Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1972 Oct;112(1):235–242. doi: 10.1128/jb.112.1.235-242.1972

Production and Purification of a Staphylococcus epidermidis Bacteriocin

A M Jetten 1, G D Vogels 1, F de Windt 1
PMCID: PMC251403  PMID: 5079063

Abstract

Liquid cultures of Staphylococcus epidermidis 1580 contained rather small amounts of a bacteriocin, staphylococcin 1580, which was found both in the supernatant fluid and in the cell pellet. It could be extracted from the cells with 5% NaCl solution. The staphylococcin production could not be induced by ultraviolet irradiation or treatment with mitomycin C. Bacteria grown on semisolid medium produced a much larger amount of the compound with a high specific activity. The staphylococcin was purified by ammonium sulfate precipitation, ultracentrifugation, and chromatography on Sephadex columns. The purified material was homogeneous on polyacrylamide gel electrophoresis. The molecular weight was between 150,000 and 400,000. The bacteriocin was composed of protein, carbohydrate, and lipid and consisted of subunits exhibiting a molecular weight of about 20,000.

Full text

PDF
235

Images in this article

Selected References

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

  1. BARROW G. I. Microbial antagonism by Staphylococcus aureus. J Gen Microbiol. 1963 Jun;31:471–481. doi: 10.1099/00221287-31-3-471. [DOI] [PubMed] [Google Scholar]
  2. BARROW G. I. THE NATURE OF INHIBITORY ACTIVITY BY STAPHYLOCOCCUS AUREUS TYPE 71. J Gen Microbiol. 1963 Aug;32:255–261. doi: 10.1099/00221287-32-2-255. [DOI] [PubMed] [Google Scholar]
  3. Bazaral M., Helinski D. R. Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J Mol Biol. 1968 Sep 14;36(2):185–194. doi: 10.1016/0022-2836(68)90374-4. [DOI] [PubMed] [Google Scholar]
  4. Dajani A. S., Gray E. D., Wannamaker L. W. Bactericidal substance from Staphylococcus aureus. Biological properties. J Exp Med. 1970 May 1;131(5):1004–1015. doi: 10.1084/jem.131.5.1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Dajani A. S., Wannamaker L. W. Demonstration of a bactericidal substance against beta-hemolytic streptococci in supernatant fluids of staphylococcal cultures. J Bacteriol. 1969 Mar;97(3):985–991. doi: 10.1128/jb.97.3.985-991.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dandeu J. P. Chemical and immunological study of colicins e(1), k, a, and q. Infect Immun. 1971 Jan;3(1):1–9. doi: 10.1128/iai.3.1.1-9.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dobardzic R., Payment P., Sonea S. Ultrastructure du bactériophage øRE agissant dans certaines conditions comme une staphylococcine. Can J Microbiol. 1971 Jul;17(7):847–849. [PubMed] [Google Scholar]
  9. Gagliano V. J., Hinsdill R. D. Characterization of a Staphylococcus aureus bacteriocin. J Bacteriol. 1970 Oct;104(1):117–125. doi: 10.1128/jb.104.1.117-125.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HAMON Y., PERON Y. QUELQUES REMARQUES SUR LES BACT'ERIOCINES PRODUITES PAR LES MICROBES GRAM-POSITIFS. C R Hebd Seances Acad Sci. 1963 Jul 29;257:1191–1193. [PubMed] [Google Scholar]
  11. HOLLAND I. B. The purification and properties of megacin, a bacteriocin from Bacillus megaterium. Biochem J. 1961 Mar;78:641–648. doi: 10.1042/bj0780641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HUTTON J. J., GOEBEL W. F. Colicine V. Proc Natl Acad Sci U S A. 1961 Sep 15;47:1498–1500. doi: 10.1073/pnas.47.9.1498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hamon Y., Péron Y. Sur la nature des bactériocines produites par Listeria monocytogenes. C R Acad Sci Hebd Seances Acad Sci D. 1966 Jul 11;263(2):198–200. [PubMed] [Google Scholar]
