This Article
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 Mayaux, J F
Right arrow Articles by Pétré, D
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mayaux, J F
Right arrow Articles by Pétré, D

 Previous Article  |  Next Article 

J Bacteriol. 1991 November; 173(21): 6694-6704

research-article

Purification, cloning, and primary structure of a new enantiomer-selective amidase from a Rhodococcus strain: structural evidence for a conserved genetic coupling with nitrile hydratase.

J F Mayaux, E Cerbelaud, F Soubrier, P Yeh, F Blanche and D Pétré

Département Biotechnologie, Centre de Recherche de Vitry-Alfortville, Vitry sur Seine, France.

ABSTRACT

A new enantiomer-selective amidase active on several 2-aryl propionamides was identified and purified from a newly isolated Rhodococcus strain. The characterized amidase is an apparent homodimer, each molecule of which has an Mr of 48,554; it has a specific activity of 16.5 mumol of S(+)-2-phenylpropionic acid formed per min per mg of enzyme from the racemic amide under our conditions. An oligonucleotide probe was deduced from limited peptide information and was used to clone the corresponding gene, named amdA. As expected, significant homologies were found between the amino acid sequences of the enantiomer-selective amidase of Rhodococcus sp., the corresponding enzyme from Brevibacterium sp. strain R312, and several known amidases, thus confirming the existence of a structural class of amidase enzymes. Genes probably coding for the two subunits of a nitrile hydratase, albeit in an inverse order, were found 39 bp downstream of amdA, suggesting that such a genetic organization might be conserved in different microorganisms. Although we failed to express an active Rhodococcus amidase in Escherichia coli, even in conditions allowing the expression of an active R312 enzyme, the high-level expression of the active recombinant enzyme could be demonstrated in Brevibacterium lactofermentum by using a pSR1-derived shuttle vector.

J Bacteriol. 1991 November; 173(21): 6694-6704




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]  
  • Yun, Y. S., Lee, W., Shin, S., Oh, B.-H., Choi, K. Y. (2006). Arg-158 Is Critical in Both Binding the Substrate and Stabilizing the Transition-state Oxyanion for the Enzymatic Reaction of Malonamidase E2. J. Biol. Chem. 281: 40057-40064 [Abstract] [Full Text]  
  • Shin, S., Yun, Y. S., Koo, H. M., Kim, Y. S., Choi, K. Y., Oh, B.-H. (2003). Characterization of a Novel Ser-cisSer-Lys Catalytic Triad in Comparison with the Classical Ser-His-Asp Triad. J. Biol. Chem. 278: 24937-24943 [Abstract] [Full Text]  
  • Trott, S., Burger, S., Calaminus, C., Stolz, A. (2002). Cloning and Heterologous Expression of an Enantioselective Amidase from Rhodococcus erythropolis Strain MP50. Appl. Environ. Microbiol. 68: 3279-3286 [Abstract] [Full Text]  
  • Trott, S., Bauer, R., Knackmuss, H.-J., Stolz, A. (2001). Genetic and biochemical characterization of an enantioselective amidase from Agrobacterium tumefaciens strain d3. Microbiology 147: 1815-1824 [Abstract] [Full Text]  
  • Soong, C.-L., Ogawa, J., Shimizu, S. (2000). A Novel Amidase (Half-Amidase) for Half-Amide Hydrolysis Involved in the Bacterial Metabolism of Cyclic Imides. Appl. Environ. Microbiol. 66: 1947-1952 [Abstract] [Full Text]  
  • Boshoff, H. I. M., Mizrahi, V. (1998). Purification, Gene Cloning, Targeted Knockout, Overexpression, and Biochemical Characterization of the Major Pyrazinamidase from Mycobacterium smegmatis. J. Bacteriol. 180: 5809-5814 [Abstract] [Full Text]  
  • Fournand, D., Bigey, F., Arnaud, A. (1998). Acyl Transfer Activity of an Amidase from Rhodococcus sp. Strain R312: Formation of a Wide Range of Hydroxamic Acids. Appl. Environ. Microbiol. 64: 2844-2852 [Abstract] [Full Text]  
  • Curnow, A. W., Hong, K.-w., Yuan, R., Kim, S.-i., Martins, O., Winkler, W., Henkin, T. M., Soll, D. (1997). Glu-tRNAGln amidotransferase: A novel heterotrimeric enzyme required for correct decoding of glutamine codons during translation. Proc. Natl. Acad. Sci. USA 94: 11819-11826 [Abstract] [Full Text]  
  • Kobayashi, M., Fujiwara, Y., Goda, M., Komeda, H., Shimizu, S. (1997). Identification of active sites in amidase: Evolutionary relationship between amide bond- and peptide bond-cleaving enzymes. Proc. Natl. Acad. Sci. USA 94: 11986-11991 [Abstract] [Full Text]  
  • Komeda, H., Kobayashi, M., Shimizu, S. (1996). A Novel Gene Cluster Including the Rhodococcus rhodochrous J1 nhlBA Genes Encoding a Low Molecular Mass Nitrile Hydratase (L-NHase) Induced by Its Reaction Product. J. Biol. Chem. 271: 15796-15802 [Abstract] [Full Text]  
  • Marti, T., Hu, Z., Pohl, N. L., Shah, A. N., Khosla, C. (2000). Cloning, Nucleotide Sequence, and Heterologous Expression of the Biosynthetic Gene Cluster for R1128, a Non-steroidal Estrogen Receptor Antagonist. INSIGHTS INTO AN UNUSUAL PRIMING MECHANISM. J. Biol. Chem. 275: 33443-33448 [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»