Purification of D-beta-hydroxybutyrate dehydrogenase from rat brain.
D-beta-Hydroxybutyrate dehydrogenase (BDH), a lipid-requiring enzyme, has been purified to homogeneity from rat brain using a new improved method. The purified rat brain BDH has a subunit molecular mass of 31 kilodaltons on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The apoenzyme, i.e., the enzyme devoid of phospholipid, has no activity, but can be activated by phospholipid to a specific activity of 125 mumol/(min.mg). This is 625-fold greater than the activity in the mitochondrial fraction. The new purification procedure involves chromatography using a quaternary amine Sepharose resin followed by a sulphonate Sepharose resin, and eliminates the need for glass bead adsorption chromatography.
Department of Biology, University of Alabama, Tuscaloosa 35487-0344.
W W
WW
Zhang
P
P
Churchill
D000818:Animals; D001921:Brain / Q000201:enzymology*; D006884:Hydroxybutyrate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D046911:Macromolecular Substances; D006884:Hydroxybutyrate Dehydrogenase