  14. Herschman H. R., Helinski D. R. Purification and characterization of colicin E2 and colicin E3. J Biol Chem. 1967 Nov 25;242(22):5360–5368. [PubMed] [Google Scholar]
  15. Higerd T. B., Baechler C. A., Berk R. S. Morphological studies on relaxed and contracted forms of purified pyocin particles. J Bacteriol. 1969 Jun;98(3):1378–1389. doi: 10.1128/jb.98.3.1378-1389.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hill C., Holland I. B. Genetic basis of colicin E susceptibility in Escherichia coli. I. Isolation and properties of refractory mutants and the preliminary mapping of their mutations. J Bacteriol. 1967 Sep;94(3):677–686. doi: 10.1128/jb.94.3.677-686.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hsu C., Wiseman G. M. Antibacterial substances from staphylococci. Can J Microbiol. 1967 Aug;13(8):947–955. doi: 10.1139/m67-127. [DOI] [PubMed] [Google Scholar]
  18. Ivanovics G. BACTERIOCINS AND BACTERIOCIN-LIKE SUBSTANCES. Bacteriol Rev. 1962 Jun;26(2 Pt 1):108–118. [PMC free article] [PubMed] [Google Scholar]
  19. Keene J. H. Preparation and chemical properties of colicine I. Can J Microbiol. 1966 Apr;12(2):425–427. doi: 10.1139/m66-061. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Lachowicz T., Walczak Z. Purification and properties of staphylococcin A. Arch Immunol Ther Exp (Warsz) 1968;16(6):855–863. [PubMed] [Google Scholar]
  22. Mennigmann H. D. Electron microscopy of the anti-bacterial agent produced by Escherichia coli 15. J Gen Microbiol. 1965 Nov;41(2):151–154. doi: 10.1099/00221287-41-2-151. [DOI] [PubMed] [Google Scholar]
  23. NOMURA M. MECHANISM OF ACTION OF COLICINES. Proc Natl Acad Sci U S A. 1964 Dec;52:1514–1521. doi: 10.1073/pnas.52.6.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nomura M. Colicins and related bacteriocins. Annu Rev Microbiol. 1967;21:257–284. doi: 10.1146/annurev.mi.21.100167.001353. [DOI] [PubMed] [Google Scholar]
  25. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  26. PARKER M. T., SIMMONS L. E. The inhibition of Corynebacterium diphtheriae and other gram-positive organisms by Staphylococcus aureus. J Gen Microbiol. 1959 Oct;21:457–476. doi: 10.1099/00221287-21-2-457. [DOI] [PubMed] [Google Scholar]
  27. REEVES P. THE BACTERIOCINS. Bacteriol Rev. 1965 Mar;29:24–45. doi: 10.1128/br.29.1.24-45.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Richmond M. H. The differential effect of arginine and canavanine on growth and enzyme formation in Staphylococcus aureus 524 SC. Biochem J. 1959 Sep;73(1):155–167. doi: 10.1042/bj0730155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tieze G. A., Stouthamer A. H., Jansz H. S., Zandberg J., van Bruggen E. F. A bacteriocinogenic factor of Enterobacter cloacae. Mol Gen Genet. 1969;106(1):48–65. [PubMed] [Google Scholar]
  30. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  31. de Graaf F. K., Goedvolk-de Groot L. E., Stouthamer A. H. Purification of a bacteriocin produced by Enterobacter cloacae DF 13. Biochim Biophys Acta. 1970 Dec 22;221(3):566–575. doi: 10.1016/0005-2795(70)90228-x. [DOI] [PubMed] [Google Scholar]
  32. de Klerk H. C., Hugo N. Phage-like structures from Lactobacillus acidophilus. J Gen Virol. 1970 Sep;8(3):231–234. doi: 10.1099/0022-1317-8-3-231. [DOI] [PubMed] [Google Scholar]
  33. de Klerk H. C., Smit J. A. Properties of a Lactobacillus fermenti bacteriocin. J Gen Microbiol. 1967 Aug;48(2):309–316. doi: 10.1099/00221287-48-2-309. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES