Preliminary acidification of the cytoplasm of E. coli cells growing at pH 6.9 by adding to the medium 50 mM of sodium acetate or propionate reduced the mutagenic effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) to almost the spontaneous level. In experiments with S. typhimurium the protective effects of cytoplasm acidification against the mutagenic effect of MNNG was observed at pH 5.5 and was absent at a medium pH of 6.9. Alkalinization of the cytoplasm by adding 80 mM of methylamine to the growth medium at pH 8.1 did not modify the effect of MNNG on the cells of E. coli and S. typhimurium. Alkalinization of the cytoplasm of E. coli B/r and K12 was followed by a reduction of the intracellular non-protein SH group level by 25 and 50%, respectively. It is supposed that the protective effect of acidification may be due to a decline in the productivity of mutagenically more active intermediates of MNNG when the pH is reduced and the associated fall of the level of intracellular non-protein thiols occurs. The above situation may serve as a model for studying the effects of MNNG and other alkylating agents on cells differing in physiological status.