The wide-spread use of laccases in industry is often limited due to the enzyme inactivation over time at conditions which exceeds the operating conditions of the enzymes, which are neutral pH and ambient temperatures (30-40 °C). Natural Deep Eutectic Solvents (NADESs) have attracted considerable attention as reaction media in biocatalysis due to their promising compatibility with enzymes and sustainable derivation. In this contribution we demonstrate the possibility of applying aqueous NADESs as incubation media to alter the activity of and inhibit thermal inactivation of laccase T. versicolor. For example we show that by incubating 0.25 g L-1 laccase in an aqueous 25 wt% betaine-xylitol based NADES at 70 °C for 15 min, the measured residual activity of laccase is a near 10 fold greater than the measured residual activity of laccase when incubated without the NADES. Moreover, the comparison of the residual activities of the enzyme in presence betaine, xylitol or NADES is clearly showing the advantage of using a NADES because a synergistic effect. The drastic enhancement of the enzyme thermostability by pre-incubation of laccase in NADES media showcases a facile, cheap and green method of boosting the stability laccase.

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.

Poly(butylene succinate-co-butylene adipate) (PBSA)-based materials are receiving growing attention in the packaging industry for their promising biodegradability. However, poor gas barrier properties and low durability of biodegradable polymers, such as PBSA, have limited their wide-spread use in food packaging applications. Here we report a scalable solution to improve gas barrier properties and stabilize PBSA against photo-aging, with minimal modifications to the biodegradable polymer backbone by using a commercially available and biocompatible layered double hydroxide (LDH) filler. We investigate and compare the mechanical, gas barrier, and photoaging properties of PBSA and PBSA-LDH nanocomposite films produced on a pilot scale. An increase in rigidity in the nanocomposite was observed upon addition of LDH fillers to neat PBSA, which direct the application of neat PBSA and PBSA-LDH nanocomposite to different food packaging applications. The addition of LDH fillers into neat PBSA improves the oxygen and water vapour barriers for the PBSA based nanocomposites, which increases the attractiveness of PBSA material in food packaging applications. Through changes in the viscoelastic behaviour, we observe an improved photo-durability of photoaged PBSA-LDH nanocomposites compared to neat PBSA. It is clear from our studies that the presence of LDH enhances the lifetime durability and modulates the photodegradation rate of the elaborated biocomposites.

Electrochemical continuous-flow reactors offer a great opportunity for enhanced and sustainable chemical syntheses. Here, we present a novel application of electrochemical continuous-flow oscillatory baffled reactors (ECOBRs) that combines advanced mixing features with electrochemical transformations to enable efficient electrochemical oxidations under continuous flow at a millimeter distance between electrodes. Different additive manufacturing techniques have been employed to rapidly fabricate reactors. The electrochemical oxidation of NADH, a very sensitive substrate key for the regeneration of enzymes in biocatalytic transformations, has been employed as a benchmark reaction. The oscillatory conditions improved bulk mixing, facilitating the contact of reagents to electrodes. Under oscillatory conditions, the ECOBR demonstrated improved performance in the electrochemical oxidation of NADH, which is attributed to improved mass transfer associated with the oscillatory regime.