Mutarotases are recently characterized group of enzymes that are involved in anomeric conversions of monosaccharides enhancing rate of metabolism. Mutarotase was extracted and purified from bovine kidney cortex by (NH4)2SO4 precipitation technique, dialysis and gel filtration chromatography. Polarimeter was used for assay of Mutarotase in this present project. Mutarotase increased activity from 0.478 to 1.711 U/ml by purification. It gained 13.4 folds purification at this final step. It is helpful in determining the glucose level and participates in glycolysis and gluconeogenesis. Activities and fold purification of Mutarotase can also be increased by much improved techniques of purification like ion exchange chromatography or FPLC hence enhancing the production of this enzyme.