[Quantitative detection of NADH by in vitro bacterial luciferase bioluminescent].

The study aimed at establishing the bacterial luciferase: FMN-NADH oxidoreductase bioluminescent system in vitro and evaluating its potential for quantitative detection of NADH. By optimizing the amount of substrates, we set up the coupled luciferase: FMN-NADH oxidoreductase bioluminescent system in vitro, based on the crude extract from Photobacterium leiognathi YL. The in vitro coupled bacterial luciferase: FMN-NADH oxidoreductase bioluminescent system was: 1 mL crude extract, 27 mmol/L Dodecane 100 microL, 10 mmol/L FMN-Na 0.5 microL and 0.14 mmol/L NADH 300 microL. Furthermore, we developed a method for quantitative detection of NADH according to the bioluminescence of NADH catalyzed of bacterial luciferase: FMN-NADH oxidoreductase system in vitro. A good linear relationship of NADH concentration was in a range of 1.0 x 10(-10) to 1.0 x 10(-8) mol/L. The bacterial luciferase: FMN-NADH oxidoreductase system used to measure NADH concentration was a good attempt to detect living bacterial cells in the fields of environment, food sanitation and other related.