A method of preparing a variant of a lipolytic enzyme

Details A method of preparing a variant of a lipolytic enzyme A method of preparing a variant of a lipolytic enzyme

C - Chemistry, Metallurgy

12

N

C12N 15/55 (2006.01) C11D 3/386 (2006.01) C12N 9/14 (2006.01) C12N 9/20 (2006.01)

Patent

CA 2183431

A method of preparing a variant of a parent lipolytic enzyme, which method comprises: (a) subjecting a DNA sequence encoding the parent lipolytic enzyme to random mutagenesis, (b) expressing the mutated DNA sequence obtained in step (a) in a host cell, and (c) screening for host cells expressing a mutated lipolytic enzyme which has a decreased dependance to calcium and/or an improved tolerance towards a detergent or a detergent component as compared to the parent lipolytic enzyme.

Cette invention se rapporte à un procédé, qui sert à préparer un variant d'une enzyme lipolytique parente et qui consiste: (a) à soumettre une séquence d'ADN codant l'enzyme lipolytique parente à une mutagenèse aléatoire; (b) à exprimer la séquence d'ADN ayant muté, obtenue à l'étape (a) dans une cellule hôte; et (c) à rechercher les cellules hôtes qui expriment une enzyme lipolytique ayant muté, qui possède une dépendance au calcium inférieure à celle de l'enzyme lipolytique parente et/ou une tolérance envers un détergent ou un constituant de détergent supérieure à celle de ladite enzyme lipolytique parente.

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