Luciferin-luciferase assay of adenosine triphosphate from bacteria: a comparison of dimethylsulphoxide (DMSO) and acetone with other solvents.
The extraction of adenosine triphosphate (ATP) from six strains of bacteria, chosen for differences in cell-wall composition and habitat, was performed. The solvents were two in common use, Tris-ethylenediaminetetraacetate (Tris-EDTA) and trichloroacetic acid (TCA), and two promising, though less utilized solvents, dimethylsulphoxide (DMSO) and acetone. ATP was determined by the luciferin-luciferase reaction. Of the solvents used, DMSO and acetone were the most effective considering the different kinds of bacteria tested and, of these two, DMSO was the most convenient to use. Tris-EDTA was not as effective as the other solvents tested and TCA, which was effective with most strains, gave low yields when used with cultures grown in artificial seawater broth. Internal standards were used to determine if there were substances present that could inhibit the reaction of released ATP with the luciferin-luciferase reagent. Extracts of ATP, stored at -20 degrees C, were stable for up to 3 weeks.
D000096:Acetone; D000255:Adenosine Triphosphate / Q000032:analysis*; D001410:Bacillus megaterium / Q000032:analysis; D001419:Bacteria / Q000032:analysis*; D004121:Dimethyl Sulfoxide; D004492:Edetic Acid; D049454:Firefly Luciferin; D008156:Luciferases; D011549:Pseudomonas / Q000032:analysis; D011551:Pseudomonas fluorescens / Q000032:analysis; D012486:Salmonella typhimurium / Q000032:analysis; D012997:Solvents; D013210:Staphylococcus / Q000032:analysis; D014238:Trichloroacetic Acid; D014325:Tromethamine
D012997:Solvents; D014325:Tromethamine; D000096:Acetone; D049454:Firefly Luciferin; D014238:Trichloroacetic Acid; D000255:Adenosine Triphosphate; D004492:Edetic Acid; D008156:Luciferases; D004121:Dimethyl Sulfoxide