Fredlund Elisabeth, Blank Lars M, Schnürer Johan, Sauer Uwe, Passoth Volkmar
Department of Microbiology, Swedish University of Agricultural Sciences, P.O. Box 7025, SE-750 07 Uppsala, Sweden.
Appl Environ Microbiol. 2004 Oct;70(10):5905-11. doi: 10.1128/AEM.70.10.5905-5911.2004.
We investigated the regulation of the central aerobic and hypoxic metabolism of the biocontrol and non-Saccharomyces wine yeast Pichia anomala. In aerobic batch culture, P. anomala grows in the respiratory mode with a high biomass yield (0.59 g [dry weight] of cells g of glucose(-1)) and marginal ethanol, glycerol, acetate, and ethyl acetate production. Oxygen limitation, but not glucose pulse, induced fermentation with substantial ethanol production and 10-fold-increased ethyl acetate production. Despite low or absent ethanol formation, the activities of pyruvate decarboxylase and alcohol dehydrogenase were high during aerobic growth on glucose or succinate. No activation of these enzyme activities was observed after a glucose pulse. However, after the shift to oxygen limitation, both enzymes were activated threefold. Metabolic flux analysis revealed that the tricarboxylic acid pathway operates as a cycle during aerobic batch culture and as a two-branched pathway under oxygen limitation. Glucose catabolism through the pentose phosphate pathway was lower during oxygen limitation than under aerobic growth. Overall, our results demonstrate that P. anomala exhibits a Pasteur effect and not a Crabtree effect, i.e., oxygen availability, but not glucose concentration, is the main stimulus for the regulation of the central carbon metabolism.
我们研究了生物防治非酿酒酵母异常毕赤酵母的中心需氧和缺氧代谢调控。在好氧分批培养中,异常毕赤酵母以呼吸模式生长,生物量产量高(每克葡萄糖产生0.59克[干重]细胞),乙醇、甘油、乙酸和乙酸乙酯产量较低。氧气限制而非葡萄糖脉冲诱导发酵,乙醇产量大幅增加,乙酸乙酯产量增加10倍。尽管乙醇生成量低或无,但在以葡萄糖或琥珀酸为底物的好氧生长过程中,丙酮酸脱羧酶和乙醇脱氢酶的活性较高。葡萄糖脉冲后未观察到这些酶活性的激活。然而,在转变为氧气限制后,两种酶均被激活三倍。代谢通量分析表明,三羧酸途径在好氧分批培养期间作为一个循环运行,在氧气限制下作为一个双分支途径运行。在氧气限制期间,通过磷酸戊糖途径的葡萄糖分解代谢低于好氧生长时。总体而言,我们的结果表明,异常毕赤酵母表现出巴斯德效应而非克奈特效应,即氧气供应而非葡萄糖浓度是调节中心碳代谢的主要刺激因素。