Premium
Biosynthesis of high specific activity 35 S‐glutathione
Author(s) -
Jayachandran N.,
Asokan K. P.,
Unny V. K. P.
Publication year - 2000
Publication title -
journal of labelled compounds and radiopharmaceuticals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 47
eISSN - 1099-1344
pISSN - 0362-4803
DOI - 10.1002/1099-1344(200009)43:10<971::aid-jlcr380>3.0.co;2-q
Subject(s) - chemistry , glutathione , cysteine , methionine , yeast , biochemistry , specific activity , amino acid , derivatization , saccharomyces cerevisiae , biosynthesis , chromatography , high performance liquid chromatography , enzyme
The biosynthesis of high specific activity 35 S‐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 35 S‐sulphate was supplemented as sole sulphur source. During growth, a major fraction of the radioactivity was incorporated into the protein as 35 S‐methionine and 35 S‐cysteine. After the growth of yeast, the cell wall was broken using a cell disrupter and the free 35 S‐glutathione present was separated from protein and other sulphur containing compounds by chromatographic procedures. The specific activity of the 35 S‐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 35 S‐cysteine. The overall radiochemical yield of 35 S‐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 35 S‐sulphate having a specific activity of 1200 Ci/mmol was used as the starting material Copyright © 2000 John Wiley & Sons, Ltd.