Pereira C I, Matos D, San Romão M V, Crespo M T Barreto
IBET-Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal.
Appl Environ Microbiol. 2009 Jan;75(2):345-52. doi: 10.1128/AEM.01958-08. Epub 2008 Nov 14.
In this work we investigated the role of the tyrosine decarboxylation pathway in the response of Enterococcus faecium E17 cells to an acid challenge. It was found that 91% of the cells were able to remain viable in the presence of tyrosine when they were incubated for 3 h in a complex medium at pH 2.5. This effect was shown to be related to the tyrosine decarboxylation pathway. Therefore, the role of tyrosine decarboxylation in pH homeostasis was studied. The membrane potential and pH gradient, the parameters that compose the proton motive force (PMF), were measured at different pHs (pH 4.5 to 7). We obtained evidence showing that the tyrosine decarboxylation pathway generates a PMF composed of a pH gradient formed due to proton consumption in the decarboxylation reaction and by a membrane potential which results from electrogenic transport of tyrosine in exchange for the corresponding biogenic amine tyramine. The properties of the tyrosine transporter were also studied in this work by using whole cells and right-side-out vesicles. The results showed that the transporter catalyzes homologous tyrosine/tyrosine antiport, as well as electrogenic heterologous tyrosine-tyramine exchange. The tyrosine transporter had properties of a typical precursor-product exchanger operating in a proton motive decarboxylation pathway. Therefore, the tyrosine decarboxylation pathway contributes to an acid response mechanism in E. faecium E17. This decarboxylation pathway gives the strain a competitive advantage in nutrient-depleted conditions, as well as in harsh acidic environments, and a better chance of survival, which contributes to higher cell counts in food fermentation products.
在本研究中,我们探究了酪氨酸脱羧途径在屎肠球菌E17细胞应对酸胁迫反应中的作用。结果发现,当在pH 2.5的复合培养基中孵育3小时时,91%的细胞在酪氨酸存在的情况下能够保持存活。这种效应被证明与酪氨酸脱羧途径有关。因此,我们研究了酪氨酸脱羧在pH稳态中的作用。在不同pH值(pH 4.5至7)下测量了构成质子动力势(PMF)的膜电位和pH梯度这两个参数。我们获得的证据表明,酪氨酸脱羧途径产生了一种PMF,它由脱羧反应中质子消耗形成的pH梯度以及酪氨酸与相应生物胺酪胺进行电中性转运所产生的膜电位组成。在本研究中,还利用全细胞和外翻小泡对酪氨酸转运体的特性进行了研究。结果表明,该转运体催化同源酪氨酸/酪氨酸反向转运以及电中性异源酪氨酸 - 酪胺交换。酪氨酸转运体具有在质子动力脱羧途径中起作用的典型前体 - 产物交换器的特性。因此,酪氨酸脱羧途径有助于屎肠球菌E17的酸应激反应机制。这种脱羧途径使该菌株在营养匮乏条件下以及恶劣的酸性环境中具有竞争优势,并有更好的存活机会,这有助于食品发酵产品中更高的细胞数量。