Purification and biochemical properties of SDS-stable low molecular weight alkaline serine protease from Citrullus colocynthis. Issue 8 (17th April 2016)

• Record Type: Journal Article
• Authors: Khan, Muhammad Bashir; Khan, Hidayatullah; Shah, Muhammad Usman; Khan, Sanaullah
• Is Part Of: Natural product research. Volume 30:Issue 8(2016)
• Page Start: 935
• Page End: 940
• Publication Date: 2016-04-17
• Subjects: Citrullus colocynthis -- serine protease -- purification -- biochemical characterisation -- thermostability -- detergent stability; Natural products -- Periodicals; Natural products -- Research -- Periodicals; Biological Factors -- Periodicals; Biological Products -- Periodicals; 547
• DOI: 10.1080/14786419.2015.1079909
• Languages: English
• ISSNs: 1478-6419

Abstract

Abstract: A low molecular weight serine protease from seeds of Citrullus colocynthis was purified to electrophoretic homogeneity with high level of catalytic efficiency (22, 945 M −1 S −1 ). The enzyme was a monomer with molecular mass of 25 kDa estimated by SDS–PAGE. The enzyme was highly active over a pH range of 6.5–9.0 and temperature range of 20–80 °C, with maximum activity at pH 7.5 and at 50 °C. The K m and K cat were 73 μg/mL and 67/s, respectively. The enzyme was strongly inhibited by PMSF, moderately by soybean trypsin inhibitor, indicating that the enzyme was a serine protease. The enzyme retained 86 and 73% of its activity in the presence of urea and DTT, respectively, and its activity was slightly enhanced in the presence of anionic detergent (SDS). Thus, the enzyme is a novel SDS-stable protease with high catalytic efficiency over wide ranges of pH and temperature which is commercially promising for various industrial applications. Graphical abstract: