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GSH assay in cell-free system GSH production was determined in a cell-free system by mixing 0.05–5 mM l-glutamate or 0.05–5 mM l-theanine with 5 mM l-cysteine, 10 mM ATP, 0.6 ng/µl glutamate cysteine ligase (GCL) recombinant protein (Novus Biologicals), 100 mM Tris-HCl (pH 8.2) followed by the addition of 5 mM glycine and 0.05 ng/µl GSH synthase recombinant protein (Syd labs) over a period of 30 min under constant shaking at 37°C