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Cholet, Orianne (2006) Etude de l’écosystème fromager par une approche biochimique et moléculaire. Doctorat Sciences des aliments, UMR de génie et microbiologie des procédés alimentaires, INAPG 2006INAP0032 p.192.
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Résumé
Identifier le rôle et la contribution de chaque flore au sein de l’écosystème
fromager a déjà fait l’objet de nombreux travaux, mais les modes d’investigation sont
restés trop souvent descriptifs par manque d’outils moléculaires. L’objectif principal de
cette étude était d’élucider les voies métaboliques impliquées dans la désacidification et
la production de composés soufrés chez trois levures (Debaryomyces hansenii,
Kluyveromyces marxianus et Yarrowia lipolytica) et une bactérie de surface
(Brevibacterium linens) par une approche transcriptomique. La conception et
l’utilisation d’une puce à ADN dédiée à ces quatre micro-organismes ont permis de
mettre en évidence une divergence des voies cataboliques de la L-méthionine entre les
levures et la bactérie d’affinage. L’ensemble des résultats obtenus a montré que la
transamination est l’étape essentielle du catabolisme de la L-méthionine chez les
levures. Elle s’accompagne d’une accumulation transitoire de l’acide α-céto-γ-
méthylthiobutyrique chez Y. lipolytica, et essentiellement de sa forme réduite, l’acide α-
hydroxy-γ-méthylthiobutyrique, chez K. marxianus et D. hansenii. La dégradation de
ces composés se traduit par une augmentation de la production de méthanethiol et des
composés soufrés volatils qui en résultent. Une étude plus approfondie réalisée sur
Y. lipolytica a montré que les gènes ARO8 et BAT2 jouent un rôle prépondérant dans
l’étape de transamination de la L-méthionine chez cette levure. En revanche, chez la
bactérie B. linens, l’enzyme clef du catabolisme de la L-méthionine est la L-méthionine
γ-lyase. L’apport de L-méthionine dans le milieu de culture induit fortement
l’expression du gène mgl et génère une large gamme de composés soufrés volatils.
L’étude des voies de dégradation du lactose et du lactate chez les levures a également
permis d’obtenir des informations sur la part fonctionnelle de chaque espèce au cours
de l’affinage. Ainsi, il semblerait qu’en culture mixte, K. marxianus soit plus impliqué
dans la consommation du lactose via l’induction des gènes LAC12 et LAC4, D. hansenii
dans le métabolisme du pyruvate puis le catabolisme du lactate en fin de culture, et
Y. lipolytica dans la dégradation de la L-méthionine via l’induction des gènes ARO8 et
BAT2. De façon plus globale, l’ensemble de nos résultats permet de mettre en évidence
la possibilité de complémentarités métaboliques entre les levures d’affinage.
| Type d'EPrint: | Thèse (Doctorat) |
|---|---|
| Directeur de Mémoire: | Bonnarme, P. |
| Date: | 13 Décembre 2006 |
| Jury de Mémoire: | Spinnler, H. e. et Lonvaud, A. et Loubière, P. et Reitz-Ausseur, J. et Casarégola, S. |
| Ecole Doctorale: | ED 435 AGRICULTURE, ALIMENTATION, BIOLOGIE, ENVIRONNEMENTS ET SANTE |
| Discipline: | Sciences des aliments |
| Fonds: | INAPG |
| Institution: | INAPG |
| Laboratoire: | UMR de génie et microbiologie des procédés alimentaires |
| Sujets: | 7. Sciences de la vie et ingénierie du vivant |
| Mots-clés libres: | Levure, Fromage, Affinage, Lactose |
| Code ID: | 3111 |
| Déposé par : | Nadine Pontal |
| Déposé le : | 20 Novembre 2007 |
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