Process for obtaining acyloins, pyruvate decarboxylases...

Details Process for obtaining acyloins, pyruvate decarboxylases... Process for obtaining acyloins, pyruvate decarboxylases...

C - Chemistry, Metallurgy

12

N

C12N 15/60 (2006.01) C12N 9/88 (2006.01) C12P 7/26 (2006.01)

Patent

CA 2222493

The invention relates to a process for obtaining a pyruvate decarboxylase by isolation from a producer organism. The pyruvate decarboxylase is capable of forming (R)-(-)-phenylacetylcarbinole (I) in >= 95 % enantiomer unit with a product ratio of (I) to 2-hydroxypropiophenone of >= 95 %. In addition, the pyruvate decarboxylase has a specific activity with regard to phenylacetylcarbinole formation of > 1 U/mg. It is the aim of the invention to obtain a pyruvate decarboxylase with improved synthesis capacity concerning the formation of (R)-(-)-phenylacetylcarbinole. The process of the invention developed for this purpose is characterized in that use is made of a producing organism with a gene coding for pyruvate decarboxylase from Zymomonas mobilis, in the DNA sequence of which the tryptophane radical coding codon TGG is replaced at position 1174-1176 by a codon which codes for an amino acid radical with a reduced volume ratio.

L'invention concerne un procédé de préparation d'une décarboxylase de pyruvate par isolement à partir d'un organisme producteur. La décarboxylase de pyruvate se prête à former du (R)-(-)-phénylacétylcarbinol (I) dans ? 95 % d'unité énantiomère avec un rapport produit de (I) à 2-hydroxypropiophénone de ? 95 %. La décarboxylase de pyruvate comporte en outre une activité spécifique en ce qui concerne la formation de phénylacétylcarbinol de > 1 U/mg. L'invention vise à obtenir une décarboxylase de pyruvate à capacité de synthèse améliorée en ce qui concerne la formation de (R)-(-)-phénylacétylcarbinol. Le procédé mis au point à cette fin se caractérise en ce qu'on utilise un organisme producteur avec un gène codant pour la décarboxylase de pyruvate, issu du Zymonas mobilis, dans la séquence d'ADN duquel le codon TGG codant le reste tryptophane est remplacé en position 1174-1176 par un codon codant un reste aminoacide à taux volumique réduit.

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