Tsai P S, Kallio P T, Bailey J E
Institut für Biotechnologie, Eidgenössische Technische Hochschule, Zürich, Switzerland.
Biotechnol Prog. 1995 May-Jun;11(3):288-93. doi: 10.1021/bp00033a008.
The oxygen-regulated promoter (Pvhb) of the Vitreoscilla hemoglobin gene has been applied to direct high-level expression of several cloned proteins, including Vitreoscilla hemoglobin (VHb), which improves productivity of many aerobic processes. In an effort to gain a better understanding of the regulation of Pvhb, and to guide further optimization of this technology, we investigated whether the Escherichia coli global regulatory molecules Fnr and the Arc system (ArcA and ArcB), which control the expression of various genes under either aerobic or anaerobic conditions, also regulate Pvhb activity in E. coli. The activity of Pvhb and the expression of VHb in E. coli were activated by Fnr, but were relatively unaffected by the Arc system under microaerobic conditions (DO less than 2% air saturation). We also examined the possibility of VHb affecting cytochrome d promoter activity during microaerobiosis. The presence of VHb increased the activity of beta-galactosidase from a cytochrome d promoter-lacZ fusion by 1.5-fold. This indicates that VHb affects oxygen-regulated transcription of E. coli genes and may contribute to the modified physiology observed in VHb-expressing E. coli.
透明颤菌血红蛋白基因的氧调节启动子(Pvhb)已被用于指导多种克隆蛋白的高水平表达,包括透明颤菌血红蛋白(VHb),它提高了许多需氧过程的生产力。为了更好地理解Pvhb的调控机制,并指导该技术的进一步优化,我们研究了在有氧或无氧条件下控制各种基因表达的大肠杆菌全局调控分子Fnr和Arc系统(ArcA和ArcB)是否也调节大肠杆菌中Pvhb的活性。在微需氧条件下(溶解氧低于2%空气饱和度),Fnr激活了大肠杆菌中Pvhb的活性和VHb的表达,但Arc系统对其影响相对较小。我们还研究了在微需氧条件下VHb影响细胞色素d启动子活性的可能性。VHb的存在使细胞色素d启动子-lacZ融合体的β-半乳糖苷酶活性提高了1.5倍。这表明VHb影响大肠杆菌基因的氧调节转录,并可能导致在表达VHb的大肠杆菌中观察到的生理变化。