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. 1992 Jan;58(1):279–284. doi: 10.1128/aem.58.1.279-284.1992

Purification and Partial Characterization of Lacticin 481, a Lanthionine-Containing Bacteriocin Produced by Lactococcus lactis subsp. lactis CNRZ 481

J-C Piard 1,*, P M Muriana 1,†, M J Desmazeaud 1, T R Klaenhammer 1
PMCID: PMC195204  PMID: 16348628

Abstract

Lacticin 481, a bacteriocin produced during the growth of Lactococcus lactis subsp. lactis CNRZ 481, was purified sequentially by ammonium sulfate precipitation, gel filtration, and preparative and analytical reversed-phase high-pressure liquid chromatography. Ammonium sulfate precipitations resulted in a 455-fold increase in total lacticin 481 activity. The entire purification protocol led to a 107, 506-fold increase in the specific activity of lacticin 481. On the basis of its electrophoretic pattern in sodium dodecyl sulfate-polyacrylamide gels, lacticin 481 appeared as a single peptide band of 1.7 kDa. However, dimers of 3.4 kDa also exhibiting lacticin activity were detected. Derivatives of the lacticin-producing strain which did not produce lacticin 481 (Bac-) were sensitive to this bacteriocin (Bacs) and failed to produce the 1.7-kDa band. Amino acid composition analysis of purified lacticin 481 revealed the presence of lanthionine residues, suggesting that lacticin 481 is a member of the lantibiotic family of antimicrobial peptides. Seven residues (K G G S G V I) were sequenced from the N-terminal portion of lacticin 481, and these did not shown any homology with nisin or other known bacteriocin sequences.

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Selected References

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  1. Allgaier H., Jung G., Werner R. G., Schneider U., Zähner H. Epidermin: sequencing of a heterodetic tetracyclic 21-peptide amide antibiotic. Eur J Biochem. 1986 Oct 1;160(1):9–22. doi: 10.1111/j.1432-1033.1986.tb09933.x. [DOI] [PubMed] [Google Scholar]
  2. BERRIDGE N. J., NEWTON G. G. F., ABRAHAM E. P. Purification and nature of the antibiotic nisin. Biochem J. 1952 Dec;52(4):529–535. doi: 10.1042/bj0520529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailey F. J., Hurst A. Preparation of a highly active form of nisin from Streptococcus lactis. Can J Microbiol. 1971 Jan;17(1):61–67. doi: 10.1139/m71-010. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. CHEESEMAN G. C., BERRIDGE N. J. An improved method of preparing nisin. Biochem J. 1957 Mar;65(3):603–608. doi: 10.1042/bj0650603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davey G. P., Richardson B. C. Purification and Some Properties of Diplococcin from Streptococcus cremoris 346. Appl Environ Microbiol. 1981 Jan;41(1):84–89. doi: 10.1128/aem.41.1.84-89.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  8. Geis A., Singh J., Teuber M. Potential of lactic streptococci to produce bacteriocin. Appl Environ Microbiol. 1983 Jan;45(1):205–211. doi: 10.1128/aem.45.1.205-211.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Holo H., Nilssen O., Nes I. F. Lactococcin A, a new bacteriocin from Lactococcus lactis subsp. cremoris: isolation and characterization of the protein and its gene. J Bacteriol. 1991 Jun;173(12):3879–3887. doi: 10.1128/jb.173.12.3879-3887.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kaletta C., Entian K. D. Nisin, a peptide antibiotic: cloning and sequencing of the nisA gene and posttranslational processing of its peptide product. J Bacteriol. 1989 Mar;171(3):1597–1601. doi: 10.1128/jb.171.3.1597-1601.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Kozak W., Bardowski J., Dobrzański W. T. Lactostrepcins--acid bacteriocins produced by lactic streptococci. J Dairy Res. 1978 Jun;45(2):247–257. doi: 10.1017/s0022029900016423. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lindgren S. E., Dobrogosz W. J. Antagonistic activities of lactic acid bacteria in food and feed fermentations. FEMS Microbiol Rev. 1990 Sep;7(1-2):149–163. doi: 10.1111/j.1574-6968.1990.tb04885.x. [DOI] [PubMed] [Google Scholar]
  17. Liu T. Y., Boykins R. A. Hydrolysis of proteins and peptides in a hermetically sealed microcapillary tube: high recovery of labile amino acids. Anal Biochem. 1989 Nov 1;182(2):383–387. doi: 10.1016/0003-2697(89)90612-x. [DOI] [PubMed] [Google Scholar]
  18. Liu W., Hansen J. N. Some chemical and physical properties of nisin, a small-protein antibiotic produced by Lactococcus lactis. Appl Environ Microbiol. 1990 Aug;56(8):2551–2558. doi: 10.1128/aem.56.8.2551-2558.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Muriana P. M., Klaenhammer T. R. Purification and partial characterization of lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088. Appl Environ Microbiol. 1991 Jan;57(1):114–121. doi: 10.1128/aem.57.1.114-121.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mørtvedt C. I., Nissen-Meyer J., Sletten K., Nes I. F. Purification and amino acid sequence of lactocin S, a bacteriocin produced by Lactobacillus sake L45. Appl Environ Microbiol. 1991 Jun;57(6):1829–1834. doi: 10.1128/aem.57.6.1829-1834.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. van Belkum M. J., Hayema B. J., Jeeninga R. E., Kok J., Venema G. Organization and nucleotide sequences of two lactococcal bacteriocin operons. Appl Environ Microbiol. 1991 Feb;57(2):492–498. doi: 10.1128/aem.57.2.492-498.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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