Author(s):
Meneses, Liane ; Gajardo-Parra, Nicolás F. ; Cea-Klapp, Esteban ; Garrido, José Matías ; Held, Christoph ; Duarte, Ana Rita ; Paiva, Alexandre
Date: 2023
Persistent ID: http://hdl.handle.net/10362/162706
Origin: Repositório Institucional da UNL
Project/scholarship:
info:eu-repo/grantAgreement/EC/H2020/725034/EU;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F50006%2F2020/PT;
info:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FEQU-EQU%2F29851%2F2017/PT;
info:eu-repo/grantAgreement/FCT/OE/SFRH%2FBD%2F148510%2F2019/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F50006%2F2020/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F50006%2F2020/PT;
Subject(s): Chemistry (miscellaneous); Analytical Chemistry; Electrochemistry; Organic Chemistry; Chemistry (miscellaneous); Chemistry (miscellaneous); Analytical Chemistry; Analytical Chemistry; Electrochemistry; Electrochemistry; Organic Chemistry; Organic Chemistry
Description
The authors thank the funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy - EXC 2033 - 390677874 - RESOLV. Nicolás Gajardo's work was supported by the German Academic Exchange Service (DAAD) under the Graduate School Scholarship Programme, 2020 (57516591). E. C.-K. acknowledges the scholarship from ANID, Chile. This research was partially supported by the supercomputing infrastructure of the Southern GPU Cluster - Fondequip EQM150134. Publisher Copyright: © 2023 RSC.
In this work, the activity of horseradish peroxidase (HRP) in betaine-based natural deep eutectic systems (NADESs) was measured and studied by molecular simulations. Focus was laid on enzymatic activity in the NADESs under thermal stress as well as under the influence of water. Furthermore, the structure of HRP under these different conditions was measured by circular dichroism (CD). As a result, HRP remains enzymatically active in all NADESs upon incubation for 24 h at 37 °C and 60 °C and after 4 h at 80 °C, especially when incubated in a NADES composed of betaine, trehalose, glycerol, and water, in a molar ratio of 2 : 1 : 3 : 5. The CD studies have shown that high activity is obtained in the systems that promoted higher α-helix contents. The molecular simulations showed that using a NADES instead of buffer solvent reduces HRP flexibility, and we found that enzymatic activity correlates with Gibbs energy of solvation of HRP. Finally, hydrophobic hydration interactions govern the stabilization mechanism of the HRP folded state as shown by a drastic enzymatic activity drop upon 5 wt% water addition using a betaine: glycerol NADES as solvent.
