Fiber optic biosensor of immobilized firefly luciferase

摘要

Luciferase from firefly lantern extract was immobilised on CNBr- activated Sepharose 4B. The kinetic properties of immobilized luciferase were extensively smdied. The K_m' for D-luciferin is 11.9 mu mol/L, the optimum pH and temperature for Sepharose-bound enzyme were 7.8 and 25 deg C respectively. A luminescence fiber optic biosensor, making use of immobilized crude luciferase, was developed for assay of Am. 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.