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Assessing the Influence of Betaine-Based Natural Deep Eutectic Systems on Horseradish Peroxidase
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Resumo(s)
To validate the use of horseradish peroxidase (HRP) in natural deep eutectic systems (NADES), five different betaine-based NADES were characterized in terms of water content, water activity, density, and viscosity experimentally and by thermodynamic modeling. The results show that the NADES under study have a water activity of about 0.4 at 37 °C for water contents between 14 and 22 wt %. The densities of the studied NADES had values between 1.2 and 1.3 g.cm-3at 20 °C. The density was modeled with a state-of-the-art equation of state; an excellent agreement with the experimental density data was achieved, allowing reasonable predictions for water activities. The system betaine:glycerol (1:2) was found to be the most viscous with a dynamic viscosity of ∼600 mPa.s at 40 °C, while all the other systems had viscosities <350 mPa.s at 40 °C. The impact of the NADES on the enzymatic activity, as well as on, conformational and thermal stability was assessed. The system betaine/sorbitol:water (1:1:3) showed the highest benefit for enzymatic activity, increasing it by two-folds. Moreover, upon NADES addition, thermal stability was increased followed by an increment in a-helix secondary structure content.
Descrição
Funding Information: The work of Nicolás Gajardo was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy - EXC 2033 - 390677874–RESOLV and by the German Academic Exchange Service (DAAD) under the Graduate School Scholarship Programme, 2020 (57516591). This work has also received funding from the European Union’s Horizon 2020 (European Research Council) under grant agreement no ERC-2016-CoG 725034. This work was further supported by the Associate Laboratory for Green Chemistry - LAQV and the project CryoDES which are financed by national funds from FCT/MCTES: UIDB/50006/2020 and PTDC/EQU–EQU/29851/2017, respectively. A.P. and L.M. also acknowledge FCT/MCTES for the financial support through IF/01146/2015 and SFRH/BD/148510/2019, respectively. The authors wish to thank Elisabete Ferreira from the BioLab, supported by the Applied Molecular Biosciences Research Unit-UCIBIO and the Associated Laboratory for Green Chemistry Research Unit–LAQV (UIDP/04378/2020, UIDB/04378/2020 and UIDB/50006/2020, UIDP/50006/2020, respectively). Publisher Copyright: © 2022 American Chemical Society. All rights reserved.
Palavras-chave
biocatalysis density natural deep eutectic systems (NADES) perturbed-chain statistical associating fluid theory (PC-SAFT) thermodynamics viscosity water activity General Chemistry Environmental Chemistry General Chemical Engineering Renewable Energy, Sustainability and the Environment SDG 7 - Affordable and Clean Energy