Visualization of enzyme-catalysed reactions using ph...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/34 (2006.01) C12N 9/18 (2006.01) C12N 15/10 (2006.01) C12N 15/12 (2006.01) C12Q 1/37 (2006.01) C12Q 1/44 (2006.01)

Patent

CA 2332638

The use of pH indicators to monitor enzyme-catalyzed reactions is described. The formation of acid following an enzyme-mediated reaction, such as hydrolysis, causes a drop in the pH that can be visualized by a change in the color of the indicator-containing solution. Preferred indicators are those showing a color transition within the operational pH range of the enzyme. Using the present system, the enantioselectivity of enzymes such as lipases and esterases can be estimated using single isomers under the same conditions and comparing the color turnover for each one. The method is also useful for application to the hierarchical screening of a library of enzymes.

Cette invention propose l'utilisation d'indicateurs de pH pour surveiller des réactions catalysées par des enzymes. La formation d'acide suivant une réaction enzymatique, telle que l'hydrolyse, entraîne une baisse du pH qui peut être visualisée par le changement de couleur de la solution contenant l'indicateur. Les indicateurs préférés sont ceux qui montrent une transition de couleur dans la plage de pH opérationnelle de l'enzyme. En utilisant le système proposé par cette invention, on peut évaluer l'énantiosélectivité d'enzymes, telles que des lipases et des estérases, en utilisant des isomères simples dans les mêmes conditions et en comparant le changement de couleur pour chacun d'eux. Ce procédé peut également s'appliquer au criblage hiérarchique d'une bibliothèque d'enzymes.

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