Matsushika Akinori, Suzuki Toshihiro, Goshima Tetsuya, Hoshino Tamotsu
Research Institute for Sustainable Chemistry (ISC), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32 Kagamiyama, Higashi-hiroshima, Hiroshima 739-0046, Japan; Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-hiroshima, Hiroshima 739-8530, Japan.
Research Institute for Sustainable Chemistry (ISC), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32 Kagamiyama, Higashi-hiroshima, Hiroshima 739-0046, Japan.
J Biosci Bioeng. 2017 Aug;124(2):164-170. doi: 10.1016/j.jbiosc.2017.03.004. Epub 2017 May 2.
We previously showed that overexpression of IoGAS1, which was isolated from the multiple stress-tolerant yeast Issatchenkia orientalis, endows Saccharomyces cerevisiae cells with the ability to grow and ferment under acidic and high-salt conditions. The deduced amino acid sequence of the IoGAS1 gene product exhibits 60% identity with the S. cerevisiae Gas1 protein, a glycosylphosphatidylinositol-anchored protein essential for maintaining cell wall integrity. However, the functional roles of ScGAS1 in stress tolerance and pH regulation remain unclear. In the present study, we characterized ScGAS1 regarding its roles in tolerance to low pH and high salt concentrations. Transcriptional analysis indicated that, as for the IoGAS1 gene, ScGAS1 expression was pH dependent, with maximum expression at pH 3.0; the presence of salt increased endogenous expression of both GAS1 genes at almost all pH levels. These results suggested that ScGAS1, like IoGAS1, is involved in a novel acid- and salt-stress adaptation mechanism in S. cerevisiae. Overexpression of ScGAS1 in S. cerevisiae improved growth and ethanol production from glucose under acid stress without added salt, although the stress tolerance of the ScGAS1-overexpressing strain was inferior to that of the IoGAS1-overexpressing strain. However, overexpression of ScGAS1 did not result in increased tolerance of S. cerevisiae to combined acid and salt stress, even though ScGAS1 appears to be a salt-responsive gene. Thus, ScGAS1 is directly implicated in tolerance to low pH but does not confer salinity tolerance, supporting the view that ScGAS1 and IoGAS1 have overlapping yet distinct roles in stress tolerance in yeast.
我们之前发现,从多重胁迫耐受型酵母东方伊萨酵母中分离出的IoGAS1的过表达,赋予了酿酒酵母细胞在酸性和高盐条件下生长和发酵的能力。IoGAS1基因产物的推导氨基酸序列与酿酒酵母Gas1蛋白具有60%的同一性,Gas1蛋白是一种糖基磷脂酰肌醇锚定蛋白,对维持细胞壁完整性至关重要。然而,ScGAS1在胁迫耐受性和pH调节中的功能作用仍不清楚。在本研究中,我们对ScGAS1在耐受低pH和高盐浓度方面的作用进行了表征。转录分析表明,与IoGAS1基因一样,ScGAS1的表达依赖于pH,在pH 3.0时表达量最高;盐的存在几乎在所有pH水平上都增加了两个GAS1基因的内源表达。这些结果表明,ScGAS1与IoGAS1一样,参与了酿酒酵母中一种新的酸和盐胁迫适应机制。在无添加盐的酸性胁迫下,ScGAS1在酿酒酵母中的过表达改善了其生长和葡萄糖乙醇产量,尽管ScGAS1过表达菌株的胁迫耐受性低于IoGAS1过表达菌株。然而,ScGAS1的过表达并没有导致酿酒酵母对酸和盐联合胁迫的耐受性增加,尽管ScGAS1似乎是一个盐响应基因。因此,ScGAS1直接参与低pH耐受性,但不赋予盐耐受性,这支持了ScGAS1和IoGAS1在酵母胁迫耐受性中具有重叠但不同作用的观点。