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Applied and Environmental Microbiology, October 2002, p. 4803-4808, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4803-4808.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Replacement of Trifluoroacetic Acid with HCl in the Hydrophobic Purification Steps of Pediocin PA-1: a Structural Effect

Hélène Gaussier,1 Hélène Morency,2 Marc C. Lavoie,2 and Muriel Subirade1*

Centre de Recherche en Sciences et Technologie du Lait, Chaire de Recherche du Canada sur les Protéines et les Aliments Fonctionnels, and Département des Sciences des Aliments et de Nutrition,1 Département de Biochimie et de Microbiologie, Faculté des Sciences et Génie, Université Laval, Sainte-Foy, Québec G1K 7P4, Canada2

Received 27 February 2002/ Accepted 4 July 2002

Trifluoroacetic acid (TFA) is a purification contaminant associated with pediocin PA-1 that interferes with Fourier transform infrared spectroscopy structural analysis. As revealed by circular dichroism, its presence affects the structural folding of pediocin. Consequently, we propose a new pediocin PA-1 purification procedure using HCl instead of TFA in all of the hydrophobic steps. This procedural change does not affect the purification yield or the amount of pediocin PA-1 purified. Furthermore, removing HCl, as opposed to TFA, after purification is an easier procedure to carry out. In fact, the removal of TFA requires more experimentation and results in protein loss. Thus, HCl is a good alternative to TFA in pediocin PA-1 purification and can be extended to the purification of other proteins. We also show that TFA-induced structural modifications do not significantly affect the antimicrobial activity of pediocin PA-1.

* Corresponding author. Mailing address: Centre STELA/INAF Université Laval, Pavillon Paul-Comtois, Québec G1K 7P4, Canada. Phone: (418) 656-2131 ext. 4278. Fax: (418) 656-3353. E-mail: Muriel.subirade{at}aln.ulaval.ca.

Applied and Environmental Microbiology, October 2002, p. 4803-4808, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4803-4808.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Drider, D., Fimland, G., Hechard, Y., McMullen, L. M., Prevost, H. (2006). The Continuing Story of Class IIa Bacteriocins. Microbiol. Mol. Biol. Rev. 70: 564-582 [Abstract] [Full Text]  
  • Mota-Meira, M., Morency, H., Lavoie, M. C. (2005). In vivo activity of mutacin B-Ny266. J Antimicrob Chemother 56: 869-871 [Abstract] [Full Text]  
  • Gaussier, H., Lefevre, T., Subirade, M. (2003). Binding of Pediocin PA-1 with Anionic Lipid Induces Model Membrane Destabilization. Appl. Environ. Microbiol. 69: 6777-6784 [Abstract] [Full Text]  


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