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Enzymatic Preparation of 14C-Labeled Phytoene, Squalene, and Geranylgeranyl Pyrophosphate from [2-14C]Mevalonic Acid

Author:
Sandmann, Gerhard, Hilgenberg, Willy, Böger, Peter
Source:
Zeitschrift für Naturforschung 2014 v.35 no.11-12 pp. 927-930
ISSN:
1865-7125
Subject:
Cucurbitaceae, Mucoraceae, Phycomyces blakesleeanus, carbon, cell free system, endosperm, isotope labeling, mevalonic acid, mutants, norflurazon, radionuclides, squalene, terpenoids, thin layer chromatography
Abstract:
Methods are described for an enzymatic preparation of ¹⁴C-labeled terpenoids. With a cell-free system of a white mutant of Phycomyces blakesleeanus (Mucoraceae) [¹⁴C]squalene and [¹⁴C- cis]phytoene can be synthesized from [2-¹⁴C]mevalonate. The application of norflurazon, a phenyl- pyridazinone herbicide, helps to increase the yield of squalene. Furthermore, the liquid endosperm of Echinocystis lobata (Cucurbitaceae) was used for the formation of either [¹⁴C(-)]kaurene from [¹⁴C]mevalonic acid or [¹⁴C-/ra/w]geranylgeranyl pyrophosphate in the presence of Amo 1618. The hydrocarbons formed were purified by alumina-column chromatography and preparative thin-layer chromatography (TLC). Geranylgeranyl pyrophosphate was separated by DE-column chromatography followed by TLC.
Agid:
5226292