INRA-AgroParisTech, Génie et Microbiologie des Procédés Alimentaires, Thiverval Grignon, France.
Proteomics. 2009 Oct;9(20):4714-25. doi: 10.1002/pmic.200900161.
The yeast Yarrowia lipolytica has to develop dynamic metabolic adaptation mechanisms to survive within the cheese habitat. The availability of amino acids (AAs) is of major importance for microbial development and/or aroma production during cheese ripening. Using 2-D protein gel electrophoresis, we analyzed the adaptation mechanisms of Y. lipolytica for AAs limitation or supplementation in a batch culture containing lactate as a carbon source. Proteome analyses allow the identification of 34 differentially expressed proteins between the culture conditions. These analyses demonstrated that prior to the AAs addition, mainly proteins involved in the oxidative stress of the yeast were induced. Following the AAs addition, yeast cells reorganize their metabolism toward AAs catabolism and also generate a higher induction of proteins related to carbon metabolism and proteins biosynthesis. Using real-time reverse transcription PCR, we re-evaluated the expression of genes encoding proteins involved in these processes. The expression levels of the genes were in accordance with the proteomic results, with the up-regulation of genes encoding a branched-chain amino transferase BAT2, a pyruvate decarboxylase PDC6 and an Hsp70 protein SSZ1 involved in protein biosynthesis. A volatile compound analysis was also performed, and increased production of dimethyldisulfide from methionine and 3-methyl-butanal from leucine was observed in media supplemented with AAs.
酵母解脂耶氏酵母必须发展动态代谢适应机制,以在奶酪栖息地中生存。氨基酸 (AA) 的可用性对微生物在奶酪成熟过程中的发育和/或香气生产至关重要。使用 2-D 蛋白质凝胶电泳,我们分析了 Y. lipolytica 在含有乳酸作为碳源的分批培养中对 AA 限制或补充的适应机制。蛋白质组分析允许鉴定出培养条件之间的 34 种差异表达蛋白。这些分析表明,在添加 AA 之前,主要诱导与酵母氧化应激相关的蛋白质。在添加 AA 后,酵母细胞将其代谢重新组织为 AA 分解代谢,同时也会更高地诱导与碳代谢和蛋白质生物合成相关的蛋白质。使用实时逆转录 PCR,我们重新评估了参与这些过程的蛋白质编码基因的表达。基因的表达水平与蛋白质组学结果一致,编码支链氨基酸转移酶 BAT2、丙酮酸脱羧酶 PDC6 和与蛋白质生物合成相关的 Hsp70 蛋白 SSZ1 的基因上调。还进行了挥发性化合物分析,发现补充 AA 的培养基中蛋氨酸产生的二甲基二硫醚和亮氨酸产生的 3-甲基丁醛增加。