Calorimetric Evaluation of the Operational Thermal Stability of Ribonuclease A in Hydrated Deep Eutectic Solvents

Kist, Jennifer A.; Henzl, Michael T.; Bañuelos, Jose; Baker, Gary A.

doi:10.1021/acssuschemeng.9b02585

Cited by 25 publications

(26 citation statements)

References 40 publications

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“…These results indicate that the specific concentration of ChCl -Lac may be able to provide a suitable microenvironment by hydrogen bonding in the active center of enzyme, and then keep its rigid confirmation and function especially at a high temperature. This result is in agreement with other’s observation and conclusion that the DES could promote the thermal stability of enzymes ( Kist et al, 2019 ). Interestingly, DESs not only can stabilize the enzyme at high temperature, but some of them also have been reported to increase the laccase stability at low temperature.…”

Section: Results and Analysissupporting

confidence: 94%

Enhancing the Thermostability of Papain by Immobilizing on Deep Eutectic Solvents-Treated Chitosan With Optimal Microporous Structure and Catalytic Microenvironment

Lin

Feng

et al. 2020

Front. Bioeng. Biotechnol.

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Deep eutectic solvents (DESs) have attracted an increasing attention in the fields of biocatalysis and biopolymer processing. In this study, papain immobilized on choline chloride- lactic acid (ChCl-Lac) DES-treated chitosan exhibited excellent thermostability as compared to the free enzyme. The properties of native or DES-treated chitosan and immobilized enzyme were characterized by FT-IR, SEM, surface area and pore property analysis. Like the common enzyme immobilization, papain immobilized on DES-treated chitosan resulted in a lower catalytic efficiency and a higher thermostability than the free enzyme due to the restricted diffusion. The results also revealed that DES could control the active group content, thus achieving the appropriate microporous structure of immobilized enzyme. Meanwhile, it could also help to construct the optimal microenvironment by hydrogen-bonding interaction between enzyme, chitosan, and residual DES, which are benefit for maintaining an active conformation and subsequently a high thermostability of papain. Moreover, it was found that trace DES (10 mM) significantly promoted the activity of free papain (145%). Deactivation thermodynamics study showed that the DES could enhance the thermostability of papain especially at high temperature (half-life of 7.4 vs. 3.5 h) because of the increased Gibbs free energy of denaturation. Secondary structure analysis by circular dichroism spectroscopy (CD) agreed well with the activity and thermostability data, further confirming the formation of rigid conformation induced by a specific amount of DES. This work provides a new way of enzyme immobilization synergistically intensified by solvents and supporting materials to achieve better microporous structure and catalytic microenvironment.

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Section: Results and Analysissupporting

confidence: 94%

Enhancing the Thermostability of Papain by Immobilizing on Deep Eutectic Solvents-Treated Chitosan With Optimal Microporous Structure and Catalytic Microenvironment

Lin

Feng

et al. 2020

Front. Bioeng. Biotechnol.

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“…Kist et al demonstrated that a single DSC thermal cycle to 80 °C in water containing 10 wt % or more reline results in the rapid and total loss of folding reversibility for bovine ribonuclease A (RNase A), an outcome partly attributed to ammonia evolution. For this reason, use of 1,3-dimethylurea in place of urea led to slightly better outcomes . In stark contrast, the inclusion of glyceline up to at least 35 wt % in aqueous media increased the T m and improved the thermodynamic stability of RNase A beyond the natural water environment.…”

Section: Selected Applications Of Deep Eutectic Solventsmentioning

confidence: 99%

“…For this reason, use of 1,3-dimethylurea in place of urea led to slightly better outcomes. 185 In stark contrast, the inclusion of glyceline up to at least 35 wt % in aqueous media increased the T m and improved the thermodynamic stability of RNase A beyond the natural water environment. Indeed, iterative thermal cycling resulted in only minor losses in the experimental heat capacity, reflecting a small degree of irreversibly unfolded RNase A in glyceline-containing systems.…”

