Enzyme Inhibition-Based Assay to Estimate the Contribution of Formulants to the Effect of Commercial Pesticide Formulations

Esimbekova, Elena N.; Kalyabina, Valeriya P.; Kopylova, Kseniya V.; Lonshakova-Mukina, Victoria I.; Antashkevich, Anna A.; Torgashina, I. G.; Lukyanenko, Kirill A.; Nemtseva, Elena V.; Kratasyuk, Valentina A.

doi:10.3390/ijms24032268

Cited by 2 publications

(2 citation statements)

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“…Thus, Red remains the only enzyme in the multi-enzyme assay systems whose activity is inhibited by the food preservative. Otherwise, we could observe an increase in the inhibitory effect of preservatives upon elongation of the series of coupled enzymes, as was the case when analyzing the inhibitory effect of a number of pesticides on enzyme assay systems of varying complexity [ 29 ]. However, an inhibition of activity of the tri-enzyme systems by 50% was observed at concentrations of the preservatives below the established maximum permissible content in food products.…”

Section: Discussionmentioning

confidence: 99%

“…The assays are based on the ability of toxic substances to specifically inhibit the activity of various enzymes or enzyme systems, with the degree of inhibition of the enzyme activity by toxic substances being proportional to the amount of toxicant in the sample [ 28 ]. Enzyme systems of different complexity have selective sensitivity to various classes of toxic substances, which allows them to be used to study the mechanisms of toxicant effects on the functioning of individual elements of metabolic pathways [ 29 ]. Thus, the enzyme inhibition assays are convenient and fast tools which allow one both to estimate the activity of potential toxicants at the molecular level and to avoid unethical procedures.…”

Section: Introductionmentioning

confidence: 99%

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Alternative Enzyme Inhibition Assay for Safety Evaluation of Food Preservatives

Esimbekova

Асанова

Kratasyuk

2023

Life

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While food additives are widely used in the modern food industry and generally are important in maintaining the ability to provide food for the increasing world population, the progress occurring in this field is much ahead of the evaluation of their possible consequences for human health. The present study suggests a set of single- and multi-enzyme assay systems for revealing toxic effects of the most widely spread food preservatives, such as sorbic acid (E200), potassium sorbate (E202), and sodium benzoate (E211) at the primary molecular level of their interaction with enzymes. The assay is based on the inhibition of enzyme activity by toxic substances proportional to the amount of the toxicants in the sample. The single-enzyme assay system based on NAD(P)H:FMN oxidoreductase (Red) proved to be most sensitive to the impact of food additives, with the IC50 values being 29, 14, and 0.02 mg/L for sodium benzoate, potassium sorbate, and sorbic acid, respectively, which is considerably lower than their acceptable daily intake (ADI). No reliable change in the degree of inhibition of the enzyme assay systems by food preservatives was observed upon elongating the series of coupled redox reactions. However, the inhibition of activity of the multi-enzyme systems by 50% was found at a preservative concentration below the maximum permissible level for food. The inhibition effect of food preservatives on the activity of butyrylcholinesterase (BChE), lactate dehydrogenase (LDH), and alcohol dehydrogenase (ADH) was either absent or found in the presence of food preservatives at concentrations significantly exceeding their ADI. Among the preservatives under study, sodium benzoate is considered to be the safest in terms of the inhibiting effect on the enzyme activity. The results show that the negative effect of the food preservatives at the molecular level of organization of living things is highly pronounced, while at the organismal level it may not be obvious.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Alternative Enzyme Inhibition Assay for Safety Evaluation of Food Preservatives

Esimbekova

Асанова

Kratasyuk

2023

Life

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Trypsin-Based Chemoenzymatic Assay for Detection of Pollutants and Safety Assessment of Food Additives

et al. 2023

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Chemoenzymatic assay systems are widely used to detect toxicants in various samples, including food and environment specimens. These methods are based on the ability of various types of toxicant to specifically inhibit/activate the functions of individual enzymes or enzyme systems. The present study examines the possibility of using the proteolytic enzyme trypsin as a specific marker to detect protease inhibitors in different samples. The study shows that trypsin activity is not affected by various heavy metals, pesticides, or quinones at levels considerably greater than their maximum allowable concentrations (MACs) in water bodies. At the same time, the IC50 value for the food preservative potassium sorbate (E202) is 15 mg/L, which is substantially lower than its acceptable daily intake (ADI). The quenching of trypsin fluorescence in the presence of potassium sorbate suggests that inhibition could occur due to the binding of the preservative to the enzyme in the region adjacent to the active center. The trypsin was immobilized in starch gel to ensure its stability in the enzyme inhibition based assay. Single-use reagents were prepared as dry starch disks that could be stored over long periods. Their sensitivity to copper (II) chloride, potassium sorbate, and chromium (III) chloride was similar to the sensitivity of the free trypsin.

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Enzyme Inhibition-Based Assay to Estimate the Contribution of Formulants to the Effect of Commercial Pesticide Formulations

Cited by 2 publications

References 66 publications

Alternative Enzyme Inhibition Assay for Safety Evaluation of Food Preservatives

Alternative Enzyme Inhibition Assay for Safety Evaluation of Food Preservatives

Trypsin-Based Chemoenzymatic Assay for Detection of Pollutants and Safety Assessment of Food Additives

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