RAPID PURIFICATION OF YEAST CYTOPLASMIC FUMARATE REDUCTASE BY AFFINITY-CHROMATOGRAPHY ON BLUE SEPHAROSE CL-6B

The rapid and effective purification of soluble fumarate reductase from baker's yeast achieved by Blue Sepharose CL-GB chromatography. Cibacron Blue F3GA the chromophore of Blue Sepharose, inhibited the activity bf fumarate reductase. The enzyme bound to the column was selectively eluted by flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN) or riboflavin. The purified enzyme was essentially homogeneous as indicated by polyacrylamide gel electrophoresis under nondenaturing conditions and under denaturing conditions in sodium dodecylsulfate. By this procedure, the enzyme could be rapidly purified with high yield from yeast cells.