Chemistry & Biochemistry
Oxalate decarboxylase (OxDC), an enzyme of the bicupin superfamily, catalyzes the decomposition of oxalate into carbon dioxide and formate at an optimal pH of 4.3 in the presence of oxygen. However, about 0.2% of all reactions occur through an oxidase mechanism that consumes oxygen while producing two equivalents of carbon dioxide and one equivalent of hydrogen peroxide. The kinetics of oxidase activity were studied by measuring 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. The temperature dependence of oxidase activity was analyzed, and preliminary results offer an estimate for the overall activation energy of the oxidase reaction within OxDC. The data reported here thus provide insights into the mechanism of the oxidase activity of OxDC.
Journal of Biochemical Technology
Molina, Laura; Goodall, Thomas; Twahir, Umar; and Moomaw, Ellen W., "Real-time Kinetic Studies of Bacillus Subtilis Oxalate Decarboxylase and Ceriporiopsis Subvermispora Oxalate Oxidase Using Luminescent Oxygen Sensor" (2014). Faculty Publications. 3718.