Immobilization of uricase on protamine bound to glass beads and its application to determination of uric acid |
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| Authors: | N Nakamura K Murayama T Kinoshita |
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| Affiliation: | 1. Key Laboratory of Nanobiosensing and Nanobioanalysis at Universities of Jilin Province, Key Laboratory of Polyoxometalate Science of Ministry of Education, National & Local United Engineering Laboratory for Power Batteries, Department of Chemistry, Northeast Normal University, Changchun, Jilin Province 130024, PR China;2. Department of Materials Science & Engineering, Southern University of Science & Technology, Shenzhen, Guangdong Province 518055, PR China |
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| Abstract: | Uricase was found to be stabilized by protamine from salmon testis. Protamine was then bound to controlled-pore glass beads aminohexyl CPG 500 using glutaraldehyde. Microbial uricase was readily immobilized on the protamine bound to glass beads. The immobilized uricase proved to be stable even at 70 degrees C, whereas free uricase was inactivated at 45 degrees C and showed activity over a broader pH range than free uricase. Automated analysis of uric acid was facilitated using the immobilized uricase. The standard curve for uric acid was linear in the range of 2 to 10 micrograms/sample and passed through the origin. This automated procedure was also applicable to the determination of uric acid in human serum. Protamine bound to glass beads is expected to be useful for the simple immobilization and stabilization of enzymes. |
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