Bioluminescent enzymatic rapid assay of water integral toxicity
A bioluminescent rapid method was developed to estimate the integral toxicity of natural and wastewater. This method is based on registering the effect of the polluted water sample on the parameters of the bioluminescent reaction catalyzed by the multi-component reagent containing NADH:FMN oxidoreductase, luciferase, and their substrates co-immobilized in a starch carrier. Several ways to increase the method's sensitivity to toxic substances were suggested; conditions were selected to make it possible to determine, with maximum efficiency, the content of toxic substances corresponding to a certain maximum permissible concentration. The sensitivity of soluble and immobilized coupled enzymatic systems to a series of organic pollutants (phenols, quinones, and salts of heavy metals) was compared. It was shown that the reagent is the most sensitive to the effect of phenols and quinones. The method was tested during analysis of the wastewater from a pulp and paper plant and can be used for biotesting in both laboratory and field conditions.
KeywordsIntegral water toxicity Ecological monitoring Bioluminescence Luciferase Immobilization
The work was financially supported by the Ministry of Education and Science Federal (government contract no. 14.A18.21.1911), the Agency for Science and Innovations (government contract no. 02.740.11.0766), the Government of the Russian Federation (contract no. 11. G34.31.058), the Russian Academy of Sciences (Program “Molecular and Cell Biology,” grant no. 6.2), and the President of the Russian Federation (Grant “Leading Scientific School” 3951.2012.4).
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