Cholet, Orianne (2006) Etude de l’écosystème fromager par une approche biochimique et moléculaire. PhD thesis Sciences des aliments, UMR de génie et microbiologie des procédés alimentaires, INAPG 2006INAP0032 p.192.

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Abstract

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.

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