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理解双歧杆菌乳亚种耐氧表型的分子线索。

Molecular clues to understand the aerotolerance phenotype of Bifidobacterium animalis subsp. lactis.

机构信息

Instituto de Productos Lácteos de Asturias-Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Department of Microbiology and Biochemistry of Dairy Products, Villaviciosa, Asturias, Spain.

出版信息

Appl Environ Microbiol. 2012 Feb;78(3):644-50. doi: 10.1128/AEM.05455-11. Epub 2011 Nov 18.

Abstract

Oxygen is one of the abiotic factors negatively affecting the survival of Bifidobacterium strains used as probiotics, mainly due to the induction of lethal oxidative damage. Aerobic conditions are present during the process of manufacture and storage of functional foods, and aerotolerance is a desired trait for bifidobacteria intended for use in industry. In the present study, the molecular response of Bifidobacterium animalis subsp. lactis IPLA4549 to aerobic conditions is presented. Molecular targets affected by oxygen were studied using two-dimensional electrophoresis (2DE) and quantitative reverse transcriptase (qRT) PCR. Globally, oxygen stress induced a shift in the glycolytic pathway toward the production of acetic acid with a concomitant increase in ATP formation. Several changes in the expression of genes coding for enzymes involved in redox reactions were detected, although the redox ratio remained unaltered. Interestingly, cells grown under aerobic conditions were characterized by higher activity of coproporphyrinogen III oxidase, which can directly detoxify molecular oxygen, and by higher NADH oxidase specific activity, which can oxidize NADH using hydrogen peroxide. In turn, this is in agreement with the glycolytic shift toward acetate production, in that more NADH molecules may be available due to the lower level of lactic acid formation. These findings further our ability to elucidate the mechanisms by which B. animalis copes with an oxygen-containing atmosphere.

摘要

氧气是影响益生菌双歧杆菌菌株存活的非生物因素之一,主要是因为它会引发致命的氧化损伤。在功能性食品的制造和储存过程中存在需氧条件,而耐氧性是用于工业的双歧杆菌的理想特性。在本研究中,介绍了乳双歧杆菌亚种。乳酸 IPLA4549 对需氧条件的分子反应。使用二维电泳(2-DE)和定量逆转录聚合酶链反应(qRT-PCR)研究了受氧气影响的分子靶标。总的来说,氧气应激导致糖酵解途径向产生乙酸的方向转变,同时伴随着 ATP 的形成增加。检测到与参与氧化还原反应的酶编码基因的表达发生了几种变化,尽管氧化还原比保持不变。有趣的是,在需氧条件下生长的细胞的特征是粪卟啉原 III 氧化酶活性更高,粪卟啉原 III 氧化酶可以直接解毒分子氧,并且 NADH 氧化酶的比活性更高,它可以使用过氧化氢氧化 NADH。反过来,这与糖酵解向乙酸产生的转变是一致的,因为由于乳酸形成水平较低,可能有更多的 NADH 分子可用。这些发现进一步阐明了动物双歧杆菌应对含氧大气的机制。

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