Abstract
The biosynthesis of high specific activity 35S-glutathione was carried out using a diploid strain of baker's yeast—“Saccharomyces cerevisiae”. Yeast cells were grown in a synthetic medium in which sodium 35S-sulphate was supplemented as sole sulphur source. During growth, a major fraction of the radioactivity was incorporated into the protein as 35S-methionine and 35S-cysteine. After the growth of yeast, the cell wall was broken using a cell disrupter and the free 35S-glutathione present was separated from protein and other sulphur containing compounds by chromatographic procedures. The specific activity of the 35S-glutathione was determined by hydrolysing into its constituent amino acids and quantifying them using an amino acid analyser employing a post-column orthophthaldehyde derivatization method, followed by radioactivity assay of 35S-cysteine. The overall radiochemical yield of 35S-glutathione was 5%. The radiochemical purity of the product was found to be greater than 95% and its specific activity, greater than 1000 Ci/mmol when sodium 35S-sulphate having a specific activity of 1200 Ci/mmol was used as the starting material Copyright © 2000 John Wiley & Sons, Ltd.
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