Principles for Construction of Bioluminescent Enzyme Biotests for Analysis of Complex Media

Kalyabina, Valeriya P.; Esimbekova, Elena N.; Torgashina, I. G.; Kopylova, Kseniya V.; Kratasyuk, Valentina A.

doi:10.1134/s1607672919020042

Cited by 5 publications

(1 citation statement)

References 9 publications

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“…Under controlled laboratory conditions, such enzyme inhibition-based assays are used to detect toxicants in the tested medium [15][16][17][18][19]. This strategy is based on the specific inhibition of the enzymes by toxic substances, which is proportional to the amounts of toxicants in the sample, and it has been successfully used to develop procedures for water, air, soil, and food monitoring [20][21][22][23]. The in vitro enzyme-based assays have a great number of advantages, including the weaker effect of external factors on the experimental procedure and the higher sensitivity of the assays; the assays are rapid and simple to perform, and their results are reproducible; and their biological relevance is retained as well [14].…”

Section: Introductionmentioning

confidence: 99%

The Effects of Commercial Pesticide Formulations on the Function of In Vitro and In Vivo Assay Systems: A Comparative Analysis

et al. 2022

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Pesticides are commonly used in agriculture and are an important factor of food security for humankind. However, the overuse of pesticides can harm non-target organisms, and, thus, it is vital to comprehensively study their effects on the different metabolic pathways of living organisms. In the present study, enzyme-inhibition-based assays have been used to investigate the effects of commercial pesticide formulations on the key enzymes of the organisms, which catalyze a wide variety of metabolic reactions (protein catabolism, lactic acid fermentation, alcohol metabolism, the conduction of nerve impulses, etc.). Assay conditions have been optimized, and the limitations of the methods used in the study, which are related to the choice of the solvent for commercial pesticide formulations and optical effects occurring when commercial pesticide formulations are mixed with solutions of enzymes and substrates of assay systems, have been revealed. The effects of commercial pesticide formulations on simple chemoenzymatic assay systems (single-enzyme reactions) have been compared to their effects on complex multicomponent molecular systems (multi-enzyme reactions) and organisms (luminescent bacterium). The in vitro assay systems have shown higher sensitivity to pesticide exposure than the in vivo assay system. The sensitivity of the in vitro assay systems increases with the elongation of the chain of conjugated chemoenzymatic reactions. The effects exerted by commercial pesticide formulations with the same active ingredient but produced by different manufacturers on assay system functions have been found to differ from each other.

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