Separation of Neurospora Crassa Myo-Inositol-1-Phosphate Synthase from Glucose-6-Phosphate Dehydrogenase by Affinity Chromatography |
| |
| Authors: | J. Aradi A. Zsindely Á. Kiss M. Szabolcs M. Schablik |
| |
| Affiliation: | 1. Departments of Biochemistry and Biology , University Medical School , Debrecen, H-4012, Hungary;2. Central Research Laboratory , University Medical School , Debrecen, H-4012, Hungary |
| |
| 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 60 μq 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. |
| |
| Keywords: | Citrus medica inactivation kinetics purification soluble peroxidase thermostable |
|
|
