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. 1983 Sep;155(3):1171–1177. doi: 10.1128/jb.155.3.1171-1177.1983

Purification and characterization of a bacteriocin from Bacteroides fragilis.

T J Hayes, K R Cundy, P B Fernandes, J K Hoober
PMCID: PMC217813  PMID: 6350264

Abstract

A bacteriocin produced by Bacteroides fragilis 1356 was purified from culture medium and characterized. The spectrum of the inhibitory activity of this bacteriocin was species specific. The bacteriocin was recovered from the initial stages of purification as a complex, greater than 2 X 10(7) daltons in mass, containing protein, lipid, and carbohydrate. The dissociation of this complex by 6.0 M guanidine hydrochloride permitted further purification of the bacteriocin by removal of lipid and carbohydrate. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the purified bacteriocin was homogeneous, with a relative molecular weight of 5,000. The activity of the purified bacteriocin was not affected by RNase, DNase, phospholipase A, pancreatic lipase, or dextranase, but was destroyed by trypsin, proteinase K, heat (80 degrees C, 30 min), or a pH below 5 or above 8. Amino acid analysis indicated a predominance of acidic and polar amino acids.

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Selected References

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

  1. BEERENS H., BARON G. MISE EN 'EVIDENCE DE BACT'ERIOCINES 'ELABOR'EES PAR LES BACT'ERIES ANA'EROBIES 'A GRAM N'EGATIF APPARTENANT AU GENRE EGGERTHELLA. Ann Inst Pasteur (Paris) 1965 Feb;108:255–256. [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Beerens H., Baron G., Tahon Mise en evidence di substances inhibitrices elaborées par les bacteries anaerobies non sporulées. Ann Inst Pasteur Lille. 1966;17:1–20. [PubMed] [Google Scholar]
  4. Booth S. J., Johnson J. L., Wilkins T. D. Bacteriocin production by strains of Bacteroides isolated from human feces and the role of these strains in the bacterial ecology of the colon. Antimicrob Agents Chemother. 1977 Apr;11(4):718–724. doi: 10.1128/aac.11.4.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fernandes P. B., Bayer M. E. Membrane-bound enterotoxin of Vibrio cholerae. J Gen Microbiol. 1977 Dec;103(2):381–387. doi: 10.1099/00221287-103-2-381. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hardy K. G. Colicinogeny and related phenomena. Bacteriol Rev. 1975 Dec;39(4):464–515. doi: 10.1128/br.39.4.464-515.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jetten A. M., Vogels G. D. Nature and properties of a Staphylococcus epidermidis bacteriocin. J Bacteriol. 1972 Oct;112(1):243–250. doi: 10.1128/jb.112.1.243-250.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jetten A. M., Vogels G. D., de Windt F. Production and purification of a Staphylococcus epidermidis bacteriocin. J Bacteriol. 1972 Oct;112(1):235–242. doi: 10.1128/jb.112.1.235-242.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Meienhofer J., Maeda H., Glaser C. B., Czombos J., Kuromizu K. Primary structure of neocarzinostatin, an antitumor protein. Science. 1972 Nov 24;178(4063):875–876. doi: 10.1126/science.178.4063.875. [DOI] [PubMed] [Google Scholar]
  12. Mossie K. G., Jones D. T., Robb F. T., Woods D. R. Characterization and mode of action of a bacteriocin produced by a Bacteroides fragilis strain. Antimicrob Agents Chemother. 1979 Dec;16(6):724–730. doi: 10.1128/aac.16.6.724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. SNYDER F., STEPHENS N. A simplified spectrophotometric determination of ester groups in lipids. Biochim Biophys Acta. 1959 Jul;34:244–245. doi: 10.1016/0006-3002(59)90255-0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Tagg J. R., Dajani A. S., Wannamaker L. W., Gray E. D. Group A streptococcal bacteriocin. Production, purification, and mode of action. J Exp Med. 1973 Nov 1;138(5):1168–1183. doi: 10.1084/jem.138.5.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Vargo V., Korzeniowski M., Spaulding E. H. Tryptic soy bile-kanamycin test for the indentification of Bacteroides fragilis. Appl Microbiol. 1974 Mar;27(3):480–483. doi: 10.1128/am.27.3.480-483.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]

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