Casal M, Cardoso H, Leão C
Department of Biology, University of Minho, Braga, Portugal.
Appl Environ Microbiol. 1998 Feb;64(2):665-8. doi: 10.1128/AEM.64.2.665-668.1998.
In glucose-grown cells of Saccharomyces cerevisiae IGC 4072, acetic acid enters only by simple diffusion of the undissociated acid. In these cells, ethanol and other alkanols enhanced the passive influx of labelled acetic acid. The influx of the acid followed first-order kinetics with a rate constant that increased exponentially with the alcohol concentration, and an exponential enhancement constant for each alkanol was estimated. The intracellular concentration of labelled acetic acid was also enhanced by alkanols, and the effect increased exponentially with alcohol concentration. Acetic acid is transported across the plasma membrane of acetic acid-, lactic acid-, and ethanol-grown cells by acetate-proton symports. We found that in these cells ethanol and butanol inhibited the transport of labelled acetic acid in a noncompetitive way; the maximum transport velocity decreased with alcohol concentration, while the affinity of the system for acetate was not significantly affected by the alcohol. Semilog plots of Vmax versus alcohol concentration yielded straight lines with negative slopes from which estimates of the inhibition constant for each alkanol could be obtained. The intracellular concentration of labelled acid was significantly reduced in the presence of ethanol or butanol, and the effect increased with the alcohol concentration. We postulate that the absence of an operational carrier for acetate in glucose-grown cells of S. cerevisiae, combined with the relatively high permeability of the plasma membrane for the undissociated acid and the inability of the organism to metabolize acetic acid, could be one of the reasons why this species exhibits low tolerance to acidic environments containing ethanol.
在酿酒酵母IGC 4072以葡萄糖为生长底物的细胞中,乙酸仅通过未解离酸的简单扩散进入细胞。在这些细胞中,乙醇和其他链烷醇增强了标记乙酸的被动内流。乙酸的内流遵循一级动力学,其速率常数随酒精浓度呈指数增加,并估算了每种链烷醇的指数增强常数。链烷醇也提高了标记乙酸的细胞内浓度,且该效应随酒精浓度呈指数增加。乙酸通过乙酸-质子同向转运体跨乙酸、乳酸和乙醇生长细胞的质膜运输。我们发现,在这些细胞中,乙醇和丁醇以非竞争性方式抑制标记乙酸的运输;最大运输速度随酒精浓度降低,而该系统对乙酸的亲和力不受酒精显著影响。Vmax对酒精浓度的半对数图产生具有负斜率的直线,由此可获得每种链烷醇抑制常数的估计值。在乙醇或丁醇存在下,标记酸的细胞内浓度显著降低,且该效应随酒精浓度增加。我们推测,酿酒酵母以葡萄糖为生长底物的细胞中不存在乙酸的有效载体,再加上质膜对未解离酸的相对高渗透性以及该生物体无法代谢乙酸,可能是该物种对含乙醇的酸性环境耐受性低的原因之一。