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在不同需氧水平下对大肠杆菌 MG1655 及其 frdA 和 sdhC 突变体的特性进行分析。

Characterization of E. coli MG1655 and frdA and sdhC mutants at various aerobiosis levels.

机构信息

MPI für Dynamik Komplexer Technischer Systeme, Germany.

出版信息

J Biotechnol. 2011 Jun 10;154(1):35-45. doi: 10.1016/j.jbiotec.2011.03.015. Epub 2011 Mar 30.

Abstract

Depending on the availability of oxygen, Escherichia coli is able to switch between aerobic respiratory metabolism and anaerobic mixed acid fermentation. An important, yet understudied, metabolic mode is the micro-aerobic metabolism at intermediate oxygen availabilities. The relationship between oxygen input, physiology and gene expression of E. coli MG1655 and two isogenic mutants lacking succinate dehydrogenase (SDH) and fumarate reductase (FRD) activities was analyzed at different aerobiosis levels. Growth rate and cell yield were very similar to the parent strain. By-product formation was altered in the sdhC mutant to higher acetic acid and glutamate production in batch cultures. In continuous cultures with defined oxygen input gene expression analysis revealed a dependency of many catabolic genes to aerobiosis. Acetate excretion was still detectable under aerobic conditions in the sdhC mutant; the frdA mutant lacked anaerobic succinate excretion. Anaerobic repression of the sdh operon was diminished in the frdA strain, possibly to allow SDH to partially replace FRD. The experiments illustrate the remarkable adaptability of E. coli physiology-to compensate for the absence of important metabolic genes by altering carbon flux and/or gene expression such that there are only minor changes in growth capability across the aerobiosis range.

摘要

根据氧气的供应情况,大肠杆菌能够在需氧呼吸代谢和厌氧混合酸发酵之间切换。一种重要但研究不足的代谢模式是中间氧气供应时的微需氧代谢。在不同需氧水平下,分析了大肠杆菌 MG1655 及其两个缺乏琥珀酸脱氢酶 (SDH) 和延胡索酸还原酶 (FRD) 活性的同基因突变体的氧气输入、生理学和基因表达之间的关系。与亲本菌株相比,生长速率和细胞产率非常相似。在分批培养中,sdhC 突变体的副产物形成发生改变,导致乙酸和谷氨酸产量更高。在具有定义氧气输入的连续培养中,基因表达分析显示许多分解代谢基因对需氧性有依赖性。在 sdhC 突变体中,仍可检测到有氧条件下的乙酸排泄;frdA 突变体缺乏厌氧琥珀酸排泄。frdA 菌株中 sdh 操纵子的厌氧抑制作用减弱,可能允许 SDH 部分替代 FRD。这些实验说明了大肠杆菌生理学的惊人适应性——通过改变碳通量和/或基因表达来补偿重要代谢基因的缺失,从而在整个需氧范围中,生长能力只有很小的变化。

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