Ring-cleaving cyanuric acid amidohydrolase activity in the atrazine-mineralizing Ralstonia basilensis M91-3 |
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| Authors: | David M Stamper Joseph A Krzycki Duongrutiai Nicomrat Samuel J Traina Olli H Tuovinen |
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| Institution: |
a Biological Sciences Group, NAVSEA Warfare Centers, Carderock Division, West Bethesda, MD, USA
b Department of Microbiology, Ohio State University, Columbus, OH, USA
c Sierra Nevada Research Institute, University of California-Merced, Merced, CA, USA |
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| Abstract: | The purpose of this study was to characterize the cyanuric acid amidohydrolase reaction in Ralstonia basilensis M91-3, an atrazine-mineralizing soil bacterium. This ring fission reaction is the last aromatic step in the degradative pathway of atrazine and other s-triazines. The products and molar stoichiometry of the cyanuric acid amidohydrolase reaction were one mol biuret (H2N·CO·NH·CO·NH2) and one mol CO2 per mol cyanuric acid hydrolyzed, as confirmed by 13C-NMR and gas chromatography. The optimum pH and temperature, substrate specificity, and kinetic parameters were also characterized for the purified enzyme. The native enzyme had two forms of different sizes, 204 kDa and 160 kDa. Each was a tetramer or pentamer of 44 kDa and 33 kDa, respectively. |
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| Keywords: | Amidohydrolase atrazine biuret cyanuric acid Ralstonia basilensis s-triazine |
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