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 Chen, Y.-S.
Right arrow Articles by Steele, J. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, Y.-S.
Right arrow Articles by Steele, J. L.
Agricola
Right arrow Articles by Chen, Y.-S.
Right arrow Articles by Steele, J. L.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, September 1998, p. 3411-3415, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Genetic Characterization and Physiological Role of Endopeptidase O from Lactobacillus helveticus CNRZ32

Yo-Shen Chen and James L. Steele*

Department of Food Science, University of Wisconsin---Madison, Madison, Wisconsin 53706

Received 6 November 1997/Accepted 11 June 1998

A previously identified insert expressing an endopeptidase from a Lactobacillus helveticus CNRZ32 genomic library was characterized. Nucleotide sequence analysis revealed an open reading frame of 1,941 bp encoding a putative protein of 71.2 kDa which contained a zinc-protease motif. Protein homology searches revealed that this enzyme has 40% similarity with endopeptidase O (PepO) from Lactococcus lactis P8-2-47. Northern hybridization revealed that pepO is monocistronic and is expressed throughout the growth phase. CNRZ32 derivatives lacking PepO activity were constructed via gene replacement. Enzyme assays revealed that the PepO mutant had significantly reduced endopeptidase activity when compared to CNRZ32 with two of the three substrates examined. Growth studies indicated that PepO has no detectable effect on growth rate or acid production by Lactobacillus helveticus CNRZ32 in amino acid defined or skim milk medium.

* Corresponding author. Mailing address: Department of Food Science, University of Wisconsin---Madison, 1605 Linden Dr., Madison, WI 53706-1565. Phone: (608) 262-5960. Fax: (608) 262-6872. E-mail: jlsteele{at}facstaff.wisc.edu.

Applied and Environmental Microbiology, September 1998, p. 3411-3415, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Liu, X., Lagenaur, L. A., Simpson, D. A., Essenmacher, K. P., Frazier-Parker, C. L., Liu, Y., Tsai, D., Rao, S. S., Hamer, D. H., Parks, T. P., Lee, P. P., Xu, Q. (2006). Engineered vaginal lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N.. Antimicrob. Agents Chemother. 50: 3250-3259 [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]  
  • Christensen, J. E., Broadbent, J. R., Steele, J. L. (2003). Hydrolysis of Casein-Derived Peptides {alpha}S1-Casein(f1-9) and {beta}-Casein(f193-209) by Lactobacillus helveticus Peptidase Deletion Mutants Indicates the Presence of a Previously Undetected Endopeptidase. Appl. Environ. Microbiol. 69: 1283-1286 [Abstract] [Full Text]  
  • Tuler, T. R., Callanan, M. J., Klaenhammer, T. R. (2002). Overexpression of Peptidases in Lactococcus and Evaluation of Their Release from Leaky Cells. J DAIRY SCI 85: 2438-2450 [Abstract] [Full Text]  
  • Christensson, C., Bratt, H., Collins, L. J., Coolbear, T., Holland, R., Lubbers, M. W., O'Toole, P. W., Reid, J. R. (2002). Cloning and Expression of an Oligopeptidase, PepO, with Novel Specificity from Lactobacillus rhamnosus HN001 (DR20). Appl. Environ. Microbiol. 68: 254-262 [Abstract] [Full Text]  
  • Dudley, E. G., Steele, J. L. (2001). Lactococcus lactis LM0230 contains a single aminotransferase involved in aspartate biosynthesis, which is essential for growth in milk. Microbiology 147: 215-224 [Abstract] [Full Text]  
  • Atiles, M. W., Dudley, E. G., Steele, J. L. (2000). Gene Cloning, Sequencing, and Inactivation of the Branched-Chain Aminotransferase of Lactococcus lactis LM0230. Appl. Environ. Microbiol. 66: 2325-2329 [Abstract] [Full Text]  
  • Froeliger, E. H., Oetjen, J., Bond, J. P., Fives-Taylor, P. (1999). Streptococcus parasanguis pepO Encodes an Endopeptidase with Structure and Activity Similar to Those of Enzymes That Modulate Peptide Receptor Signaling in Eukaryotic Cells. Infect. Immun. 67: 5206-5214 [Abstract] [Full Text]  
  • Pederson, J. A., Mileski, G. J., Weimer, B. C., Steele, J. L. (1999). Genetic Characterization of a Cell Envelope-Associated Proteinase from Lactobacillus helveticus CNRZ32. J. Bacteriol. 181: 4592-4597 [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»