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CLC number: Q814.3

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Received: 2002-03-06

Revision Accepted: 2002-06-30

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Journal of Zhejiang University SCIENCE A 2002 Vol.3 No.5 P.563~566

http://doi.org/10.1631/jzus.2002.0563


Fiber optic biosensor of immobilized firefly luciferase


Author(s):  CAI Jin, MENG Wen-fang, JI Xin-song

Affiliation(s):  College of Material & Chemical Engineering, Zhejiang University, Hangzhou 310027, China; more

Corresponding email(s):   caij@che.zju.edu.cn

Key Words:  Firefly luciferase, ATP, Immobilized enzyme, Fiber optic biosensor


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CAI Jin, MENG Wen-fang, JI Xin-song. Fiber optic biosensor of immobilized firefly luciferase[J]. Journal of Zhejiang University Science A, 2002, 3(5): 563~566.

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%I Zhejiang University Press & Springer
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T1 - Fiber optic biosensor of immobilized firefly luciferase
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Abstract: 
Luciferase from firefly lantern extract was immobilized on CNBr-activated Sepharose 4B. The kinetic properties of immobilized luciferase were extensively studied. The Km′ 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.

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Reference

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[3] Leach, F.R., 1981. ATP determination with firefly luciferase. J. Appl. Biochem., 3: 473-478.

[4] Lee,Y., Jablonski, I., Deluca, M., 1981. Immobilization of firefly luciferase on glass rods: Properties of the immobilized enzyme. Anal. Biochem., 80:496-500.

[5] Ji, X.S., Li, H.X., Yuan, Z.Y., Liu S.H., 1984. Immobilization of NAD kinase. Enzyme Engineering 7, Anal. N. Y. Acad. Sci., 434:264-266.

[6] Maria, P. X., Begona, V., Maria, D. M., 2000. Fiber optic monitoring of carbamate pesticides using porous glass with covalently bound chlorophenol red. Biosensors & Bioelectronics, 14:895-905.

[7] Michel, P.E., Gautier-Sauvigne, S.M., Blum, L.J., 1998. A transient enzymatic inhibition as an efficient tool for the discriminating bioluminescent analysis of three adenylic nucleotides with a fiberoptic sensor based on a compartmentalized tri-enzymatic sensing layer. Analytica Chimica Acta, 360: 89-99.

[8] Neuberg, C., 1944. Convenient method for deproteinization. Arch. Biochem. 4:101-105.

[9] Tservistas, M., Koneke, R., Comte, A. Scheper T., 2001. Oxygen monitoring in supercritical carbon dioxide using a fibre optic sensor. Enzyme and Microbial Technology, 28:637-634.

[10] Vangelis, G. A., Yannis, D. C., 2002. A portable fiber-optic pesticide biosensor based on immobilized cholinesterase and sol-gel entrapped bromcresol purple for in-field use. Biosensor & Bioelectronics, 17:61-69.

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