This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Shapir, N.
Right arrow Articles by Wackett, L. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shapir, N.
Right arrow Articles by Wackett, L. P.

 Previous Article  |  Next Article 

Journal of Bacteriology, August 2006, p. 5859-5864, Vol. 188, No. 16
0021-9193/06/$08.00+0     doi:10.1128/JB.00517-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

TrzN from Arthrobacter aurescens TC1 Is a Zinc Amidohydrolase

Nir Shapir,1,2,4 Charlotte Pedersen,5 Omer Gil,2,3 Lisa Strong,2 Jennifer Seffernick,1,2,3 Michael J. Sadowsky,2,3,4 and Lawrence P. Wackett1,2,3*

Department of Biochemistry, Molecular Biology and Biophysics,1 BioTechnology Institute,2 Center for Microbial and Plant Genomics,3 Department of Soil, Water & Climate, University of Minnesota, St. Paul, Minnesota 55108,4 Department of Biology, Southern Utah State University, Cedar City, Utah 847205

Received 11 April 2006/ Accepted 6 June 2006

TrzN, the broad-specificity triazine hydrolase from Arthrobacter and Nocardioides spp., is reportedly in the amidohydrolase superfamily of metalloenzymes, but previous studies suggested that a metal was not required for activity. To help resolve that conundrum, a double chaperone expression system was used to produce multimilligram quantities of functionally folded, recombinant TrzN. The TrzN obtained from Escherichia coli (trzN) cells cultured with increasing zinc in the growth medium showed corresponding increases in specific activity, and enzyme obtained from cells grown with 500 µM zinc showed maximum activity. Recombinant TrzN contained 1 mole of Zn per mole of TrzN subunit. Maximally active TrzN was not affected by supplementation with most metals nor by EDTA, consistent with previous observations (E. Topp, W. M. Mulbry, H. Zhu, S. M. Nour, and D. Cuppels, Appl. Environ. Microbiol. 66:3134-3141, 2000) which had led to the conclusion that TrzN is not a metalloenzyme. Fully active native TrzN showed a loss of greater than 90% of enzyme activity and bound zinc when treated with the metal chelator 8-hydroxyquinoline-5-sulfonic acid. While exogenously added zinc or cobalt restored activity to metal-depleted TrzN, cobalt supported lower activity than did zinc. Iron, manganese, nickel, and copper did not support TrzN activity. Both Zn- and Co-TrzN showed different relative activities with different s-triazine substrates. Co-TrzN showed a visible absorption spectrum characteristic of other members of the amidohydrolase superfamily replaced with cobalt.

* Corresponding author. Mailing address: Department of Biochemistry, Molecular Biology and Biophysics, 140 Gortner Lab, 1479 Gortner Ave., University of Minnesota, St. Paul, MN 55108. Phone: (612) 625-3785. Fax: (612) 625-5780. E-mail: wackett{at}biosci.cbs.umn.edu.

Journal of Bacteriology, August 2006, p. 5859-5864, Vol. 188, No. 16
0021-9193/06/$08.00+0     doi:10.1128/JB.00517-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Li, Q., Seffernick, J. L., Sadowsky, M. J., Wackett, L. P. (2009). Thermostable Cyanuric Acid Hydrolase from Moorella thermoacetica ATCC 39073. Appl. Environ. Microbiol. 75: 6986-6991 [Abstract] [Full Text]  
  • Seffernick, J. L., Aleem, A., Osborne, J. P., Johnson, G., Sadowsky, M. J., Wackett, L. P. (2007). Hydroxyatrazine N-Ethylaminohydrolase (AtzB): an Amidohydrolase Superfamily Enzyme Catalyzing Deamination and Dechlorination. J. Bacteriol. 189: 6989-6997 [Abstract] [Full Text]  
  • Shapir, N., Mongodin, E. F., Sadowsky, M. J., Daugherty, S. C., Nelson, K. E., Wackett, L. P. (2007). Evolution of Catabolic Pathways: Genomic Insights into Microbial s-Triazine Metabolism. J. Bacteriol. 189: 674-682 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»