Immobilization of enzymes is a good field of study to extend the life of enzyme and reduce the cost of the chemical processes, such as separation processes. Urease is an important enzyme with medical and industrial applications. The aim of the present study is to prepare an immobilized urease on a strong cation exchange resin (Amberlite IR120 Na) and study its activity and stability. We monitored the release of Na ions in the collected fractions and searching for enzyme in the fractions as indicators of immobilization by ion exchange phenomenon. Sodium is determined by using atomic absorption spectroscopy technique, while the enzyme concentration was tested by Bradford’s method. Immobilized urease activity was evaluated by salicylate-hypochlorite method. The results indicated a complete immobilization of urease enzyme on the resin surface with reserving 92% of the activity of free enzyme. The immobilized urease enzyme on resin showed good stability and it has a 62% of its activity after 154 days of storage at room temperature. It is concluded that a new immobilized urease enzyme system is prepared with good enzyme activity and stability.
If the inline PDF is not rendering correctly, you can download the PDF file here.
. R.P. Pogorilyi, I.V. Melnyk, Y.L. Zub, G.A. Seisenbaeva, V.G. Kessler, Enzyme immobilization on a nanoadsorbent for improved stability against heavy metal poisoning, Colloids Surf. B 144 (2016) 135-142.
. G. Pozniak, B. Krajewska, W. Trochimczuk, Urease immobilized on modified polysulphone membrane: Preparation and properties, Biomaterials 16 (1995) 129-134.
. A.K. De Brito, C.S. Nordi, L. Caseli, Algal polysaccharides as matrices for the immobilization of urease in lipid ultrathin films studied with tensiometry and vibrational spectroscopy: Physical-chemical properties and implications in the enzyme activity, Colloids Surf. B 135 (2015) 639-645.
. Y. I. Doğaç, I. Deveci, M. Teke, B. Mercimek, TiO2 beads and TiO2-chitosan beads for urease immobilization, Mater. Sci. Eng. C 42 (2014) 429-435.
. F.S. Alatawi, M. Monier, N.H. Elsayed, Amino functionalization of carboxymethyl cellulose for efficient immobilization of urease, Int. J. Biol. Macromol. 114 (2018) 1018-1025.
. S.H. Baysal, R. Karagöz, Preparation and characterization of kappa-carrageenan immobilized urease, Prep. Biochem. Biotechnol. 35 (2005) 135-143.
. F.Y. Mahlicli, S.A. Altinkaya, The effects of urease immobilization on the transport characteristics and protein adsorption capacity of cellulose acetate based hemodialysis membranes, J. Mater. Sci. Mater. Med. 20 (2009) 2167-2179.
. D.E. Anderson, S. Balapangu, H.N. Fleischer, R. Viade, F. Krampa, P. Kanyong, G. Awandare, E. Tiburu, Investigating the influence of temperature on the kaolinite-base synthesis of zeolite and urease immobilization for the potential fabrication of electrochemical urea biosensors, Sensors (Basel) 17 (2017) 1831.
. A. Tiwari, S. Aryal, S. Pilla, S. Gong, An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles, Talanta 78 (2009) 1401-1407.
. E. György, F. Sima, I. Mihailescu, T. Smausz, G. Megyeri, R. Kékesi, B. Hopp, L. Zdrentu, S. Petrescu, Immobilization of urease by laser techniques: synthesis and application to urea biosensors, J. Biomed. Mater. Res. A 89 (2009) 186-191.
. N. Mohamad, N. Marzuki, N. Buang, F. Huyop, R. Wahab, An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes, Biotechnol. Biotechnol. Equip. 29 (2015) 205-220.
. L. Zhang, Y. Du, J. Song, H. Qi, Biocompatible magnetic nanoparticles grafted by poly(carboxybetaine acrylamide) for enzyme immobilization, Int. J. Biol. Macromol. 118 (2018) 1004-1012.
. H.K. Al-Hakeim, M. K. Kadhem, E.A. Grulke, Immobilization of urease enzyme on nanoceria modifies secondary and tertiary protein structures, Acta Chimica Slovaka 9 (2016) 44-53.
. H.K. Al-Hakeim, I.M. Al-Dahan. R. Bustan, Interaction of prolactin hormone with the surfaces of two new azo compounds, Int. J. Pharm. Pharm. Sci. 6 (2014) 383-387.
. H.K. Al-Hakeim, R.S. Al-Zabeba, E.A. Grulke, E.A. Al-Mulla, Interaction of calcium phosphate nanoparticles with human chorionic gonadotropin modifies secondary and tertiary protein structure, Nova Biotechnol. Chim. 14 (2015) 141-157.
. H.K. Al-Hakeim, K. Jasem, S. Moustafa, Antipepsin activity of silicon dioxide nanoparticles, Rev. Colomb. Quim. 45 (2016) 5-11.
. H.K. Al-Hakeim, J.K. Al-Shams, M.A. Kadhem, Immobilization of urease enzyme on ion-exchange resin, J. Univ. Babylon 20 (2012) 1231-1236.
. M.M. Bradford, A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem. 72 (1976) 248-254.
. C.E. Bower, T. Holm-Hansen, A salicylate–hypochlorite method for determining ammonia in seawater, Can. Fish Aquat. Sci. 37 (1980) 794-798.
. M.J. Fishman, S.C. Downs, Methods for analysis of selected metals in water by atomic absorption, U.S. Geological Survey Water-Supply Paper 1540-C)1966(38-41.
. C. Vaghela, M. Kulkarni, S. Haram, R. Aiyer, M. Karve, A novel inhibition based biosensor using urease nanoconjugate entrapped biocomposite membrane for potentiometric glyphosate detection, Int. J. Biol. Macromol. 108 (2018) 32-40.
. S.F. D’Souza, J. Kumar, S.K. Jha, B.S. Kubal, Immobilization of the urease on eggshell membrane and its application in biosensor, Mater. Sci. Eng. C 33 (2013) 850-854.
. R.P. Pogorilyĭ, V.P. Goncharik, L.I. Kozhara, I. Zub, Covalent immobilization of urease on polysiloxane templates containing 3-aminopropyl and 3-mercaptopropyl groups, Prikl. Biokhim. Mikrobiol. 44 (2008) 621-625.
. J. Zhou, S. Chen, J. Wang, Research on the orientedly immobilized urease via concanavalin A, Chinese J. Biotechnol. 24 (2008) 617-621.
. S. Sungur, M. Elcin, U. Akbulut, Studies on immobilization of urease in gelatin by cross-linking, Biomaterials 13 (1992) 795-800.
. M.V. Cattaneo, T.M. Chang, The potential of a microencapsulated urease-zeolite oral sorbent for the removal of urea in uremia, ASAIO Trans 37 (1991) 80-87.
. H.K. Al-Hakeim, K. Jasem, High ionic strength enhances the anti-pepsin activity of titanium dioxide nanoparticles, Nano Biomed. Eng. 8 (2016) 136-143.
. S. Dumitriu, M. Popa, V. Artenie, F. Dan, Bioactive polymers. 56: Urease immobilization on carboxymethylcellulose, Biotechnol. Bioeng. 34 (1989) 283-290.