Тип публикации: статья из журнала

Год издания: 2022

Идентификатор DOI: 10.3390/mi13071146

Ключевые слова: bioluminescence, chemoenzymatic system, droplet microfluidics, luciferase, sensing

Аннотация: The rapid detection of pollutants in water can be performed with enzymatic probes, the catalytic light-emitting activity of which decreases in the presence of many types of pollutants. Herein, we present a microfluidic system for continuous chemoenzymatic biosensing that generates emulsion droplets containing two enzymes of the bacПоказать полностьюterial bioluminescent system (luciferase and NAD(P)H:FMN–oxidoreductase) with substrates required for the reaction. The developed chip generates “water-in-oil” emulsion droplets with a volume of 0.1 (Formula presented.) L and a frequency of up to 12 drops per minute as well as provides the efficient mixing of reagents in droplets and their distancing. The bioluminescent signal from each individual droplet was measured by a photomultiplier tube with a signal-to-noise ratio of up to 3000/1. The intensity of the luminescence depended on the concentration of the copper sulfate with the limit of its detection of 5 (Formula presented.) M. It was shown that bioluminescent enzymatic reactions could be carried out in droplet reactors in dispersed streams. The parameters and limitations required for the bioluminescent reaction to proceed were also studied. Hereby, chemoenzymatic sensing capabilities powered by a droplet microfluidics manipulation technique may serve as the basis for early-warning online water pollution systems. © 2022 by the authors. The rapid detection of pollutants in water can be performed with enzymatic probes, the catalytic light-emitting activity of which decreases in the presence of many types of pollutants. Herein, we present a microfluidic system for continuous chemoenzymatic biosensing that generates emulsion droplets containing two enzymes of the bacterial bioluminescent system (luciferase and NAD(P)H:FMN–oxidoreductase) with substrates required for the reaction. The developed chip generates “water-in-oil” emulsion droplets with a volume of 0.1 (Formula presented.) L and a frequency of up to 12 drops per minute as well as provides the efficient mixing of reagents in droplets and their distancing. The bioluminescent signal from each individual droplet was measured by a photomultiplier tube with a signal-to-noise ratio of up to 3000/1. The intensity of the luminescence depended on the concentration of the copper sulfate with the limit of its detection of 5 (Formula presented.) M. It was shown that bioluminescent enzymatic reactions could be carried out in droplet reactors in dispersed streams. The parameters and limitations required for the bioluminescent reaction to proceed were also studied. Hereby, chemoenzymatic sensing capabilities powered by a droplet microfluidics manipulation technique may serve as the basis for early-warning online water pollution systems. © 2022 by the authors.

Журнал: Micromachines

Выпуск журнала: Vol. 13, Is. 7

Номера страниц: 1146

ISSN журнала: 2072666X

Издатель: MDPI

• Yakimov A.S. (Laboratory of Physical and Chemical Technologies for the Development of Hard-to-Recover Hydrocarbon Reserves, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation)
• Denisov I.A. (Laboratory of Physical and Chemical Technologies for the Development of Hard-to-Recover Hydrocarbon Reserves, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation; Laboratory of Bioluminescent Biotechnologies, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation)
• Bukatin A.S. (Laboratory of Renewable Energy Sources, Alferov University, Saint Petersburg, 194021, Russian Federation; Institute for Analytical Instrumentation RAS, Saint Petersburg, 194021, Russian Federation)
• Lukyanenko K.A. (Laboratory of Bioluminescent Biotechnologies, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation; Laboratory for Biomolecular and Medical Technologies, Prof. V.F. Voino-Yasenetsky Krasnoyarsk State Medical University, Krasnoyarsk, 660022, Russian Federation; Laboratory for Digital Controlled Drugs and Theranostics, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, 660036, Russian Federation)
• Belousov K.I. (Laboratory of Renewable Energy Sources, Alferov University, Saint Petersburg, 194021, Russian Federation)
• Kukhtevich I.V. (Institute of Silicate Chemistry of RAS, Saint Petersburg, 199034, Russian Federation; Institute of Functional Epigenetics, Helmholtz Zentrum München, Neuherberg, 85764, Germany)
• Esimbekova E.N. (Institute of Biophysics SB RAS, Krasnoyarsk, 660036, Russian Federation; Department of Biophysics, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation)
• Evstrapov A.A. (Institute for Analytical Instrumentation RAS, Saint Petersburg, 194021, Russian Federation)
• Belobrov P.I. (Department of Biophysics, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation)

• Scopus