Enhancing efficiency of ultrasound-assisted biodiesel production catalyzed by Eversa® Transform 2.0 at low lipase concentration: Enzyme characterization and process optimization.

This study focused on the ultrasound-assisted transesterification of simulated low-quality feedstocks using a low-cost liquid lipase Eversa® Transform 2.0 (ET2). Enzyme characterization was also performed to investigate the effect of ultrasound parameters on enzyme structure. The optimal ultrasound parameters, 40 % amplitude, and 5 % duty cycle effectively enhanced the reaction rate compared to the conventional stirring method while retaining 95 % of the enzyme activity. Analysis of circular dichroism (CD) spectra revealed the preservation of the secondary structure of ET2 under the optimal ultrasound intensities, while fluorescence spectra indicated a slight change in its tertiary structure. The implementation of a two-stage methanol dosing strategy in the ultrasound-assisted reaction effectively mitigated lipase inhibition, yielding a remarkable fatty acid methyl ester (FAME) content of 92.2 % achieved within a 12-h reaction time. Notable, this high FAME content was achieved with only a 4:1 methanol-to-oil molar ratio and a 0.5 wt% enzyme concentration. Under these optimized conditions, the ultrasound-assisted reaction also demonstrated a 15 % improvement in the final FAME content compared to the conventional stirring method. These promising results hold significant potential for advancing the field of biodiesel production via ultrasound technology, contributing substantively to sustainable energy sources.

Stabilization of Eversa® Transform 2.0 lipase with sorbitol to enhance the efficiency of ultrasound-assisted biodiesel production.

Ultrasound technology has emerged as a promising tool for enhancing enzymatic biodiesel production, yet the cavitation effect induced can compromise enzyme stability. This study explored the efficiency of polyols in enhancing lipase stability under ultrasound conditions to further improve biodiesel yield. The incorporation of sorbitol resulted in the highest fatty acid methyl ester (FAME) content in the ultrasound-assisted biodiesel production catalyzed by Eversa® Transform 2.0 among the investigated polyols. Furthermore, sorbitol enhanced the stability of the lipase, allowing it to tolerate up to 100 % ultrasound amplitude, compared to 60 % amplitude in its absence. Enzyme activity assays revealed that sorbitol preserved 99 % of the lipase activity, in contrast to 84 % retention observed without sorbitol under an 80 % ultrasound amplitude. Circular dichroism (CD) and fluorescence spectroscopy analyses confirmed that sorbitol enhanced lipase rigidity and preserved its conformational structure under ultrasound exposure. Furthermore, employing a stepwise methanol addition strategy in ultrasound-assisted reactions with sorbitol achieved an 81.2 wt% FAME content in 8 h with only 0.2 wt% enzyme concentration. This promising result highlights the potential of sorbitol as a stabilizing agent in ultrasound-assisted enzymatic biodiesel production, offering a viable approach for enhancing biodiesel yield and enzyme stability in industrial applications.