Recombinant materials and methods for the production of...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/02 (2006.01) C07H 21/04 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/63 (2006.01)

Patent

CA 2294475

cDNA encoding spearmint (-)-limonene-6-hydroxylase and peppermint (-)-limonene- 3-hydroxylase have been isolated and sequenced, and the corresponding amino acid sequences determined. DNA sequences are provided which code for the expression of these enzymes (SEQ ID NO:1, from Mentha spicata and SEQ ID NO:8 from Mentha piperita. Systems and methods are provided for recombinant expression of limonene hydroxylases that may be used to facilitate the production, isolation and purification of significant quantities of the enzymes (or of the primary enzyme products, trans-carveol or trans-isopiperitenol as shown in the Figure) for subsequent use, to obtain expression or enhanced expression of the enzymes in plants to attain enhanced production of the primary enzyme products as a predator or pathogen defense mechanism, or for the regulation or expression of the enzymes or their primary products.

Un ADN complémentaire codant une (-)-limonène-6-hydroxylase de la menthe verte et une (-)-limonène-3-hydroxylase de la menthe poivrée a été isolé et séquencé et les séquences aminoacides correspondantes déterminées. Il est fourni, dans le cadre de cette invention, des séquences d'ADN codant l'expression de ces enzymes (SEQ ID N DEG 1, issue de Mentha spicata et SEQ ID N DEG 8, issue de Mentha piperita). L'invention concerne également des systèmes, ainsi que les méthodes correspondantes, aux fins de l'expression de recombinaison de limonène hydroxylases. Ces systèmes et ces méthodes sont susceptibles d'être employées pour faciliter la production, l'isolation et la purification de quantités importantes de ces enzymes (ou des produits enzymatiques primaires, trans-carvéol ou trans-isopipériténol, comme présenté dans la figure) utilisables ultérieurement, et ce, pour obtenir une expression desdites enzymes dans des végétaux ou la renforcer en vue d'augmenter la production des produits enzymatiques primaires, agissant en tant que mécanismes prédateurs ou mécanisme de défense contre des agents pathogènes, ou bien pour réguler l'expression de ces enzymes ou celle de leurs produits primaires.

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