Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Biotechnol J. 2020 Jun;15(6):e1900353. doi: 10.1002/biot.201900353.
Oxygen supply is one of the major factors determining the metabolic state in microorganisms, and it affects the productivity of various compounds during bioproduction. In Escherichia coli (E. coli), the expression levels of numerous metabolic genes are regulated by transcription factors in response to changes in environmental oxygen conditions. Even at a same dissolved oxygen concentration, the amount of available oxygen changes depending on the oxygen transfer coefficient. However, it is not known whether E. coli is able to sense differences in the oxygen consumption rate. Therefore, the present study, is focused on the role of the transcription factor ArcA in the oxygen response of E. coli and investigated the relationship between ArcA activity and the oxygen consumption rate. To evaluate the activity of ArcA, a sensor plasmid expressing fluorescent protein under the control of the icd promoter, which is regulated by ArcA, is designed. E. coli containing the sensor plasmid is grown in continuous cultures with different oxygen supplies under different dilution rates. Although there is no correlation between ArcA activity and dissolved oxygen concentration, a strong negative correlation between ArcA activity and the specific oxygen consumption rate (R > 0.93) is observed.
供氧是决定微生物代谢状态的主要因素之一,它会影响生物生产过程中各种化合物的产量。在大肠杆菌(E. coli)中,许多代谢基因的表达水平受转录因子的调控,以响应环境氧气条件的变化。即使在相同的溶解氧浓度下,可用氧气的量也会因氧气传递系数的不同而发生变化。然而,目前尚不清楚大肠杆菌是否能够感知耗氧量的差异。因此,本研究专注于转录因子 ArcA 在大肠杆菌氧气响应中的作用,并研究了 ArcA 活性与耗氧量之间的关系。为了评估 ArcA 的活性,设计了一个表达受 ArcA 调控的 icd 启动子控制的荧光蛋白的传感器质粒。含有该传感器质粒的大肠杆菌在不同稀释率下的不同供氧条件下进行连续培养。虽然 ArcA 活性与溶解氧浓度之间没有相关性,但观察到 ArcA 活性与比耗氧速率(R > 0.93)之间存在很强的负相关性。