{"rendered":"https:\/\/www.toulouse-biotechnology-institute.fr\/?p=14885"},"modified":"2024-09-11T18:42:35","modified_gmt":"2024-09-11T16:42:35","slug":"soutenance-de-these-xiaoqian-li","status":"publish","type":"post","link":"https:\/\/www.toulouse-biotechnology-institute.fr\/en\/soutenance-de-these-xiaoqian-li\/","title":{"rendered":"PhD Defence Xiaoqian LI"},"content":{"rendered":"
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Th\u00e8se : Characterization and Engineering of Oligosaccharide Transporters<\/strong><\/p>

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Composition du jury propos\u00e9<\/b><\/p>

Mme Gabrielle\u00a0POTOCKI-VERONESE<\/td>INRAE Occitanie-Toulouse<\/td>Directrice de th\u00e8se<\/td><\/tr>
Mme Aurore\u00a0LABOUREL<\/td>INRAE Occitanie-Toulouse<\/td>Co-directrice de th\u00e8se<\/td><\/tr>
M. Fran\u00e7ois\u00a0THOMAS<\/td>CNRS Bretagne et Pays de la Loire<\/td>Examinateur<\/td><\/tr>
Mme Isabelle\u00a0MEYNIAL-SALLES<\/td>INSA Toulouse<\/td>Examinatrice<\/td><\/tr>
M. Roland\u00a0MARMEISSE<\/td>CNRS Paris-Centre<\/td>Rapporteur<\/td><\/tr>
M. Henri-Pierre\u00a0FIEROBE<\/td>CNRS C\u00f4te d’Azur<\/td>Rapporteur<\/td><\/tr><\/tbody><\/table><\/blockquote>

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Mots-cl\u00e9s\u00a0:<\/b><\/td>Transporteurs,Oligosaccharides,Ing\u00e9nierie des prot\u00e9ines,SusC\/D,Relations structure-Fonction,Enzymes actives sur les sucres<\/td><\/tr><\/tbody><\/table>

