Marine Biotechnology

, Volume 6, Issue 6, pp 633–641 | Cite as

Purification and Partial Identification of Novel Antimicrobial Protein from Marine Bacterium Pseudoalteromonas Species Strain X153

  • Arlette Longeon
  • Jean Peduzzi
  • Michel Barthelemy
  • Sophie Corre
  • Jean-Louis Nicolas
  • Michele Guyot
Article
Received:
Accepted:

Abstract

A marine bacterium, X153, was isolated from a pebble collected at St. Anne du Portzic (France). By 16S ribosomal DNA gene sequence analysis, X153 strain was identified as a Pseudoalteromonas sp. close to P. piscicida. The crude culture of X153 was highly active against human pathogenic strains involved in dermatologic diseases, and marine bacteria including various ichthyopathogenic Vibrio strains. The active substance occurred both in bacterial cells and in culture supernatant. An antimicrobial protein was purified to homogeneity by a 4-step procedure using size-exclusion and ion-exchange chromatography. The highly purified P-153 protein is anionic, and sodium dodecylsulfate polyacrylamide gel electrophoresis gives an apparent molecular mass of 87 kDa. The X153 bacterium protected bivalve larvae against mortality, following experimental challenges with ichthyopathogenic Vibrio. Pseudoalteromonas sp. X153 may be useful in aquaculture as a probiotic bacterium.

Keywords

antimicrobial protein Pseudoalteromonas probiotic bacteria 
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Notes

Acknowledgments

We thank C. Deregnaucourt (USM 504, Museum National d’Histoire Naturelle) for SDS-PAGE. This work was supported in part by the program 97.C.0111 “Recherche de nouveaux agents antibiotiques et probiotiques dans les bacteries marines” of the French ministry for Research and Technology.

