Production and purification of laccase from Trametes versicolor

Abstract

This study describes the production and purification of laccase from Trametes versicolor isolate. The activated fungus was grown in 50 mL YpSs broth medium for 3 days at 30 °C and 120 rpm shaking speed. Then, pellets were transferred to the laccase production medium including 5 g/L glucose, 1 g/L yeast extract, 1 g/L beef extract, 0.2 g/L pepton, 1 g/L KH2PO4, 0.5 g/L MgSO4, 0.1 g/L CuSO4, 3 g/L (NH4)2SO4, 1 g/L CaCl2, and 1 g/L Na2HPO4. The highest enzyme activity was found after the sixth day of cell growth. For purification of laccase from crude extract, three-phase partitioning was used for the first time. The enzyme was purified by 9.5-fold with an enzyme activity recovery of 93% from the TPP system prepared at pH 7.0 with a ratio of 1:1 crude extract:t-butanol and containing 30% (w/w) ammonium sulphate. As a result, it is shown that the laccase enzyme from T. versicolor can be purified using three-phase systems, which are timesaving, inexpensive, and easy to use, instead of the traditional chromatography method.