SEARCH

SEARCH BY CITATION

Keywords:

  • chemically modified nisin;
  • lantibiotic;
  • mass spectrometry;
  • nisin;
  • NMR

Abstract

  1. Top of page
  2. Abstract
  3. References

Nisin, a 34-residue peptide bacteriocin, contains the less common amino acids lanthionine, β-methyl-lanthionine, dehydroalanine (Dha), and dehydrobutyrine (Dhb). Several chemically modified nisin A species were purified by reverse-phase HPLC and characterized by two-dimensional NMR and electrospray mass spectrometry. Five constituents, [2-hydroxy-Ala5]nisin, [Ile4-amide,pyruvyl-Leu6]des-Dha5-nisin, [Met(O)21]nisin, [Ser33]nisin, and nisin-(1–32)-peptide amide, were found in a commercial nisin sample. A further species, [2-hydroxy-Ala5]nisin-(1–32)-peptide amide, was obtained by freeze drying an acidic nisin solution. These compounds are formed by chemical modification of nisin: the addition of a water molecule to the dehydroalanine residues, which can lead to the cleavage of the polypeptide chain, or the oxidation of methionine residues.

The 2-hydroxyalanine-containing products have a limited stability; they are spontaneously converted into the corresponding des-dehydroalanine derivatives. The growth-inhibiting activity of the modified nisins towards different bacteria was determined. The 2-hydroxyalanine-containing species and the des-dehydroalanine derivative show a strong reduction in biological activity as compared to native nisin. [Met(O)21]nisin and [Ser33]nisin show moderate or no reduction in biological activity.

Abbreviations.
Dha

dehydroalanine

Dhb

dehydrobutyrine

ESI

electrospray ionization

MIC

minimal inhibitory concentration

RP

reverse phase

References

  1. Top of page
  2. Abstract
  3. References
  • Brot, N. & Weissbach, H. (1991) Biochemistry of methionine sulfoxide residues in proteins, Biofactors 3, 9196.
  • Chan, W. C., Bycroft, B. W., Lian, L.-Y. & Roberts, G. C. K. (1989) Isolation and characterization of two degradation products derived from the peptide antibiotic nisin, FEBS Lett. 252, 2936.
  • Delves-Broughton, J. (1990) Nisin and its uses as a food preservative, Food Technol. 44, 100117.
  • Dodd, H. M. & Gasson, M. J. (1994) Bacteriocins of lactic acid bacteria, in Genetics and biotechnology of lactic acid bacteria (Gasson, M. J. & deVos, W. M., eds) pp. 211251, Chapman & Hall, London .
  • Driessen, A. J. M., van den Hooven, H. W., Kuiper, W., van de Kamp, M., Sahl, H.-G., Konings, R. N. H. & Konings, W. N. (1995) Mechanistic studies of lantibiotic-induced permeabilization of phospholipid vesicles, Biochemistry 34, 16061614.
  • Gao, F. H., Abee, T. & Konings, W. N. (1991) Mechanism of action of the peptide antibiotic nisin in liposomes and cytochrome c oxidase-containing proteoliposomes, Appl. Environ. Microbiol. 57, 21642170.
  • Gross, E. (1977) α,β-Unsaturated and related amino acids in peptides and proteins, in Protein crosslinking Nutritional and medical consequences (Friedman, M., ed.) pp. 131153, Plenum Press, New York .
  • Hurst, A. (1981) Nisin, Adv. Appl. Microbiol. 27, 85123.
  • Kuipers, O. P., Rollema, H. S., Yap, W. M. G. J., Boot, H. J., Siezen, R. J. & de Vos, W. M. (1992) Engineering dehydrated amino acid residues in the antimicrobial peptide nisin, J. Biol. Chem. 267, 2434024346.
  • Kuipers, O. P., Bierbaum, G., Ottenwälder, B., Dodd, H. M., Horn, N., Metzger, J., Kupke, T., Gnau, V., Bongers, R., van den Bogaard, P., Kosters, H., Rollema, H. S., de Vos, W. M., Siezen, R. J., Jung, G., Götz, F., Sahl, H.-G. & Gasson, M. J. (1996) Protein engineering of lantibiotics, Antonie van Leeuwenhoek 69, 161169.
  • Lian, L.-Y., Chan, W. C., Morley, S. D., Roberts, G. C. K., Bycroft, B. W. & Jackson, D. (1992) Solution structures of nisin A and its two major degradation products determined by n.m.r., Biochem. J. 283, 413420.
  • Liu, W. & Hansen, J. N. (1992) Enhancement of the chemical and antimicrobial properties of subtilin by site-directed mutagenesis, J. Biol. Chem. 267, 2507825085.
  • Rollema, H. S., Both, P. & Siezen, R. J. (1991) NMR and activity studies of nisin degradation products, in Nisin and novel lantibiotics (Jung, G. & Sahl, H.-G., eds) pp. 123130, ESCOM, Leiden , The Netherlands .
  • Rollema, H. S., Kuipers, O. P., Both, P., de Vos, W. M. & Siezen, R. J. (1995) Improvement of solubility and stability of the antimicrobial peptide nisin by protein engineering, Appl. Environ. Microbiol. 61. 28732878.
  • Sahl, H.-G., Jack, R. W. & Bierbaum, C. (1995) lantibiotics: biosynthesis and biological activities of peptides with unique post-translational modifications, Eur. J. Biochem. 230, 827853.
    Direct Link:
  • Slijper, M., Hilbers, C. W., Konings, R. N. H. & van de Ven, F. J. M. (1989) NMR studies of lantibiotics. Assignment of the 1H-NMR spectrum of nisin and identification of interresidual contacts, FEBS Lett. 252, 2228.
  • Tramer, J. J. & Fowler, G. G. (1964) Estimation of nisin in foods, J. Sci. Food Agric. 15, 522528.
  • Van de Ven, F. J. M., van den Hooven, H. W., Konings, R. N. H. & Hilbers, C. W. (1991) NMR studies of lantibiotics. The structure of nisin in aqueous solution, Eur. J. Biochem. 202, 11811188.
  • Van den Hooven, H. W., Doeland, C. C. M., van de Kamp, M., Konings, R. N. H., Hilbers, C. W. & van de Ven, F. J. M. (1996a) Three-dimensional structure of the lantibiotic nisin in the presence of membrane-mimetic micelles of dodecylphosphycholine and of sodium dodecylsulphate, Eur. J. Biochem. 235, 382393.
  • Van den Hooven, H. W., Spronk, C. A. E. M., van de Kamp, M., Konings, R. N. H., Hilbers, C. W. & van de Ven, F. J. M. (1996b) Surface location and orientation of the lantibiotic nisin bound to membrane-mimicking micelles of dodecylphosphocholine and of sodium dodecylsulphate, Eur. J. Biochem. 235, 394403.