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.
Highlights
IntroductionMultimetric enzyme–cyclooxygenase (prostaglandin endoperoxide G/H synthase, COX) is a heme-containing enzyme that catalyzes the reaction of arachidonic acid (AA) to produce prostaglandins (PGs) [1]
Multimetric enzyme–cyclooxygenase is a heme-containing enzyme that catalyzes the reaction of arachidonic acid (AA) to produce prostaglandins (PGs) [1]
Undesirable PGs generated from arachidonic acid are mainly catalyzed by COX-2, which is an inducible COX isozyme that plays an important role in inflammation and tumor formation [2,3]
Summary
Multimetric enzyme–cyclooxygenase (prostaglandin endoperoxide G/H synthase, COX) is a heme-containing enzyme that catalyzes the reaction of arachidonic acid (AA) to produce prostaglandins (PGs) [1]. Over the last several decades, techniques for discovering COX-2-selective inhibitors, including high performance liquid chromatography [10], liquid chromatography-mass spectrometry (LC-MS) [7,8,9], ultrafiltration LC-MS [11,12], virtual screening methods [13,14], bioassays [13,15], and so on, have been increasingly reported Some of these methods [10,15] used cultured cell lines and were time consuming and required special experimentation skills. In the present study, functionalized porous SiO2 microspheres were prepared and COX-2 was immobilized on the surface of these microspheres with GA as crosslinker Immobilization conditions, such as GA dosage, reaction pH, reaction temperature and enzyme dosage were optimized, according to the relative activity of the immobilized COX-2, and the properties of the immobilized COX-2 were investigated
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
