Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: A structural effect
Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: A structural effect
复制标题
DOI：
10.1128/aem.68.10.4803-4808.2002
复制
发表时间：
2002-10-01
影响因子：
4.4
通讯作者：
Subirade, M
中科院分区：
文献类型：
--
作者：
Gaussier, H;Morency, H;Subirade, M
三氟乙酸（TFA）是一种与pediocin PA-1相关的纯化污染物，会干扰傅里叶变换红外光谱结构分析。圆二色性表明，它的存在影响了pediocin的结构折叠。因此，我们提出了一种新的pediocin PA-1的纯化程序，使用盐酸代替TFA在所有的疏水步骤。该程序变更不影响纯化收率或纯化的pediocin PA-1的量。此外，与TFA相反，在纯化后除去HCl是更容易进行的程序。事实上，去除TFA需要更多的实验，并导致蛋白质损失。因此，盐酸是一个很好的替代三氟乙酸在pediocin PA-1的纯化，并可以扩展到其他蛋白质的纯化。我们还表明，TFA诱导的结构修饰不显着影响抗菌活性的pediocin PA-1。
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.