The paper presents a method for producing highly purified glucosoisomerase from Actinomyces olivocinereus 154. The scheme of purification includes enzyme extraction from dry biomass, acetone fractionation, ammonium sulfate precicipation, and Sephadex G-200 chromatography. The highly purified enzyme is homogeneous as shown by polyacryl amide gel electrophoresis, analytical ultracentrifugation, and gel filtration. The highly purified enzyme has been prepared in a crystalline form. The molecular weight of the enzyme has been estimated by sedimentation equilibrium to be 162 000 and by gel filtration to be 158 000. The sedimentation constant of glucosoisomerase has been found to be 8.52 S.

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