Rosenfeld Eric, Beauvoit Bertrand, Blondin Bruno, Salmon Jean-Michel
Laboratoire de Microbiologie et de Technologie des Fermentations, Unité Mixte de Recherches Sciences pour l'oenologie, Institut National de la Recherche Agronomique, F-34060 Montpellier Cedex 1, France.
Appl Environ Microbiol. 2003 Jan;69(1):113-21. doi: 10.1128/AEM.69.1.113-121.2003.
The anaerobic growth of the yeast Saccharomyces cerevisiae normally requires the addition of molecular oxygen, which is used to synthesize sterols and unsaturated fatty acids (UFAs). A single oxygen pulse can stimulate enological fermentation, but the biochemical pathways involved in this phenomenon remain to be elucidated. We showed that the addition of oxygen (0.3 to 1.5 mg/g [dry mass] of yeast) to a lipid-depleted medium mainly resulted in the synthesis of the sterols and UFAs required for cell growth. However, the addition of oxygen during the stationary phase in a medium containing excess ergosterol and oleic acid increased the specific fermentation rate, increased cell viability, and shortened the fermentation period. Neither the respiratory chain nor de novo protein synthesis was required for these medium- and long-term effects. As de novo lipid synthesis may be involved in ethanol tolerance, we studied the effect of oxygen addition on sterol and UFA auxotrophs (erg1 and ole1 mutants, respectively). Both mutants exhibited normal anaerobic fermentation kinetics. However, only the ole1 mutant strain responded to the oxygen pulse during the stationary phase, suggesting that de novo sterol synthesis is required for the oxygen-induced increase of the specific fermentation rate. In conclusion, the sterol pathway appears to contribute significantly to the oxygen consumption capacities of cells under anaerobic conditions. Nevertheless, we demonstrated the existence of alternative oxygen consumption pathways that are neither linked to the respiratory chain nor linked to heme, sterol, or UFA synthesis. These pathways dissipate the oxygen added during the stationary phase, without affecting the fermentation kinetics.
酿酒酵母的厌氧生长通常需要添加分子氧,用于合成甾醇和不饱和脂肪酸(UFA)。单次氧脉冲可刺激葡萄酒发酵,但这种现象所涉及的生化途径仍有待阐明。我们发现,向脂质缺乏的培养基中添加氧气(0.3至1.5毫克/克[干重]酵母)主要导致细胞生长所需的甾醇和UFA的合成。然而,在含有过量麦角固醇和油酸的培养基的稳定期添加氧气,可提高比发酵速率、提高细胞活力并缩短发酵周期。这些中、长期效应既不需要呼吸链也不需要从头合成蛋白质。由于从头合成脂质可能与乙醇耐受性有关,我们研究了添加氧气对甾醇和UFA营养缺陷型(分别为erg1和ole1突变体)的影响。两种突变体均表现出正常的厌氧发酵动力学。然而,只有ole1突变株在稳定期对氧脉冲有反应,这表明甾醇的从头合成是氧诱导比发酵速率增加所必需的。总之,甾醇途径似乎对厌氧条件下细胞的耗氧能力有显著贡献。尽管如此,我们证明了存在既不与呼吸链相连也不与血红素、甾醇或UFA合成相关的替代耗氧途径。这些途径消耗稳定期添加的氧气,而不会影响发酵动力学。