The effect of cavited water on physicochemical properties of horseradish peroxidase as studied at the level of single enzyme molecules. Change of enzyme properties under the influence of cavitation

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Abstract

The effect of water subjected to cavitation on horseradish peroxidase (HRP) enzyme is studied using atomic force microscopy (AFM) and spectrophotometry (SP). By AFM, a significant change in the adsorption of HRP on freshly cleaved mica after keeping the enzyme solution in water subjected to cavitation, is found – as compared to the control enzyme sample. According to SP, the enzymatic activity of HRP does not change. The effect observed is useful to take into account when working with enzymes on industrial and research equipment, in which cavitation may occur in the flow of an aqueous medium.

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About the authors

V. S. Ziborov

Institute of Biomedical Chemistry (IBMC); Joint Institute for High Temperatures of the Russian Academy of Sciences (JIHT RAS)

Email: shum230988@yandex.ru
ORCID iD: 0000-0001-7942-3337

Cand. of Sci. (Physics and Mathematics), Senior Researcher

Russian Federation, Moscow; Moscow

I. D. Shumov

Institute of Biomedical Chemistry (IBMC)

Author for correspondence.
Email: shum230988@yandex.ru
ORCID iD: 0000-0002-9795-7065

Cand. of Sci. (Biology), Researcher

Russian Federation, Moscow

E. E. Vazhenkova

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0009-0001-4224-8907

Laboratory assistant

Russian Federation, Moscow

A. N. Ableev

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0009-0004-3096-107X

Leading Engineer

Russian Federation, Moscow

A. F. Kozlov

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0000-0002-2117-8743

Leading Engineer

Russian Federation, Moscow

A. V. Vinogradova

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0009-0001-6044-3490

Junior Researcher

Russian Federation, Moscow

E. D. Nevedrova

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0000-0003-2767-2299

Junior Researcher

Russian Federation, Moscow

O. N. Afonin

Institute of Biomedical Chemistry (IBMC)

Email: shum230988@yandex.ru
ORCID iD: 0009-0008-7947-3674

Cand. of Sci. (Tech), Senior Researcher

Russian Federation, Moscow

V. Yu. Tatur

Foundation of Perspective Technologies and Novations (FPTN)

Email: shum230988@yandex.ru
ORCID iD: 0000-0002-6415-5189

Executive Director

Russian Federation, Moscow

A. A. Lukyanitsa

Foundation of Perspective Technologies and Novations (FPTN); Moscow State University

Email: shum230988@yandex.ru
ORCID iD: 0000-0002-0517-0602

Doct. of Sci. (Tech), Leading Researcher

Russian Federation, Moscow; Moscow

A. V. Shcherbakov

LLC “Scientific and Production Association “KURS”

Email: shum230988@yandex.ru
ORCID iD: 0000-0002-8067-4624

Director General

Russian Federation, Moscow Region, Dolgoprudny

N. D. Ivanova

Moscow State Academy of Veterinary Medicine and Biotechnology Named after Skryabin

Email: shum230988@yandex.ru
ORCID iD: 0000-0001-5942-8050

Lecturer

Russian Federation, Moscow

E. S. Yushkov

National Research Nuclear University MEPhI

Email: shum230988@yandex.ru
ORCID iD: 0009-0002-9161-0877

Cand. of Sci. (Tech), Prof.

Russian Federation, Moscow

D. V. Enikeev

Institute for Urology and Reproductive Health, Sechenov University

Email: shum230988@yandex.ru
ORCID iD: 0000-0001-7169-2209

Doct. of Sci. (Medicine), Prof., Urologist surgeon

Russian Federation, Moscow

Yu. D. Ivanov

Institute of Biomedical Chemistry (IBMC); Joint Institute for High Temperatures of the Russian Academy of Sciences (JIHT RAS)

Email: shum230988@yandex.ru
ORCID iD: 0000-0001-5041-1914

Doct. of Sci. (Biology), Prof., Head of Laboratory

Russian Federation, Moscow; Moscow

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Supplementary files

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2. Fig.1. Photographic image of the (hydrodynamic generator)-based setup used for obtaining cavitational water

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3. Fig.2. Schematic representation of the experiment on the incubation of HRP solution to study effect and post-effect of interaction of cavitational water on the enzyme. The tube with the control sample of HRP solution not shown

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4. Fig.3. Typical AFM images of the surface of freshly cleaved mica substrates incubated in the studied samples of HRP solution. The sample was kept for 3 hours in a beaker with cavitational water (a), at the same place as the beaker with cavitational water (b), or 10 m away from the beaker with cavitational water (c, control sample). Experimental conditions: 10–7 M HRP in 2 mM PBS-D, pH 7.4, temperature 23 °C, incubation time 3 hours. The size of each scan is 2×2 μm

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5. Fig.4. Histograms of the absolute number of visualized objects (normalized to an area of 400 μm2) depending on the heights of objects N400(h), obtained for the studied samples of the HRP solution. The samples were kept for 3 hours in a beaker with cavitational water (red columns), at the place of the glass with cavitational water (green columns) or 10 m from the glass with cavitational water (blue columns). Experimental conditions: 10–7 M HRP in 2 mM PBS-D, pH 7.4, Т = 23 °C, incubation time 3 hours

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6. Fig.5. Dependences of absorption at 405 nm on time A405(t), obtained for the studied samples of the HRP solution. The sample was kept for 3 hours in a glass with cavitation water (red curve), at the place of the glass with cavitation water (green curve) or 10 m from the glass with cavitation water (blue curve). Experimental conditions: the concentration of HRP in the SP cell was 10–9 M, pH 5.0, Т = 23 °C, the optical path length of the SP cell was 1 cm

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Copyright (c) 2025 Ziborov V.S., Shumov I.D., Vazhenkova E.E., Ableev A.N., Kozlov A.F., Vinogradova A.V., Nevedrova E.D., Afonin O.N., Tatur V.Y., Lukyanitsa A.A., Shcherbakov A.V., Ivanova N.D., Yushkov E.S., Enikeev D.V., Ivanov Y.D.