Liu Pei-Han, Urban Pawel L
Department of Applied Chemistry, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan.
Department of Applied Chemistry, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 30013, Taiwan.
Anal Biochem. 2017 Dec 15;539:54-59. doi: 10.1016/j.ab.2017.10.007. Epub 2017 Oct 13.
The temporal effects of luciferase reaction luminescence have only been discussed in the context of light intensity (flash vs. glow). However, alterations in the color of the light emitted over the course of the luciferase reaction have not been reported. Here, we show a temporal change in the light color emitted during the reaction catalyzed by unmodified firefly luciferase when concentrations of one of the substrates, adenosine triphosphate (ATP), are gradually increased. The temporal color change from green to red occurs within the first few minutes of the luciferase reaction when an ATP-containing solution is either added or synthesized in situ with the aid of an autocatalytic reaction occurring simultaneously. This color change is not accompanied by pH changes. An analysis of the red and green channels demonstrates dissimilar kinetics, suggesting the co-existence of two or more temporally shifted luminescence pathways. The implications of these findings might improve dual-color biosensing/imaging protocols and influence the engineering of biophotonic systems.
荧光素酶反应发光的时间效应仅在光强度（闪光与辉光）的背景下进行过讨论。然而，尚未有关于荧光素酶反应过程中发射光颜色变化的报道。在此，我们展示了在未修饰的萤火虫荧光素酶催化的反应中，当其中一种底物三磷酸腺苷（ATP）的浓度逐渐增加时，发射光颜色的时间变化。当添加含ATP的溶液或借助同时发生的自催化反应原位合成ATP时，荧光素酶反应的最初几分钟内会出现从绿色到红色的时间颜色变化。这种颜色变化不伴随pH值变化。对红色和绿色通道的分析表明动力学不同，这表明存在两条或更多条时间上有偏移的发光途径。这些发现的意义可能会改进双色生物传感/成像方案，并影响生物光子系统的工程设计。
