Boehringer Mannheim Gmbh

Erich Elstner

Hans Seidel

Albert Roder

Hans Mollering

Friedrich Gotz

Karl-Heinz Schleifer

Barbara Sedewitz

The present invention provides a new pyruvate oxidase which decarboxylates pyruvate with the formation of hydrogen peroxide, characterized in that it is active without the addition of FAD, TPP and divalent metal ions. The present invention also provides a process for preparing this new enzyme and a reagent containing it.

Pyruvate oxidase

Process for the selective production of the reduced oxygen species superoxide, hydrogen peroxide and hydroxyl radicals, wherein oxygen is reduced with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an appropriate autoxidisable redox partner and optionally of an appropriate buffer system. Also, process for the determination of superoxide dismutase in which a superoxide-yielding reaction is coupled with two competing superoxide-consuming reactions, one superoxide-consuming reaction being a conventional indicator reaction and the other superoxide-consuming reaction being the superoxide dismutase reaction, wherein, as superoxide-yielding reaction, there is used the reduction of oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an autoxidisable one-electron step-inducing redox partner with a one-electron redox potential in the range of from -150 mV to -500 mV and optionally of an appropriate buffer system. Also, process for the determination of NAD(P)H and of NAD(P)H-yielding reactions in which oxygen is reduced to hydrogen peroxide by NAD(P)H and the hydrogen peroxide is determined in knowm manner, wherein the reduction of the oxygen by NAD(P)H is carried out in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an appropriate two-electron redox partner with a two-electron redox potential in the range of from 0 mV to 150 mV and of an appropriate buffer system. Also, reagents for carrying out these processes.

Process for the selective production of reduced oxygen species

Growing community of inventors

Grobenzell, Germany

Erich Elstner