In recent years,
natural deep eutectic solvents (NADESs) have gained
increasing attention as promising nontoxic solvents for biotechnological
applications, due to their compatibility with enzymes and ability
to enhance their activity. Betaine-based NADESs at a concentration
of 25 wt % in a buffered aqueous solution were used as media to inhibit
thermal inactivation of POXA1b laccase and its five variants when
incubated at 70 and 90 °C. All the tested laccases showed higher
residual activity when incubated in NADES solutions, with a further
enhancement achieved also for the most thermostable variant. Furthermore,
the residual activity of laccases in the presence of NADESs showed
a clear advantage over the use of NADESs’ individual components.
Molecular docking simulations were performed to understand the role
of NADESs in the stabilization of laccases toward thermal inactivation,
evaluating the interaction between each enzyme and NADESs’
individual components. A correlation within the binding energies between
laccases and NADES components and the stabilization of the enzymes
was demonstrated. These findings establish the possibility of preincubating
enzymes in NADESs as a facile and cost-effective solution to inhibit
thermal inactivation of enzymes when exposed to high temperatures.
This computer-aided approach can assist the tailoring of NADES composition
for every enzyme of interest.