Kinetic analysis of the NADH-dehydrogenase activity of the NAD-dependent hydrogenase (hydrogen:NAD+oxidoreductase, EC 220.127.116.11) from the hydrogen-oxidizing bacterium Alcaligenes eutrophus Z1 has been carried out. The enzyme followed a ping-pong reaction mechanism with all the substrates tested. The inhibition analysis revealed an additional coenzyme binding site in the hydrogenase molecule. The presence of the spatially independent substrate-binding sites for different electron acceptors was verified by the alternative substrate method. © 1985.
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