Droplet Reactors with Bioluminescent Enzymes for Real-Time Water Pollution Monitoring
The 1st International Electronic Conference on Biosensors(2020)
摘要
Early detection of pollutants in wastewater, water coming out of treatment facilities, drinking water, and water for agricultural needs is a challenging problem. Effective water quality monitoring requires development of new methods for express detection of pollutants. Enzymes from bioluminescent bacteria can be used for the development of new express enzyme-based bioassay systems. This work demonstrates, for the first time, a microfluidic chip to generate emulsion droplets containing two enzymes of the bacterial bioluminescent system (luciferase and NAD(P)H:FMN-oxidoreductase) with reaction substrates. The developed chip generated “water-in-oil” emulsion droplets with a volume of 0.1 μL and a frequency of up to 12 droplets per second. A portable photomultiplier tube (PMT) was used to measure the bioluminescent signal in each individual droplet; the signal-to-noise ratio was 3000/1. The intensity of luminescence in droplets depended on the concentration of copper ions. The limit of detection (LOD) for copper sulfate was 1 mg/L. We showed that bioluminescent enzymatic reactions can be carried out in droplet reactors that can be applied for online monitoring of water quality. Thus, the suggested method of biological measurements has a good perspective for biosensing in general.
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关键词
early warning systems,luciferase,microfluidic droplet generator,emulsion,bioassay
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