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 Sanz, Y.
Right arrow Articles by Toldrá, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sanz, Y.
Right arrow Articles by Toldrá, F.
Agricola
Right arrow Articles by Sanz, Y.
Right arrow Articles by Toldrá, F.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, April 2002, p. 1980-1987, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1980-1987.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Purification and Characterization of an Arginine Aminopeptidase from Lactobacillus sakei

Yolanda Sanz* and Fidel Toldrá

Instituto de Agroquímica y Tecnología de Alimentos (CSIC), 46100 Burjasot (Valencia), Spain

Received 9 July 2001/ Accepted 21 January 2002

An arginine aminopeptidase (EC 3.4.11.6) that exclusively hydrolyzes basic amino acids from the amino (N) termini of peptide substrates has been purified from Lactobacillus sakei. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included hydrophobic interaction, gel filtration, and anion-exchange chromatography. This procedure resulted in a recovery rate of 4.2% and a 500-fold increase in specific activity. The aminopeptidase appeared to be a trimeric enzyme with a molecular mass of 180 kDa. The activity was optimal at pH 5.0 and 37°C. The enzyme was inhibited by sulfhydryl group reagents and several divalent cations (Cu2+, Hg2+, and Zn2+) but was activated by reducing agents, metal-chelating agents, and sodium chloride. The enzyme showed a preference for arginine at the N termini of aminoacyl derivatives and peptides. The Km values for Arg-7-amido-4-methylcoumarin (AMC) and Lys-AMC were 15.9 and 26.0 µM, respectively. The nature of the amino acid residue at the C terminus of dipeptides has an effect on hydrolysis rates. The activity was maximal toward dipeptides with Arg, Lys, or Ala as the C-terminal residue. The properties of the purified enzyme, its potential function in the release of arginine, and its further metabolism are discussed because, as a whole, it could constitute a survival mechanism for L. sakei in the meat environment.

* Corresponding author. Mailing address: Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Apartado 73, 46100 Burjasot (Valencia), Spain. Phone: 34 96 3900022. Fax: 34 96 3636301. E-mail: yolsanz{at}iata.csic.es.

Applied and Environmental Microbiology, April 2002, p. 1980-1987, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1980-1987.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Matthews, A., Grimaldi, A., Walker, M., Bartowsky, E., Grbin, P., Jiranek, V. (2004). Lactic Acid Bacteria as a Potential Source of Enzymes for Use in Vinification. Appl. Environ. Microbiol. 70: 5715-5731 [Full Text]  
  • Bolumar, T., Sanz, Y., Aristoy, M.-C., Toldra, F. (2003). Purification and Characterization of a Prolyl Aminopeptidase from Debaryomyces hansenii. Appl. Environ. Microbiol. 69: 227-232 [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»