Three-Phase Partitioning for Purification of Laccase Produced by Coriolopsis trogii under Solid Fermentation

摘要

In order to explore separation and purification methods of Coriolopsis trogii laccase production by solid fermentation, we used a three-phase separation technique and wherein the main factors (ammonium sulfate concentration, t butanol concentration, temperature and pH) were optimized. The results showed that when the three-phase partitioning optimum conditions were as follows: Ammonium sulfate concentration was 40%, tertbutanol water phase volume ratio of 1:1, the temperature is 20°C, pH 5, laccase purification factor and recovery can be up to 20 times and 75%, respectively, SDS-PAGE electrophoresis showed that the purified protein is substantially a single band and a molecular weight of about 38 kD, smaller than the known fungal laccase molecular weight; the results prove that the three-phase separation from the crude enzyme solution Coriolopsis trogii solid obtained after fermentation relatively pure laccase, eliminating the need for complex chromatography step, the method is simple and low cost, for a lot of separation and purification of laccase provide practical foundation.