A Minimized Chemoenzymatic Cascade for Bacterial Luciferase in Bioreporter Applications

Abstract

Bacterial luciferase (Lux) catalyzes a bioluminescence reaction using long‐chain aldehyde, reduced flavin and molecular oxygen as substrates. The reaction can be applied in reporter gene systems for biomolecular detection in both prokaryotic and eukaryotic organisms. Because reduced flavin is unstable under aerobic conditions, another enzyme, flavin reductase, is needed for supplying reduced flavin to the Lux‐catalyzed reaction. To create a minimized cascade for Lux that would have greater ease of use, a chemo‐enzymatic reaction using the biomimetic nicotinamide (BNAH) was used in place of the flavin reductase reaction in the Lux system. The results showed that the minimized cascade reaction can be applied to monitor bioluminescence of the Lux reporter in eukaryotic cells effectively, and that it can achieve higher efficiencies than the system using flavin reductase. This development is useful for future applications as high throughput detection tools for drug screening applications.