Determination of the activity of some antioxidant enzymes during the germination periods of cereals and legumes

. As a result of the exchange of substances in a living organism, "products" "free radicals" and peroxide compounds of organic and inorganic substances are formed. It is known that free radicals damage cells. Enzymes of the body's antioxidant defense system, superoxide dismutase, catalase, glutathione reductase, glutathione peroxidases, and toxic products of lipid peroxide oxidation turn free radicals into harmless substances. In this work, the activity of one of such enzymes - peroxidase, catalase was studied. The activity of catalase enzyme during the germination periods of cereal and leguminous plants is different, the highest activity of the catalase enzyme during the germination periods of cereal plants was determined on the 5th day of the germination period: in wheat – 5,644 mg H 2 O 2 /g, in barley – 4,08 mg H 2 O 2 /g, in oats – 5,134 mg H 2 O 2 /g. In leguminous plants, the highest activity was determined on the 3rd day of the germination period: it was 11,832 mg H 2 O 2 /g in soybean, and 8,126 mg H 2 O 2 /g in mash. The highest activity of peroxidase enzyme is on the first day of wheat and is 1,158 µmol/min.g mass. In leguminous plants, the highest activity of the peroxidase enzyme is on the 9th day of the soybean germination period, and its activity is 0,687 µmol/min.g. constituted the mass.

As a result of metabolism in a living organism, oxidized products -"free radicals" and peroxide compounds of organic and inorganic substances are formed.The influence of adverse conditions accelerates this process.It is known that free radicals damage the cell [3], there are several specific anti-oxidation mechanisms in the cell, including superoxide dismutase, catalase, peroxidase, and glutathione reductase.

Materials and methods
These antioxidant systems neutralize the amount of free radicals in organisms, that is, active forms of oxygen: O 2 − , H 2 O 2 , HO 0 binds and increases resistance to various unfavorable external influences [6].
Catalase (from the Greek katalysis) is an enzyme belonging to the class of oxidoreductases, which catalyzes the decomposition of hydrogen peroxide by water (H2O) into oxygen: Catalase is present in all animal and plant tissues as well as in aerobic microorganisms.Catalase enzymes have been studied in many plants and animals, and it has been determined that their amount varies depending on time, conditions, and type of organism [15].
Peroxidase enzyme controls oxidation-reduction processes in plant cells.Peroxidase enzyme is widespread in both animals and plants.The toxic substance formed in the process of oxidation of peroxidase prevents the accumulation of H202, that is, the enzyme acts as an acceptor on hydrogen peroxide [5].Peroxidase is an enzyme that shows sensitivity to adverse environmental effects in plants.Various biologically active compounds with antioxidant properties have been identified in cultivated cereal plants [10,11,15].
The activity of the peroxidase enzyme has been studied in many plants, and it has been determined that its activity depends on time, conditions, and the type of organism [15].It has been found that it is important in protection against damage caused by pathogenic microorganisms [1,2,6].
The main goal of our scientific research is to determine the activity of catalase and peroxidase enzymes, which have an antioxidant effect, during the germination of the seeds of wheat, barley, oat, soybean, mash plants.
Research object: collected wheat, barley, oats, soybeans, mash.The essence of the method.The method of determining the activity of the catalase enzyme in the object under study is based on the determination of the dissolved amount of hydrogen peroxide by titration with a solution of potassium permanganate.
The activity of the catalase enzyme is determined by the amount of milligrams of hydrogen peroxide decomposed by the enzyme during 30 minutes in 1 gram of the tested material.Enzyme activity is determined using the following formula: Here A is catalase activity, mg H2O2/g; a -the amount of KMnO4 used for the titration of the control sample in ml; b -the amount of KMnO4 used for the titration of the experimental sample, in ml (1 ml of KMnO4 solution is equivalent to 1.7 mg H2O2); V1 -the volume of the plant taken for analysis in ml; V2 -the volume of the solution taken for titration; m is the mass of the sample taken for research, grams [15].
In the reaction catalyzed by peroxidase, the product formed by benzidine with diphenoquinonediimine produces a blue color (benzidine blue).Enzyme activity is determined by the following formula: Activity of enzyme (μ.mol/min.gmass) E -extension; a-received buffer mixture (ml), for extracting the plant; b-volume after additional centrifugation; v-dilution of the extract placed in the cuvette; c-cuvette thickness (2cm); t-time in seconds.
Research results: the obtained results show that the activity of the catalase enzyme, which enters the system of antioxidant enzymes, depends on the germination period of the collected grain and leguminous plants, and its activity was determined to be different.
The obtained results showed that the high activity of catalase was 5,644 mg H2O2/g in wheat and 5,134 mg H2O2/g in oat after 5 days of germination.(Table 1, Fig. 1), catalase activity decreased in the later days of germination.On the 9th day of the germination period, it was found to be equal to 3,06 mg H2O2/g in wheat and 4,726 mg in oats.It was found that the activity of catalase enzyme during the germination periods of barley is lower than that of harvested wheat and oats (fig.1).The highest activity of catalase was on the 5th day of the germination period and was 4,08 mg H2O2/g.It was 2,108 mg H2O2/g on the 7th day and 2,856 mg H2O2/g on the 9th day [15].was found that the activity of catalase enzyme is different during the germination periods of leguminous plants.In soybean, the highest activity of catalase was on the 3rd day of the germination period and was 11,832 mg H2O2/g.Enzyme activity decreased on days 5,7,9.On the 9th day, its activity was 7.31 mg H2O2/g (Fig. 2) [15].

