Determination of intracellular pyridine nucleotide levels by bioluminescence using anaerobic bacteria as a model

An assay for the determination of NAD(P) + and NAD(P)H in extracts from the obligate anaerobe bacterium Thermoanaerobacter finnii is developed and the strategy for this development is described. This assay performed with constant FMN reductase (EC 1.6.8.1) and luciferase (EC 1.14.14.3) concentration...

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Veröffentlicht in:	Analytical biochemistry 1989-07, Vol.180 (1), p.17-23
Hauptverfasser:	Schmid, Ulrike, Schimz, Karl-Ludwig, Sahm, Hermann
Format:	Artikel
Sprache:	eng
Schlagworte:	Adenine Nucleotides - analysis Analytical, structural and metabolic biochemistry Bacteria, Anaerobic - metabolism Biological and medical sciences Fermentation FMN Reductase Fundamental and applied biological sciences. Psychology Luciferases - metabolism Luminescent Measurements Models, Biological NAD - analysis NADH, NADPH Oxidoreductases - metabolism NADP - analysis Nucleic acids Nucleic bases, nucleotides Xylose - metabolism
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Zusammenfassung:	An assay for the determination of NAD(P) + and NAD(P)H in extracts from the obligate anaerobe bacterium Thermoanaerobacter finnii is developed and the strategy for this development is described. This assay performed with constant FMN reductase (EC 1.6.8.1) and luciferase (EC 1.14.14.3) concentrations has been shown to detect as low as 1 pmol pyridine nucleotide. With this assay recovery, efficiency of extraction, reliability, and detection limit of the procedures were determined. To our knowledge this is the first bioluminometric assay for the determination of pyridine nucleotide levels in bacterial extracts.
ISSN:	0003-2697 1096-0309
DOI:	10.1016/0003-2697(89)90082-1