An intracellular, soluble 1,4-benzoquinone reductase was purified from Baker's Yeast by ammonium sulfate precipitation, DEAE-Sephacel anion exchange chromatography, and Sephacryl S-200 gel filtration chromatography. 1,4-Benzoquinone reductase was achieved 123.8 fold purification from crude homogenate with a yield of 11.1%.