Optimization of Laccase Production and Purification from Penicillium chrysogenum

Laccase (benzenediol:oxygen oxidoreductase, EC 1.10.3.2) is one of the best-known multicopper enzymes and catalyzes the oxidation of a variety of aromatic compounds, in particular phenolic substrates, coupled to the reduction of molecular oxygen to water. Using molecular oxygen as an electron acceptor, laccase oxidizes its substrates by an electron transfer mechanism that generates unstable free radical intermediates and causes non-enzymatic reactions that break down substrate molecules. The enzyme was isolated from Penicillium chrysogenum. The study discovered that laccase production reached its highest value occurred after 72 hours of incubation at 35°C and pH 7.0. Laccase acted on various phenolic compounds including ABTS, 2,6-DMP, syringaldazine, guaiacol, catechol in addition to pyrogallol and BTS was the best substrate. Laccase was purified using ammonium sulfate fractionation, Sephadex G-200 and DEAE-Sepharose with final specific activity of 113.5 units mg-1 protein, 103.2-fold of purification and recovery of 26.8%.Laccase enzyme is very important to many applications in our life.