Catalase as a sulfide-sulfur oxido-reductase: An ancient (and modern?) regulator of reactive sulfur species (RSS)

Catalase is well-known as an antioxidant dismutating H₂O₂ to O₂ and H₂O. However, catalases evolved when metabolism was largely sulfur-based, long before O₂ and reactive oxygen species (ROS) became abundant, suggesting catalase metabolizes reactive sulfide species (RSS). Here we examine catalase metabolism of H₂S_n, the sulfur analog of H₂O₂, hydrogen sulfide (H₂S) and other sulfur-bearing molecules using H₂S-specific amperometric electrodes and fluorophores to measure polysulfides (H₂S_n; SSP4) and ROS (dichlorofluorescein, DCF). Catalase eliminated H₂S_n, but did not anaerobically generate H₂S, the expected product of dismutation. Instead, catalase concentration- and oxygen-dependently metabolized H₂S and in so doing acted as a sulfide oxidase with a P₅₀ of 20mmHg. H₂O₂ had little effect on catalase-mediated H₂S metabolism but in the presence of the catalase inhibitor, sodium azide (Az), H₂O₂ rapidly and efficiently expedited H₂S metabolism in both normoxia and hypoxia suggesting H₂O₂ is an effective electron acceptor in this reaction. Unexpectedly, catalase concentration-dependently generated H₂S from dithiothreitol (DTT) in both normoxia and hypoxia, concomitantly oxidizing H₂S in the presence of O₂. H₂S production from DTT was inhibited by carbon monoxide and augmented by NADPH suggesting that catalase heme-iron is the catalytic site and that NADPH provides reducing equivalents. Catalase also generated H₂S from garlic oil, diallyltrisulfide, thioredoxin and sulfur dioxide, but not from sulfite, metabisulfite, carbonyl sulfide, cysteine, cystine, glutathione or oxidized glutathione. Oxidase activity was also present in catalase from Aspergillus niger. These results show that catalase can act as either a sulfide oxidase or sulfur reductase and they suggest that these activities likely played a prominent role in sulfur metabolism during evolution and may continue do so in modern cells as well. This also appears to be the first observation of catalase reductase activity independent of peroxide dismutation.