Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction
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Procedures and conditions have been established such that the unstable enzyme-bound flavin intermediate produced in the bacterial luciferase reaction can be isolated as approximately 70% of the flavin product, the remaining being the final product, FMN. The structure of the intermediate is proposed to be that of a luciferase-bound 4a,5-dihydroflavin-4a-hydroxide. The intermediate has a half-life of 33 min at 2°C and decays spontaneously to give H2O and luciferase-bound FMN with an activation enthalpy of about 120 kJ/mol. It has an absorption spectrum (λmax = 360 nm) that is consistent with the proposed structure, and a fluorescence emission (λmax = 485 nm) that matches the bioluminescence emission closely.
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KURFÜRST, Manfred, Peter MACHEROUX, Sandro GHISLA, J. Woodland HASTINGS, 1987. Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction. In: Biochimica et Biophysica Acta. 1987, 924(1), pp. 104-110. ISSN 0304-4165. Available under: doi: 10.1016/0304-4165(87)90076-6BibTex
@article{Kurfurst1987Isola-7258, year={1987}, doi={10.1016/0304-4165(87)90076-6}, title={Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction}, number={1}, volume={924}, issn={0304-4165}, journal={Biochimica et Biophysica Acta}, pages={104--110}, author={Kurfürst, Manfred and Macheroux, Peter and Ghisla, Sandro and Hastings, J. Woodland} }
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