A Comparative Kinetic Study of Free and Immobilized Urease on Commercial and Glutaraldehyde Activated Cotton


 Urease, cotton, immobilization, glutaraldehyde, hydrolysis.

How to Cite

M.M. Yousaf, & Umer Nasar. (2017). A Comparative Kinetic Study of Free and Immobilized Urease on Commercial and Glutaraldehyde Activated Cotton. Journal of Basic & Applied Sciences, 13, 321–325. https://doi.org/10.6000/1927-5129.2017.13.53


Urease was immobilized on commercial cotton and commercial cotton activated with glutaraldehyde. The kinetic of urea hydrolysis by free and immobilized urease was studied as a function of pH, temperature and time of hydrolysis. High concentration of ammonia was released at pH 10.0 for free enzyme (0.097 mg/mL) and urease immobilized on cotton (0.092 mg/mL) and pH 4.0 for enzyme, immobilized on activated commercial cotton (0.083 mg/mL). High concentration of ammonia was released at 30 °C for free enzyme (0.006 mg/mL), 25 °C for urease immobilized on cotton (0.043 mg/mL) and urease immobilized on cotton activated with glutaraldehyde (0.015 mg/mL). High concentration of ammonia was released after 10 minutes for free enzyme (0.0016 mg/mL), 60 minutes for inactivated (0.043 mg/mL) and for activated cotton (0.015 mg/ml). The result show that immobilized urease is less effective than the free enzyme and is more active in acid medium than the basic medium.



Sujoy B, Aparna A. Potential clinical significance of urease enzyme. Eur Sci J 2013; 9: 94-102.

Mobley HL, Island MD, Hausinger RP. Molecular biology of microbial ureases. Microbiol Rev 1995; 95: 451-480.

Bairoch A. The Enzyme database in 2000. Nucl Acids Res 2000; 28: 304-305. https://doi.org/10.1093/nar/28.1.304

Banerjee S, Aggarwal A. Enzymology, Immobilization and applications of Urease Enzyme. Intern Res J Biol Sci 2013; 2: 51-56.

Banerjee S, Aggarwal A. Isolation, partial purification, Characterization and inhibition of urease (E.C. enzyme from the Cajanus cajan seeds. Asian J Biol Science 2012; 7: 203-209.

Becker-Ritt AB, Martinelli AH, Mitidieri S, Feder V, Wassermann GE, Santi L, Vainstein MH, Oliveira JT, Fiuza LM, Pasquali G, Carlini CR. Antifungal activity of plant and bacterial ureases. Toxicon 2007; 41: 821-827. https://doi.org/10.1016/j.toxicon.2007.07.008

Benoit SL, Zbell AL, Maier RJ. Nickel enzyme maturation in Helicobacter hepaticus, roles of accessory proteins in hydrogenase and urease activities. Microbiology 2007; 153: 3748-3756. https://doi.org/10.1099/mic.0.2007/010520-0

Krajewska B, Van-Eldik R, Brindell M. Temperature- and Pressure-dependent stopped-flow kinetic studies of Jack Bean urease. Implications for the catalytic mechanism. J Biol Inorganic Chem 2012; 17: 1123-1134. https://doi.org/10.1007/s00775-012-0926-8

Menegassi A, Wassermann GE, Olivera-Severo D, Becker-Ritt AB, Martinelli AHS, Feder V, Carlini CR. Urease from Cotton (Gossypium hirsutum) Seeds, Isolation, Physicochemical Characterization, and Antifungal Properties of the Protein. J Agric Food Chem 2008; 56: 4399-4405. https://doi.org/10.1021/jf0735275

El-Sherif H, De Maio A, Di Martino S, Zito E. et al. Urease immobilization on chemically grafted nylon membranes - Part 2. Non-isothermal characterization. J Mol Catal B Enz 2001; 14(1): 31-43. https://doi.org/10.1016/S1381-1177(00)00240-X

Barth A, Rollka W, Michel HJ. Phosphoric acid ester diamides as urease inhibitors. A contribution to the research of active compounds for development of tight-binding inhibitors of selected enzymes, Wiss.Beitr. Mal1in-Luther-Univ. Halle-Wittenberg 1980; (2): 5-10.

Krajewska B, Zaborska W. Jack bean urease. The effect of active-site binding inhibitors on the reactivity of enzyme thiol groups. Bioorg Chem 2007; 35: 355-365. https://doi.org/10.1016/j.bioorg.2007.02.002

DeGroot AR, Neufeld RJ. Encapsulation of urease in alginate beads and protection from ?- chymotrypsin with chitosan membranes. Enz Microb Technol 2001; 29: 321-327. https://doi.org/10.1016/S0141-0229(01)00393-3

Alther. Preparing organoclay-enzyme complexes using a quaternary ionic compound and mineral. US Pat 6534296. 2003.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein Measurement with the Folin Phenol Reagent. J Biol Chem 1951; 193: 265-275.

Robertson GP, Vitousek PM. Nitrogen in agriculture: balancing the cost of an essential resource. Annual Review of Environment and Resources 2009; 34: 97-125. https://doi.org/10.1146/annurev.environ.032108.105046

GayBayrmi L, Emine Y, Yakup M. immobilization of urease through adsorption onto L-histidine-Ni Complexed poly (HEMA-MAH microspheres: Preparation and characterization. Process Biochem 2005; 23: 3503-3513.

Guo Z, Bai S, Sun Y. Preparation and characterization of immobilized Lipase on magnetic nanoparticles Biotechnol Prog 2003; 3: 1095-1100.

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