Enzymatic properties of a novel highly active and chelator resistant protease from a Pseudomonas aeruginosa PD100

• Published in: Enzyme and microbial technology 2006-11, Vol.39 (7), p.1433-1440
• Main Authors: Najafi, Mohsen Fathi, Deobagkar, Dileep N., Mehrvarz, Mohsen, Deobagkar, Deepti D.
• Format: Article
• Language: English
• Subjects: Acid-PAGE; Biological and medical sciences; Biotechnology; Collagen; Disulfide bonds; Fundamental and applied biological sciences. Psychology; Inhibitors; P. aeruginosa; Protease; Pseudomonas aeruginosa
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2006.03.031

Pseudomonas aeruginosa PD100 capable of producing an extracellular protease was isolated from the soil collected from local area (garbage site) from Shivage market in Pune, India. The purified protease showed a single band on native and SDS-PAGE with a molecular weight of 36 kDa on SDS-PAGE. The optimum pH value and temperature range were found to be 8 and 55–60 °C, respectively. The enzyme exhibited broad range of substrate specificity with higher activity for collagen. The enzyme was inhibited with low concentration of Ag 2+, Ni 2+, and Cu 2+. β-Mercaptoethanol was able to inactivate the enzyme at 2.5 mM, suggesting that disulfide bond(s) play a critical role in the enzyme activity. Studies with inhibitors showed that different classes of protease inhibitors, known to inhibit specific proteases, could not inhibit the activity of this protease. Amino acid modification studies data and p K a values showed that Cys, His and Trp were involved in the protease activity. P. aeruginosa PD100 produces one form of protease with some different properties as compared to other reported proteases from P. aeruginosa strains. With respect to properties of the purified protease such as pH optimum, temperature stability with capability to degrade different proteins, high stability in the presences of detergents and chemicals, and metal ions independency, suggesting that it has great potential for different applications.