Scheme 1.
Alternate pathways to fluorescence enhancement. (A) FlamB oxidation followed by Cu2+ binding and spirolactam ring opening to give modestly fluorescent [CuCl(FlamS)] (drawn as the neutral monomer unit of the tetranuclear arrangement seen in the X-ray crystal structure), or (B) hydrolysis of FlamB to FH followed by reaction with Cu2+ to yield highly fluorescent fluorescein. FH also converts to fluorescein upon interaction with H2O2.31