Fig. 2. Activity of catalase enzyme during germination of legumes
We found out that the highest activity in mosh was 8,126 mg H2O2/g on the 3rd day of the germination period, and decreased on the 7,9 days of the germination period, and it was 3,128 mg H2O2/g on the 9th day of the germination period (Fig. 2).

Result and discussion
In a number of scientific studies, it has been shown that the change in the activity of antioxidant enzymes depends on the metabolic processes in the cell during the germination of grain plants [7,15].
The results showed that the activity of the peroxidase enzyme, which is a part of the system of antioxidant enzymes, depends on the germination period of the collected cereal and leguminous plants, and its activity was determined to be different.
The high activity of peroxidase was 1,158 μmol/min g on the first day of germination in wheat from cereal plants (Table 2).

Table 2. Activity of peroxidase during germination of cereal and leguminous plants
(μ.mol/min.g.mass)
It was found that the activity of the peroxidase enzyme during the germination periods of barley is lower than that of the harvested wheat (Figure 3).The highest activity of peroxidase was on the 3rd day of the germination period and was 0,725 μmol/min.gmass [15].

Fig. 3. The highest activity of the peroxidase enzyme in cereal plants is the germination period
During the germination periods of legumes, the highest activity of peroxidase in soybeans was determined on the 9th day of the germination period and was 0,687 μmol/min.gmass (Table 2).Fig. 4. The highest activity of the enzyme peroxidase in legumes is the period of germination It was found that the highest activity of the peroxidase enzyme was on the 1st day of the germination period and was 0,523 μ.mol/min.g.mass, and it decreased by the 9th day of the germination period and was 0,156 μ.mol/min.g.mass (Fig. 4).
In a number of scientific works, it was noted that the change in the activity of antioxidant enzymes depends on the metabolic processes in the cell during the germination of grain plants [8,15].

Conclusion
The obtained results show that the activity of catalase and peroxidase enzymes during the germination periods of cereal and leguminous plants is different.The highest activity of catalase was determined on the 5th day of germination of harvested cereal plants.The high activity of catalase was 5,644 mg H2O2/g in wheat, 5,134 mg H2O2/g in oats, and 4,08 mg H2O2/g in barley [15].
In legumes, the highest activity of catalase was determined on the 3rd day of the germination period.In soybean it was -11,832 mg H2O2/g, in mash -8,126 mg H2O2/g [15].
The high activity of peroxidase during the germination period of cereal and leguminous plants was 1,158 μ.mol/min.g.mass on the 1st day of the germination period of the cereal plant wheat, and on the 9th day of the germination period of the soybean plant in the shade lib, the activity was equal to 0,687 μ.mol/min.g.mass.

Table 1 .
Activity of catalase during germination of cereal and leguminous plants (mg H2O2/g)