Membrane chromatography for the purification of laccase from the supernatant of Pleurotus sapidus

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 57.4 kDa. The fermentation broth had a low product titre of 80 μg/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.

► Laccase from fermentation broth purified using membrane chromatography. ► Membrane chromatography modules with different functionalisation. ► Pre-treatment of the fermentation broth not required. ► Enzyme activity is retained and enrichment up to fifteen fold observed.