An Efficient Probe for Bacterial Nitroreductase Imaging and Detection Based on NanoLuc‐Furimazine Bioluminescent Pair

Comprehensive Summary The detection of critical endogenous species, such as bacteria in microenvironments in the body, requires better imaging tools for visualization and monitoring of biological events. Bioluminescence imaging is the most popular strategy for obtaining real‐time in living cells and organisms. Herein, we introduced a nitroaryl group on the C‐3 position and a hydroxy group at the C‐6 phenyl ring on furimazine to report the first bioluminescent probe ( 7 ) based on NanoLuc‐furimazine bioluminescent pair for the detection of nitroreductase in bacteria. The probe, which possessed up to 560‐fold intensity increase with a low detection limit of 16 ng/mL of nitroreductase, has the most efficient uncage efficiency in comparison with other bioluminescent congeners, thus enabling highly selective and sensitive visualization of NTR activity in a panel of clinical priority pathogens. Additionally, imaging of the recombinant strain as well as the NTR from mouse feces indicated the potential of this probe in the application of different mouse disease models.