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. 1993 Jul;59(7):2014–2021. doi: 10.1128/aem.59.7.2014-2021.1993

Isolation and characterization of the lantibiotic salivaricin A and its structural gene salA from Streptococcus salivarius 20P3.

K F Ross 1, C W Ronson 1, J R Tagg 1
PMCID: PMC182229  PMID: 8357242

Abstract

A bacteriocin-like inhibitory substance, salivaricin A, was purified from cultures of Streptococcus salivarius 20P3 and was shown by ion spray mass spectrometry to have a molecular mass of 2,315 +/- 1.1 Da. Amino acid composition analysis demonstrated the presence of lanthionine, indicating that salivaricin A may be a member of the lantibiotic class of antibiotic substances. The sequence of eight amino acids at the N terminus of the molecule was determined by Edman degradation, and mixed oligonucleotide probes based on part of this sequence (GSGWIA) were used to detect the salivaricin A structural gene. A 6.2-kb EcoRI fragment of chromosomal DNA from strain 20P3 that hybridized with the probes was cloned, and the hybridizing region was further localized to a 379-bp DraI-AluI fragment. Analysis of the nucleotide sequence of this fragment indicated that salivaricin A is synthesized as a 51-amino-acid prepeptide that is posttranslationally modified and cleaved to give a biologically active 22-residue peptide containing one lanthionine and two beta-methyllanthionine residues. The secondary structure of presalivaricin A was predicted to be similar to that of type A lantibiotics, with a hydrophilic alpha-helical leader sequence and a propeptide region with potential for beta-turn formation and a lack of alpha-helicity. The sequence around the cleavage site of presalivaricin A differed from that of other type A lantibiotics but was similar to that of several bacteriocin-like inhibitory substances produced by lactic acid bacteria.