Section: Biomolecular Structure Folding and Stabilitymentioning

confidence: 99%

Deep Eutectic Solvents: A Review of Fundamentals and Applications

et al. 2020

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Deep eutectic solvents (DESs) are an emerging class of mixtures characterized by significant depressions in melting points compared to those of the neat constituent components. These materials are promising for applications as inexpensive "designer" solvents exhibiting a host of tunable physicochemical properties. A detailed review of the current literature reveals the lack of predictive understanding of the microscopic mechanisms that govern the structure−property relationships in this class of solvents. Complex hydrogen bonding is postulated as the root cause of their melting point depressions and physicochemical properties; to understand these hydrogen bonded networks, it is imperative to study these systems as dynamic entities using both simulations and experiments. This review emphasizes recent research efforts in order to elucidate the next steps needed to develop a fundamental framework needed for a deeper understanding of DESs. It covers recent developments in DES research, frames outstanding scientific questions, and identifies promising research thrusts aligned with the advancement of the field toward predictive models and fundamental understanding of these solvents.

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“…25 In addition to this, DESs are also suitable for protein extraction providing enhanced yield. 18,26 Kist et al 27 have presented work on thermodynamic refolding of RNase by using hydrated cholinium chloride-based DESs. Thus, DESs are currently attracting noteworthy recognition as "greener" media for a variety of biological as well as biochemical processes.…”

Section: ■ Introductionmentioning

confidence: 99%

“…It was found that B-form helical structures of DNA were well preserved in DESs along with exhibiting enhanced thermal and pH stability of regenerated DNA. , Zhao further deduced that G-quadruplexes of DNA were more stable in neat DESs compared to aqueous media, even at temperatures greater than 110 °C . In addition to this, DESs are also suitable for protein extraction providing enhanced yield. , Kist et al have presented work on thermodynamic refolding of RNase by using hydrated cholinium chloride-based DESs. Thus, DESs are currently attracting noteworthy recognition as “greener” media for a variety of biological as well as biochemical processes.…”

Section: Introductionmentioning

confidence: 99%

Expanding the Potential Role of Deep Eutectic Solvents toward Facilitating the Structural and Thermal Stability of α-Chymotrypsin

Yadav

Bhakuni

Bisht

et al. 2020

ACS Sustainable Chem. Eng.

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Since the introduction of deep eutectic solvents (DESs), numerous reports have surfaced describing its tunable properties and environmentally benign nature. Due to such favorable properties of DESs, they have found a wide variety of applications. Moreover, in order to harness the potential of proteins in numerous industries, there is an emergent need to find a suitable cosolvent that is biocompatible with protein and is also environmentally safe. In this context, this work presents a systematic evaluation of effect of two deep eutectic solvents (DESs), namely, choline chloride-urea (ChCl-urea) and choline chloride-glycerol (ChCl-gly) on the structural and thermal stability along with activity of enzyme α-chymotrypsin (CT) using circular dichroism (CD), UV–visible, steady state, and thermal fluorescence spectroscopy. It was observed that the presence of DESs does lead to enhancement in the thermal stability of CT along with the preservation of activity. The enzymatic activity was well maintained in both the DESs, and the deleterious effect of urea was overcome by ChCl-urea on the enzyme. Also, desirable results were observed for ChCl-urea, despite having urea as one of its major components. Thus, the negative outcome of urea was overpowered by the combination of ChCl and urea. Furthermore, all the biomolecular studies were also performed with the individual constituents of DESs. It was found that the effect imparted by both the ChCl-based DESs on CT is by the virtue of DES itself rather than its individual constituent. Overall, both the DESs can be described as potential biocompatible, sustainable, and promising cosolvents for CT with enhanced structural and thermal stability along with preservation of its activity.

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Calorimetric Evaluation of the Operational Thermal Stability of Ribonuclease A in Hydrated Deep Eutectic Solvents

Cited by 25 publications

References 40 publications

Enhancing the Thermostability of Papain by Immobilizing on Deep Eutectic Solvents-Treated Chitosan With Optimal Microporous Structure and Catalytic Microenvironment

Enhancing the Thermostability of Papain by Immobilizing on Deep Eutectic Solvents-Treated Chitosan With Optimal Microporous Structure and Catalytic Microenvironment

Deep Eutectic Solvents: A Review of Fundamentals and Applications

Expanding the Potential Role of Deep Eutectic Solvents toward Facilitating the Structural and Thermal Stability of α-Chymotrypsin

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