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Review

Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing

by
Vadim R. Viviani
1,2,*,
Gabriel F. Pelentir
2 and
Vanessa R. Bevilaqua
3
1
Department of Physics, Chemistry and Mathematics, Federal University of São Carlos (UFSCar), Sorocaba 18052-780, Brazil
2
Graduate Program of Biotechnology, Federal University of São Carlos (UFSCar), Sorocaba 18052-780, Brazil
3
Faculty of Medical and Health Sciences, Pontifical Catholic University of São Paulo (PUC), Sorocaba 05014-901, Brazil
*
Author to whom correspondence should be addressed.
Biosensors 2022, 12(6), 400; https://doi.org/10.3390/bios12060400
Submission received: 22 April 2022 / Revised: 18 May 2022 / Accepted: 25 May 2022 / Published: 10 June 2022
(This article belongs to the Special Issue Biosensing and Drug Delivery)

Abstract

Firefly luciferases catalyze the efficient production of yellow-green light under normal physiological conditions, having been extensively used for bioanalytical purposes for over 5 decades. Under acidic conditions, high temperatures and the presence of heavy metals, they produce red light, a property that is called pH-sensitivity or pH-dependency. Despite the demand for physiological intracellular biosensors for pH and heavy metals, firefly luciferase pH and metal sensitivities were considered drawbacks in analytical assays. We first demonstrated that firefly luciferases and their pH and metal sensitivities can be harnessed to estimate intracellular pH variations and toxic metal concentrations through ratiometric analysis. Using Macrolampis sp2 firefly luciferase, the intracellular pH could be ratiometrically estimated in bacteria and then in mammalian cells. The luciferases of Macrolampis sp2 and Cratomorphus distinctus fireflies were also harnessed to ratiometrically estimate zinc, mercury and other toxic metal concentrations in the micromolar range. The temperature was also ratiometrically estimated using firefly luciferases. The identification and engineering of metal-binding sites have allowed the development of novel luciferases that are more specific to certain metals. The luciferase of the Amydetes viviani firefly was selected for its special sensitivity to cadmium and mercury, and for its stability at higher temperatures. These color-tuning luciferases can potentially be used with smartphones for hands-on field analysis of water contamination and biochemistry teaching assays. Thus, firefly luciferases are novel color-tuning sensors for intracellular pH and toxic metals. Furthermore, a single luciferase gene is potentially useful as a dual bioluminescent reporter to simultaneously report intracellular ATP and/or luciferase concentrations luminometrically, and pH or metal concentrations ratiometrically, providing a useful tool for real-time imaging of intracellular dynamics and stress.
Keywords: bioluminescence; pH indication; heavy metals; cadmium; mercury; ratiometric biosensors; bioimaging bioluminescence; pH indication; heavy metals; cadmium; mercury; ratiometric biosensors; bioimaging

Share and Cite

MDPI and ACS Style

Viviani, V.R.; Pelentir, G.F.; Bevilaqua, V.R. Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing. Biosensors 2022, 12, 400. https://doi.org/10.3390/bios12060400

AMA Style

Viviani VR, Pelentir GF, Bevilaqua VR. Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing. Biosensors. 2022; 12(6):400. https://doi.org/10.3390/bios12060400

Chicago/Turabian Style

Viviani, Vadim R., Gabriel F. Pelentir, and Vanessa R. Bevilaqua. 2022. "Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing" Biosensors 12, no. 6: 400. https://doi.org/10.3390/bios12060400

APA Style

Viviani, V. R., Pelentir, G. F., & Bevilaqua, V. R. (2022). Bioluminescence Color-Tuning Firefly Luciferases: Engineering and Prospects for Real-Time Intracellular pH Imaging and Heavy Metal Biosensing. Biosensors, 12(6), 400. https://doi.org/10.3390/bios12060400

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