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A positive colony was grown in Luria broth (LB) medium supplemented with 100 mg L −1 ampicillin at 37 °C with shaking at 180 rpm till the optical density reaches to 0.6

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In addition, genetic glutathione synthetase defects (Figure 2, reaction 5), also an autosomal recessive trait, reduce glutathione levels and generate overproduction of 5-oxoproline