Biotechnology & Biotechnological Equipment (Jan 2021)
Ultrasound-assisted lipase-catalyzed synthesis of ethyl acetate: process optimization and kinetic study
Abstract
Synthesis of ethyl acetate (EA) using lipase as a catalyst, as an environmentally-friendly process, has attracted considerable attention. However, the slow rate of enzymatic reaction and higher cost limit its applications. Ultrasound irradiation (UI) is known as an effective tool for accelerating the rate of enzymatic reaction under mild conditions. To attain maximum yield, it is crucial to understand the effect of UI on EA synthesis and the underlying mechanism. Therefore, the present study investigated the effect of UI on lipase-catalyzed ethyl acetate synthesis. The results clearly demonstrated that UI could remarkably accelerate the process of lipase-catalyzed synthesis of ethyl acetate compared with mechanical shaking. The optimal conditions of ultrasound-assisted reaction by using Novozyme 435® were 6% enzyme loading, 4:1 molar ratio of ethanol to acetic acid, 150 W & 28 kHz ultrasonic power and frequency, respectively. Bi-bi ping-pong model was employed to give a deep insight into the underlying mechanism of the beneficial effect of UI. It was found that Vmax under UI was elevated by 143% compared with that under mechanical shaking. Application of UI could decrease Kacetic acid and Ki ethanol while increase Kethanol and Ki acetic acid. In conclusion, the present study addresses that UI could remarkably accelerate the process of lipase-catalyzed synthesis of EA compared with mechanical shaking, and kinetic model results explained why ultrasound can significantly speed up lipase catalyzed synthesis of ethyl acetate, which may help to improve the industrial application potential of ethyl acetate biosynthesis.
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