CHARACTERIZATION AND SOLUBILIZATION OF KAURENOIC ACID HYDROXYLASE FROM GIBBERELLA-FUJIKUROI

被引:24
作者
JENNINGS, JC [1 ]
COOLBAUGH, RC [1 ]
NAKATA, DA [1 ]
WEST, CA [1 ]
机构
[1] UNIV CALIF LOS ANGELES,DEPT CHEM,LOS ANGELES,CA 90024
关键词
D O I
10.1104/pp.101.3.925
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
A key step in gibberellin biosynthesis is the conversion of ent-kaurenoic acid to ent-7alpha-hydroxykaurenoic acid, mediated by the enzyme kaurenoic acid hydroxylase. A cell-free system obtained from Gibberella fujikuroi (Saw.) Wr. was used to characterize kaurenoic acid hydroxylase activity. Microsomal preparations f rom disrupted fungal cells, in the presence of O2 and NADPH, converted [17-C-14]ent-kaurenoic acid to oxidation products that were separated by high-performance liquid chromatography and identified as ent-7alpha-hydroxykaurenoic acid and gibberellin A14 by combined gas chromatography-mass spectrometry. Flavin adenine dinucleotide and the chloride salts of several monovalent cations stimulated the conversion of ent-kaurenoic acid to these products, whereas CO and a number of known inhibitors of cytochrome P-450-dependent reactions, including paclobutrazol, tetcyclacis, BAS lll..W, flurprimidol, triarimol, metyrapone, and 1-phenylimidazole, significantly reduced kaurenoic acid hydroxylase activity. Kaurenoic acid hydroxylase was solubilized from fungal microsomes by treatment with 1 M KCl. The properties of the enzyme noted above suggest that kaurenoic acid hydroxylase from G. fujikuroi is a cytochrome P-450-dependent monooxygenase.
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页码:925 / 930
页数:6
相关论文
共 32 条
[1]   CHARACTERIZATION OF ENT-KAURENE OXIDASE ACTIVITY FROM GIBBERELLA-FUJIKUROI [J].
ASHMAN, PJ ;
MACKENZIE, A ;
BRAMLEY, PM .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1036 (02) :151-157
[2]  
Bearder JR, 1983, BIOCH PHYSL GIBBEREL, V1, P251
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   RESOLUTION AND RECONSTITUTION OF CYTOCHROME-P-450 CONTAINING STEROID HYDROXYLATION SYSTEM OF RHIZOPUS-NIGRICANS [J].
BRESKVAR, K ;
CRESNAR, B ;
HUDNIKPLEVNIK, T .
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1987, 26 (04) :499-501
[5]   STUDIES ON SPECIFICITY AND SITE OF ACTION OF ALPHA-CYCLOPROPYL-ALPHA-[P-METHOXYPHENYL]-5-PYRIMIDINE METHYL-ALCOHOL (ANCYMIDOL), A PLANT-GROWTH REGULATOR [J].
COOLBAUGH, RC ;
HIRANO, SS ;
WEST, CA .
PLANT PHYSIOLOGY, 1978, 62 (04) :571-576
[6]   COMPARATIVE EFFECTS OF ANCYMIDOL AND ITS ANALOGS ON GROWTH OF PEAS AND ENT-KAURENE OXIDATION IN CELL-FREE-EXTRACTS OF IMMATURE MARAH-MACROCARPUS ENDOSPERM [J].
COOLBAUGH, RC ;
SWANSON, DI ;
WEST, CA .
PLANT PHYSIOLOGY, 1982, 69 (03) :707-711
[7]  
CROZIER A, 1983, BIOCH PHYSL GIBBEREL, V1, P485
[8]  
DEMONTELLANO PRO, 1986, CYTOCHROME P450 STRU, P273
[9]  
DENNIS DT, 1967, J BIOL CHEM, V242, P3293
[10]   MULTIPLE FORMS OF PLANT CYTOCHROMES-P-450 [J].
DONALDSON, RP ;
LUSTER, DG .
PLANT PHYSIOLOGY, 1991, 96 (03) :669-674