C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/26 (2006.01) C12Q 1/00 (2006.01) C12Q 1/32 (2006.01)
Patent
CA 2400660
The present invention relates to screening methods for oxidation enzymes, particularly mono- and dioxygenases. According to the methods of the invention, a product of an oxidation reaction is converted into a phenol or a catechol, which is easily detected by a Gibbs assay. This conversion allows for a sensitive and efficient assay. Both high-throughput liquid-phase and solid-phase methods using these principles are provided. Also described are methods for detecting phenolic ether-products and sulfhydryl products from oxidation reactions, also using a Gibbs assay. Figure (1) shows this athway for a generic, monosubstituted benzene being degraded by the toluene dioxygenase system.
L'invention porte sur des procédés de criblage d'enzymes d'oxydation, en particulier de mono et dioxygénases selon lesquels on convertit le produit d'une réaction d'oxydation en un phénol ou un catéchol, facilement détectable par un essai de Gibbs rendu sensible et efficace du fait de la conversion. On utilise à la fois dans ce cadre des méthodes en phase liquide et des méthodes en phase solide à fort débit. L'invention porte également sur un procédé de détection des produits d'éther phénoliques et des produits sulfhydryliques de réactions d'oxydation en recourant également à un essai de Gibbs. La fig (1) représente le mécanisme d'obtention d'un benzène générique monosubstitué dégradé par le système de toluène dioxygénases.
Arnold Frances F.
Joern John
Sakamoto Takeshi
Schwaneberg Ulrich
California Institute Of Technology
Smart & Biggar
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