Purification and characterization of an arginine aminopeptidase from Lactobacillus sakei.

Applied and Environmental Microbiology
Yolanda Sanz, Fidel Toldrá

Abstract

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 degrees C. The enzyme was inhibited by sulfhydryl group reagents and several divalent cations (Cu(2+), Hg(2+), and Zn(2+)) 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 K(m) values for Arg-7-amido-4-methylcoumarin (AMC) and Lys-AMC were 15.9 and 26.0 microM, 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...Continue Reading

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Citations

Aug 2, 2003·International Journal of Food Microbiology·Tomás BolumarFidel Toldrá
Jan 7, 2003·Applied and Environmental Microbiology·Tomás BolumarFidel Toldrá
Oct 7, 2004·Applied and Environmental Microbiology·Angela MatthewsVladimir Jiranek
Aug 23, 2003·Infection and Immunity·Araceli Contreras-RodriguezAhide Lopez-Merino
Mar 1, 2011·Prikladnaia biokhimiia i mikrobiologiia·U Jankiewicz, A Wnuk
Dec 29, 2011·Food Microbiology·Quirin Sinz, Wilfried Schwab
Feb 22, 2005·Oral Microbiology and Immunology·K OhishiT Watanabe
Aug 21, 2010·Journal of Food Science·Antonio TraniAldo Di Luccia
Sep 12, 2012·Journal of Applied Microbiology·P CastellanoF Toldrá
Feb 1, 2005·International Journal of Food Microbiology·Tomás BolumarFidel Toldrá
Dec 29, 2005·Protein Expression and Purification·Frank C GolichMichael W Crowder
Nov 12, 2014·Journal of Basic Microbiology·Hui-qing XuYu Chen
May 7, 2015·International Journal of Food Microbiology·Leticia MoraFidel Toldrá
Jul 3, 2021·Animals : an Open Access Journal From MDPI·Maria J XavierSofia Engrola

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