Both Escherichia coli and Streptococcus faecalis grew in an alkaline medium containing 100 microM carbonyl cyanide m-chlorophenylhydrazone (CCCP). Our data suggested that CCCP functioned as a protonophore at a high pH and that the proton-motive force was dissipated almost completely under such conditions. The pH gradient measured with dimethyloxazolidine-2,4-dione and acetylsalicylic acid was very small in both bacteria at a high pH above 8, and was not affected significantly by the addition of CCCP. Based on these findings, we propose that the maintenance of neutral cytoplasmic pH is not obligatory for the growth of E. coli and S. faecalis in an alkaline medium.