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Selected References

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  1. Banerjee S., Hansen J. N. Structure and expression of a gene encoding the precursor of subtilin, a small protein antibiotic. J Biol Chem. 1988 Jul 5;263(19):9508–9514. [PubMed] [Google Scholar]
  2. Bill N. J., Washington J. A. Bacterial interference by Streptococcus salivarius. Am J Clin Pathol. 1975 Jul;64(1):116–120. doi: 10.1093/ajcp/64.1.116. [DOI] [PubMed] [Google Scholar]
  3. Buchman G. W., Banerjee S., Hansen J. N. Structure, expression, and evolution of a gene encoding the precursor of nisin, a small protein antibiotic. J Biol Chem. 1988 Nov 5;263(31):16260–16266. [PubMed] [Google Scholar]
  4. Carlsson J., Grahnén H., Jonsson G., Wikner S. Early establishment of Streptococcus salivarius in the mouth of infants. J Dent Res. 1970 Mar-Apr;49(2):415–418. doi: 10.1177/00220345700490023601. [DOI] [PubMed] [Google Scholar]
  5. Chou P. Y., Fasman G. D. Prediction of protein conformation. Biochemistry. 1974 Jan 15;13(2):222–245. doi: 10.1021/bi00699a002. [DOI] [PubMed] [Google Scholar]
  6. Darling C. L., Hart G. D. Antagonistic action of Streptococcus salivarius and Streptococcus faecalis to Mycobacterium tuberculosis. J Clin Microbiol. 1976 Oct;4(4):375–378. doi: 10.1128/jcm.4.4.375-378.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dempster R. P., Tagg J. R. The production of bacteriocin-like substances by the oral bacterium Streptococcus salivarius. Arch Oral Biol. 1982;27(2):151–157. doi: 10.1016/0003-9969(82)90136-4. [DOI] [PubMed] [Google Scholar]
  8. Dodd H. M., Horn N., Gasson M. J. Analysis of the genetic determinant for production of the peptide antibiotic nisin. J Gen Microbiol. 1990 Mar;136(3):555–566. doi: 10.1099/00221287-136-3-555. [DOI] [PubMed] [Google Scholar]
  9. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  10. Hastings J. W., Sailer M., Johnson K., Roy K. L., Vederas J. C., Stiles M. E. Characterization of leucocin A-UAL 187 and cloning of the bacteriocin gene from Leuconostoc gelidum. J Bacteriol. 1991 Dec;173(23):7491–7500. doi: 10.1128/jb.173.23.7491-7500.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Jack R. W., Tagg J. R. Factors affecting production of the group A streptococcus bacteriocin SA-FF22. J Med Microbiol. 1992 Feb;36(2):132–138. doi: 10.1099/00222615-36-2-132. [DOI] [PubMed] [Google Scholar]
  13. Johanson W. G., Jr, Blackstock R., Pierce A. K., Sanford J. P. The role of bacterial antagonism in pneumococcal colonization of the human pharynx. J Lab Clin Med. 1970 Jun;75(6):946–952. [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  18. Marugg J. D., Gonzalez C. F., Kunka B. S., Ledeboer A. M., Pucci M. J., Toonen M. Y., Walker S. A., Zoetmulder L. C., Vandenbergh P. A. Cloning, expression, and nucleotide sequence of genes involved in production of pediocin PA-1, and bacteriocin from Pediococcus acidilactici PAC1.0. Appl Environ Microbiol. 1992 Aug;58(8):2360–2367. doi: 10.1128/aem.58.8.2360-2367.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  20. Muriana P. M., Klaenhammer T. R. Cloning, phenotypic expression, and DNA sequence of the gene for lactacin F, an antimicrobial peptide produced by Lactobacillus spp. J Bacteriol. 1991 Mar;173(5):1779–1788. doi: 10.1128/jb.173.5.1779-1788.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  22. Pearson W. R. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol. 1990;183:63–98. doi: 10.1016/0076-6879(90)83007-v. [DOI] [PubMed] [Google Scholar]
  23. Piard J. C., Muriana P. M., Desmazeaud M. J., Klaenhammer T. R. Purification and Partial Characterization of Lacticin 481, a Lanthionine-Containing Bacteriocin Produced by Lactococcus lactis subsp. lactis CNRZ 481. Appl Environ Microbiol. 1992 Jan;58(1):279–284. doi: 10.1128/aem.58.1.279-284.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rawlings N., Ashman K., Wittmann-Liebold B. Computerised version of the Chou and Fasman protein secondary structure predictive method. Int J Pept Protein Res. 1983 Nov;22(5):515–524. doi: 10.1111/j.1399-3011.1983.tb02124.x. [DOI] [PubMed] [Google Scholar]
  25. Russell C., Tagg J. R. Role of bacteriocin during plaque formation by Streptococcus salivarius and Streptococcus sanguis on a tooth in an artificial mouth. J Appl Bacteriol. 1981 Apr;50(2):305–313. doi: 10.1111/j.1365-2672.1981.tb00895.x. [DOI] [PubMed] [Google Scholar]
  26. Sahl H. G., Grossgarten M., Widger W. R., Cramer W. A., Brandis H. Structural similarities of the staphylococcin-like peptide Pep-5 to the peptide antibiotic nisin. Antimicrob Agents Chemother. 1985 May;27(5):836–840. doi: 10.1128/aac.27.5.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanders C. C., Sanders W. E., Jr Enocin: an antibiotic produced by Streptococcus salivarius that may contribute to protection against infections due to group A streptococci. J Infect Dis. 1982 Nov;146(5):683–690. doi: 10.1093/infdis/146.5.683. [DOI] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schnell N., Entian K. D., Götz F., Hörner T., Kellner R., Jung G. Structural gene isolation and prepeptide sequence of gallidermin, a new lanthionine containing antibiotic. FEMS Microbiol Lett. 1989 Apr;49(2-3):263–267. doi: 10.1016/0378-1097(89)90050-5. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Spanier J. G., Cleary P. P. A DNA substitution in the group A streptococcal bacteriophage SP24. Virology. 1983 Oct 30;130(2):514–522. doi: 10.1016/0042-6822(83)90104-6. [DOI] [PubMed] [Google Scholar]
  32. Tagg J. R., Bannister L. V. "Fingerprinting" beta-haemolytic streptococci by their production of and sensitivity to bacteriocine-like inhibitors. J Med Microbiol. 1979 Nov;12(4):397–411. doi: 10.1099/00222615-12-4-397. [DOI] [PubMed] [Google Scholar]
  33. Tagg J. R., Dajani A. S., Wannamaker L. W. Bacteriocin of a group B streptococcus: partial purification and characterization. Antimicrob Agents Chemother. 1975 Jun;7(6):764–772. doi: 10.1128/aac.7.6.764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Tagg J. R., Pybus V., Phillips L. V., Fiddes T. M. Application of inhibitor typing in a study of the transmission and retention in the human mouth of the bacterium Streptococcus salivarius. Arch Oral Biol. 1983;28(10):911–915. doi: 10.1016/0003-9969(83)90086-9. [DOI] [PubMed] [Google Scholar]
  36. Tagg J. R., Russell C. Bacteriocin production by Streptococcus salivarius strain P. Can J Microbiol. 1981 Sep;27(9):918–923. doi: 10.1139/m81-144. [DOI] [PubMed] [Google Scholar]
  37. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  38. 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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