Bioluminescence of Monolayers of Firefly Luciferase Immobilized on Graphite

We report on the immobilization of the firefly protein luciferase on the hydrophobic surface of graphite. Observation by liquid-phase atomic force microscopy of islands with a height consistent with the size of a single molecule confirmed that the protein was contained within a monomolecular layer....

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Bibliographic Details
Published in:Langmuir 2006-06, Vol.22 (12), p.5451-5454
Main Authors: Palomba, S, Berovic, N, Palmer, R. E
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - chemistry
Catalysis
Chemistry
Electrochemistry
Enzyme Activation
Enzymes, Immobilized - chemistry
Exact sciences and technology
Firefly Luciferin - chemistry
General and physical chemistry
Graphite - chemistry
Luciferases, Firefly - chemistry
Luminescence
Theory of reactions, general kinetics. Catalysis. Nomenclature, chemical documentation, computer chemistry
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Summary:We report on the immobilization of the firefly protein luciferase on the hydrophobic surface of graphite. Observation by liquid-phase atomic force microscopy of islands with a height consistent with the size of a single molecule confirmed that the protein was contained within a monomolecular layer. The enzyme activity was assayed by single-photon counting of the bioluminescence, which is the catalytic product of luciferase. Attachment to the surface modified the efficiency of the enzyme, but the introduction of the substrates luciferin and ATP resulted in the reactivation of the enzyme. The functionalized graphite surface was employed as a cathode in a bioelectrochemical cell. This demonstrated that the electric field caused a substantial loss of enzyme catalytic activity.
ISSN:0743-7463
1520-5827
DOI:10.1021/la060597h