Abstract：A novel reversed micellar system had been developed to investigate the purification of lignin peroxidase (LiP). The anionic biosurfactant rhamnolipid (RL) was used to construct the reversed micellar system, while the isooctane and n-hexanol (1:1, V/V) were used as the apolar organic solvent and cosurfactant, respectively. In both the forward and backward extraction processes, the effects of RL concentration, shaking time, the pH value and ionic strength of water phase were studied. The results showed that the best condition for forward extraction was at 2.75mmol/L RL, 0.04mol/L KCl, pH 3.0 and with the shaking time of 30min (200r/min). As regards backward extraction process, 0.5mol/L KCl and pH 6.0 were suggested. In the above optimum conditions, the LiP activity recovery (AR) and purification fold (PF) were up to 93% and 2.9 times, respectively. The SDS-PAGE analysis further proved that the LiP was successfully purified.
