Abstract Extracellular laccase from the fermentation broth of Pleurotus sapidus was purified using membrane chromatography. Five laccase iso-enzymes produced in submerged cultures were purified; the enzymes had isoelectric points (pI) ranging from 3.3 to 4.7 and a calculated molecular weight of 57.4kDa. The fermentation broth had a low product titre of 80μg/l. Hydrophobic interaction (HIC) and ion exchange (IEX) membranes chromatography modules were used in an ÄKTApurifier 100 liquid chromatography system for the dynamic experiments. The pre-treatment of the fermentation broth was not required. The enzyme activity was retained, and enrichment up to 15-fold was observed via activity assays and SDS-Page analysis.