Natural deep eutectic solvents as thermostabilizer for Humicola insolens cutinase

As a new generation of green solvents, deep eutectic solvents (DESs) are considered a promising alternative to
current harsh organic solvents and find application in many chemical processing methods such as extraction and
synthesis. DESs, normally formed by two or more components via various hydrogen bond interactions, offer high
potential as medium for biocatalysis reactions where they can improve efficiency by enhancing substrate solubility
and the activity and stability of the enzymes. In the current study, the stabilization of Humicola insolens
cutinase (HiC) in natural deep eutectic solvents (NADESs) was assessed. The best hydrogen bond donor among
sorbitol, xylitol, erythritol, glycerol and ethylene glycol, and the best acceptor among betaine, choline chloride,
choline acetate, choline dihydrogen citrate and tetramethylammonium chloride, were selected, evaluating
binding energies and molecular orientations through molecular docking simulations, and finally used to prepare
NADES aqueous solutions. The effects of component ratio and NADES concentration on HiC thermostability at
90 °C were also investigated. The choline dihydrogen citrate:xylitol, in a 1:1 ratio with a 20 wt% concentration,
was selected as the best combination in stabilizing HiC, increasing its half-life three-fold.