SpringerProtocols: Abstract: Strategies for the Isolation and Characterization of Antibacterial Lantibiotics

3. Strategies for the Isolation and Characterization of Antibacterial Lantibiotics

By: Daniela Jabes¹, Stefano Donadio¹

Abstract

Full Text

Download PDF (570K)

Lantibiotics are biologically active peptides produced by several strains from the phyla Firmicutes and Actinobacteria. They are ribosomally synthesized and undergo posttranslational modifications that endow them with the characteristic (methyl)-lanthionine residues. As a result, lantibiotics contain a variable number of rings, each carrying one thioether link. Many lantibiotics inhibit growth of Gram-positive bacterial strains by interfering with peptidoglycan formation. Because they bind to the key intermediate lipid II at a site not affected by clinically used antibiotics, they are effective against multidrug-resistant strains. We describe a bioassay-based method suitable for finding antibacterial lantibiotics from actinomycete strains and provide selected procedures for characterizing newly discovered lantibiotics for their antibacterial properties.

Affiliation(s): (1) NAICONS Scrl, Milano, Italy

Book Title: Antimicrobial Peptides: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 618 | Pub. Date: Jan-22-2010 | Page Range: 31-45 | DOI: 10.1007/978-1-60761-594-1_3

Subject: Immunology

Key Words: Actinomycetes - antibiotics - bioactive peptides - lantibiotics - MRSA - screening

References

Download References

1.	Chatterjee, C., Paul, M., Xie, L., and van der Donk, W. A. (2005) Biosynthesis and mode of action of lantibiotics. Chem. Rev. 105, 633–684.

2.	Willey, J. M. and van der Donk, W. A. (2007) Lantibiotics: peptides of diverse structure and function. Annu. Rev. Microbiol. 61, 477–501.

3.	Märki, F., Hänni, E., Fredenhagen, A., and van Oostrum, J. (1991) Mode of action of the lanthionine-containing peptide antibiotics duramycin, duramycin B and C, and cinnamycin as indirect inhibitors of phospholipase A2. Biochem. Pharmacol. 42, 2027–2035.

4.	Cloutier, M. M., Guernsey, L., Mattes, P., and Koeppen, B. (1990) Duramycin enhances chloride secretion in airway epithelium. Am. J. Physiol. 259, C450–C454.

5.	Molina y Vedia, L. M., Stutts, M. J., Boucher, J. R. C., and Henke, D. C. (1995) U.S. Patent No. 5,716,931.

6.	Cox, C. R., Coburn, P. S., and Gilmore, M. S. (2005) Enterococcal cytolysin: a novel two component peptide system that serves as a bacterial defense against eukaryotic and prokaryotic cells. Curr. Protein Pept. Sci. 6, 77–84.

7.	Kodani, S., Hudson, M. E., Durrant, M. C., Buttner, M. J., Nodwell, J. R., and Willey, J. M. (2004) The SapB morphogen is a lantibiotic-like peptide derived from the product of the developmental gene ramS in Streptomyces coelicolor. Proc. Natl. Acad. Sci. USA 101, 11448–11453.

8.	Kodani, S., Lodato, M. A., Durrant, M. C., Picart, F., and Willey, J. M. (2005) SapT, a lanthionine-containing peptide involved in aerial hyphae formation in the streptomycetes. Mol. Microbiol. 58, 1368–1380.

9.	Tillotson, R. D., Wösten, H. A., Richter, M., and Willey, J. M. (1998) A surface active protein involved in aerial hyphae formation in the filamentous fungus Schizophyllum commune restores the capacity of a bald mutant of the filamentous bacterium Streptomyces coelicolor to erect aerial structures. Mol. Microbiol. 30, 595–602.

10.	Kellner, R., Jung, G., Josten, M., Kaletta, C., Entian, K. D., and Sahl, H. G. (1989) Pep5: structure elucidation of a large lantibiotic. Angew. Chem. 101, 618–621.

11.	Pag, U. and Sahl, H. G. (2002) Multiple activities in lantibiotics – models for the design of novel antibiotics? Curr. Pharm. Des. 8, 815–833.

