For the overproduction of glutathione, Candida sp. mutant was isolated by the treatment with U.V. light. The highest glutathione production of Candida sp. mutant was obtained after shaking culture for 48 hours in the cullture medium containing glucose 1.5%(w/v), yeast extract 4.0% (w/v), KH2PO4 0.04%(w/v), biotin 5 ng/ml, and L-cysteine 0.04%(w/v). The optimal pH and temperature for the glutathione production were pH 6.0 and 25C, respectively.