This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Häußler, S.
Right arrow Articles by Steinmetz, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Häußler, S.
Right arrow Articles by Steinmetz, I.

 Previous Article  |  Next Article 

Infect Immun, April 1998, p. 1588-1593, Vol. 66, No. 4
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Purification and Characterization of a Cytotoxic Exolipid of Burkholderia pseudomallei

S. Häußler,1 M. Nimtz,2 T. Domke,2 V. Wray,2 and I. Steinmetz1,*

Institute of Medical Microbiology, Hannover Medical School, 30625 Hannover,1 and Department of Molecular Structure Research, Gesellschaft für Biotechnologische Forschung mbH, 38124 Braunschweig,2 Germany

Received 22 September 1997/Returned for modification 24 November 1997/Accepted 6 January 1998

Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease, which is increasingly recognized as an important public health problem in various tropical regions. This study describes the identification and characterization of a heat-stable extracellular toxin of B. pseudomallei. After cultivation of B. pseudomallei in liquid media, the heated cell-free supernatant was concentrated by ultrafiltration. The concentrate exhibited a cytotoxic and hemolytic activity which showed remarkable resistance against alkaline and acidic treatments. For further purification, reversed-phase chromatography using a fast-performance liquid chromatography system was performed. After elution with an acetonitrile gradient, a single cytotoxic and hemolytic peak was detected. Structural characterization of the toxin was performed by a combination of mass spectrometric and nuclear magnetic resonance spectroscopic techniques. A highly purified glycolipid, 2-O-alpha -L-rhamnopyranosyl-alpha -L-rhamnopyranosyl-beta -hydroxytetradecanoyl-beta -hydroxytetradecanoate (Rha-Rha-C14-C14), with a molecular mass of 762 Da was identified. The purified exolipid showed a time- and dose-dependent cytotoxic effect on phagocytic (HL60) and nonphagocytic (HeLa) cell lines. In addition, a time- and dose-dependent hemolysis of erythrocytes from various species was observed. The toxin structure makes a detergentlike action most probable. Interestingly, the cytotoxic and hemolytic activities of the glycolipid could be neutralized by albumin. Future studies will concentrate on the role of this exolipid as a virulence factor in the pathogenesis of melioidosis.

* Corresponding author. Mailing address: Medical Microbiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Phone: 0511-532-4352. Fax: 0511-532-4366. E-mail: Steinmetz.Ivo{at}mh-hannover.de.




This article has been cited by other articles:

  • Schell, M. A., Lipscomb, L., DeShazer, D. (2008). Comparative Genomics and an Insect Model Rapidly Identify Novel Virulence Genes of Burkholderia mallei. J. Bacteriol. 190: 2306-2313 [Abstract] [Full Text]  
  • Gunther, N. W. IV, Nunez, A., Fett, W., Solaiman, D. K. Y. (2005). Production of Rhamnolipids by Pseudomonas chlororaphis, a Nonpathogenic Bacterium. Appl. Environ. Microbiol. 71: 2288-2293 [Abstract] [Full Text]  
  • Ong, C., Ooi, C. H., Wang, D., Chong, H., Ng, K. C., Rodrigues, F., Lee, M. A., Tan, P. (2004). Patterns of large-scale genomic variation in virulent and avirulent Burkholderia species. Genome Res 14: 2295-2307 [Abstract] [Full Text]  
  • Ulrich, R. L. (2004). Quorum Quenching: Enzymatic Disruption of N-Acylhomoserine Lactone-Mediated Bacterial Communication in Burkholderia thailandensis. Appl. Environ. Microbiol. 70: 6173-6180 [Abstract] [Full Text]  
  • Ulrich, R. L., Hines, H. B., Parthasarathy, N., Jeddeloh, J. A. (2004). Mutational Analysis and Biochemical Characterization of the Burkholderia thailandensis DW503 Quorum-Sensing Network. J. Bacteriol. 186: 4350-4360 [Abstract] [Full Text]  
  • Inglis, T. J. J., Aravena-Roman, M., Ching, S., Croft, K., Wuthiekanun, V., Mee, B. J. (2003). Cellular Fatty Acid Profile Distinguishes Burkholderia pseudomallei from Avirulent Burkholderia thailandensis. J. Clin. Microbiol. 41: 4812-4814 [Abstract] [Full Text]  
  • Haussler, S., Rohde, M., von Neuhoff, N., Nimtz, M., Steinmetz, I. (2003). Structural and Functional Cellular Changes Induced by Burkholderia pseudomallei Rhamnolipid. Infect. Immun. 71: 2970-2975 [Abstract] [Full Text]  
  • ATKINS, T., PRIOR, R., MACK, K., RUSSELL, P., NELSON, M., PRIOR, J., ELLIS, J., OYSTON, P. C. F., DOUGAN, G., TITBALL, R. W. (2002). Characterisation of an acapsular mutant of Burkholderia pseudomallei identified by signature tagged mutagenesis. J Med Microbiol 51: 539-553 [Abstract] [Full Text]  
  • BROWN, N. F., BEACHAM, I. R. (2000). Cloning and analysis of genomic differences unique to Burkholderia pseudomallei by comparison with B. thailandensis. J Med Microbiol 49: 993-1001 [Abstract] [Full Text]  
  • HART, C A, WINSTANLEY, C (1999). Pseudomonas and all that. Arch. Dis. Child. 81: 2-4 [Full Text]  
  • Hoppe, I., Brenneke, B., Rohde, M., Kreft, A., Haussler, S., Reganzerowski, A., Steinmetz, I. (1999). Characterization of a Murine Model of Melioidosis: Comparison of Different Strains of Mice. Infect. Immun. 67: 2891-2900 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»