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

• Published in: Molecular biology reports Vol. 36; no. 8; pp. 2169 - 2174
• Main Authors: Zhu, Hui-Yuan, Tian, Yong, Hou, Yun-Hua, Wang, Tian-hong
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01-11-2009; Springer Netherlands; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Bacteria; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Biochemistry; Biomedical and Life Sciences; Endopeptidases - chemistry; Endopeptidases - isolation & purification; Endopeptidases - metabolism; Enzyme Stability; Enzymes; Fungal Proteins - chemistry; Fungal Proteins - isolation & purification; Fungal Proteins - metabolism; Histology; Hydrogen-Ion Concentration; Life Sciences; Metals - chemistry; Molecular biology; Morphology; Penicillium chrysogenum - chemistry; Penicillium chrysogenum - enzymology; Proteases; Temperature; Enzymatic characterization; Cold-active alkaline protease; Purification
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-008-9431-0

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.