Oxalate decarboxylase (OxDC), an enzyme of the bicupinsuperfamily, catalyzes the decomposition of oxalate into carbondioxide and formate at an optimal pH of 4.3 in the presence ofoxygen. However, about 0.2% of all reactions occur through anoxidase mechanism that consumes oxygen while producing twoequivalents of carbon dioxide and one equivalent of hydrogenperoxide. The kinetics of oxidase activity were studied bymeasuring the consumption of dissolved oxygen over time using a luminescent oxygen sensor. We describe the implementation of and improvements to the oxygen consumption assay. The oxidase activity of wild type OxDC was compared to that of the T165V OxDC mutant, which contains an impaired flexible loop covering the active site. The effects of various carboxylic acid-based buffers on the rate of oxidase activity were also studied. These results were compared to the oxidase activity of oxalate oxidase (OxOx), a similar bicupin enzyme that only carries out oxalate oxidation. Thetemperature dependence of oxidase activity was analyzed, andpreliminary results offer an estimate for the overall activationenergy of the oxidase reaction within OxDC. The data reported here thus provide insights into the mechanism of the oxidase activity of OxDC.