Feria-Gervasio David, Mouret Jean-Roch, Gorret Nathalie, Goma Gérard, Guillouet Stéphane E
UMR5504, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, CNRS, INRA, INSA, 135 Avenue de Rangueil, 31077, Toulouse Cedex, France.
Appl Microbiol Biotechnol. 2008 Feb;78(2):319-31. doi: 10.1007/s00253-007-1161-z. Epub 2007 Oct 2.
This work aimed to study the transition from respiratory to fermentative metabolism in Saccharomyces cerevisiae CEN.PK 113-7D and more specifically to evaluate the implication of the acetyl-coenzymeA-derived carbon transport from cytosol to mitochondria in the onset of the metabolic shift. The strategy consisted in introducing, during aerobic glucose-limited chemostat (D = 0.16 h(-1)), [corrected] a local perturbation around the step to be studied by the addition of cosubstrate and in analyzing the consequences of such a perturbation on the metabolic transition. Oleic acid and L: -carnitine were among the tested cosubstrates because they were known to stimulate enzymes implicated in the acetyl-coenzymeA transport between the different cell compartments, such as the carnitine acetyl transferases. The metabolic transition was then comparatively quantified in sole glucose and in glucose/oleic acid chemostats in presence/absence of L: -carnitine after a pulse of glucose. Feeding the culture with oleic acid (D (ole) = 0.0041 and 0.0073 h(-1)) [corrected] led to a delay in the onset of the metabolic shift (up to 15 min), a 33% decrease in the ethanol production and a redirection of the carbon flux toward biomass production. The data clearly showed a modulation of the carbon distribution among respiration and fermentation, in favor of a decrease in the "short-term" Crabtree effect by the oleic acid.
这项工作旨在研究酿酒酵母CEN.PK 113 - 7D中从呼吸代谢到发酵代谢的转变,更具体地说是评估在代谢转变开始时,乙酰辅酶A衍生的碳从细胞质向线粒体转运的影响。该策略包括在有氧葡萄糖限制恒化器(D = 0.16 h⁻¹)期间,通过添加共底物在要研究的步骤周围引入局部扰动,并分析这种扰动对代谢转变的影响。油酸和L-肉碱是测试的共底物之一,因为已知它们能刺激参与不同细胞区室间乙酰辅酶A转运的酶,如肉碱乙酰转移酶。在葡萄糖脉冲后,分别在单独葡萄糖以及添加/不添加L-肉碱的葡萄糖/油酸恒化器中,对代谢转变进行了比较定量。用油酸(D(ole)=0.0041和0.0073 h⁻¹)培养导致代谢转变开始延迟(长达15分钟),乙醇产量降低33%,并且碳通量重新导向生物量生产。数据清楚地表明了呼吸和发酵之间碳分布的调节,有利于油酸降低“短期”克奈特效应。