1887

Microbiology Society logo

Volume 148, Issue 11

NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin Free

Abstract

Nisin produced by subsp. is a 34-residue antibacterial polypeptide and belongs to a group of post-translationally modified peptides, lantibiotics, with dehydrated residues and cyclic amino acids, lanthionines. These modifications are supposed to be made by enzymes encoded by and genes, found only in biosynthetic operons encoding lantibiotics. To analyse the extent of modification, His-tagged nisin precursors were expressed in and mutant strains. The His-tagged nisin precursors were purified from the cytoplasm of the cells, as lack of NisB or NisC activity impaired translocation of the nisin precursor. The purified His-tagged polypeptides were analysed with trypsin digestion followed by nisin bioassay, SDS-PAGE, N-terminal sequencing and mass spectroscopy. According to the results, nisin precursors from the strain lacking NisB activity were totally unmodified, whereas nisin precursors from the strain lacking NisC activity, but having NisB activity, were dehydrated and devoid of normal lanthionine formation. This is the first experimental evidence showing that NisB is required for dehydration and NisC for correct lanthionine formation in nisin maturation.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): dehydroalanine , dehydrobutyrine , lantibiotic , MALDI-TOF, matrix-assisted laser desorption ionization/time-of-flight and nisin biosynthesis
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-11-3561
2002-11-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/11/1483561a.html?itemId=/content/journal/micro/10.1099/00221287-148-11-3561&mimeType=html&fmt=ahah

