Preprint Communication Version 1 Preserved in Portico This version is not peer-reviewed

Monitoring Horseradish Peroxidase Activity with a Nanopore Detector

Yuri Ivanov
* ORCID logo ,
Alexander Ableev
,
Ivan Shumov
,
Irina Ivanova
,
Nikita Vaulin
,
Denis Lebedev
ORCID logo ,
Anton Bukatin
ORCID logo ,
Ivan Mukhin
ORCID logo ,
Alexander Archakov
Version 1 : Received: 18 August 2023 / Approved: 22 August 2023 / Online: 22 August 2023 (08:03:02 CEST)

How to cite: Ivanov, Y.; Ableev, A.; Shumov, I.; Ivanova, I.; Vaulin, N.; Lebedev, D.; Bukatin, A.; Mukhin, I.; Archakov, A. Monitoring Horseradish Peroxidase Activity with a Nanopore Detector. Preprints 2023, 2023081511. https://doi.org/10.20944/preprints202308.1511.v1 Ivanov, Y.; Ableev, A.; Shumov, I.; Ivanova, I.; Vaulin, N.; Lebedev, D.; Bukatin, A.; Mukhin, I.; Archakov, A. Monitoring Horseradish Peroxidase Activity with a Nanopore Detector. Preprints 2023, 2023081511. https://doi.org/10.20944/preprints202308.1511.v1

Abstract

We demonstrate the use of a nanotechnology-based approach for the investigation of enzymatic activity of a single molecule of horseradish peroxidase with a solid-state nanopore. The artificial 5 nm solid-state nanopore has been formed in a 40-nm-thick silicon nitride structure. Single molecule of HRP has been entrapped into the nanopore. The activity of horseradish peroxidase (HRP) enzyme molecule inserted in the nanopore has been monitored by recording the time dependence of the ion current through the nanopore in the course of the reaction of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) oxidation reaction. We have found that in the process of ABTS oxidation in the presence of 2.5 mM hydrogen peroxide, individual HRP enzyme molecules are able to retain activity for approximately 700 seconds before a decrease in the ion current through the nanopore, which can be explained by structural changes of the enzyme.

Keywords

nanopore detector; solid-state nanopore; horseradish peroxidase; enzymatic activity

Subject

Biology and Life Sciences, Biophysics

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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