Metabolically engineered cells for the production of pinosylvin

Details Metabolically engineered cells for the production of pinosylvin Metabolically engineered cells for the production of pinosylvin

C - Chemistry, Metallurgy

12

P

C12P 7/22 (2006.01)

Patent

CA 2658294

A genetically engineered micro-organism having an operative metabolic pathway producing cinnamoyl-CoA and producing pinosylvin therefrom by the action of a stilbene synthase is used for pinosylvin production. Said cinnamic acid may be formed from L-phenylalanine by a L-phenylalanine ammonia lyase (PAL) which is one accepting phenylalanine as a substrate and producing cinammic acid therefrom, preferably such that if the PAL also accepts tyrosine as a substrate and forms coumaric acid therefrom, the ratio Km(phenylalanine)/Km(tyrosine) for said PAL is less than 1:1 and if said micro-organism produces a cinammate-4- hydroxylase enzyme (C4H), the ratio Kcat(PAL)/Kcat(C4H) is at least 2:1.

Selon l'invention, le micro-organisme génétiquement modifié, doté d'une voie métabolique fonctionnelle produisant du cinnamoyl-CoA et synthétisant de la pinosylvine à partir de ce substrat grâce à une stilbène synthase, est utilisé pour la production de pinosylvine. Ledit acide cinnamique peut être formé à partir de L-phénylalanine à l'aide d'une L-phénylalanine ammoniac lyase (PAL) qui est une enzyme acceptant la phénylalanine à titre de substrat et qui produit de l'acide cinammique à partir d'un tel substrat, de sorte que si la PAL accepte également la tyrosine à titre de substrat et forme de l'acide coumarique à partir d'un tel substrat, le rapport Km(phénylalanine)/Km(tyrosine) pour ladite PAL soit de préférence inférieur à 1:1, et si ledit micro-organisme produit une enzyme cinammate-4-hydroxylase (C4H), le rapport Kcat(PAL)/Kcat(C4H) soit au moins de 2:1.

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