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2014 Volume 5 Issue 4

Real-Time kinetic studies of Bacillus subtilis oxalate decarboxylase and Ceriporiopsis subvermispora oxalate oxidase using a luminescent oxygen sensor


Abstract
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.

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J BIOCHEM TECH
J BIOCHEM TECH
Journal of Biochemical Technology is a peer reviewed International Journal published by the Sevas Publishing on behalf of the Biochemical Technology Society, a Registered Charity Organization from India

AREA OF INTEREST
AREA OF INTEREST
Areas of high interest, cover new advances in enzymatic and protein mechanims; applied molecular genetics and biotechnology; genomics and proteomics; metabolic; medical, environmental, food and agro biotechnology.

FOCUS AND SCOPE
FOCUS AND SCOPE
Journal Of Biochemical Technology” Provides A Medium For The Rapid Publication Of Full-Length Articles, Mini-Reviews Of New And Emerging Products And Short Communications On All Aspects Of ...

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This journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. Keywords include, Biochemical Research: Endo/exocytosis, Trafficking, Membrane Biology, Cell Migration, Cell-Matrix Organelle Biogenesis, Cytoskeleton Proteolysis, Cell Death, Cell Cycle, Cancer, Cell Growth/Death, Differentiation, Drug Targets, Gene Therapy, Models of Disease, Proteomics, Stem Cells, Bioenergetics, Mitochondria, Free Radicals, Redox Signaling, Ion Transport/Channels, Oxidative