Method of modifying enzyme and oxidoreductase variant

Details Method of modifying enzyme and oxidoreductase variant Method of modifying enzyme and oxidoreductase variant

C - Chemistry, Metallurgy

12

N

C12N 15/53 (2006.01) C12N 1/21 (2006.01) C12N 9/04 (2006.01) C12N 9/06 (2006.01) C12P 7/62 (2006.01)

Patent

CA 2450867

A method of modifying an enzyme whereby the coenzyme-dependency of an oxidoreductase is changed is developed. Using this method, a novel carbonyl reductase variant capable of using NADH as a coenzyme is provided. It is also intended to provide a process of enzymatically producing an optically active (S)-4-halo-3-hydroxybutyric acid ester by using the carbonyl reductase variant. A method of modifying an enzyme itself so as to change the coenzyme- dependency of carbonyl reductase which asymmetrically reduce a carbonyl compound to give an optically active alcohol; a carbonyl reductase having been converted in the coenzyme-dependency from NADPH to NADH which is obtained by the above method; DNA encoding this enzyme variant; a plasmid having this DNA; transformant cells transformed by this plasmid; and a process for producing an optically active alcohol by using the above enzyme variant and/or the above transformed cells.

L'invention concerne une méthode qui permet de modifier un enzyme et qui consiste à changer la dépendance coenzymatique d'une oxydoréductase. Ce procédé permet également d'obtenir un nouveau variant de carbonyl réductase capable d'utiliser NADH en tant que coenzyme. L'invention concerne également un procédé qui permet de produire enzymatiquement un ester d'acide (S)-4-halo-3-hydroxybutyrique optiquement actif au moyen du variant de carbonyl réductase. L'invention concerne en outre : une méthode qui permet de modifier un enzyme afin de changer la dépendance coenzymatique d'une oxydoréductase, ce qui réduit asymétriquement un composé carbonyle afin d'obtenir un alcool optiquement actif ; une carbonyl réductase convertie par la dépendance coenzymatique de NADPH en NADH, laquelle carbonyl réductase est obtenue au moyen de ladite méthode ; un ADN codant pour ce variant enzymatique ; un plasmide contenant cet ADN ; des cellules transformantes transformées par ce plasmide ; et un procédé qui permet de produire un alcool optiquement actif au moyen dudit variant enzymatique et/ou desdites cellules transformées.

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