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R\u00e9sum\u00e9\u00a0:<\/b>Les mammif\u00e8res h\u00e9bergent des communaut\u00e9s microbiennes tr\u00e8s diverses. Parmi celles-ci, le microbiote intestinal joue un r\u00f4le crucial dans la sant\u00e9 de l\u2019h\u00f4te. Les bact\u00e9ries intestinales utilisent des polysaccharides et oligosaccharides vari\u00e9s comme principales sources de carbone, en utilisant des syst\u00e8mes prot\u00e9iques complexes pour d\u00e9grader ces substrats en monosaccharides assimilables. Les bact\u00e9ries du phylum Bacteroidota sont parmi les plus abondantes dans les microbiomes intestinaux des mammif\u00e8res. Elles produisent une s\u00e9rie de prot\u00e9ines n\u00e9cessaires \u00e0 la d\u00e9tection, \u00e0 la capture, au transport et \u00e0 la d\u00e9gradation des polysaccharides et oligosaccharides. Ces syst\u00e8mes complexes sont cod\u00e9s par des loci d\u00e9di\u00e9s \u00e0 l’utilisation des polysaccharides (PUL). A la surface cellulaire, les prot\u00e9ines de liaison aux glycanes (SGBPs) jouent un r\u00f4le majeur dans l\u2019efficacit\u00e9 de capture des polysaccharides et des oligosaccharides entourant les cellules, offrant ainsi aux Bacteroidota un avantage comp\u00e9titif dans la colonisation de leurs habitats. . Dans ce travail de th\u00e8se, nous nous sommes concentr\u00e9s sur la caract\u00e9risation et l’ing\u00e9nierie des transporteurs de bact\u00e9ries du phylum Bacteroidota, en particulier leurs composants SGBPs, isses des microbiomes digestifs humains et bovins. L’objectif \u00e9tait d’\u00e9lucider les relations structure-fonction des diff\u00e9rents \u00e9l\u00e9ments prot\u00e9iques de ces transporteurs impliqu\u00e9s dans l’utilisation des polysaccharides d\u2019origine alimentaire ou microbiennne. Nous avons tout d\u2019abord caract\u00e9ris\u00e9 la fonction et la structure cristallographique d\u2019une prot\u00e9ine de type SusD (appel\u00e9e ici F5_SusD-like) cod\u00e9e par un PUL provenant d\u2019un Bacteroides non cultiv\u00e9 de l\u2019intestin humain, et qui est conserv\u00e9 chez Bacteroides vulgatus, une esp\u00e8ce dominante dans le microbiote intestinal. Malgr\u00e9 son incapacit\u00e9 \u00e0 se lier aux xylooligosaccharides (XOS), qui sont pourtant les substrats cibl\u00e9s par le F5_PUL, F5_SusD est essentiel \u00e0 la fonctionnalit\u00e9 du transport. L’analyse structurale a r\u00e9v\u00e9l\u00e9 des boucles d\u00e9sordonn\u00e9es et des r\u00e9sidus cl\u00e9s mal align\u00e9s, qui pourraient \u00eatre responsables de l’incapacit\u00e9 de cette prot\u00e9ine de type SGBP \u00e0 se lier aux XOS. Dans un second temps, nous avons \u00e9tudi\u00e9 la sp\u00e9cificit\u00e9 des transporteurs F5_SusC\/D et F5_MFS vis-\u00e0-vis des XOS en introduisant une \u03b2-mannosidase dans le p\u00e9riplasme des souches recombinantes d\u2019E. coli produisant ces transporteurs. En utilisant des variants du locus F5 pr\u00e9c\u00e9demment construits par Tauzin et al., nous avons compl\u00e9t\u00e9 le syst\u00e8me d’utilisation des oligosaccharides de F5 avec l’enzyme BT_0458 de la famille CAZy GH2. Nos donn\u00e9es pr\u00e9liminaires sugg\u00e8rent que les transporteurs du clone m\u00e9tag\u00e9nomique F5 ne peuvent pas internaliser le mannotriose, indiquant une probable stricte sp\u00e9cificit\u00e9 de ces syst\u00e8mes vis-\u00e0-vis des XOS. Par la suite, nous avons explor\u00e9 le potentiel de l\u2019\u00e9volution adaptative en laboratoire (ALE) pour l\u2019ing\u00e9nierie du syst\u00e8me recombinant d\u2019utilisation des XOS chez E. coli. Cette \u00e9tude pr\u00e9liminaire indique que l\u2019\u00e9volution in cellulo pourrait servir \u00e0 am\u00e9liorer les fonctions de transporteurs recombinants chez E. coli et \u00e0 approfondir l\u2019\u00e9tude de leurs relations structure-fonction sous pression \u00e9volutive. Enfin, nous avons caract\u00e9ris\u00e9 une autre prot\u00e9ine de type SusD (appel\u00e9e 41O1_SusD) cod\u00e9e par un locus d’utilisation de \u03b2-glucanes et provenant d’un clone m\u00e9tag\u00e9nomique isol\u00e9 du microbiome du rumen bovin. La caract\u00e9risation fonctionnelle de cette prot\u00e9ine a montr\u00e9 que 41O1_SusD pr\u00e9sente une affinit\u00e9 de liaison pour les \u03b2-1,3-glucanes ramifi\u00e9s par des liaisons \u03b2-1,6. Une comparaison structurale avec des prot\u00e9ines homologues a permis de mettre en \u00e9vidence des motifs similaires de reconnaissance du substrat, impliquant trois r\u00e9sidus tryptophane. Ces r\u00e9sultats ont permis de progresser dans la compr\u00e9hension du r\u00f4le jou\u00e9 par les prot\u00e9ines de type SusD dans l\u2019utilisation des substrats issus de la paroi cellulaire v\u00e9g\u00e9tale et\/ou d\u2019origine fongique dans le rumen bovin.<\/p>

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September 26, 2024<\/p>","protected":false},"author":1,"featured_media":14886,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[10,75],"tags":[35],"class_list":["post-14885","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-evenement","category-soutenance","tag-equipe-cimes"],"yoast_head":"\nSoutenance de th\u00e8se Xiaoqian LI | TBI, Toulouse Biotechnology Institute<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.toulouse-biotechnology-institute.fr\/en\/soutenance-de-these-xiaoqian-li\/\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Soutenance de th\u00e8se Xiaoqian LI | TBI, Toulouse Biotechnology Institute\" \/>\n<meta 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