Investigation of static magnetic field effect on horseradish peroxidase enzyme activity and stability in enzymatic oxidation process.

The activity of Horseradish Peroxidase (HRP) Enzyme exposed to a static magnetic field (SMF) during the oxidation reaction of pyrogallol (PGL) and the epigallocatechin gallate (EPCG) flavonoid was recorded at different times. As the data showed, the enzyme activity increased by 77.17% with increasing incubation time up to 30 min. The kinetic parameters KM and Vmax for PGL sample incubated in SMF for 30 min were 5.641 × 10-3 mM, 4.424 × 10-2 mmol/min, respectively, and for EPCG sample with the same condition were 8.65 × 10-4 mM, 2.37 × 10-3 mmol/min, respectively. Exposure of HRP enzyme to SMF changed the optimum pH from 7.0 to 6.0 in 10 min, but did not create any change in the optimum temperature of the enzyme. After 120 h, the residual activity of normal enzyme was 17% higher than that of the incubated enzyme. The structural changes of the control and HRP enzyme incubated in SMF were investigated by relative viscosity, fluorescence and CD, UV-Vis spectrophotometry. The structural changes in the presence of SMF were found to cause changes in the enzyme activity. In fact, changes in the amount of hydrogen bonds between enzymes and solvents can be a reason for this behavior from a molecular point of view. Using a static magnetic field can provide a new approach to control and direct enzyme-based biological processes.

The effect of magnetized water on the oxidation reaction of phenol derivatives and aromatic amines by horseradish peroxidase enzyme.

The present study aimed to investigate, for the first time, the rate of the oxidation reaction of some derivatives of phenol and aromatic amines, that is, pyrogallol, catechol, resorcinol, ortho-aminophenol, meta-aminophenol, para-aminophenol, ortho-phenylenediamine, and para-phenylenediamine, in the presence of hydrogen peroxide in pure and magnetized solvents using horseradish peroxidase enzyme. The reaction was studied in the absence and presence of a magnetized solvent under completely identical conditions. The results showed that magnetized solvent could change the structure of the enzyme and reduce its activity. In addition, it affected the rate of oxidation of the selected derivatives through altering the strength of the hydrogen bonds of the system. The changes in the structure and activity of the enzyme were examined using UV-Vis and fluorescence spectroscopy as well as viscosity measurement technique. Examination of the secondary structure via the far UV-CD spectrum indicated the increase in the alpha helical structure in the magnetized solvent. When dissolved in a magnetized solvent, hydrogen peroxide as an enzyme substrate reduced the rate of enzymatic reaction and provided lower saturation conditions for the enzyme compared with when it was dissolved in the pure solvent.