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氧气对大肠杆菌ArcA和FNR调控系统及代谢反应的影响。

Effect of oxygen on the Escherichia coli ArcA and FNR regulation systems and metabolic responses.

作者信息

Levanon Sagit Shalel, San Ka-Yiu, Bennett George N

机构信息

Department of Biochemistry and Cell Biology, MS 140, 6100 Main St., Rice University, Houston, TX 77005-1892, USA.

出版信息

Biotechnol Bioeng. 2005 Mar 5;89(5):556-64. doi: 10.1002/bit.20381.

Abstract

Escherichia coli has several elaborate sensing mechanisms for response to the availability of oxygen and the presence of other electron acceptors. The adaptive responses are coordinated by a group of global regulators, which include the one-component Fnr protein, and the two-component Arc system. To quantitate the contribution of Arc and Fnr-dependent regulation in catabolism, arcA and fnr mutant strains were constructed using the recently developed lambda derived recombination system. The metabolic activity of wildtype E. coli, an arcA mutant, an fnr mutant, and a double arcA-fnr mutant, via the fermentative pathways in glucose-limited cultures and different oxygen concentrations was studied in chemostat cultures at steady state. It was found that the most significant role of ArcA is under microaerobic conditions, while that of FNR is under more strictly anaerobic conditions. The FNR protein is normally inactive during microaerobic conditions. However, our results indicate that in the arcA mutant strain the cells behave as if a higher level of the FNR regulator is in the activated form compared to the wildtype strain during the transition from aerobic to microanaerobic growth. The results show a significant increase in the flux through pyruvate formate lyase (PFL) in the presence of oxygen. The activity of FNR-regulated pathways in the arcA mutant strain is correlated with the high redox potential obtained under microaerobic growth.

摘要

大肠杆菌具有多种精细的传感机制,用于响应氧气的可利用性以及其他电子受体的存在。这些适应性反应由一组全局调节因子协调,其中包括单组分Fnr蛋白和双组分Arc系统。为了定量Arc和Fnr依赖性调节在分解代谢中的作用,利用最近开发的λ衍生重组系统构建了arcA和fnr突变株。在恒化器培养的稳态条件下,研究了野生型大肠杆菌、arcA突变体、fnr突变体和arcA-fnr双突变体在葡萄糖限制培养物中以及不同氧气浓度下通过发酵途径的代谢活性。结果发现,ArcA的最重要作用是在微需氧条件下,而FNR的作用是在更严格的厌氧条件下。FNR蛋白在微需氧条件下通常无活性。然而,我们的结果表明,在从有氧生长向微厌氧生长转变过程中,与野生型菌株相比,arcA突变株中的细胞表现得就好像更高水平的FNR调节因子处于激活形式。结果表明在有氧条件下,通过丙酮酸甲酸裂解酶(PFL)的通量显著增加。arcA突变株中FNR调节途径的活性与微需氧生长下获得的高氧化还原电位相关。

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