Purification, catalytic, kinetic and thermodynamic characteristics of a novel ficin from Ficus johannis. (April 2017)

• Record Type: Journal Article
• Authors: Homaei, Ahmad; Stevanato, Roberto; Etemadipour, Rasoul; Hemmati, Roohullah
• Is Part Of: Biocatalysis and agricultural biotechnology. Number 10(2017)
• Page Start: 360
• Page End: 366
• Publication Date: 2017-04
• Subjects: Enzyme purification -- Thermodynamic parameters -- Kinetic properties -- Homag Mountain; Agricultural biotechnology -- Periodicals; Enzymes -- Biotechnology -- Periodicals; 660.6
• DOI: 10.1016/j.bcab.2017.04.008
• Languages: English
• ISSNs: 1878-8181

Abstract

Abstract: Ficin from Ficus johannis was purified to homogeneity by (NH4 )2 SO4 precipitation, CM-Sepharose and SP-Sepharose chromatography, and gel filtration Sephadex G-25 chromatography. The enzyme, that is characterized by a molecular weight of 25 kDa measured by SDS-PAGE, showed higher affinity towards casein and the highest catalytic efficiency at pH 7.0 and 60 °C. Catalytic hydrolysis rate follows the Michaelis–Menten equation and values of 32.9 s −1 and 13.7 µM for k cat and K m respectively, by Michaelis–Menten, were found. The pK a1 and pK a2 values of the active site ionizable groups controlling V max were 4.2 and 8.7, respectively. Thermodynamic parameters for soluble casein hydrolysis and for irreversible inactivation of ficin at different temperatures in the range from 45 to 60 °C were also determined. This is the first report on the thermodynamic parameters of ficin produced by Ficus johannis. These results indicate that Ficin from Ficus johannis may have potential applications in the industrial biotechnology.