Polydisulfides of substituted phenols as effective protectors of peroxidase against inactivation by ultrasonic cavitation

• Published in: Biochemistry (Moscow) Vol. 66; no. 7; pp. 740 - 746
• Main Authors: Grintsevich, E E, Adzerikho, I E, Mrochek, A G, Metelitza, D I
• Format: Journal Article
• Language: English
• Published: United States Springer Nature B.V 01.07.2001
• Subjects: Aqueous solutions; Cavitation; Disulfides - chemistry; Disulfides - pharmacology; Free Radical Scavengers - pharmacology; Free radicals; Gallic Acid - chemistry; Humans; Hydrogen-Ion Concentration; Inactivation; Kinetics; Peroxidase - drug effects; Peroxidase - radiation effects; Phenols; Phenols - chemistry; Serum Albumin - pharmacology; Ultrasonics
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1023/A:1010256511200

Kinetics of inactivation of horseradish peroxidase (HP) induced by low-frequency ultrasonic (US) treatment (27 kHz) with the specific power of 60 W/cm2 were studied in phosphate (pH 7.4) and acetate (pH 5.2) buffers within the temperature range of 36.0 to 50.0 degrees C and characterized by effective first-order rate constants of US inactivation k(in)(us) in min(-1). Values of k(in)(us) depend on the specific ultrasonic power within the range of 20-60 W/cm2, on the concentration of HP, and on pH and temperature of the solutions. The activation energy of US inactivation of HP is 9.4 kcal/mole. Scavengers of HO* radicals, mannitol and dimethylformamide, significantly inhibit the US inactivation of HP at 36.0 degrees C, whereas micromolar concentrations of polydisulfide of gallic acid (poly(DSG)) and of poly(2-aminodisulfide-4-nitrophenol) (poly(ADSNP)) virtually completely suppress the US inactivation of peroxidase at the ultrasonic power of 60 W/cm2 on the sonication of the enzyme solutions for more than 1 h at pH 5.2. Various complexes of poly(DSG) with human serum albumin effectively protect HP against the US inactivation in phosphate buffer (pH 7.4). The findings unambiguously confirm a free radical mechanism of the US inactivation of HP in aqueous solutions. Polydisulfides of substituted phenols are very effective protectors of peroxidase against inactivation caused by US cavitation.