{"id":10370,"date":"2022-03-01T11:34:00","date_gmt":"2022-03-01T10:34:00","guid":{"rendered":"https:\/\/www.toulouse-biotechnology-institute.fr\/?p=10370"},"modified":"2022-07-21T11:36:10","modified_gmt":"2022-07-21T09:36:10","slug":"controle-de-la-filamentation-de-yarrowia-lipolytica-par-la-maitrise-de-la-conduite-de-fermentation","status":"publish","type":"post","link":"https:\/\/www.toulouse-biotechnology-institute.fr\/en\/controle-de-la-filamentation-de-yarrowia-lipolytica-par-la-maitrise-de-la-conduite-de-fermentation\/","title":{"rendered":"Contr\u00f4le de la filamentation de Yarrowia lipolytica par la ma\u00eetrise de la conduite de fermentation"},"content":{"rendered":"
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La levure Yarrowia lipolytica<\/em> suscite un int\u00e9r\u00eat grandissant de part de sa capacit\u00e9 \u00e0 utiliser des sources de carbone diverses et \u00e0 produire un large spectre de m\u00e9tabolites d\u2019int\u00e9r\u00eat (lipides, prot\u00e9ines, acides organiques, ar\u00f4mes). Cependant, en r\u00e9ponse aux fluctuations de son environnement, Y. lipolytica<\/em> met en place des transitions m\u00e9taboliques et dimorphiques qui peuvent conduire \u00e0 des difficult\u00e9s de mise en \u0153uvre des bioproc\u00e9d\u00e9s. Nos travaux pr\u00e9c\u00e9dents (projet ANR-PIA Probio3) ont montr\u00e9 que la r\u00e9ponse de Y. lipolytica<\/em> \u00e0 des conditions de stress (pH, oxyg\u00e8ne dissous) \u00e9tait d\u00e9pendante du mode de culture et ont permis d\u2019\u00e9mettre l\u2019hypoth\u00e8se d\u2019une relation entre la concentration en sucre r\u00e9siduel (i. e.<\/em> exc\u00e8s pour le mode discontinu et limitant pour le mode continu) et l\u2019induction de la filamentation en pr\u00e9sence de stress. Par une approche couplant le g\u00e9nie microbiologique, la microfluidique et l\u2019analyse \u00e0 l\u2019\u00e9chelle de la cellule unique, nous avons pu d\u00e9montrer le lien entre la concentration en sucre r\u00e9siduel, l\u2019AMPc et la filamentation. Des \u00e9tudes transcriptomiques devraient nous permettre de mieux comprendre les m\u00e9canismes de r\u00e9gulation mis en jeu. N\u00e9anmoins, le contr\u00f4le de la filamentation par la ma\u00eetrise de la conduite du bioproc\u00e9d\u00e9 est un r\u00e9sultat-cl\u00e9 pour le d\u00e9veloppement de proc\u00e9d\u00e9s biotechnologiques avec la levure Y. lipolytica<\/em>.<\/p>

La levure Yarrowia lipolytica<\/em> suscite un int\u00e9r\u00eat grandissant pour des applications en biotechnologie industrielle de part sa capacit\u00e9 \u00e0 utiliser des sources de carbone diverses et \u00e0 produire un large spectre de m\u00e9tabolites d\u2019int\u00e9r\u00eat. Cependant, en r\u00e9ponse aux fluctuations environnementales, Y. lipolytica<\/em> subit des transitions dimorphiques amenant des difficult\u00e9s de mise en \u0153uvre des bioproc\u00e9d\u00e9s (perte de rendement, limitation des transferts, impact sur la rh\u00e9ologie des milieux).<\/p>

Dans nos travaux pr\u00e9c\u00e9dents r\u00e9alis\u00e9s dans le cadre du projet Probio3 (ANR-PIA), il a \u00e9t\u00e9 d\u00e9montr\u00e9 qu’en culture discontinue (exc\u00e8s de sucre), Y. lipolytica<\/em> filamente en r\u00e9ponse \u00e0 des fluctuations de pH ou d\u2019oxyg\u00e8ne dissous. En revanche, elle conserve sa forme ovo\u00efde en culture continue (sucre r\u00e9siduel nul) sous des conditions identiques de stress. Ainsi la morphologie de la levure peut \u00eatre control\u00e9e par la conduite du proc\u00e9d\u00e9. Jusqu\u2019\u00e0 pr\u00e9sentle d\u00e9clenchement de la filamentation \u00e9tait d\u00e9crit comme induit par des conditions de stress et d\u00e9pendant de la souche mise en \u0153uvre. L\u2019hypoth\u00e8se soulev\u00e9e par nos travaux \u00e9tait l\u2019existence d\u2019une relation entre la concentration en sucre r\u00e9siduel dans la culture, et l\u2019induction de la filamentation en pr\u00e9sence de stress.<\/p>

