EXTRACTION, PARTIAL PURIFICATION, AND CHARACTERIZATION OF UREASE FROM CHICKPEA SEEDS (Phaseolus vulgaris L.)

Abstract

Urease is an enzyme which as an essential role as a catalyst in the hydrolysis reaction of urea into ammonia and CO₂. Urease has been extracted from legumes, and chickpea seeds standing out among the legume varieties with the potential as a urease source. This research aims to delineate the attributes of urease derived from chickpea seeds. The urease enzyme was isolated from the chickpea seeds, resulting in a crude enzyme extract. Subsequently, this crude extract underwent purification utilizing ammonium sulfate salt through the precipitation principle. Fractions exhibiting the highest activity were subject to characterization encompassing pH, temperature, substrate concentration, and the addition of metallic elements. The urease activity assay employed the Nessler method, with the absorbance measurements conducted using a spectrophotometer at the wavelength of 500 nm. Urease originating from chickpea seeds demonstrated an optimal activity at the pH of 7, incubation temperature of 35 °C, and substrate concentration of 0.2 M, resulting in the F30 activity of 17.720 U/mL. Notably, the metal ions of Ni²⁺ and Hg²⁺ acted as inhibitors for urease