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Fiber optic biosensor of immobilized firefly luciferase |
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| Authors: | |
| Institution: | (1) College of Material & Chemical Engineering, Zhejiang University, 310027 Hangzhou, China;(2) Sir Run Run Shaw Hospital, Zhejiang University, 310016 Hangzhou, China;(3) Institute of Biochemistry and Cell Biology, The Chinese Academy of Sciences, 200031 Shanghai, China |
| Abstract: | Luciferase from firefly lantern extract was immobilized on CNBr-activated Sepharose 4B. The kinetic properties of immobilized luciferase were extensively studied. TheK m′ for D-luciferin is 11.9 μmol/L, the optimum pH and temperature for Sepharose-bound enzyme were 7.8 and 25°C respectively. A luminescence fiber optic biosensor, making use of immobilized crude luciferase, was developed for assay of ATP. The peak light intensity was linear with respect to ATP concentration in range of 10−9–10−5 mol/L. A biological application was also demonstrated with the determination of serum ATP from rats bred in low versus normal oxygen environments. |
| Keywords: | Firefly luciferase ATP Immobilized enzyme Fiber optic biosensor |
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