Inactivation method

摘要

The present invention relates to the field of thermostable enzymes, methods for their inactivation, and related uses, kits and reagents. In particular, the invention relates to a method for reversible inactivation of thermophilic enzymes by chemical modification under aqueous or non-aqueous conditions.

主权项

1. An enzymatically inactivated derivatized thermostable DNA polymerase obtained by a method comprising reacting a thermostable DNA polymerase with a dicarboxylic acid imide compound of formula I: wherein: X is an electron withdrawing group; Y is a linker group and represents a radical of formula IIa or IIb: wherein: R1, R2, R3 and R4 are each independently hydrogen or alkyl, or R1 and R2 together with the carbon atoms to which they are attached form a cycloalkylene, cycloalkenylene, cycloalkyarylene or arylene group; provided that when Y represents a radical of formula IIb at least one of R1 and R2 is alkyl, and each α is a bond linking the radical to a carbon atom of a carbonyl group shown in formula I; wherein the linkage to the derivatized thermostable DNA polymerase is at one or more amino groups of the derivatized thermostable DNA polymerase structure and wherein the derivatized thermostable DNA comprises a moiety of formula IIIa or IIIb: wherein X, Y, R1, R2, R3 and R4 are defined as above and β is a bond linking the moiety to the enzyme; and wherein inactivation of the enzymatic activity is reversible and wherein the derivatized thermostable DNA polymerase is stable to heat at 50° C.