Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: A structural effect

被引:85
作者
Gaussier, H
Morency, H
Lavoie, MC
Subirade, M
机构
[1] Univ Laval, Ctr Rech Sci & Technol Lait, Chaire Rech Canada Prot & Aliments Fonctionnels, Ste Foy, PQ G1K 7P4, Canada
[2] Univ Laval, Fac Sci & Genie, Dept Sci Aliments & Nutr, Ste Foy, PQ G1K 7P4, Canada
[3] Univ Laval, Fac Sci & Genie, Dept Biochim & Microbiol, Ste Foy, PQ G1K 7P4, Canada
关键词
D O I
10.1128/AEM.68.10.4803-4808.2002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:4803 / 4808
页数:6
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