Screening for lipolytic enzyme or amidase activity

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/26 (2006.01) C12Q 1/34 (2006.01) C12Q 1/44 (2006.01)

Patent

CA 2540379

A method of testing samples for their enzymatic activity for hydrolysis of a particular ester or amide bond in a substrate uses a test substrate with one or more polyunsaturated fatty acyl groups linked through amide or ester bonds. The release of the polyunsaturated fatty acid is detected by the use of a lipoxygenase to convert the polyunsaturated fatty acid into a hy-droperoxide which is then detected, e.g. through a color reaction.

L'invention concerne un procédé de test d'échantillons permettant de révéler leur activité enzymatique pour l'hydrolyse d'une liaison ester ou amide particulière dans un substrat. Le procédé selon l'invention met en oeuvre un substrat de test contenant au moins un groupe acyle gras polyinsaturé à liaisons amide ou ester. La libération de l'acide gras polyinsaturé est détectée au moyen d'une lipoxygénase destinée à convertir l'acide gras polyinsaturé en peroxyde d'hydrogène. Le peroxyde d'hydrogène est ensuite détecté, au moyen d'une réaction colorée par exemple.

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