[Quantitative detection of NADH by in vitro bacterial luciferase bioluminescent].
OBJECTIVE
The study aimed at establishing the bacterial luciferase: FMN-NADH oxidoreductase bioluminescent system in vitro and evaluating its potential for quantitative detection of NADH.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Mei
Cexia
C
Food Safety Laboratory, Food Science and Engineering Department, Ocean University of China, Qingdao 266003. mcx_106@163.com
Wang
Jingxue
J
Lin
Hong
H
Wang
Jing
J
D038181:FMN Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D049453:Luciferases, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D009243:NAD / Q000378:metabolism*; D010776:Photobacterium / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D009243:NAD; D049453:Luciferases, Bacterial; D038181:FMN Reductase