References

  1. Axelsson L. T, Ahrne S. E. J, Andersson M. C., Ståhl S. R. 1988; Identification and cloning of a plasmid-encoded erythromycin resistance determinant from Lactobacillus reuteri . Plasmid 20:171–174
     [Google Scholar]
  2. Chan W. C, Dodd H. M, Horn N., Maclean K. 1996; Structure-activity relationships in the peptide antibiotic nisin: role of dehydroalanine 5. Appl Environ Microbiol 62:2966–2969
     [Google Scholar]
  3. Delves-Broughton J, Blackburn P, Evans R. J., Hugenholtz J. 1996; Applications of the bacteriocin, nisin. Antonie Leeuwenhoek 69:193–202
     [Google Scholar]
  4. Engelke G, Gutowski-Eckel Z, Kiesau P, Siegers K, Hammelmann M., Entian K.-D. 1994; Regulation of nisin biosynthesis and immunity in Lactococcus lactis 6F3. Appl Environ Microbiol 60:814–825
     [Google Scholar]
  5. Gasson M. J. 1983; Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing. J Bacteriol 154:1–9
     [Google Scholar]
  6. Graeffe T, Rintala H, Paulin L., Saris P. E. J. 1991; A natural nisin variant. In Nisin and Novel Lantibiotics pp 260–268 Edited by Jung G., Sahl H.-G. Leiden: ESCOM Science Publishers;
     [Google Scholar]
  7. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
     [Google Scholar]
  8. Holo H., Nes I. F. 1989; High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl Environ Microbiol 55:3119–3123
     [Google Scholar]
  9. Immonen T., Saris P. E. J. 1998; Characterization of the nisFEG operon of the nisin Z producing Lactococcus lactis subsp. lactis N8 strain. DNA Seq 9:263–274
     [Google Scholar]
  10. Immonen T, Ye S, Ra R, Qiao M, Paulin L., Saris P. E. J. 1995; The codon usage of the nisZ operon in Lactococcus lactis N8 suggests a non-lactococcal origin of the conjugative nisin-sucrose transposon. DNA Seq 5:203–218
     [Google Scholar]
  11. Jacobs M. F, Tynkkynen S., Sibakov M. 1995; Highly bioluminescent Streptococcus thermophilus strain for the detection of dairy-relevant antibiotics in milk. Appl Environ Microbiol 44:405–412
     [Google Scholar]
  12. Kalkkinen N., Tilgman C. 1988; A gas-pulsed liquid-phase sequencer constructed from a Beckman 890D instrument by using Applied Biosystems delivery and cartridge blocks. J Protein Chem 7:242–243
     [Google Scholar]
  13. Kuipers O. P, Beerthuyzen M. M, Siezen R. J., de Vos W. M. 1993; Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis . Requirement of expression of the nisA and nisI genes for development of immunity. Eur J Biochem 216:281–291
     [Google Scholar]
  14. Kuipers O. P, Beerthuyzen M. M, de Ruyter P. G. G. A, Luesink E. J., de Vos W. M. 1995; Autoregulation of nisin biosynthesis in Lactococcus lactis by signal transduction. J Biol Chem 270:27299–27304
     [Google Scholar]
  15. Kupke T., Götz F. 1996; Posttranslational modifications of lantibiotics. Antonie Leeuwenhoek 69:139–150
     [Google Scholar]
  16. Liu W., Hansen J. N. 1993; The antimicrobial effect of a structural variant of subtilin against outgrowing Bacillus cereus T spores and vegetative cells occurs by different mechanisms. Appl Environ Microbiol 59:648–651
     [Google Scholar]
  17. Maniatis T, Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  18. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acids from micro-organisms. J Mol Biol 3:208–218
     [Google Scholar]
  19. McAuliffe O, Ross R. P., Hill C. 2001; Lantibiotics: structure, biosynthesis and mode of action. FEMS Microbiol Rev 25:285–308
     [Google Scholar]
  20. Meyer C, Bierbaum G, Heidrich C, Reis M, Suling J, Iglesias-Wind M. I, Kempter C, Molitor E., Sahl H.-G. 1995; Nucleotide sequence of the lantibiotic Pep5 biosynthetic gene cluster and functional analysis of PepB and PepC. Eur J Biochem 232:478–489
     [Google Scholar]
  21. Mulders J. W, Boerrighter I. J, Rollema H. S, Siezen R. J., de Vos W. M. 1991; Identification and characterization of the lantibiotic nisin Z, a natural nisin variant. Eur J Biochem 201:581–584
     [Google Scholar]
  22. Nes I. F, Diep D. B, Havarstein L. S, Brurberg M. B, Eijsink V., Holo H. 1996; Biosynthesis of bacteriocins in lactic acid bacteria. Antonie Leeuwenhoek 70:113–128
     [Google Scholar]
  23. Qiao M., Saris P. E. J. 1996; Evidence for a role of NisT in transport of the lantibiotic nisin produced by Lactococcus lactis N8. FEMS Microbiol Lett 144:89–93
     [Google Scholar]
  24. Qiao M, Immonen T, Koponen O., Saris P. E. J. 1995; The cellular location and effect on nisin immunity of the NisI protein from Lactococcus lactis N8 expressed in Escherichia coli and L. lactis. FEMS Microbiol Lett 131:75–80
     [Google Scholar]
  25. Qiao M, Ye S, Koponen O, Ra R, Immonen T., Saris P. E. J. 1996; Regulation of the nisin operons in Lactococcus lactis N8. J Appl Bacteriol 80:626–634
     [Google Scholar]
  26. Ra R, Qiao M, Immonen T, Pujana I., Saris P. E. J. 1996; Genes responsible for nisin synthesis and immunity form a regulon of two operons and are induced by nisin in Lactococcus lactis N8. Microbiology 142:1281–1288
     [Google Scholar]
  27. Ra R, Beerthuyzen M. M, de Vos W. M, Saris P. E. J., Kuipers O. P. 1999; Differential contributions of NisI and NisFEG to producer-immunity analysed by in-frame disruptions in the nisin gene cluster of Lactococcus lactis . Microbiology 145:1227–1233
     [Google Scholar]
  28. Saarinen J, Welgus H. G, Flizar C. A, Kalkkinen N., Helin J. 1999; N-glycan structures of matrix metalloproteinase-1 derived from human fibroblasts and from HT-1080 fibrosarcoma cells. Eur J Biochem 259:829–840
     [Google Scholar]
  29. Sahl H.-G, Jack R. W., Bierbaum G. 1995; Biosynthesis and biological activities of lantibiotics with unique post-translational modifications. Eur J Biochem 230:827–853
     [Google Scholar]
  30. Sen A. K, Narbad A, Horn N, Dodd H. M, Parr A. J, Colquhoun I., Gasson M. J. 1999; Post-translational modification of nisin: the involvement of NisB in the dehydration process. Eur J Biochem 261:524–532
     [Google Scholar]
  31. Siegers K, Heinzman S., Entian K.-D. 1996; Biosynthesis of lantibiotic nisin – posttranslational modification of its prepeptide occurs at a multimeric membrane-associated lanthionine synthetase complex. J Biol Chem 271:12294–12301
     [Google Scholar]
  32. Terzaghi B. E., Sandine W. E. 1975; Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol 29:807–813
     [Google Scholar]
  33. van der Meer J. R, Polman J, Beerthuyzen M. M, Siezen R. J, Kuipers O. P., de Vos W. M. 1993; Characterization of the Lactococcus lactis nisin A operon genes nisP encoding a subtilisin-like serine protease involved in precursor processing, and nisR , encoding a regulatory protein involved in nisin biosynthesis. J Bacteriol 175:2578–2588
     [Google Scholar]
  34. Vieira J., Messing J. 1991; New pUC-derived cloning vectors with different selectable markers and DNA replication origins. Gene 100:189–194
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-11-3561
Loading
NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin
Microbiology 148, 3561 (2002); https://doi.org/10.1099/00221287-148-11-3561
/content/journal/micro/10.1099/00221287-148-11-3561
/content/journal/micro/10.1099/00221287-148-11-3561
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error