References

  1. Almeida, M.J., Milet, C., Peduzzi, J., Pereira, L., Haigle, J., Barthelemy, M., Lopez, E. 2000Effect of water-soluble matrix fraction extracted from the nacre of Pinctada maxima on the alkaline phosphatase activity of cultured fibroblastsJ Exp Zool288327334PubMedGoogle Scholar
  2. Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J. 1990Basic local alignment search toolJ Mol Biol215403410CrossRefPubMedGoogle Scholar
  3. Austin, B., Stuckey, L.F., Robertson, P.A.W., Effendi, I, Griffith, D.R.W. 1995A probiotic strain of Vibrio alginolyticus effective in reducing diseases caused by Aeromonas salmonicida, Vibrio anguillarum and Vibrio ordaliiJ Fish Dis189396Google Scholar
  4. Baam, B.R., Gandhi, N.M., Freitas, Y.M. 1966Antibiotic activity of marine microorganismsHelgol Wiss Meeresunters13181185Google Scholar
  5. Barja, J.L., Lemos, M.L., Toranzo, A. 1989Purification and characterization of an antibacterial substance produced by a marine Alteromonas speciesAntimicrob Agents Chemother3316741679PubMedGoogle Scholar
  6. Baumann, P., Baumann, L. 1981The marine gram negative eubacteria: genera Photobacterium, Benekea, Alteromonas, Pseudomonsa and AlcaligenesRuiz-Ponte, C.Samain, J. F.Sanchez, J. L.J. L., Nicolas eds. The Prokayotes vol. 2, StarrSpringer-VerlagBerlin, Germany13021331Google Scholar
  7. Cohen, S.A., Strydom, D.J. 1988Amino acid analyses utilizing phenylisothiocyanate derivativesAnal Biochem174116PubMedGoogle Scholar
  8. Delves-Broughton, J., Blackburn, P., Evans, R.J., Hugenholtz, J. 1996Applications of the bacteriocin, nisinAntonie Van Leeuwenhoek69193202PubMedGoogle Scholar
  9. Faulkner, D.J. 2001Marine natural productsNat Prod Rep18149PubMedGoogle Scholar
  10. Fenical, W. 1993Chemical studies of marine bacteria: developing a new resourceChem Rev9316731683Google Scholar
  11. Fenical, W. 1997New pharmaceuticals from marine organismsTrends Biotechnol15339341PubMedGoogle Scholar
  12. Galtier, N., Gouy, M., Gautier, C. 1996SEAVIEW and PHYLO_WIN: two graphic tools for sequence alignment and molecular phylogenyComput Appl Biosci12543548PubMedGoogle Scholar
  13. Gatesoupe, F.J. 1999The use of probiotics in aquacultureAquaculture180147165Google Scholar
  14. Gibson, L.F., Woodworth, J., George, A.M. 1998Probiotic activity of Aeromonas media on the Pacific oyster, Crassostrea gigas, when challenged with Vibrio tubiashiiAquaculture169111120Google Scholar
  15. Gomez-Gil, B., Roque, A., Turnbull, J. 2001The use and selection of probiotic bacteria for use in the culture of larval aquatic organismsAquaculture191259270Google Scholar
  16. Ivanova, E.P., Shevchenko, L.S., Sawabe, T., Lysenko, A.M., Svetashev, V.I., Gorshkova, N.M., Satomi, M., Christen, R., Mikhailov, V.V. 2002Pseudoalteromonas maricaloris sp. nov., isolated from an Australian sponge, and reclassification of [Pseudoalteromonas aurantia] NCIMB 2033 as Pseudoalteromonas flavipulchra sp. novInt J Syst Evol Microbiol52263271PubMedGoogle Scholar
  17. James, S.G., Holmstrom, C., Kjelleberg, S. 1996Purification and characterization of a novel antibacterial protein from the marine bacterium D2Appl Environ Microbiol6227832788PubMedGoogle Scholar
  18. Jensen, P.R., Fenical, W. 1994Strategies for the discovery of secondary metabolites from marine bacteria: ecological perspectivesAnnu Rev Microbiol48559584PubMedGoogle Scholar
  19. Konisky, J. 1982Colicins and other bacteriocins with established modes of actionAnnu Rev Microbiol36125144PubMedGoogle Scholar
  20. Klaenhammer, T.R. 1988Bacteriocins of lactic acid bacteriaBiochimie70337349PubMedGoogle Scholar
  21. Laemmli, U.K. 1970Cleavage of structural proteins during the assembly of the head of bacteriophage T4Nature227680685PubMedGoogle Scholar
  22. Lovejoy, C., Bowman, J.P., Hallegraeff, G.M. 1998Algicidal effects of a novel marine Pseudoalteromonas isolate (class Proteobacteria, gamma subdivision) on harmful algal bloom species of the genera Chattonella, Gymnodinium, and HeterosigmaAppl Environ Microbiol6428062813PubMedGoogle Scholar
  23. Mandal, S., Manda, M.D., Pal, N.K. 2003Ofloxacin minimum inhibitory concentration versus disk diffusion zone diameter for Salmonella enterica serovar Typhi isolates: problems of the detection of oflaxin resistanceJpn J Infect Dis56210212PubMedGoogle Scholar
  24. McCarthy, S.A., Johnson, R.M., Kakimoto, D. 1994Characterization of an antibiotic produced by an Alteromonas luteoviolacea Gauthier 1982, 85 isolated from Kinko Bay, JapanJ Appl Bacteriol77426432PubMedGoogle Scholar
  25. Olafsen, J.A. 2001Interactions between fish larvae and bacteria in marine aquacultureAquaculture200223247Google Scholar
  26. Rayman, M.K., Aris, B., Hurst, A. 1981Nisin: a possible alternative or adjunct to nitrite in the preservation of meatsAppl Environ Microbiol41375380PubMedGoogle Scholar
  27. Ringo, E., Vadstein, O. 1998Colonization of Vibrio pelagius and Aeromonas caviae in early developing turbot (Scophthalmus maximus L.) larvaeJ Appl Microbiol84227233PubMedGoogle Scholar
  28. Robert, R., Nicolas, J.L, Miner, P. 1996Mortality control of scallop (Pecten maximus) larvae in the hatcheryAquacult Int4305313Google Scholar
  29. Rosenfeld, D.W., Zobell, C.E. 1947Antibiotic production by marine microorganismsJ Bacteriol54393398Google Scholar
  30. Ruiz-Ponte, C., Samain, J.F., Sanchez, J.L., Nicolas, J.L. 1999The benefit of a Roseobacter species on the survival of scallop larvaeMar Biotechnol15259PubMedGoogle Scholar
  31. Schilinger, U., Lucke, F. 1989Antibacterial activity of Lactobacillus sake isolated from meatAppl Environ Microbiol5519011907PubMedGoogle Scholar
  32. Tagg, J.R., Dajani, A.S., Wannamaker, L.W. 1976Bacteriocins of gram-positive bacteriaBacterial Rev40722756Google Scholar
  33. Thomas, X., Destoumieux-Garzon, D., Peduzzi, J., Afonso, C., Blond, A., Birlirakis, N., Goulard, C., Dubost, L., Thai, R., Tabet, J.-C., Rebuffat, S. 2004Siderophore peptide, a new type of post-translationally modified antibacterial peptide with potent activityJ Biol Chem2792823328242PubMedGoogle Scholar
  34. Thompson, J.D., Higgins, D.G., Gibson, T.J. 1994CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucleic Acids Res1146734680Google Scholar
  35. Venkateswaran, K., Dohmoto, N. 2000Pseudoalteromonas peptidolytica sp. nov., a novel marine mussel-thread-degrading bacterium isolated from the Sea of JapanInt J Syst Evol Microbiol50565574PubMedGoogle Scholar
  36. Verschuere, L., Rombaut, G., Sorgeloos, P., Verstraete, W. 2000Probiotic bacteria as biological control agents in aquaculrureMicrobiol Mol Biol Rev64655671PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2004

Authors and Affiliations

  • Arlette Longeon
    • 1
  • Jean Peduzzi
    • 1
  • Michel Barthelemy
    • 1
  • Sophie Corre
    • 2
  • Jean-Louis Nicolas
    • 3
  • Michele Guyot
    • 1
  1. 1.Laboratoire de Chimie et Biochimie des Substances NaturellesUMR 5154 CNRS, Museum National d’Histoire NaturelleParisFrance
  2. 2.Micromer Technopole Brest-IroiseBrestFrance
  3. 3.Laboratoire de Physiologie des InvertebresDRV, IFREMERPlouzaneFrance

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