Document type
ArticleVersion
Accepted versionPublication date
All rights reserved
Please use this identifier to cite or link to this item: https://hdl.handle.net/2445/104311
Influence of macromolecular crowding on the oxidation of ABTS by hydrogen peroxide catalyzed by HRP
Journal Title
Director/Tutor
Journal ISSN
Volume Title
Related resource
Abstract
The interior of the living cell is highly concentrated and structured with molecules having different shapes and sizes. However, almost all experimental biochemical data have been obtained working in dilute solutions that do not reflect in vivo conditions. In this paper, we study in vitro the effect of macromolecular crowding on the reaction rates of the oxidation of 2,2'-azino-bis(3- ethylbenzothiazoline-6-sulfonate) (ABTS) by hydrogen peroxide (H2O2) catalyzed by Horseradish Peroxidase (HRP), by adding dextrans of various molecular weights to the reaction solutions as crowding agents. The results indicate that the volume occupied by the crowding agent, regardless its size, plays an important role in the rate of this reaction. Both Michaelis-Menten parameters, vmax and Km, decrease when the dextran concentration in the sample increases, which might be due to a crowding-induced effect in the catalytic constant, kcat of this enzymatic reaction. Thus, our results suggest that there is an activation control of the enzymatic reaction in this particular system. In our opinion, this work could facilitate the understanding of biomolecules behavior in vivo and be useful for biotechnology in vitro applications, since HRP is widely used in the development of biosensors.
Subject
Subject (English)
Citation
Citation
PITULICE, Laura, et al. Influence of macromolecular crowding on the oxidation of ABTS by hydrogen peroxide catalyzed by HRP. Journal of Biocatalysis & Biotransformation. 2013. Vol. 2, num. 1, pags. 1000107-1000107. ISSN 2324-9099. [consulted: 14 of August of 2026]. Available at: https://hdl.handle.net/2445/104311