12.	Castiglione, F., Lazzarini, A., Carrano, L., Corti, E., Ciciliato, I., Gastaldo, L., Candiani, P., Losi, D., Marinelli, F., Selva, E., and Parenti, F. (2008) Determining the structure and mode of action of microbisporicin, a potent lantibiotic active against multiresistant pathogens. Chem. Biol. 15, 22–31.

13.	Maffioli, S. I., Potenza, D., Vasile, F., De Matteo, M., Sosio, M., Marsiglia, B., Rizzo, V., Scolastico, C., and Donadio, S. (2009) Structure revision of the lantibiotic 97518. J. Nat. Prod. DOI: 10.1021/np800794y.

14.	Li, B., Cooper, L. E., and van der Donk, W. A. (2009) In vitro studies of lantibiotic biosynthesis. Meth. Enzymol. 458, 533–558.

15.	Cortés, J., Appleyard, A. N., and Dawson, M. J. (2009) Whole-cell generation of lantibiotic variants. Meth. Enzymol. 458, 559–574.

16.	Somma, S., Merati, W., and Parenti, F. (1977) Gardimycin, a new antibiotic inhibiting peptidoglycan synthesis. Antimicrob. Agents Chemother. 11, 396–401.

17.	Dodd, H., Gasson, M., Mayer, M., and Narbad, A. (2006) Identifying lantibiotic gene clusters and novel lantibiotic genes. WO 2006/111743.

18.	Donadio, S., Monciardini, P., and Sosio, M. (2009) Approaches to discovering novel antibacterial and antifungal agents. Meth. Enzymol. 458, 3–28.

19.	Meyer, H. E., Heber, M., Eisermann, B., Korte, H., Metzger, J. W., and Jung, G. (1994) Sequence analysis of lantibiotics: chemical derivatization procedures allow a fast access to complete Edman degradation. Anal. Biochem. 223, 185–190.

20.	Brumfitt, W., Salton, M. R., and Hamilton-Miller, J. M. (2002) Nisin, alone and combined with peptidoglycan-modulating antibiotics: activity against methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. J. Antimicrob. Chemother. 50, 731–734.

21.	Goldstein, B. P., Wei, J., Greenberg, K., and Novick, R. (1998) Activity of nisin against Streptococcus pneumoniae, in vitro, and in a mouse infection model. J. Antimicrob. Chemother. 42, 277–278.

22.	Brötz, H., Bierbaum, G., Markus, A., Molitor, E., and Sahl, H. G. (1995) Mode of action of the lantibiotic mersacidin: inhibition of peptidoglycan biosynthesis via a novel mechanism? Antimicrob. Agents Chemother. 39, 714–719.

23.	Brötz, H., Bierbaum, G., Reynolds, P. E., and Sahl, H. G. (1997) The lantibiotic mersacidin inhibits peptidoglycan biosynthesis at the level of transglycosylation. Eur. J. Biochem. 246, 193–199.

24.	Brötz, H., Bierbaum, G., Leopold, K., Reynolds, P. E., and Sahl, H. G. (1998) The lantibiotic mersacidin inhibits peptidoglycan synthesis by targeting lipid II. Antimicrob. Agents Chemother. 42, 154–160.

25.	Bavin, E. M., Beach, A. S., Falconer, R., and Friedman, R. (1952) Nisin in experimental tuberculosis. Lancet 259, 127–129.

26.	NCCLS document M100-S16. (2006) Performance standards for antimicrobial susceptibility testing.

27.	NCCLS document M7-A7. (2008) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically.

28.	NCCLS document M26-A. (2005) Methods for determining bactericidal activity of antimicrobial agents.

29.	Zhang, J. H., Chung, T. D., and Oldenburg, K. R. (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screen 4, 67–73.

30.	Barrett, M. S., Wenzel, R. P., and Jones, R. N. (1992) In vitro activity of mersacidin (M87-1551), an investigational peptide antibiotic tested against Gram-positive bloodstream isolates. Diagn. Microbiol. Infect Dis. 15, 641–644.

31.	Schneider, T. R., Kärcher, J., Pohl, E., Lubini, P., and Sheldrick, G. M. (2000) Ab initio structure determination of the lantibiotic mersacidin. Acta Crystallogr. D Biol. Crystallogr. 56, 705–713.

Comments (Loading...)

Post comment/View all comments