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Antioxidant Capacity of Poly(Ethylene Glycol) (PEG) as Protection Mechanism Against Hydrogen Peroxide Inactivation of Peroxidases

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Abstract

The ability of poly(ethylene glycol) (PEG) to protect enzymatic peroxidase activity was determined for horseradish peroxidase (HRP), versatile peroxidase (VP), commercial Coprinus peroxidase (BP), and chloroperoxidase (CPO). The operational stability measured as the total turnover number was determined for the four peroxidases. The presence of PEG significantly increased the operational stability of VP and HRP up to 123 and 195 %, respectively, and dramatically increased the total turnover number of BP up to 597 %. Chloroperoxidase was not protected by PEG, which may be due to the different oxidation mechanism, in which the oxidation is mediated by hypochlorous ion instead of free radicals as in the other peroxidases. The presence of PEG does not protect the enzyme when incubated only in the presence of H2O2 without reducing substrate. The catalytic constants (k cat) are insensitive to the presence of PEG, suggesting that the protection mechanism is not due to a competition between the PEG and the substrate as electron donors. On the other hand, PEG showed to have a significant antioxidant capacity. Thus, we conclude that the protection mechanism for peroxidases of PEG is based in its antioxidant capacity with which it is able scavenge or drain radicals that are harmful to the protein.

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Acknowledgments

The authors wish to thank Dr. Katrin Quester for her expert technical assistance. K. Juarez-Moreno was awarded with a DGAPA-UNAM postdoctoral fellowship, and this work was funded by the National Council of Science and Technology of Mexico (CONACyT  -165633).

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Authors and Affiliations

  1. Centro de Nanociencias y Nanotecnología, UNAM, Km. 107 Carretera Tijuana-Ensenada, Ensenada, Baja California, 22860, Mexico

    Karla Juarez-Moreno & Rafael Vazquez-Duhalt

  2. Instituto de Biotecnología UNAM, Av. Universidad 2001, Cuernavaca, Morelos, 62250, Mexico

    Marcela Ayala

Authors
  1. Karla Juarez-Moreno
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  2. Marcela Ayala
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  3. Rafael Vazquez-Duhalt
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Correspondence to Rafael Vazquez-Duhalt.

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Juarez-Moreno, K., Ayala, M. & Vazquez-Duhalt, R. Antioxidant Capacity of Poly(Ethylene Glycol) (PEG) as Protection Mechanism Against Hydrogen Peroxide Inactivation of Peroxidases. Appl Biochem Biotechnol 177, 1364–1373 (2015). https://doi.org/10.1007/s12010-015-1820-y

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  • DOI: https://doi.org/10.1007/s12010-015-1820-y

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