Department of Chemical and Bioprocess Engineering, School of Engineering, Pontificia Universidad Católica de Chile, Santiago, Chile.
Appl Environ Microbiol. 2012 Dec;78(23):8340-52. doi: 10.1128/AEM.02305-12. Epub 2012 Sep 21.
Discrete additions of oxygen play a critical role in alcoholic fermentation. However, few studies have quantitated the fate of dissolved oxygen and its impact on wine yeast cell physiology under enological conditions. We simulated the range of dissolved oxygen concentrations that occur after a pump-over during the winemaking process by sparging nitrogen-limited continuous cultures with oxygen-nitrogen gaseous mixtures. When the dissolved oxygen concentration increased from 1.2 to 2.7 μM, yeast cells changed from a fully fermentative to a mixed respirofermentative metabolism. This transition is characterized by a switch in the operation of the tricarboxylic acid cycle (TCA) and an activation of NADH shuttling from the cytosol to mitochondria. Nevertheless, fermentative ethanol production remained the major cytosolic NADH sink under all oxygen conditions, suggesting that the limitation of mitochondrial NADH reoxidation is the major cause of the Crabtree effect. This is reinforced by the induction of several key respiratory genes by oxygen, despite the high sugar concentration, indicating that oxygen overrides glucose repression. Genes associated with other processes, such as proline uptake, cell wall remodeling, and oxidative stress, were also significantly affected by oxygen. The results of this study indicate that respiration is responsible for a substantial part of the oxygen response in yeast cells during alcoholic fermentation. This information will facilitate the development of temporal oxygen addition strategies to optimize yeast performance in industrial fermentations.
氧气的离散添加在酒精发酵中起着关键作用。然而,很少有研究定量研究溶解氧的命运及其在酿造条件下对葡萄酒酵母细胞生理学的影响。我们通过用含氧氮混合气体对氮限制连续培养物进行鼓泡来模拟酿造过程中泵入后发生的溶解氧浓度范围。当溶解氧浓度从 1.2μM 增加到 2.7μM 时,酵母细胞从完全发酵代谢转变为混合呼吸发酵代谢。这种转变的特征是三羧酸循环(TCA)的操作发生转变,以及 NADH 从细胞质穿梭到线粒体的激活。然而,在所有氧气条件下,发酵乙醇生产仍然是细胞质 NADH 的主要汇,这表明线粒体 NADH 再氧化的限制是 Crabtree 效应的主要原因。尽管糖浓度很高,但氧气诱导了几个关键呼吸基因的表达,这证实了这一点,表明氧气可以克服葡萄糖抑制。与其他过程相关的基因,如脯氨酸摄取、细胞壁重塑和氧化应激,也受到氧气的显著影响。这项研究的结果表明,呼吸是酵母细胞在酒精发酵过程中对氧气反应的重要组成部分。这些信息将有助于开发时间性氧气添加策略,以优化工业发酵中酵母的性能。