Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1991 Jan;57(1):114–121. doi: 10.1128/aem.57.1.114-121.1991

Purification and partial characterization of lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088.

P M Muriana 1, T R Klaenhammer 1
PMCID: PMC182671  PMID: 1903624

Abstract

Lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088 (NCK88), was purified and characterized. Lactacin F is heat stable, proteinaceous, and inhibitory to other lactobacilli as well as Enterococcus faecalis. The bacteriocin was isolated as a floating pellet from culture supernatants brought to 35 to 40% saturation with ammonium sulfate. Native lactacin F was sized at approximately 180 kDa by gel filtration. Column fractions having lactacin F activity were examined by electron microscopy and contained micelle-like globular particles. Purification by ammonium sulfate precipitation, gel filtration, and high-performance liquid chromatography resulted in a 474-fold increase in specific activity of lactacin F. The purified bacteriocin was identified as a 2.5-kDa peptide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The lactacin F peptide retained activity after extraction from SDS-PAGE gel slices, confirming the identity of the 2.5-kDa peptide. Variants of NCK88 that failed to exhibit lactacin F activity did not produce the 2.5-kDa band. Sequence analysis of purified lactacin F identified 25 N-terminal amino acids containing an arginine residue at the N terminus. Composition analysis indicates that lactacin F may contain as many as 56 amino acid residues.

Full text

PDF
114

Images in this article

117Image
on p.117
118Image
on p.118
119Image
on p.119

Selected References

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

  1. Austin-Prather S. L., Booth S. J. Evidence for a membrane-bound form of a bacteriocin of Bacteroides uniformis Tl-1. Can J Microbiol. 1984 Feb;30(2):268–272. doi: 10.1139/m84-040. [DOI] [PubMed] [Google Scholar]
  2. Barefoot S. F., Klaenhammer T. R. Detection and activity of lactacin B, a bacteriocin produced by Lactobacillus acidophilus. Appl Environ Microbiol. 1983 Jun;45(6):1808–1815. doi: 10.1128/aem.45.6.1808-1815.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barefoot S. F., Klaenhammer T. R. Purification and characterization of the Lactobacillus acidophilus bacteriocin lactacin B. Antimicrob Agents Chemother. 1984 Sep;26(3):328–334. doi: 10.1128/aac.26.3.328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Deh M. E., Dzandu J. K., Wise G. E. Sialoglycoproteins with a high amount of O-glycosidically linked carbohydrate moieties stain yellow with silver in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1985 Oct;150(1):166–173. doi: 10.1016/0003-2697(85)90456-7. [DOI] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dzandu J. K., Deh M. E., Barratt D. L., Wise G. E. Detection of erythrocyte membrane proteins, sialoglycoproteins, and lipids in the same polyacrylamide gel using a double-staining technique. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1733–1737. doi: 10.1073/pnas.81.6.1733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giulian G. G., Moss R. L., Greaser M. Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels. Anal Biochem. 1983 Mar;129(2):277–287. doi: 10.1016/0003-2697(83)90551-1. [DOI] [PubMed] [Google Scholar]
  9. Gross E., Kiltz H. H. The number and nature of , -unsaturated amino acids in subtilin. Biochem Biophys Res Commun. 1973 Jan 23;50(2):559–565. doi: 10.1016/0006-291x(73)90876-0. [DOI] [PubMed] [Google Scholar]
  10. Gross E., Morell J. L. The structure of nisin. J Am Chem Soc. 1971 Sep 8;93(18):4634–4635. doi: 10.1021/ja00747a073. [DOI] [PubMed] [Google Scholar]
  11. Joerger M. C., Klaenhammer T. R. Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481. J Bacteriol. 1986 Aug;167(2):439–446. doi: 10.1128/jb.167.2.439-446.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kaletta C., Entian K. D., Kellner R., Jung G., Reis M., Sahl H. G. Pep5, a new lantibiotic: structural gene isolation and prepeptide sequence. Arch Microbiol. 1989;152(1):16–19. doi: 10.1007/BF00447005. [DOI] [PubMed] [Google Scholar]
  13. Katsu T., Kuroko M., Morikawa T., Sanchika K., Fujita Y., Yamamura H., Uda M. Mechanism of membrane damage induced by the amphipathic peptides gramicidin S and melittin. Biochim Biophys Acta. 1989 Aug 7;983(2):135–141. doi: 10.1016/0005-2736(89)90226-5. [DOI] [PubMed] [Google Scholar]
  14. Kellner R., Jung G., Hörner T., Zähner H., Schnell N., Entian K. D., Götz F. Gallidermin: a new lanthionine-containing polypeptide antibiotic. Eur J Biochem. 1988 Oct 15;177(1):53–59. doi: 10.1111/j.1432-1033.1988.tb14344.x. [DOI] [PubMed] [Google Scholar]
  15. Klaenhammer T. R. Bacteriocins of lactic acid bacteria. Biochimie. 1988 Mar;70(3):337–349. doi: 10.1016/0300-9084(88)90206-4. [DOI] [PubMed] [Google Scholar]
  16. Kordel M., Schüller F., Sahl H. G. Interaction of the pore forming-peptide antibiotics Pep 5, nisin and subtilin with non-energized liposomes. FEBS Lett. 1989 Feb 13;244(1):99–102. doi: 10.1016/0014-5793(89)81171-8. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Muriana P. M., Klaenhammer T. R. Conjugal Transfer of Plasmid-Encoded Determinants for Bacteriocin Production and Immunity in Lactobacillus acidophilus 88. Appl Environ Microbiol. 1987 Mar;53(3):553–560. doi: 10.1128/aem.53.3.553-560.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schnell N., Entian K. D., Schneider U., Götz F., Zähner H., Kellner R., Jung G. Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulphide-rings. Nature. 1988 May 19;333(6170):276–278. doi: 10.1038/333276a0. [DOI] [PubMed] [Google Scholar]
  21. Schüller F., Benz R., Sahl H. G. The peptide antibiotic subtilin acts by formation of voltage-dependent multi-state pores in bacterial and artificial membranes. Eur J Biochem. 1989 Jun 1;182(1):181–186. doi: 10.1111/j.1432-1033.1989.tb14815.x. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Upreti G. C., Hinsdill R. D. Production and mode of action of lactocin 27: bacteriocin from a homofermentative Lactobacillus. Antimicrob Agents Chemother. 1975 Feb;7(2):139–145. doi: 10.1128/aac.7.2.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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 Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES