Three-phase partitioning for concentration and purification of laccase produced by submerged cultures of Ganoderma sp. WR-1

Three-phase partitioning (TPP), an efficient bioseparation technique, was used to purify laccase from fermentation broths of Ganoderma sp. WR-1. In a two-step procedure, the physiochemical parameters of ammonium sulfate concentration, the aqueous phase-to-t-butanol ratio, temperature and pH were optimized for laccase separation. In step 1 with 20% ammonium sulfate and a 1:0.5 (v/v) aqueous phase-to-t-butanol ratio at 35 °C and pH 7, most of the laccase remained in the lower aqueous phase. Step 2, with 90% ammonium sulfate and the other variables the same as in step 1, resulted in laccase concentrated at the interface. The two-step TPP led to 60% recovery of laccase with 13.2-fold purification. The TPP protocol developed in our study for the concentration and purification of laccase is rapid, simple and highly efficient.

Research highlights► The paper provides a study of downstream operations for laccase recovery. ► This is probably the first report on laccase separation using TPP. ► TPP was optimized to concentrate and purify laccase effectively. ► Laccase purity using TPP was better than ion exchange chromatography. ► This work helps select recovery protocols with a trade between purity and yield.