Purification and properties of the NAD(P)H:nitrate reductase of the yeast Rhodotorula glutinis.
The assimilatory nitrate reductase from the yeast Rhodotorula glutinus has been purified 740-fold, its different catalytic activities have been characterized and some inhibitors studied. The purified enzyme (150 units per mg protein) contains a cytochrome of the b-557 type. An S20,w of 7.9 S was found by the use of sucrose density gradient centrifugation, and a Stokes radius of 7.05 nm was determined by gel filtration. From these values, a molecular weight of 230 000 was estimated for the native enzyme. The purified preparation consisted of two electrophoretically distinguishable proteins, both of which exhibited nitrate reductase activity. The species with the higher electrophoretic mobility which represented the great majority of the total nitrate reductase gave a positive stain for heme and was shown to be composed of subunits with a molecular weight of about 118 000. Thus the molecule contains two subunits of the same size.
Purification and properties of the NAD(P)H:nitrate reductase of the yeast Rhodotorula glutinis. The assimilatory nitrate reductase from the yeast Rhodotorula glutinus has been purified 740-fold, its different catalytic activities have been characterized and some inhibitors studied. The purified enzyme (150 units per mg protein) contains a cytochrome of the b-557 type. An S20,w of 7.9 S was found by the use of sucrose density gradient centrifugation, and a Stokes radius of 7.05 nm was determined by gel filtration. From these values, a molecular weight of 230 000 was estimated for the native enzyme. The purified preparation consisted of two electrophoretically distinguishable proteins, both of which exhibited nitrate reductase activity. The species with the higher electrophoretic mobility which represented the great majority of the total nitrate reductase gave a positive stain for heme and was shown to be composed of subunits with a molecular weight of about 118 000. Thus the molecule contains two subunits of the same size.