Protein film voltammetry of arsenite oxidase from the chemolithoautotrophic arsenite-oxidizing bacterium NT-26.
The chemolithoautotrophic bacterium NT-26 (isolated from a gold mine in the Northern Territory of Australia) is unusual in that it acquires energy by oxidizing arsenite to arsenate while most other arsenic-oxidizing organisms perform this reaction as part of a detoxification mechanism against the potentially harmful arsenite [present as As(OH)(3) at neutral pH]. The enzyme that performs this reaction in NT-26 is the molybdoenzyme arsenite oxidase, and it has been previously isolated and characterized. Here we report the direct (unmediated) electrochemistry of NT-26 arsenite oxidase confined to the surface of a pyrolytic graphite working electrode. We have been able to demonstrate that the enzyme functions natively while adsorbed on the electrode where it displays stable and reproducible catalytic electrochemistry in the presence of arsenite. We report a pH dependence of the catalytic electrochemical potential of -33 mV/pH unit that is indicative of proton-coupled electron transfer. We also have performed catalytic voltammetry at a number of temperatures between 5 and 25 degrees C, and the catalytic current (proportional to the turnover number) follows simple Arrhenius behavior.
Bernhardt
Paul V
PV
Centre for Metals in Biology, Department of Chemistry, University of Queensland, Brisbane, 4072 Queensland, Australia. P.Bernhardt@uq.edu.au
Santini
Joanne M
JM
D042481:Alcaligenes faecalis / Q000201:enzymology / Q000378:metabolism; D002384:Catalysis; D004563:Electrochemistry; D004579:Electron Transport; D006091:Gram-Negative Chemolithotrophic Bacteria / Q000201:enzymology* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D010088:Oxidoreductases; C078179:arsenite oxidase