Biosynthesis of [14C]geranylgeranyldiphosphate by a prenyl transferase system from a mutant strain of Gibberella fujikuroi.
[14C]Geranylgeranyldiphosphate was produced by a prenyl transferase system extracted from the mutant strain SG4 of Gibberella fujikuroi (wild-type strain IMI 58289), which is blocked in carotenoid biosynthesis. The fungus was grown in liquid medium containing only potato-dextrose broth. Crude extracts of 12- to 14-day-old mycelia in Tris buffer, pH 8.5, were centrifuged at 100,000g. The supernatant was desalted and used to produce [4,8,12,16-14C]geranylgeranyldiphosphate from R,S-[2-14C]mevalonic acid. Maximum yield of [14C]geranylgeranyldiphosphate was obtained using mycelia from the stationary growth phase with incubation of the enzyme preparation at pH 8.5 for 5 h. Routinely, 65% of the biogenetic available R-mevalonic acid was converted to [14C]geranylgeranyldiphosphate, which was identified by cochromatography with authentic [1-3H]geranylgeranyldiphosphate by means of radio-TLC, radio-HPLC, and hydrolysis to geranylgeraniol.
Knöss
W
W
Institute of Pharmaceutical Biology, University of Bonn, Nussallee 6, Bonn, D-53115, Germany.
Reuter
B
B
D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D004224:Diterpenes / Q000378:metabolism; D005874:Gibberella / Q000201:enzymology* / Q000235:genetics; D006868:Hydrolysis; D008798:Mevalonic Acid / Q000378:metabolism; D016296:Mutagenesis; D010088:Oxidoreductases / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis*
D004224:Diterpenes; D011106:Polyisoprenyl Phosphates; C017338:geranylgeraniol; D010088:Oxidoreductases; C022398:phytoene dehydrogenase; D004122:Dimethylallyltranstransferase; C002963:geranylgeranyl pyrophosphate; D008798:Mevalonic Acid