Immobilization of Cyclooxygenase-2 on Silica Gel Microspheres: Optimization and CharacterizationReportar como inadecuado




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1

College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, China

2

Research Center for Marine Ecology, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China





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Authors to whom correspondence should be addressed.



Academic Editor: Roberto Fernandez-Lafuente

Abstract 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





Autor: Qian Shi 1,2, Junhui Chen 2,* , Yanlong Wang 2, Zhaoyong Li 2, Xianguo Li 1,* , Chengjun Sun 2 and Li Zheng 2

Fuente: http://mdpi.com/



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