College of Life Science and Technology, Beijing University of Chemical Technology, No. 15 Beisanhuan East Road, Chaoyang District, Beijing 100029, People's Republic of China.
Can J Microbiol. 2013 Sep;59(9):589-97. doi: 10.1139/cjm-2012-0745. Epub 2013 Jul 9.
In the fermentative process using Saccharomyces cerevisiae to produce bioethanol, the performance of cells is often compromised by the accumulation of ethanol. However, the mechanism of how S. cerevisiae responds against ethanol stress remains elusive. In the current study, S. cerevisiae cells were cultured in YPD (yeast extract - peptone - dextrose) medium containing various concentrations of ethanol (0%, 2.5%, 5%, 7.5%, 10%, and 15% (v/v)). Compared with the control group without ethanol, the mean cell volume of S. cerevisiae decreased significantly in the presence of 7.5% and 10% ethanol after incubation for 16 h (P < 0.05), and in the presence of 15% ethanol at all 3 sampling time points (1, 8, and 16 h) (P < 0.05). The exposure of S. cerevisiae cells to ethanol also led to an increase in malonyldialdehyde content (P < 0.05) and a decrease in sulfhydryl group content (P < 0.05). Moreover, the observations through transmission electron microscopy enabled us to relate ultrastructural changes elicited by ethanol with the cellular stress physiology. Under ethanol stress, the integrity of the cell membrane was compromised. The swelling or distortion of mitochondria together with the occurrence of a single and large vacuole was correlated with the addition of ethanol. These results suggested that the cell membrane is one of the targets of ethanol, and the degeneration of mitochondria promoted the accumulation of intracellular reactive oxygen species.
在利用酿酒酵母发酵生产生物乙醇的过程中,细胞的性能常常会受到乙醇积累的影响。然而,酵母细胞如何应对乙醇胁迫的机制仍然难以捉摸。在本研究中,将酿酒酵母细胞在含有不同浓度乙醇(0%、2.5%、5%、7.5%、10%和 15%(v/v))的 YPD(酵母提取物-蛋白胨-葡萄糖)培养基中培养。与不含乙醇的对照组相比,在孵育 16 小时后,7.5%和 10%乙醇存在时,酿酒酵母的平均细胞体积明显减小(P<0.05),而在所有 3 个采样时间点(1、8 和 16 小时),15%乙醇存在时体积也明显减小(P<0.05)。酿酒酵母细胞暴露于乙醇还导致丙二醛含量增加(P<0.05)和巯基含量减少(P<0.05)。此外,通过透射电子显微镜观察,我们将乙醇引起的超微结构变化与细胞应激生理学联系起来。在乙醇胁迫下,细胞膜的完整性受到损害。线粒体的肿胀或变形以及单个大液泡的出现与乙醇的添加有关。这些结果表明,细胞膜是乙醇的靶标之一,线粒体的退化促进了细胞内活性氧的积累。