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Proteome Analyses of Heme-Dependent Respiration in Lactococcus lactis: Involvement of the Proteolytic System

机译:乳酸乳球菌血红素依赖性呼吸的蛋白质组分析:蛋白水解系统的参与。

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摘要

Sugar fermentation was long considered the sole means of energy metabolism available to lactic acid bacteria. We recently showed that metabolism of Lactococcus lactis shifts progressively from fermentation to respiration during growth when oxygen and heme are available. To provide insights into this phenomenon, we compared the proteomic profiles of L. lactis under fermentative and respiratory growth conditions in rich medium. We identified 21 proteins whose levels differed significantly between these conditions. Two major groups of proteins were distinguished, one involved in carbon metabolism and the second in nitrogen metabolism. Unexpectedly, enzymes of the proteolytic system (PepO1 and PepC) which are repressed in rich medium in fermentation growth were induced under respiratory conditions despite the availability of free amino acids. A triple mutant (dtpT dtpP oppA) deficient in oligopeptide transport displayed normal respiration, showing that increased proteolytic activity is not an absolute requirement for respiratory metabolism. Transcriptional analysis confirmed that pepO1 is induced under respiration-permissive conditions. This induction was independent of CodY, the major regulator of proteolytic functions in L. lactis. We also observed that pepO1 induction is redox sensitive. In a codY mutant, pepO1 expression was increased twofold in aeration and eightfold in respiration-permissive conditions compared to static conditions. These observations suggest that new regulators activate proteolysis in L. lactis, which help to maintain the energetic needs of L. lactis during respiration.
机译:长期以来,糖发酵一直被认为是乳酸菌可利用的能量代谢的唯一手段。我们最近表明,乳酸和乳酸乳球菌的代谢在生长过程中从发酵到呼吸在氧气和血红素可用时逐渐转移。为了提供对该现象的见解,我们比较了乳酸菌在富营养培养基中在发酵和呼吸生长条件下的蛋白质组学特征。我们鉴定了21种蛋白质,其水平在这些条件之间差异显着。区分了两大类蛋白质,一类涉及碳代谢,另一类涉及氮代谢。出乎意料的是,尽管有游离氨基酸,但在呼吸条件下仍会在发酵条件下诱导在发酵培养基中被抑制的蛋白水解系统的酶(PepO1和PepC)。缺乏寡肽转运的三重突变体(dtpT dtpP oppA)显示正常的呼吸作用,表明蛋白水解活性的提高并不是呼吸代谢的绝对要求。转录分析证实pepO1是在允许呼吸的条件下诱导的。这种诱导独立于CodY,后者是乳酸乳球菌中蛋白水解功能的主要调节剂。我们还观察到pepO1诱导对氧化还原敏感。与静态条件相比,在codY突变体中,pepO1表达在通气条件下增加了两倍,在呼吸允许条件下增加了八倍。这些观察结果表明,新的调节剂可激活乳酸乳球菌的蛋白水解作用,从而有助于维持呼吸过程中乳酸乳球菌的能量需求。

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