Methodology and measurement of adenylate energy charge ratios in environmental samples

A method for measuring ATP, ADP and AMP levels in environmental samples was devised, and applied to seawater and bacterial cell extracts. This procedure is specifically designed for measuring the extremely low concentrations of total adenine nucleotides (A_T]=ATP]+ADP]+AMP]) that are apt to occur in most natural ecosystems (i.e., 10 ng A_T ml^-1 of sample extract). Although the current assay methodology can be used with purified firefly luciferase reagents, it has been suitably modified to accept crude luciferase preparations as well. ATP, ADP and AMP levels have been measured, and the corresponding energy charge (EC) ratios determined for seawater samples collected off the Southern California coast. The EC ratios ranged from 0.50 to 0.89, with peak values corresponding to the subsurface maxima in ATP and chlorophyll a concentrations, and the minimum values corresponding to the deepest water sampled (1500 m). The measurement of adenylate energy charge ratios in environmental samples can be a useful indicator of mean community metabolic activity and potential for cell growth.