Abstract
► The biosurfactant rhamnolipid was employed in reversed micellar extraction for purification of laccase. ► Parameters in both forward and backward extraction process were optimized. ► The laccase after extraction was purified 4.79 fold and confirmed by SDS-PAGE profiles. ► The revered micellar system can be reused after backward extraction.
Biosurfactant-based reversed micellar extraction (RME) is an innovative method for separation and purification of biomolecules. In this study, rhamnolipid (RL), a kind of biosurfactant, was firstly adopted to form a novel reversed micellar system for extracting and purifying laccase from Coriolus versicolor crude extract. Several significant factors affecting both forward and backward extraction processes were studied. The appropriate conditions for forward extraction process were: 3.3mM RL, 50mM KCl, pH 5.5 and extracting time 40min. As regards backward extraction process, 250mM KCl, pH 7.0 and extracting time 40min were suggested. The corresponding activity recovery (AR) and purification fold (PF) were 92.7% and 4.79, respectively. Electrophoresis analysis indicated that the laccase was successfully purified. After this reversed micellar system was reused three times, the AR and PF declined to 70.8% and 4.35, respectively, indicating that the reversed micellar system could be reused. Comparisons results of synthetic surfactant-based RME and RL-based RME further verified the superiority of RL.