Development of New Affinity Material for Urease Purification: Synthesis and Characterization

Abstract

Urease (urea amidohydrolase, E.C.3.5.1.5) is an enzyme that catalyzes the hydrolyzation of urea to form ammonium and carbon dioxide. This is an essential reaction in industrial and medical applications. Urease has a vital role in accelerating the hydrolysis of the urea in the agricultural gut, calculating the amount of urea in biological fluids, urine removal from artificial kidneys, and urea removal in wastewater and fruit juices. Purifying urease is also crucial for treating some diseases, such as gastrointestinal infection and hypertension. This research aimed at developing an environmentally friendly green biocomposite with high mechanical stability by suspension polymerization of chitosan (CTS), 2-hydroxyethyl methacrylate (HEMA), and N-methacryloyl- (L) -glutamic acid (MAGA) and to determine its optimum conditions for urease purification. The synthesized p (HEMA-MAGA) -CTS sorbent was investigated by FTIR, XPS, XRD, TGA, SEM, and particle size analysis. After the characterization, the parameters affecting the adsorption of urease, such as pH, sorbent amount, contact time, initial urease concentration, heat, and ionic strength, were optimized. In this study, equilibrium adsorption isotherms of urease on the synthesized affinity sorbent were examined and The Freundlich isotherm was found to provide a much better fit than the Langmuir and Dubinin-Radushkevich isotherms. The urease adsorbed on the p(HEMA-MAGA)-CTS affinity sorbent was desorbed in over 75% yield with 1.0 M NaSCN (pH 8.0) in 2 hours. The maximum adsorption capacity was found to be 9.47 mg mg-1 at pH 4.0 and 25°C. p(HEMA-MAGA)-CTS for urease purification was successfully developed.