title: Separation of Neurospora crassa myo-inositol-1-phosphate synthase from glucose-6-phosphate dehydrogenase by affinity chromatography.
abstract: The purification of Neurospora crassa myo-inositol-1-phosphate synthase (EC 5.5.1.4) was studied by affinity chromatography using the substrate (glucose-6-phosphate), the inhibitor (pyrophosphate), the coenzyme (NAD+) and the coenzyme analogues (5'AMP and Cibacron Blue F3G-A) of the enzyme as adsorbents attached to agarose gel. Myo-inositol-1-phosphate synthase could be separated completely from the contaminating substance, glucose-6-phosphate dehydrogenase (EC 1.1.1.49), on Blue Sepharose CL-6B and on pyrophosphate-Sepharose. The purified enzyme had a specific activity of 16 400 U/mg. The sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the 60 micrograms of this purified enzyme gave a homogenous band. The enzyme was found to be composed of four identical subunits having a molecular weight of 65 000.
mesh_terms: D002238:Carbohydrate Epimerases / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D008970:Molecular Weight; D007296:Myo-Inositol-1-Phosphate Synthase / Q000302:isolation & purification*; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D012685:Sepharose / Q000031:analogs & derivatives
chemical_list: C029483:Blue-Sepharose; D012685:Sepharose; D005954:Glucosephosphate Dehydrogenase; D002238:Carbohydrate Epimerases; D007296:Myo-Inositol-1-Phosphate Synthase