Effect of ultrasonic treatment on enzymes: Decoupling the relation between the ultrasonic driven conformational change and enzyme activity

Considering the growing interest in enzyme-based extraction technology as a safe and eco-friendly extraction technique, along with the relatively high cost associated with enzymatic applications, it became necessary to explore novel strategies aimed to improve enzyme activity. In this study, the impact of ultrasonic treatment on commercial cellulase and pectinase was investigated. As this effect may be influenced by various ultrasonic and enzyme-related parameters, changes in enzyme conformation were explored under optimal and non-optimal enzyme conditions. The intrinsic fluorescence spectrum was utilized as a tool for monitoring these changes. Additionally, the enzyme's catalytic potential was also assessed under the same conditions. Results indicated that the impact of ultrasonic treatment on enzyme conformation primarily depends on the total ultrasonic energy delivered to the system, rather than other ultrasonic parameters such as power, sample volume, treatment time, or duty cycle. The maximum relative decrease in intrinsic fluorescence intensity of Pectinex® Ultra Clear (PUC) and Pectinex® Ultra SPL (PUS) after ultrasonic treatment was approximately 51% and 55%, respectively, while the decrease induced by thermal denaturation was 25% and 30% respectively. Furthermore, a blue shift in the fluorescence spectrum of both pectinases was observed upon sonication for all process conditions indicating a change in enzyme conformation. However, ultrasonic treatment did not result in a significant change in enzyme activity, suggesting that these conformational adjustments may occur in regions other than the active sites. Moreover, ultrasonicated pectinases and cellulases did not exhibit any improvement in their catalytic potential under either optimal or non-optimal conditions.