Molecular Understanding of Activity Changes of Alcohol Dehydrogenase in Deep Eutectic Solvents

verfasst von: Jan Philipp Bittner, Ningning Zhang, Pablo Domínguez de María, Irina Smirnova, Selin Kara, Sven Jakobtorweihen
Abstract: Deep eutectic solvents (DESs) have emerged as promising solvents for biocatalysis. While their impact on enzyme solvation and stabilization has been studied for several enzyme classes, their role in substrate binding is yet to be investigated. Herein, molecular dynamics (MD) simulations of horse-liver alcohol dehydrogenase (HLADH) are performed in choline chloride-ethylene glycol (ChCl-EG) and choline chloride-glycerol (ChCl-Gly) at varying water concentrations. In the DES solutions, the active site was significantly constricted, and its flexibility reduced when compared to the aqueous medium. Importantly, the cavity size follows a similar trend as the catalytic activity of HLADH and as such explains previously observed activity changes. To understand the impact on the binding of the substrate (cyclohexanone), an umbrella sampling (US) setup was established to calculate the free energy changes along the substrate binding tunnel of HLADH. The US combined with replica exchange and NADH in its cofactor pocket provided the best sampling of the entire active site, explaining why the cyclohexanone binding on HLADH is reduced with increasing DES content. As different components in these multicomponent mixtures influence the substrate binding, we additionally applied the US setup to study the ability of the DES components to be present inside the substrate tunnel. The presented approach may become useful to understand enzyme behaviors in DESs and to enable the design of more enzyme-compatible and tunable solvents.
Organisationseinheit(en): Institut für Technische Chemie
Externe Organisation(en): Technische Universität Hamburg (TUHH)
Sustainable Momentum SL
Aarhus University
Typ: Artikel
Journal: Journal of Physical Chemistry B
Band: 129
Seiten: 1197–1213
Anzahl der Seiten: 17
ISSN: 1520-6106
Publikationsdatum: 17.01.2025
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Physikalische und Theoretische Chemie, Oberflächen, Beschichtungen und Folien, Werkstoffchemie
Elektronische Version(en): https://doi.org/10.1021/acs.jpcb.4c06523 (Zugang: Geschlossen)

BibTeX

@article{6c7154d1eb834dc1b8fb95c0808079b5,
title = "Molecular Understanding of Activity Changes of Alcohol Dehydrogenase in Deep Eutectic Solvents",
abstract = "Deep eutectic solvents (DESs) have emerged as promising solvents for biocatalysis. While their impact on enzyme solvation and stabilization has been studied for several enzyme classes, their role in substrate binding is yet to be investigated. Herein, molecular dynamics (MD) simulations of horse-liver alcohol dehydrogenase (HLADH) are performed in choline chloride-ethylene glycol (ChCl-EG) and choline chloride-glycerol (ChCl-Gly) at varying water concentrations. In the DES solutions, the active site was significantly constricted, and its flexibility reduced when compared to the aqueous medium. Importantly, the cavity size follows a similar trend as the catalytic activity of HLADH and as such explains previously observed activity changes. To understand the impact on the binding of the substrate (cyclohexanone), an umbrella sampling (US) setup was established to calculate the free energy changes along the substrate binding tunnel of HLADH. The US combined with replica exchange and NADH in its cofactor pocket provided the best sampling of the entire active site, explaining why the cyclohexanone binding on HLADH is reduced with increasing DES content. As different components in these multicomponent mixtures influence the substrate binding, we additionally applied the US setup to study the ability of the DES components to be present inside the substrate tunnel. The presented approach may become useful to understand enzyme behaviors in DESs and to enable the design of more enzyme-compatible and tunable solvents.",
author = "Bittner, {Jan Philipp} and Ningning Zhang and {Dom{\'i}nguez de Mar{\'i}a}, Pablo and Irina Smirnova and Selin Kara and Sven Jakobtorweihen",
note = "Publisher Copyright: {\textcopyright} 2025 American Chemical Society.",
year = "2025",
month = jan,
day = "17",
doi = "10.1021/acs.jpcb.4c06523",
language = "English",
volume = "129",
pages = "1197–1213",
journal = "Journal of Physical Chemistry B",
issn = "1520-6106",
publisher = "American Chemical Society",
number = "4",
}