Partial purification and characterization of esterase from the seeds of Phaseolus vulgaris

Abstract

Carboxylesterases are hydrolases which catalyze the hydrolysis of various types of esters. In this study, carboxylesterases enzyme was isolated from the soaked seeds of Phaseolus vulgaris. Partial purification of esterases from Phaseolus vulgaris was done by conventional protein purification techniques such as CM-Cellulose ion exchange chromatography and Sephadex G-75 gel filtration Chromatography. Kinetic parameters such as pH, temperature, Km and Vmax of the purified enzyme was studied. Optimum pH value was determined as 6.5 and was found to be stable between pH range of 6-7. Optimum temperature was examined as 37°C with a ranging from 15 ⁰C – 37 ⁰C. The Km and Vmax values were found to be 0.15mM and 0.013µmoles/min, respectively. Finally, half-maximal inhibitory concentration IC50 was found to be 2.8×10-6.