The Cu,Zn superoxide dismutase catalyzes HCO(3)(-) -dependent oxidations by H(2)O(2). This activity has been shown to depend on the creation of a bound oxidant at the Cu(II) by interactions with H(2)O(2). The bound oxidant was then thought to oxidize HCO(3)(-) to CO(3)(.-), which diffuses into the bulk solution and there oxidizes diverse substrates. We now find that CO(2) rather than HCO(3)(-) facilitates the peroxidations catalyzed by Cu,Zn superoxide dismutase. This fact was shown by a lag in the rate of peroxidation of NADPH when NaHCO(3)(-) was added last and by a burst in the rate when aqueous CO(2) was added last. Both the lag and the burst were eliminated by carbonic anhydrase.