Cloning and expression of the gene encoding...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/53 (2006.01) C12N 9/04 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2247038

The adhB gene encoding Thermoanaerobacter ethanolicus 39E secondary-alcohol dehydrogenase (2-Adh) was cloned, sequenced, and expressed in Escherichia coli. Both the nucleic acid sequence coding for the enzyme and the amino acid sequence are reported. The purified recombinant enzyme is homotetrameric consisting of 37.7 kDa subunits and optimally active above 90°C with a half-life of approximately of 1.7 h at 90°C. The enzyme has a substantial preference in the oxidation of secondary alcohols over primary alcohols.

Le gène adhB codant la déshydrogénase d'alcool secondaire (2-Adh) de thermoanaerobacter ethanolicus 39E, a été cloné, séquencé et exprimé chez Escherichia coli. L'invention porte à la fois sur la séquence d'acides nucléiques codant l'enzyme et sur la séquence d'acides aminés. L'enzyme recombinante purifiée est de type homotétramère, composé de sous-unités de 37,7 kDa; elle présente une activité optimale au-dessus de 90 degrés C et possède une demi-vie d'environ 1,7 h à 90 degrés C. Cette enzyme oxyde de façon nettement préférentielle les alcools secondaires, plutôt que les alcools primaires.

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