This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fenster, K. M. Articles by Steele, J. L. Search for Related Content PubMed PubMed Citation Articles by Fenster, K. M. Articles by Steele, J. L.

 Previous Article  |  Next Article 

J. Bacteriol., 04 1997, 2529-2533, Vol 179, No. 8
Copyright © 1997, American Society for Microbiology

Characterization of a thiol-dependent endopeptidase from Lactobacillus helveticus CNRZ32

KM Fenster, KL Parkin and JL Steele
Department of Food Science, University of Wisconsin-Madison 53706, USA.

An endopeptidase gene (pepE) was isolated from a previously constructed genomic library of Lactobacillus helveticus CNRZ32. The pepE gene consisted of a 1,314-bp open reading frame encoding a putative peptide of 52.1 kDa. Significant identity was found between the deduced amino acid sequence of pepE and the sequences for aminopeptidase C from Lactobacillus delbrueckii subsp. lactis DSM7290, L. helveticus CNRZ32, Streptococcus thermophilus CNRZ302, and Lactococcus lactis subsp. cremoris AM2. A recombinant PepE fusion protein containing an N- terminal six-histidine tag was constructed and purified to electrophoretic homogeneity. Characterization of PepE revealed that it was a thiol-dependent protease having a monomeric mass of 50 kDa, with optimum temperature, NaCl concentration, and pH for activity at 32 to 37 degrees C, 0.5%, and 4.5, respectively. PepE had significant activity under conditions which simulate those of ripening cheese (10 degrees C, 4% NaCl, pH 5.1). PepE hydrolyzed internal peptide bonds in Met-enkephalin and bradykinin; however, hydrolysis of alpha-, beta-, and kappa-caseins was not detected.

This article has been cited by other articles:

• Scolari, G., Vescovo, M., Zacconi, C., Vescovi, F. (2006). Extraction and Partial Characterization of Proteolytic Activities from the Cell Surface of Lactobacillus helveticus Zuc2.. J DAIRY SCI 89: 3800-3809 [Abstract] [Full Text]  
• Janer, C., Arigoni, F., Lee, B. H., Pelaez, C., Requena, T. (2005). Enzymatic Ability of Bifidobacterium animalis subsp. lactis To Hydrolyze Milk Proteins: Identification and Characterization of Endopeptidase O. Appl. Environ. Microbiol. 71: 8460-8465 [Abstract] [Full Text]  
• Sridhar, V. R., Hughes, J. E., Welker, D. L., Broadbent, J. R., Steele, J. L. (2005). Identification of Endopeptidase Genes from the Genomic Sequence of Lactobacillus helveticus CNRZ32 and the Role of These Genes in Hydrolysis of Model Bitter Peptides. Appl. Environ. Microbiol. 71: 3025-3032 [Abstract] [Full Text]  
• Chen, Y.-S., Christensen, J. E., Broadbent, J. R., Steele, J. L. (2003). Identification and Characterization of Lactobacillus helveticus PepO2, an Endopeptidase with Post-Proline Specificity. Appl. Environ. Microbiol. 69: 1276-1282 [Abstract] [Full Text]  
• Deutsch, S.-M., Molle, D., Gagnaire, V., Piot, M., Atlan, D., Lortal, S. (2000). Hydrolysis of Sequenced beta -Casein Peptides Provides New Insight into Peptidase Activity from Thermophilic Lactic Acid Bacteria and Highlights Intrinsic Resistance of Phosphopeptides. Appl. Environ. Microbiol. 66: 5360-5367 [Abstract] [Full Text]  
• Chen, Y.-S., Steele, J. L. (1998). Genetic Characterization and Physiological Role of Endopeptidase O from Lactobacillus helveticus CNRZ32. Appl. Environ. Microbiol. 64: 3411-3415 [Abstract] [Full Text]