Related Experiment Video
Updated: Jun 10, 2025

Preparation of Binary and Ternary Deep Eutectic Systems
Published on: October 31, 2019
Assessing the Effect of Deep Eutectic Solvents on α-Chymotrypsin Thermal Stability and Activity
Nicolás F Gajardo-Parra1, Esteban Cea-Klapp2, Anshu Chandra3
1Escuela de Ingeniería Industrial, Facultad de Ingeniería y Ciencias, Universidad Diego Portales, Santiago, 8370191, Chile.
None:
Optimizing the liquid reaction phase holds significant potential for enhancing the efficiency of biocatalytic processes since it determines reaction equilibrium and kinetics. This study investigates the influence of the addition of deep eutectic solvents on the stability and activity of α-chymotrypsin, a proteolytic enzyme with industrial relevance. Deep eutectic solvents, composed of choline chloride or betaine mixed with glycerol or sorbitol, were added in the reaction phase at various concentrations. Experimental techniques, including kinetic and fluorometry, were employed to assess the α-chymotrypsin activity, thermal stability, and unfolding reversibility. Atomistic molecular dynamics simulations were also conducted to assess the interactions and provide molecular-level insights between α-chymotrypsin and the solvent. The results showed that among all studied mixtures, adding choline chloride + sorbitol improved thermal stability up to 18 °C and reaction kinetic efficiency up to two-fold upon adding choline chloride + glycerol. Notably, the choline chloride + sorbitol system exhibited the most substantial stabilization effect, attributed to the surface preferential accumulation of sorbitol, as corroborated by the computational analyses. This work highlights the potential of tailoring liquid reaction phase of α-chymotrypsin catalyzed reaction using neoteric solvents such as deep eutectic solvents to enhance α-chymotrypsin performance and stability in industrial applications.
Related Concept Videos
Solvating Effects
E2 Reaction: Kinetics and Mechanism
E1 Reaction: Stereochemistry and Regiochemistry
Introduction to Mechanisms of Enzyme Catalysis
Factors Affecting α-Alkylation of Ketones: Choice of Base
The reaction involving bases like EtO− whose conjugate acid EtOH (pKa = 15.9) is stronger than the ketone (pKa = 19.2) results in an equilibrium mixture with higher ketone concentration. As a consequence,...
E1 Reaction: Kinetics and Mechanism