Des cultures de Y. lipolytica<\/em> en mode acc\u00e9lerostat ont \u00e9t\u00e9 r\u00e9alis\u00e9es en condition de stress. Le mode acc\u00e9l\u00e9rostat permet d’augmenter progressivement la vitesse de croissance cellulaire , qui s’accompagne d’une augmentation progressive de la concentration en glucose r\u00e9siduel. Les r\u00e9sultats obtenus ont mis clairement en \u00e9vidence que Y. lipolytica<\/em> conservait une morphologie ovo\u00efde lorsque la concentration r\u00e9siduelle de glucose restait inf\u00e9rieure \u00e0 une valeur seuil d’environ 0,35mg L-1<\/sup>. Une transition vers des formes plus allong\u00e9es se d\u00e9clenchent \u00e0 ce seuil, et s’intensifient progressivement avec l’augmentation de la concentration en sucre r\u00e9siduel.\u00a0 Gr\u00e2ce \u00e0 la mise en \u0153uvre d\u2019un micror\u00e9acteur instrument\u00e9, il nous a \u00e9t\u00e9 possible d\u2019explorer le m\u00e9canisme de r\u00e9gulation de la transition dimorphique en lien avec la concentration en sucre r\u00e9siduel et la concentration en AMPc impliqu\u00e9e dans les cascades de r\u00e9ponse au stress chez les levures. L’effet de l’AMPc sur la transition a \u00e9t\u00e9 \u00e9valu\u00e9 par l’ajout d’AMPc et la quantification de ses niveaux intracellulaires au cours d\u2019un acc\u00e9l\u00e9rostat. Nous avons confirm\u00e9 que l’ajout exog\u00e8ne d’AMPc inhibait la morphologie myc\u00e9lienne de Y. lipolytica<\/em>, m\u00eame avec des concentrations de glucose d\u00e9passant le niveau seuil de 0,35mg.L-1<\/sup>. Tous ces r\u00e9sultats sugg\u00e8rent que les r\u00e9ponses dimorphiques chez Y. lipolytica<\/em> sont r\u00e9gul\u00e9es par les voies de signalisation du sucre, tr\u00e8s probablement via la voie d\u00e9pendante de l’AMPc-PKA, pour lequel l\u2019AMPc a \u00e9t\u00e9 d\u00e9crit comme activateur.<\/p>

Des \u00e9tudes transcriptomiques en cours de traitement devraient nous permettre de mieux comprendre les m\u00e9canismes de r\u00e9gulation mis en en jeu.<\/p>

Valorization<\/strong> :<\/p>

D\u2019ores et d\u00e9j\u00e0 la possibilit\u00e9 de contr\u00f4ler la filamentation par la ma\u00eetrise de la conduite du bioproc\u00e9d\u00e9 est un r\u00e9sultat cl\u00e9 pour le d\u00e9veloppement de proc\u00e9d\u00e9s biotechnologiques avec la levure Y. lipolytica<\/em>.<\/p>

\u00a0<\/p>

R\u00e9f\u00e9rences bibliographiques<\/strong> :<\/p>

  1. Lesage J, Timoumi A, Cenard S, Lombard E, Lee HLT, Guillouet SE, Gorret N (2021). Accelerostat study in conventional and microfluidic bioreactors to assess the key role of residual glucose in the dimorphic transition of Yarrowia lipolytica<\/em> in response to environmental stimuli<\/strong>. New Biotechnology 64:37-45 doi:https:\/\/doi.org\/10.1016\/j.nbt.2021.05.004<\/em><\/li><\/ol>
    1. Timoumi A, Guillouet SE, Molina-Jouve C, Fillaudeau L, Gorret N. (2018). Impacts of environmental conditions on product formation and morphology of Yarrowia lipolytica<\/em><\/strong>. Appl Microbiol Biotechnol 102:3831-3848. doi: 10.1007\/s00253-018-8870-3<\/em><\/li><\/ol>
      1. Timoumi A, Bideaux C, Guillouet SE, Allouche Y, Molina-Jouve C, Fillaudeau L, Gorret N. (2017). <\/strong>Influence of oxygen availability on the metabolism and morphology of Yarrowia lipolytica<\/em>: insights into the impact of glucose levels on dimorphism<\/strong>. Appl Microbiol Biotechnol 101:7317-7333.<\/em><\/li><\/ol>
        1. Timoumi A, Cl\u00e9ret M, Bideaux C, Guillouet SE, Allouche Y, Molina-Jouve C, Fillaudeau L, Gorret N (2016). Dynamic behavior of Yarrowia lipolytica<\/em> in response to pH perturbations: dependence of the stress response on the culture mode<\/strong>. Applied Microbiology and Biotechnology:1-16 doi:10.1007\/s00253-016-7856-2<\/em><\/li><\/ol>

          \u00a0<\/p>

          Contact : ngorret@insa-toulouse.fr<\/strong><\/a> (Nathalie Gorret \u2013 TBI-FAME group)<\/strong><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<\/div>","protected":false},"excerpt":{"rendered":"

          La levure Yarrowia lipolytica suscite un int\u00e9r\u00eat grandissant de part de sa capacit\u00e9 \u00e0 utiliser des sources de carbone diverses et \u00e0 produire un large spectre de m\u00e9tabolites d\u2019int\u00e9r\u00eat (lipides, prot\u00e9ines, acides organiques, ar\u00f4mes). Cependant, en r\u00e9ponse aux fluctuations de son environnement, Y. lipolytica met en place des transitions m\u00e9taboliques et dimorphiques qui peuvent conduire […]<\/p>\n","protected":false},"author":1,"featured_media":10371,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[18],"tags":[71],"class_list":["post-10370","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-actus","tag-equipe-fame"],"yoast_head":"\nContr\u00f4le de la filamentation de Yarrowia lipolytica par la ma\u00eetrise de la conduite de fermentation | 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\/controle-de-la-filamentation-de-yarrowia-lipolytica-par-la-maitrise-de-la-conduite-de-fermentation\/\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Contr\u00f4le de la filamentation de Yarrowia lipolytica par la ma\u00eetrise de la conduite de fermentation | TBI, Toulouse Biotechnology Institute\" \/>\n<meta property=\"og:description\" content=\"La levure Yarrowia lipolytica suscite un int\u00e9r\u00eat grandissant de part de sa capacit\u00e9 \u00e0 utiliser des sources de carbone diverses et \u00e0 produire un large spectre de m\u00e9tabolites d\u2019int\u00e9r\u00eat (lipides, prot\u00e9ines, acides organiques, ar\u00f4mes). 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