ISOLATION, PURIFICATION AND PARTIAL CHARACTERIZATION OF PEROXIDASE FROM SNAKE GOURD (TRICHOSANTHES CUCUMERINA L.)

ABSTRACT This study focused on the isolation, purification, and partial characterization of peroxidase from snake gourd (Trichosanthes cucumerina), an underexplored plant species with potential industrial enzyme applications. Peroxidase was extracted using 10 mM sodium phosphate buffer (pH 6.0) containing 10% glycerol and 0.2% ascorbic acid. The peroxidase was purified using aqueous two-phase partitioning. The partially purified T. cucumerina peroxidase was thereafter characterized. The crude extract showed an activity of 135.1 2.0 U/mL, with a protein concentration of 0.81 0.1 mg/mL, yielding a specific activity of 167 3.0 U/mg protein. Purification using an aqueous two-phase partitioning system resulted in a 45% yield and a 2-fold purification. The enzyme exhibited preference for catechol. Kinetic characterization of the purified enzyme revealed Michaelis-Menten constants (Km) of 7.5 0.2 mM for catechol and 17.4 0.3 mM for hydrogen peroxide, with a maximum reaction velocity (Vmax) of 476.1 0.5 U/mg protein. The enzyme exhibited optimal activity at pH 6.0 and was stable within a pH range of 5.0±7.0. Temperature profiling indicated an optimum activity at 50 ƒ& with stability maintained between 30 ƒ& and 60 ƒ& The enzyme retained approximately 75% of its activity after 1 hour of incubation at 40 ƒ& These findings highlight T. cucumerina as a promising plant source of peroxidase with favorable biochemical properties, including good thermal and pH stability. This work contributes new knowledge on the enzymatic potential of snake gourd and lays a foundation for future industrial and biotechnological applications of its peroxidase.