Biochemical, Thermodynamic And Kinetic Characterization Of Glucose Oxidase Purified From Pseudomonas And Actinomyces Spp. From Natural Sources

Glucose Oxidase (GOX) is naturally produced by several microorganisms. In this article, the enzymes isolated from Pseudomonas and Actinomyces species were studied for enzyme characteristics, activity, stability and kinetic parameters. Enzyme extracted from Strain 1 (ES1), has shown optimum activity at 27 degrees C and pH 5, half-life at 30 degrees C. The enzyme was highly tolerant to AgNO3 (1.4mM) and less tolerant to NaCl; but was stable at 2.4mM of NaCl. 96% of activity was observed at 1.7mmol of Mg2+. 94% and 83% of activity were seen for Co and Cu when used as chelating agents. Denaturation of the enzyme occurred when DTAB was tested for its denaturing effect. 92% of enzyme activity was recorded by D-glucose when used as a substrate. The activation energy of 23.95 kJmol/l, 27 degrees C, Vmax of 1.2U and Km of 6.91mM were recorded. Whereas Enzyme extracted from Strain 2 (ES2), reported optimum activity at 30 degrees C and 5 pH, attained half-life after 30 minutes at 45 degrees C. ES2 exhibited tolerance CoCl2 at 1.6mM and HgCl2 at 0.6mM. Stability of the enzyme observed at 3mM concentration for all salts used in experimentation. Enzyme activity of 98% for Mg2+ and 0% for Fe2+ were recorded among other metal ions. Enzyme activity of 87% for Co and 78% for Cu when used as chelating agents. Denaturation of the enzyme occurred when urea was used. 96% for D-glucose and 20% for sucrose were calculated as enzyme activity. Activation energy of 39.5 kJmol, 30 degrees C, Vmax of 0.7U and Km of 72mM were recorded.