Viability and Metabolic Capability Are Maintained by Escherichia Coli, Pseudomonas Aeruginosa, and Streptococcus Lactis at Very Low Adenylate Energy Charge

Metabolic regulation by nucleotides has been examined in several bacteria within the context of the adenylate energy charge (EC) concept. The ECs of bacteria capable of only fermentative metabolism (Streptococcus lactis and the ATPase-less mutant Escherichia coli AN718) fell to less than 0.2 under carbon-limiting conditions, but the bacteria were able to step up the EC to greater than 0.8 upon exposure to nutrient sugars. Similarly, nongrowing E. coli 25922, whose EC had been artificially lowered to less than 0.1 by the addition of the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), was able to immediately step up the EC to 0.8 to 0.9 upon the addition of glucose but was unable to respond to respiratory substrates. The EC of respiring bacteria (E. coli 25922 and Pseudomonas aeruginosa 27853) fell to 0.3 to 0.4 under certain limiting growth conditions, but the bacteria also responded immediately when challenged with succinate to give EC values greater than 0.8. These bacteria could not step up the EC with respiratory substrates in the presence of CCCP. For all bacteria, the loss of the ability to step up the EC was attributable to the loss of nutrient transport function. Mixtures of viable and HOCl-killed E. coli 25922 were able to step up the EC in proportion to the fraction of surviving cells. The data indicate that nucleotide phosphorylation levels are not regulatory in nongrowing bacteria but that the EC step-up achievable upon nutrient addition may be an accurate index of viability.