| Abstract: | Ergothioneine was synthesized and accumulated in growing cultures of Mycobacterium smegmatis when the medium was adequately supplied with sulfur. In a low sulfur medium, the accumulation was sharply limited although growth of the organism was apparently normal. Synthesis of hercynine, the precursor of ergothioneine, was unaffected by low sulfur levels and was markedly increased by addition of l-histidine, the precursor of hercynine. Resting-cell pellicle experiments, performed with cells grown on the low sulfur high histidine medium, showed that ergothioniene was synthesized from endogenous hercynine, when cysteine or compounds readily converted to cysteine (such as cystine, lanthionine, cystathionine, and thiazolidine carboxylic acid) were added. Homocysteine and djenkolic acid allowed for minimal synthesis of betaine, whereas methionine, S-methylcysteine, sodium sulfate, and sodium thiosulfate were unable to donate sulfur for ergothioniene synthesis under the experimental conditions employed. Addition of cysteine to a resting pellicle preparation caused the formation of 100 to 200 mug of ergothioneine per g of dry cells in 2.5 to 3 hr. A modified procedure for isolating ergothioneine and hercynine, employing a 75% ethyl alcohol extraction of wet organisms, followed by a single alumina column separation of the compounds, is described. |
