DETECTION OF LIGANDS VIA BACTERIAL LUCIFERASE

Abstract

Bacterial luciferase is a widely applied enzyme in medical diagnosis. The applications harness the FMNH,-dependency of the luciferase. Via FMNH,it is possible to couple other FMN-dependent enzymes or enzyme systems. With the help of FMN/NADHoxidoreductase coupling with NAD/NADH-dependent enzymes and enzymesystems it is possible to broaden the range of application”. luciferase FMNH,+ O, + aldehyde ------------------ » FMN + H,O + acid + light The amount of emitted luminescent light is proportional to the substrates and can be easily monitored with a luminometer. In addition to FMNH,, other substrates are converted, for example an aldehyde into the corresponding acid. Very low concentrations of these aldehydes can be detected and the reaction has a large linear range (5 to 100000 nM). Maximal activity (luminescence) is achieved through nalkanes with a chain length of C-10 to 14. Hereit will be shown that modified aldehydes are as effective substrates as unmodified ones. Furthermore, the interaction of both will be characterized and a general scheme of ligand detection presented.