This Article Full Text 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 Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Franz, C. M. A. P. Articles by Stiles, M. E. Search for Related Content PubMed PubMed Citation Articles by Franz, C. M. A. P. Articles by Stiles, M. E. Agricola Articles by Franz, C. M. A. P. Articles by Stiles, M. E.

Applied and Environmental Microbiology, May 1999, p. 2170-2178, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Atypical Genetic Locus Associated with Constitutive Production of Enterocin B by Enterococcus faecium BFE 900

Charles M. A. P. Franz,¹ Randy W. Worobo,^1, Luis E. N. Quadri,^1, Ulrich Schillinger,² Wilhelm H. Holzapfel,² John C. Vederas,³ and Michael E. Stiles^1,*

Departments of Agricultural, Food and Nutritional Science¹ and Chemistry,³ University of Alberta, Edmonton, Alberta T6G 2P5, Canada, and Federal Research Center for Nutrition, Institute of Hygiene and Toxicology, D-76131 Karlsruhe, Germany²

Received 5 November 1998/Accepted 9 March 1999

A purified bacteriocin produced by Enterococcus faecium BFE 900 isolated from black olives was shown by Edman degradation and mass spectrometric analyses to be identical to enterocin B produced by E. faecium T136 from meat (P. Casaus, T. Nilsen, L. M. Cintas, I. F. Nes, P. E. Hernández, and H. Holo, Microbiology 143:2287-2294, 1997). The structural gene was located on a 2.2-kb HindIII fragment and a 12.0-kb EcoRI chromosomal fragment. The genetic characteristics and production of EntB by E. faecium BFE 900 differed from that described so far by the presence of a conserved sequence like a regulatory box upstream of the EntB gene, and its production was constitutive and not regulated. The 2.2-kb chromosomal fragment contained the hitherto undetected immunity gene for EntB in an atypical orientation that is the reverse of that of the structural gene. Typical transport and other genes associated with bacteriocin production were not detected on the 12.0-kb chromosomal fragment containing the EntB structural gene. This makes the EntB genetic system different from most other bacteriocin systems, where transport and possible regulatory genes are clustered. EntB was subcloned and expressed by the dedicated secretion machinery of Carnobacterium piscicola LV17A. The structural gene was amplified by PCR, fused to the divergicin A signal peptide, and expressed by the general secretory pathway in Enterococcus faecalis ATCC 19433.

---

^* Corresponding author. Mailing address: Department of Agricultural, Food and Nutritional Science, 4-10 Agriculture Forestry Centre, University of Alberta, Edmonton, Alberta T6G 2P5, Canada. Phone: (780) 492-2386. Fax: (780) 492-8914. E-mail: mstiles{at}afns.ualberta.ca.

Present address: Cornell University, Department of Food Science and Technology, New York State Agricultural Experiment Station, Geneva, NY 14456-0462.

Present address: Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Boston, MA 02146.

---

Applied and Environmental Microbiology, May 1999, p. 2170-2178, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• Nes, I. F., Diep, D. B., Holo, H. (2007). Bacteriocin Diversity in Streptococcus and Enterococcus. J. Bacteriol. 189: 1189-1198 [Full Text]  
• Criado, R., Diep, D. B., Aakra, A., Gutierrez, J., Nes, I. F., Hernandez, P. E., Cintas, L. M. (2006). Complete Sequence of the Enterocin Q-Encoding Plasmid pCIZ2 from the Multiple Bacteriocin Producer Enterococcus faecium L50 and Genetic Characterization of Enterocin Q Production and Immunity.. Appl. Environ. Microbiol. 72: 6653-6666 [Abstract] [Full Text]  
• van Belkum, M. J., Stiles, M. E. (2006). Characterization of the theta-type plasmid pCD3.4 from Carnobacterium divergens, and modulation of its host range by RepA mutation. Microbiology 152: 171-178 [Abstract] [Full Text]  
• Abriouel, H., Herrmann, A., Starke, J., Yousif, N. M. K., Wijaya, A., Tauscher, B., Holzapfel, W., Franz, C. M. A. P. (2004). Cloning and Heterologous Expression of Hematin-Dependent Catalase Produced by Lactobacillus plantarum CNRZ 1228. Appl. Environ. Microbiol. 70: 603-606 [Abstract] [Full Text]  
• Vaughan, A., Eijsink, V. G. H., van Sinderen, D. (2003). Functional Characterization of a Composite Bacteriocin Locus from Malt Isolate Lactobacillus sakei 5. Appl. Environ. Microbiol. 69: 7194-7203 [Abstract] [Full Text]  
• Cote, C. K., Honeyman, A. L. (2003). The LicT protein acts as both a positive and a negative regulator of loci within the bgl regulon of Streptococcus mutans. Microbiology 149: 1333-1340 [Abstract] [Full Text]  
• Franz, C. M. A. P., Grube, A., Herrmann, A., Abriouel, H., Starke, J., Lombardi, A., Tauscher, B., Holzapfel, W. H. (2002). Biochemical and Genetic Characterization of the Two-Peptide Bacteriocin Enterocin 1071 Produced by Enterococcus faecalis FAIR-E 309. Appl. Environ. Microbiol. 68: 2550-2554 [Abstract] [Full Text]  
• Cintas, L. M., Casaus, P., Herranz, C., Håvarstein, L. S., Holo, H., Hernández, P. E., Nes, I. F. (2000). Biochemical and Genetic Evidence that Enterococcus faecium L50 Produces Enterocins L50A and L50B, the sec-Dependent Enterocin P, and a Novel Bacteriocin Secreted without an N-Terminal Extension Termed Enterocin Q. J. Bacteriol. 182: 6806-6814 [Abstract] [Full Text]  
• Franz, C. M. A. P., van Belkum, M. J., Worobo, R. W., Vederas, J. C., Stiles, M. E. (2000). Characterization of the genetic locus responsible for production and immunity of carnobacteriocin A: the immunity gene confers cross-protection to enterocin B. Microbiology 146: 621-631 [Abstract] [Full Text]  
• Rose, N. L., Sporns, P., McMullen, L. M. (1999). Detection of Bacteriocins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. Appl. Environ. Microbiol. 65: 2238-2242 [Abstract] [Full Text]