Energy transfer to gold nanoparticles from firefly luciferase reaction

Photinus pyralis Luciferase enzyme is a monomeric protein that emits bioluminescence light by substrate including D-luciferin, ATP my MgSO۴. The molecular weight of this enzyme is ۶۲ kDa and the light emitted from it is green-yellow in the range between ۵۵۰-۵۷۰ nm. The ability to measure the light emitted by the enzyme has led to its use in various fields such as microbial contamination detection, ATP assay, biological assessments and reporter genes. This enzyme is very sensitive and changing environmental conditions cause a sharp and rapid decline in its activity, so the use of analytes to improve this issue is very useful. Gold is a stable metal and gold nanoparticles have valuable applications in the fields of biosensors, drug release and medical diagnosis, disease treatment and antimicrobial effects. In this study, we evaluate the effect of gold nanoparticles on luciferase enzyme. For this purpose, recombinant luciferase enzyme was expressed and then purification of the enzyme based on histidine tags of N-terminal head of the protein was performed using a chromatographic column. Then, the effect of gold nanoparticles on the third structure of luciferase protein, assuming no change in the intrinsic fluorescence emission of luciferase enzyme, was performed using a fluorescence device. The results showed that increasing the concentration of gold nanoparticles in the presence of luciferase enzyme reduces the emission of intrinsic fluorescence of protein, which is similar to the published study in which the effect of quantum dot cadmium tellurium on the emission of intrinsic fluorescence luciferase was reported. It is possible to change the structure of luciferase protein in the presence of gold nanoparticles.

کلیدواژه ها:

Luciferase enzyme ، Gold nanoparticles ، ATP assay ، Bioluminescence ، Intrinsic fluorescence