The effect of penicillin on fatty acid synthesis and excretion inStreptococcus mutans BHT

Abstract

Treatment of exponentially growing cultures ofStreptococcus mutans BHT with growth-inhibitory concentrations (0.2 μg/ml) of benzylpenicillin stimulates the incorporation of [2-¹⁴C] acetate into lipids excreted by the cells by as much as 69-fold, but does not change the amount of¹⁴C incorporated into intracellular lipids. At this concentration of penicillin cellular lysis does not occur. The radioactive label is incorporated exclusively into the fatty acid moieties of the glycerolipids. The increase in the radioactive content of the extracellular lipids reflects an actual net increase in the total fatty acid content as determined by a chemical assay. During a 4-hr incubation in the presence of penicillin, the extracellular fatty acid ester concentration (per mg cell dry weight) increases 1.5 fold, even though there is no growth or cellular lysis. No change is observed in the intracellular fatty acid ester content.

An indication of the relative rate of fatty acid synthesis was most readily obtained by placingS. mutans BHT in a buffer containing¹⁴C-acetate. Under these nongrowing conditions free fatty acids are the only lipids labeled, a factor which simplifies the assay. The addition of glycerol to the buffer causes all of the nonesterified fatty acids to be incorporated into glycerolipid. The cells excrete much of the lipid whether glycerol is present or not. Addition of penicillin to the nongrowth supporting buffer system does not stimulate the incorporation of [¹⁴C]-acetate into fatty acids. However, if cells are exposed to penicillin in a growth-supporting medium and then are transferred to the nongrowing buffer system containing no penicillin, the previously exposed cells retain the ability to incorporate [¹⁴C]-acetate into fatty acid at a higher rate than untreated cells over a prolonged period of time. The stimulation of [¹⁴C]-acetate into fatty acids in this system parallels but is not dependent on the stimulation by penicillin of the incorporation of [¹⁴C]-glycerol into glycerolipid and lipoteichoic acid synthesis previously demonstrated by our laboratory.