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 McLeish, M. J.
Right arrow Articles by Kenyon, G. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McLeish, M. J.
Right arrow Articles by Kenyon, G. L.

 Previous Article  |  Next Article 

Journal of Bacteriology, April 2003, p. 2451-2456, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2451-2456.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of a Mandelamide Hydrolase and an NAD(P)+-Dependent Benzaldehyde Dehydrogenase from Pseudomonas putida ATCC 12633

Michael J. McLeish,1,2* Malea M. Kneen,1 Kota N. Gopalakrishna,1 Carolyn W. Koo,2,{dagger} Patricia C. Babbitt,2 John A. Gerlt,3 and George L. Kenyon1,2

College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109,1 Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143,2 Department of Biochemistry, University of Illinois, Urbana, Illinois 618013

Received 11 September 2002/ Accepted 21 January 2003

The enzymes of the mandelate metabolic pathway permit Pseudomonas putida ATCC 12633 to utilize either or both enantiomers of mandelate as the sole carbon source. The genes encoding the mandelate pathway were found to lie on a single 10.5-kb restriction fragment. Part of that fragment was shown to contain the genes coding for mandelate racemase, mandelate dehydrogenase, and benzoylformate decarboxylase arranged in an operon. Here we report the sequencing of the remainder of the restriction fragment, which revealed three further open reading frames, denoted mdlX, mdlY, and mdlD. All were transcribed in the opposite direction from the genes of the mdlABC operon. Sequence alignments suggested that the open reading frames encoded a regulatory protein (mdlX), a member of the amidase signature family (mdlY), and an NAD(P)+-dependent dehydrogenase (mdlD). The mdlY and mdlD genes were isolated and expressed in Escherichia coli, and the purified gene products were characterized as a mandelamide hydrolase and an NAD(P)+-dependent benzaldehyde dehydrogenase, respectively.

* Corresponding author. Mailing address: College of Pharmacy, University of Michigan, 428 Church St., Ann Arbor, MI 48109. Phone: (734) 615-1787. Fax: (734) 615-3079. E-mail: mcleish{at}umich.edu.

{dagger} Present address: Genencor, Inc., South San Francisco, CA 94080.

Journal of Bacteriology, April 2003, p. 2451-2456, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2451-2456.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wang, P.-F., Yep, A., Kenyon, G. L., McLeish, M. J. (2009). Using directed evolution to probe the substrate specificity of mandelamide hydrolase. Protein Eng Des Sel 22: 103-110 [Abstract] [Full Text]  
  • Kiziak, C., Conradt, D., Stolz, A., Mattes, R., Klein, J. (2005). Nitrilase from Pseudomonas fluorescens EBC191: cloning and heterologous expression of the gene and biochemical characterization of the recombinant enzyme. Microbiology 151: 3639-3648 [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»