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 Kerbarh, O.
Right arrow Articles by Abell, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kerbarh, O.
Right arrow Articles by Abell, C.

Next Article 

Journal of Bacteriology, August 2005, p. 5061-5066, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5061-5066.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Salicylate Biosynthesis: Overexpression, Purification, and Characterization of Irp9, a Bifunctional Salicylate Synthase from Yersinia enterocolitica

Olivier Kerbarh, Alessio Ciulli, Nigel I. Howard, and Chris Abell*

Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom

Received 13 April 2005/ Accepted 14 May 2005

In some bacteria, salicylate is synthesized using the enzymes isochorismate synthase and isochorismate pyruvate lyase. In contrast, gene inactivation and complementation experiments with Yersinia enterocolitica suggest the synthesis of salicylate in the biosynthesis of the siderophore yersiniabactin involves a single protein, Irp9, which converts chorismate directly into salicylate. In the present study, Irp9 was for the first time heterologously expressed in Escherichia coli as a hexahistidine fusion protein, purified to near homogeneity, and characterized biochemically. The recombinant protein was found to be a dimer, each subunit of which has a molecular mass of 50 kDa. Enzyme assays, reverse-phase high-pressure liquid chromatography and 1H nuclear magnetic resonance (NMR) spectroscopic analyses confirmed that Irp9 is a salicylate synthase and converts chorismate to salicylate with a Km for chorismate of 4.2 µM and a kcat of 8 min–1. The reaction was shown to proceed through the intermediate isochorismate, which was detected directly using 1H NMR spectroscopy.

* Corresponding author. Mailing address: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom. Phone: 44 1223 336405. Fax: 44 1223 336362. E-mail: ca26{at}cam.ac.uk.

Journal of Bacteriology, August 2005, p. 5061-5066, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5061-5066.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Van Lanen, S. G., Lin, S., Shen, B. (2008). Biosynthesis of the enediyne antitumor antibiotic C-1027 involves a new branching point in chorismate metabolism. Proc. Natl. Acad. Sci. USA 105: 494-499 [Abstract] [Full Text]  
  • Miethke, M., Marahiel, M. A. (2007). Siderophore-Based Iron Acquisition and Pathogen Control. Microbiol. Mol. Biol. Rev. 71: 413-451 [Abstract] [Full Text]  
  • Strawn, M. A., Marr, S. K., Inoue, K., Inada, N., Zubieta, C., Wildermuth, M. C. (2007). Arabidopsis Isochorismate Synthase Functional in Pathogen-induced Salicylate Biosynthesis Exhibits Properties Consistent with a Role in Diverse Stress Responses. J. Biol. Chem. 282: 5919-5933 [Abstract] [Full Text]  
  • Zaitseva, J., Lu, J., Olechoski, K. L., Lamb, A. L. (2006). Two Crystal Structures of the Isochorismate Pyruvate Lyase from Pseudomonas aeruginosa. J. Biol. Chem. 281: 33441-33449 [Abstract] [Full Text]  
  • Harrison, A. J., Yu, M., Gardenborg, T., Middleditch, M., Ramsay, R. J., Baker, E. N., Lott, J. S. (2006). The Structure of MbtI from Mycobacterium tuberculosis, the First Enzyme in the Biosynthesis of the Siderophore Mycobactin, Reveals It To Be a Salicylate Synthase.. J. Bacteriol. 188: 6081-6091 [Abstract] [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»