Purification and characterization of the cold-active alkaline protease from marine cold-adaptive Penicillium chrysogenum FS010

摘要

An extracellular cold-active alkaline serine protease from Penicillium chrysogenum FS010 has been purified. The purification procedure involved: ammonium sulfate precipitation, DEAE ion-exchange chromatography and sephadex G-100 gel chromatography. SDS–PAGE of the purified enzyme indicated a molecular weight of 41,000 ± 1,000 Da. The protease is stable in a pH range of 7.0–9.0 and has a maximum activity at pH 9.0. Compared with other industrial proteases, the enzyme shows a high hydrolytic activities at lower temperatures and a high sensitivity at a temperature over 50°C. The isoelectric point of the enzyme is approximate to 6.0. Enzymatic activity is enhanced by the addition of divalent cations such as Mg2+ and Ca2+ and inhibited by addition of Cu2+and Co2+. PMSF and DFP are its specific inhibitors. The application of the cold-active alkaline protease is extremely extensive, and widely used in detergents, feed, food, leather and many other industries.