[Bioluminescent method of determining picomolar amounts of nicotinamide-adenine dinucleotide using an immobilized extract of the luminescent bacterium Beneckea harveyi].
The luminous bacteria Beneckea Harveyi were immobilized on BrCN-sepharose and cellulose films activated with cyanuric chloride. Preparations with high luciferase and FMN-reductase activities were obtained, which showed no background luminescence without NADH being added. The storage conditions for the preparations obtained were optimized, and their kinetic parameters and thermostability were studied. Standard curves for NADH determining within the concentration range 1 nM-1 microM were plotted with the detection level of 1 picomol NADH. The preparations are very promising for bioluminescent assay due to their high activity, simple production, high stability during storage and a possibility for the repeated use.
D001681:Biological Assay / Q000379:methods*; D038181:FMN Reductase; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D009243:NAD / Q000032:analysis*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D013696:Temperature; D014733:Vibrio / Q000201:enzymology*; D014737:Vibrionaceae / Q000201:enzymology*
D009243:NAD; D008156:Luciferases; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases
Bioliuminestsentyĭ metod opredeleniia nikotinamidadenindinukleotida s pomoshch'iu svetiashchikhsia bakteriĭ Beneckea harveyi.