Method of designing alpha-amylase mutants with predetermined...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/56 (2006.01) C11D 3/386 (2006.01) C12N 9/28 (2006.01) C12P 19/14 (2006.01) C12S 11/00 (2006.01)

Patent

CA 2211316

A method of constructing a variant of a parent Termamyl-like .alpha.-amylase, which variant has .alpha.-amylase activity and at least one altered property as compared to the parent .alpha.-amylase, comprises i) analysing the structure of the parent Termamyl-like .alpha.-amylase to identify at least one amino acid residue or at least one structural part of the Termamyl-like .alpha.-amylase structure, which amino acid residue or structural part is believed to be of relevance for altering the property of the parent Termamyl-like .alpha.-amylase (as evaluated on the basis of structural or functional considerations), ii) constructing a Termamyl-like .alpha.-amylase variant, which as compared to the parent Termamyl-like .alpha.-amylase, has been modified in the amino acid residue or structural part identified in i) so as to alter the property, and iii) testing the resulting Termamyl-like .alpha.-amylase variant for the property in question.

L'invention porte sur une technique de construction d'un allèle d'une amylase-alpha du type Termamyl. Cet allèle, qui possède une activité diastasique, est doté d'au moins une propriété modifiée eu égard à l'amylase-alpha parente. Cette technique comporte plusieurs étapes, à savoir, i) analyser la structure de l'amylase-alpha parente du type Termamyl afin d'identifier au moins un reste aminoacide ou, à tout le moins une partie structurelle de la structure de l'amylase-alpha du type Termamyl, reste aminoacide dont on pense qu'il est de nature à modifier la propriété de l'amylase-alpha du type Termamyl (comme estimé à partir d'observations structurelles ou fonctionnelles); ii) construire un allèle de l'amylase-alpha du type Termamyl qui, en comparaison de l'amylase-alpha du type Termamyl, a été modifié dans le reste aminoacide ou la partie structurelle identifiée en i, de manière à modifier la propriété susmentionnée et, iii) analyser l'allèle de l'amylase-alpha du type Termamyl qui en est issu, pour évaluer la propriété en question.

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