Purification and characterisation of laccase from lactarius volemus and its application in removal of phenolic compounds from fruit juice

Laccases are multicopper oxidases (EC 1.10.3.2.), widely distributed in fungi, higher plants and bacteria. In this research, a laccase from Lactarius volemus (Russulaceae) was purified by 73.08 fold using precipitate of saturation (NH4)2SO4, DEAE-cellulose and Sephacryl S200. It was shown that purified enzyme was homogeneous in term of SDS-PAGE with molecular mass estimated of 30 kDa. The pH was optimal at 4.0, enzyme was stable at pH 3.0-5.0 and optimal temperature was at 50°C. Cu2+ and Fe2+ were activated to the purified laccase enzyme, and it was partially inhibited by 5 mM concentration of some metal salts and EDTA, urea, SDS, L-cysteine, dithioeritritol and β-mercaptoethanol. In addition, it was investigated whether the purified and characterized laccase enzyme would remove the phenolic compound of some fruit juices which are responsible for haze formation and browning during storage. Their application led to reduction of total phenolic compounds for apple, apricot, cherry and grape juice. As a result, the effect of these enzymatic treatments on total phenol content and clarity of the juice were evaluated using a central composite design. This laccase enzyme had potential application in food processing.