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长双歧杆菌NCIMB 8809在胆盐暴露期间蛋白质表达全局变化的蛋白质组学分析。

Proteomic analysis of global changes in protein expression during bile salt exposure of Bifidobacterium longum NCIMB 8809.

作者信息

Sánchez Borja, Champomier-Vergès Marie-Christine, Anglade Patricia, Baraige Fabienne, de Los Reyes-Gavilán Clara G, Margolles Abelardo, Zagorec Monique

机构信息

Unité Flore Lactique et Environnement Carné, INRA, Domaine de Vilvert, 78350 Jouy-en-Josas, France.

出版信息

J Bacteriol. 2005 Aug;187(16):5799-808. doi: 10.1128/JB.187.16.5799-5808.2005.

Abstract

Adaptation to and tolerance of bile stress are among the main limiting factors to ensure survival of bifidobacteria in the intestinal environment of humans. The effect of bile salts on protein expression patterns of Bifidobacterium longum was examined. Protein pattern comparison of strains grown with or without bile extract allowed us to identify 34 different proteins whose expression was regulated. The majority of these proteins were induced after both a minor (0.6 g liter(-1)) and a major (1.2 g liter(-1)) exposure to bile. These include general stress response chaperones, proteins involved in transcription and translation and in the metabolism of amino acids and nucleotides, and several enzymes of glycolysis and pyruvate catabolism. Remarkably, xylulose 5-phosphate/fructose 6-phosphate phosphoketolase, the key enzyme of the so-called bifidobacterial shunt, was found to be upregulated, and the activity on fructose 6-phosphate was significantly higher for protein extracts of cells grown in the presence of bile. Changes in the levels of metabolic end products (acetate and lactate) were also detected. These results suggest that bile salts, to which bifidobacteria are naturally exposed, induce a complex physiological response rather than a single event in which proteins from many different functional categories take part. This study has extended our understanding of the molecular mechanism underlying the capacity of intestinal bifidobacteria to tolerate bile.

摘要

适应和耐受胆汁应激是确保双歧杆菌在人类肠道环境中存活的主要限制因素。研究了胆盐对长双歧杆菌蛋白质表达模式的影响。通过比较在有无胆汁提取物条件下生长的菌株的蛋白质模式,我们鉴定出34种表达受到调控的不同蛋白质。这些蛋白质中的大多数在暴露于少量(0.6 g/L)和大量(1.2 g/L)胆汁后均被诱导表达。其中包括一般应激反应伴侣蛋白、参与转录和翻译以及氨基酸和核苷酸代谢的蛋白质,以及糖酵解和丙酮酸分解代谢的几种酶。值得注意的是,发现所谓双歧分流途径的关键酶木酮糖5-磷酸/果糖6-磷酸磷酸酮醇酶被上调,并且在有胆汁存在的情况下生长的细胞的蛋白质提取物对果糖6-磷酸的活性显著更高。还检测到代谢终产物(乙酸和乳酸)水平的变化。这些结果表明,双歧杆菌自然暴露于其中的胆盐会诱导一种复杂的生理反应,而不是许多不同功能类别的蛋白质参与的单一事件。这项研究扩展了我们对肠道双歧杆菌耐受胆汁能力的分子机制的理解。

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