Landolfo Sara, Politi Huguette, Angelozzi Daniele, Mannazzu Ilaria
Dipartimento di Scienze degli Alimenti, Università Politecnica delle Marche, Via Brecce Bianche, 60131, Ancona, Italy.
Biochim Biophys Acta. 2008 Jun;1780(6):892-8. doi: 10.1016/j.bbagen.2008.03.008. Epub 2008 Mar 18.
To further elucidate the impact of fermentative stress on Saccharomyces cerevisiae wine strains, we have here evaluated markers of oxidative stress, oxidative damage and antioxidant response in four oenological strains of S. cerevisiae, relating these to membrane integrity, ethanol production and cell viability during fermentation in high-sugar-containing medium. The cells were sampled at different fermentation stages and analysed by flow cytometry to evaluate membrane integrity and accumulation of reactive oxygen species (ROS). At the same time, catalase and superoxide dismutase activities, trehalose accumulation, and protein carbonylation and degradation were measured. The results indicate that the stress conditions occurring during hypoxic fermentation in high-sugar-containing medium result in the production of ROS and trigger an antioxidant response. This involves superoxide dismutase and trehalose for the protection of cell structures from oxidative damage, and protein catabolism for the removal of damaged proteins. Cell viability, membrane integrity and ethanol production depend on the extent of oxidative damage to cellular components. This is, in turn, related to the 'fitness' of each strain, which depends on the contribution of individual cells to ROS accumulation and scavenging. These findings highlight that the differences in individual cell resistances to ROS contribute to the persistence of wine strains during growth under unfavourable culture conditions, and they provide further insights into our understanding of yeast behaviour during industrial fermentation.
为了进一步阐明发酵应激对酿酒酵母葡萄酒菌株的影响,我们在此评估了四株酿酒酵母葡萄酒菌株中的氧化应激、氧化损伤和抗氧化反应标志物,并将这些与含高糖培养基发酵过程中的膜完整性、乙醇产生和细胞活力相关联。在不同发酵阶段对细胞进行取样,并通过流式细胞术分析以评估膜完整性和活性氧(ROS)的积累。同时,测定过氧化氢酶和超氧化物歧化酶活性、海藻糖积累以及蛋白质羰基化和降解情况。结果表明,在含高糖培养基中的缺氧发酵过程中出现的应激条件会导致ROS的产生并引发抗氧化反应。这涉及超氧化物歧化酶和海藻糖来保护细胞结构免受氧化损伤,以及蛋白质分解代谢来去除受损蛋白质。细胞活力、膜完整性和乙醇产生取决于细胞成分氧化损伤的程度。反过来,这又与每个菌株的“适应性”相关,而这又取决于单个细胞对ROS积累和清除的贡献。这些发现突出表明,单个细胞对ROS的抗性差异有助于葡萄酒菌株在不利培养条件下生长期间的存活,并且它们为我们理解工业发酵过程中的酵母行为提供了进一步的见解。