Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Nov;63(11):4185–4190. doi: 10.1128/aem.63.11.4185-4190.1997

Purification and characterization of staphylococcin BacR1, a broad-spectrum bacteriocin.

S S Crupper 1, A J Gies 1, J J Iandolo 1
PMCID: PMC168735  PMID: 9361402

Abstract

The bacteriocin BacR1 was purified from culture supernatant of Staphylococcus aureus UT0007 by sequential ammonium sulfate precipitation, cation-exchange chromatography, and C4 reverse-phase chromatography steps. Mass spectrographic analysis indicated that the purified peptide has a molecular mass of 3,338 Da. It is resistant to environmental conditions, retaining full biological activity after exposure to pH extremes (pHs 3 to 11), heating at 95 degrees C for 15 min, and exposure to strong chaotropic agents. BacR1 was destroyed with a complete loss of biological activity after digestion with trypsin and proteinase K. Amino acid sequence analysis revealed a high concentration of Asx, Gly, and Pro residues and a high proportion of hydrophobic amino acids. The peptide is bactericidal and kills in a dose-dependent manner, but it does not lyse log-phase cells of Corynebacterium renale, the routine indicator organism for bacteriocin assay. A specific receptor for binding was detected on sensitive cells but not on insensitive cells. Competition assays showed that UV-inactivated cells could protect susceptible cells from antibacterial action. A partial inhibitory spectrum revealed that organisms from the following genera are susceptible: Staphylococcus, Streptococcus, Corynebacterium, Haemophilus, Bordetella, Moraxella, Pasteurella, Neisseria, and Bacillus.

Full Text

The Full Text of this article is available as a PDF (527.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Augustin J., Rosenstein R., Wieland B., Schneider U., Schnell N., Engelke G., Entian K. D., Götz F. Genetic analysis of epidermin biosynthetic genes and epidermin-negative mutants of Staphylococcus epidermidis. Eur J Biochem. 1992 Mar 15;204(3):1149–1154. doi: 10.1111/j.1432-1033.1992.tb16740.x. [DOI] [PubMed] [Google Scholar]
  2. Crupper S. S., Iandolo J. J. Purification and partial characterization of a novel antibacterial agent (Bac1829) Produced by Staphylococcus aureus KSI1829. Appl Environ Microbiol. 1996 Sep;62(9):3171–3175. doi: 10.1128/aem.62.9.3171-3175.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dajani A. S., Gray E. D., Wannamaker L. W. Effect of Bactericidal Substance from Staphylococcus aureus on Group A Streptococci I. Biochemical Alterations. Infect Immun. 1970 May;1(5):485–490. doi: 10.1128/iai.1.5.485-490.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Engelke G., Gutowski-Eckel Z., Hammelmann M., Entian K. D. Biosynthesis of the lantibiotic nisin: genomic organization and membrane localization of the NisB protein. Appl Environ Microbiol. 1992 Nov;58(11):3730–3743. doi: 10.1128/aem.58.11.3730-3743.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Engelke G., Gutowski-Eckel Z., Kiesau P., Siegers K., Hammelmann M., Entian K. D. Regulation of nisin biosynthesis and immunity in Lactococcus lactis 6F3. Appl Environ Microbiol. 1994 Mar;60(3):814–825. doi: 10.1128/aem.60.3.814-825.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. González B., Arca P., Mayo B., Suárez J. E. Detection, purification, and partial characterization of plantaricin C, a bacteriocin produced by a Lactobacillus plantarum strain of dairy origin. Appl Environ Microbiol. 1994 Jun;60(6):2158–2163. doi: 10.1128/aem.60.6.2158-2163.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hale E. M., Hinsdill R. D. Characterization of a bacteriocin from Staphylococcus aureus strain 462. Antimicrob Agents Chemother. 1973 Dec;4(6):634–640. doi: 10.1128/aac.4.6.634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hughes D. E., Pugh G. W., Jr A five-year study of infectious bovine keratoconjunctivitis in a beef herd. J Am Vet Med Assoc. 1970 Aug 15;157(4):443–451. [PubMed] [Google Scholar]
  9. Jack R. W., Tagg J. R., Ray B. Bacteriocins of gram-positive bacteria. Microbiol Rev. 1995 Jun;59(2):171–200. doi: 10.1128/mr.59.2.171-200.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kader O. A., Sahl H. G., Brandis H. Isolation and mode of action of a staphylococcin-like substance active against gram-positive and gram-negative bacteria. J Gen Microbiol. 1984 Sep;130(9):2291–2300. doi: 10.1099/00221287-130-9-2291. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Melish M. E., Glasgow L. A. Staphylococcal scalded skin syndrome: the expanded clinical syndrome. J Pediatr. 1971 Jun;78(6):958–967. doi: 10.1016/s0022-3476(71)80425-0. [DOI] [PubMed] [Google Scholar]
  13. Morriss D. M., Lawson J. W., Rogolsky M. Effect of a staphylococcin on Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1978 Aug;14(2):218–223. doi: 10.1128/aac.14.2.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakamura T., Yamazaki N., Taniguchi H., Fujimura S. Production, purification, and properties of a bacteriocin from Staphylococcus aureus isolated from saliva. Infect Immun. 1983 Feb;39(2):609–614. doi: 10.1128/iai.39.2.609-614.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Peschel A., Augustin J., Kupke T., Stevanovic S., Götz F. Regulation of epidermin biosynthetic genes by EpiQ. Mol Microbiol. 1993 Jul;9(1):31–39. doi: 10.1111/j.1365-2958.1993.tb01666.x. [DOI] [PubMed] [Google Scholar]
  16. Rogolsky M., Wiley B. B. Production and properties of a staphylococcin genetically controlled by the staphylococcal plasmid for exfoliative toxin synthesis. Infect Immun. 1977 Mar;15(3):726–732. doi: 10.1128/iai.15.3.726-732.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tagg J. R., Dajani A. S., Wannamaker L. W. Bacteriocins of gram-positive bacteria. Bacteriol Rev. 1976 Sep;40(3):722–756. doi: 10.1128/br.40.3.722-756.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tagg J. R., McGiven A. R. Assay system for bacteriocins. Appl Microbiol. 1971 May;21(5):943–943. doi: 10.1128/am.21.5.943-943.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Warren R. L. Exfoliative toxin plasmids of bacteriophage group 2 Staphylococcus aureus: sequence homology. Infect Immun. 1980 Nov;30(2):601–606. doi: 10.1128/iai.30.2.601-606.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES