title: Purification of geranylgeranyl diphosphate synthase from Phycomyces blakesleanus.
abstract: Geranylgeranyl diphosphate synthase has been purified to homogeneity from the carotene-overproducing strain M1 of Phycomyces blakesleanus. Usually two activity peaks with molecular weights of 60,000 and 30,000 eluted on gel exclusion chromatography, suggesting that the enzyme consists of two subunits, with a tendency to dissociate. With homogeneous protein, a single-staining band with molecular weight of 30,000 appeared on sodium dodecyl sulfate gel electrophoresis, confirming a subunit molecular weight of 30,000. Only isopentenyl diphosphate and farnesyl diphosphate were accepted by this enzyme for geranylgeranyl diphosphate formation. The smaller allylic compounds, dimethylallyl and geranyl diphosphate, were utilized at less than 1/20th the rate of farnesyl diphosphate. Michaelis constants of 9 microM for isopentenyl diphosphate and 60 microM for farnesyl diphosphate were found. The isoelectric point is 4.8.
authors.affiliation: Department of Biochemistry, University of Utah School of Medicine, Salt Lake City 84108.
authors.forename: F L
authors.initials: FL
authors.lastname: Brinkhaus
authors.forename: H C
authors.initials: HC
authors.lastname: Rilling
mesh_terms: D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008970:Molecular Weight; D009090:Mucorales / Q000201:enzymology*; D009154:Mutation; D010800:Phycomyces / Q000201:enzymology*; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis; D013045:Species Specificity; D014166:Transferases / Q000302:isolation & purification*
chemical_list: D011106:Polyisoprenyl Phosphates; D014166:Transferases; D004122:Dimethylallyltranstransferase; C002963:geranylgeranyl pyrophosphate