An alkaline protease from fresh fruiting bodies of the edible mushroom Pleurotus citrinopileatus

A protease was purified from fresh fruiting bodies of the edible mushroom Pleurotus citrinopileatus. The isolation procedure included ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose and fast protein liquid chromatography-gel filtration on Superdex 75. The protease was unadsorbed on DEAE-cellulose and Q-Sepharose, but adsorbed on CM-cellulose. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protease demonstrated a single band with a molecular mass of 28 kDa. The protease showed an optimal pH at 10 and an optimal temperature at 50°C. The activity of the protease was not affected by EDTA, indicating that it is not a metalloprotease. The protease exhibited a higher activity in the presence of K+ and Li+, but its activity was potently inhibited by Al3+, Cu2+, and Hg2+ ions. It manifested a Km of 3.44 mg/ml and a Vmax of 0.139 mg ml−1 min−1. It was devoid of ribonuclease and antifungal activities.