Abstract:A mixed reversed micelle system had been developed to investigate the catalytic performance of laccase. The anionic biosurfactant Rhamnolipid and nonionic surfactant Tween-80 were used to construct the mixed reversed micelle system, while the isooctane and short-chain alcohols (1:1, V/V) were used as oil phase and cosurfactant, respectively. UV spectrum was analysed to find out how the molar fraction Tween-80, water content, pH, ionic strength and cosurfactant stype affected enzyme activity of laccase in the mixed reversed micelle system. And it has been found that while the molar fraction Tween-80 was 30%, water content was 20, pH was 4.5, the concentration of KCl was 70mmol/L, cosurfactant stype was n-hexanol, the optimal condition of mixed reversed micelle system could be reached. In optimum catalytic conditions, the catalytic activity was higher 1.44 times in the mixed reversed micelle than that in the single Rhamnolipid reversed micelle system, and higher 4.35 times than that in water. The research indicated mixed reversed micelle has huge potential in micellar enzymology.