Abstract: In order to increase the ability of bio-concentration and organic selenium transform of yeast, and provide safe organic selenium resource for the production of selenium-rich agriculture and animal husbandry, selenium-enriched yeast was screened and the culture condition was optimized. Selenium-enriched yeast strain was obtained through UV mutagenesis and resistance screening test, culture medium and ferment condition were optimized through single factor orthogonal test. The results showed that a strain which could resist 5 mg/mL sodium selenite and 10 mg/kg ethionine was isolated and identified as Saccharomyces cerevisiae. The optimum ferment formula consisted of glucose, peptone, yeast extract, adding a small amount of inorganic salt. The optimum culture medium condition was initial pH value of 4, liquid volume of 50 mL/500 mL, shaking speed of 160 r/min, seed inoculation volume of 10% (V/V), and cultured for 28 h at 30℃, the ability of selenium-enriched strain was up to 26.75 mg/g dry cell under the optimum condition.