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Molecules 2015, 20(11), 19971-19983;

Immobilization of Cyclooxygenase-2 on Silica Gel Microspheres: Optimization and Characterization

College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, China
Research Center for Marine Ecology, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China
Authors to whom correspondence should be addressed.
Academic Editor: Roberto Fernandez-Lafuente
Received: 4 August 2015 / Accepted: 28 August 2015 / Published: 5 November 2015
(This article belongs to the Section Molecular Diversity)
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In this study, immobilized COX-2 was successfully constructed through glutaraldehyde-mediated covalent coupling on functional silica gel microspheres. The optimum conditions, properties, and morphological characteristics of the immobilized COX-2 were investigated. The optimal immobilization process was as follows: about 0.02 g of aminated silica gel microspheres was activated by 0.25% GA solution for 6 h and mixed with 5 U of free recombinant COX-2 solution. Then, the mixture was shaken for 8 h at 20 °C. Results showed that the immobilized COX-2 produced by this method exhibited excellent biocatalytic activity, equivalent to that of free COX-2 under the test conditions employed. The best biocatalytic activity of immobilized COX-2 appeared at pH 8.0 and still maintained at about 84% (RSD < 7.39%, n = 3) at pH 10.0. For temperature tolerance, immobilized COX-2 exhibited its maximum biocatalytic activity at 40 °C and about 68% (RSD < 6.99%, n = 3) of the activity was maintained at 60 °C. The immobilized COX-2 retained over 85% (RSD < 7.26%, n = 3) of its initial biocatalytic activity after five cycles, and after 10 days storage, the catalytic activity of immobilized COX-2 still maintained at about 95% (RSD < 3.08%, n = 3). These characteristics ensured the convenient use of the immobilized COX-2 and reduced its production cost. View Full-Text
Keywords: cyclooxyenase 2; immobilized enzyme; aminated silica gel; cross linking cyclooxyenase 2; immobilized enzyme; aminated silica gel; cross linking

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Shi, Q.; Chen, J.; Wang, Y.; Li, Z.; Li, X.; Sun, C.; Zheng, L. Immobilization of Cyclooxygenase-2 on Silica Gel Microspheres: Optimization and Characterization. Molecules 2015, 20, 19971-19983.

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