Volume 26, Issue 2 (3-2022)                   IBJ 2022, 26(2): 132-141 | Back to browse issues page

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Morovvat F, Samsam Shariat* S Z, Davoudi M, Norouzian D. Immobilization of Urease onto Modified Egg Shell Membrane through Cross Linking. IBJ 2022; 26 (2) :132-141
URL: http://ibj.pasteur.ac.ir/article-1-2869-en.html
Background: Immobilization is an approach in industry to improve stability and reusability of urease. The efficiency of this technique depends on the type of membrane and the method of stabilization.
Methods: The PEI-modified egg shell membrane was used to immobilize urease by absorption and glutaraldehyde cross-linking methods. The membranes were characterized by Fourier-transform infrared spectroscopy (FTIR) and AFM, and Nessler method was applied to measure the kinetic of the immobilized enzymes. Finally, the storage stability (6 °C for 21 days) and reusability (until enzyme activity reached to zero) of the immobilized enzymes were investigated.  
Results: Based on FTIR, three new peaks were observed in both the absorption- (at 1389.7, 1230.8, and 1074.2 cm-1) and the cross-linking (at 1615-1690, 1392.7, 1450 cm-1) immobilized enzymes. The surface roughness of the native membrane was altered after PEI treatment and enzyme immobilization. The optimal pH of cross-linking immobilized enzymes was shifted to a more neutral pH, while it was alkaline in adsorption-immobilized and free enzymes. The reaction time decreased in all immobilized enzymes (100 min for free enzyme vs. 60 and 30 min after immobilizing by adsorption and cross-linking methods, respectively). The optimal temperature for all enzymes was 70 °C and they had a higher Km and a lower Vmax than free enzyme. The stability and reusability of urease were improved by both methods.
Conclusion: Our findings propose these approaches as promising ways to enhance the urease efficiency for its applications in industries and medicines.

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