Solubilization of kaurenoic acid oxidase from microsomal preparations of Gibberella fujikuroi

Conference · Tue May 01 04:00:00 EDT 1990 · Plant Physiology, Supplement; (USA)

OSTI ID:5725680

Jennings, J C; Coolbaugh, R C ^[1]

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1. Purdue Univ., West Lafayette, IN (USA)

The gibberellin biosynthetic pathway from ent-kaurenoic acid was investigated using cell-free preparations from G. fujikuroi. Fungal cells were disrupted under 20,000 psi and centrifuged at 150,000 {times} g. Enzyme activity was assayed using chemically synthesized {sup 14}C-kaurenoic acid, and reaction products were separated and quantified by reverse phase HPLC. Products were identified as ent-7{alpha}-hydroxykaurenoic acid and GA{sub 14} by co-chromatography with standards and GC/MS. The 150,000 {times} g pellet was found to contain kaurenoic acid oxidase which was subsequently solubilized using cholic acid. Further separation of solubilized proteins was obtained by HPLC and FPLC anion exchange and Sepharose 4B-octylamine chromatography.

• OSTI ID: 5725680
• Report Number(s): CONF-9007196--
• Conference Information: Journal Name: Plant Physiology, Supplement; (USA) Journal Volume: 93:1
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
ESTERS
FUNGI
GIBBERELLIC ACID
HETEROCYCLIC COMPOUNDS
HYDROXY ACIDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LACTONES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
SYNTHESIS
TRACER TECHNIQUES