682
C. Troncoso et al. / Phytochemistry 69 (2008) 672–683
Keller, N., Hohn, T., 1997. Metabolic Pathway gene cluster in filamentous
fungi. Fungal Genet. Biol. 21, 17–29.
temperatures were 220 ꢁC, 270 ꢁC and 210 ꢁC, respectively.
Full scans were obtained from 50 to 650 amu at 1 scan sÀ1
.
Lamb, D.C., Kelly, D.E., Manning, N.J., Kaderbhai, M.A., Kelly,
S.L., 1999. Biodiversity of the P450 catalytic cycle: yeast cyto-
chrome b5/NADH cytochrome b5 reductase complex efficiently
drives the entire sterol 14-demethylation (CYP51) reaction. FEBS
Lett. 462, 283–288.
Lange, T., Hedden, P., Graebe, J.E., 1994. Expression, cloning of a
gibberellin 20 oxidase, a multifunctional enzyme involved in gibber-
ellin biosynthesis. Proc. Natl. Acad. Sci. USA 91, 8552–8556.
Lange, T., Kegler, K., Hedden, P., Phillips, A., Graebe, J., 1997.
Molecular characterization of gibberellin 20-oxidases. Structure–func-
tion studies on recombinant enzymes and chimaeric proteins. Physiol.
Plantarum 100, 543–549.
Linnemansto¨ns, P., Voss, T., Hedden, P., Gaskin, P., Tudzynski, B., 1999.
Deletions in the gibberellin biosynthesis gene cluster of Gibberella
fujikuroi by restriction enzyme-mediated integration and conventional
transformation-mediated mutagenesis. Appl. Environ. Microbiol. 65,
2558–2564.
Compounds were identified by comparison of their mass
spectra with those of authentic samples and/or with pub-
lished spectra (Gaskin and MacMillan, 1992). For incuba-
tions with deuterated precursors the EtOAc extract was
derivatized directly for GC–MS analysis.
Acknowledgements
This work was supported by Fondo Nacional de Ciencia
´
y Tecnologıa (Grant 1020140) and by the CONICYT/
DAAD Cooperation Program. Sponsorship of the British
Council is gratefully acknowledged. Rothamsted Research
receives grant-aided support from the Biotechnology and
Biological Sciences Research Council of the United
Kingdom.
MacMillan, J., 1997. Biosynthesis of the gibberellin plant hormones. Nat.
Prod. Res. 14, 221–243.
Malonek, S., Rojas, M.C., Hedden, P., Gaskin, P., Hopkins, P.,
Tudzynski, B., 2004. The NADPH–cytochrome P450 reductase gene
from Gibberella fujikuroi is essential for gibberellin biosynthesis. J.
Biol. Chem. 279, 25075–25084.
References
Miller, W., 2005. Regulation of steroidogenesis by electron transfer.
Endocrinology 146, 2544–2550.
Bearder, J.R., MacMillan, J., Phinney, B.O., 1975. Fungal products Part
XIV. Metabolic pathways from ent-kaurenoic acid to the fungal
gibberellins in mutant B1-41a of Gibberella fujikuroi. J. Chem. Soc.,
Perkin Trans. I, 721–726.
Bearder, J.R., MacMillan, J., Phinney, B.O., 1976. Origin of the oxygen
atoms in the lactone bridge of C19-gibberellins. J. Chem. Soc., Chem.
Commun., 834–835.
Bewley, M.C., Marohnic, C.C., Barber, M.J., 2001. The structure and
biochemistry of NADH-dependent cytochrome b5 reductase are now
consistent. Biochemistry 40, 13574–13582.
Davidson, S.E., Elliot, R.C., Helliwell, C.A., Poole, A.T., Reid, J.B., 2003.
The pea gene NA encodes ent-kaurenoic acid oxidase. Plant Physiol.
131, 335–344.
Mokashi, V., Li, L., Porter, T.D., 2003. Cytochrome b5 reductase and
cytochrome b5 support CYP2E1-mediated activation of nitrosamines
in a recombinant Ames test. Arch. Biochem. Biophys. 412, 147–152.
O’Donnell, K., Cigelnik, E., Nirenberg, H.I., 1998. Molecular systematics
and phylogeography of the Gibberella fujikuroi species complex.
Mycologia 90, 465–493.
Pandey, A.V., Miller, W.L., 2005. Regulation of 17,20 lyase activity by
cytochrome b5 and by serine phosphorylation of P450c17. J. Biol.
Chem. 280, 13265–13271.
Porter, T.D., 2002. The roles of cytochrome b5 in cytochrome P450
reactions. J. Biochem. Mol. Toxicol. 16, 311–316.
Prosser, D.E., Jones, G., 2004. Enzymes involved in the activation and
inactivation of vitamin D. Trends Biochem. Sci. 29, 664–673.
Rademacher, W., 1992. Inhibition of gibberellin production in the fungi
Gibberella fujikuroi and Sphaceloma manihoticola by plant growth
retardants. Plant Physiol. 100, 625–629.
Dockerill, B., Hanson, J.R., 1978. The fate of C20 in C19 gibberellin
biosynthesis. Phytochemistry 17, 701–704.
Edebo, L., 1983. Disintegration of cells by extrusion under pressure. In:
Lafferty, R.M. (Ed.), Enzyme Technology. Springer Verlag, Berlin.
Gaskin, P., MacMillan, J., 1992. GC–MS of Gibberellins and Related
Compounds: Methodology and a Library of Reference Spectra.
Cantocks Enterprises, Bristol, UK.
Geissmann, T.A., Verbiscar, A.J., Phinney, B.O., Cragg, G., 1966. Studies
on the biosynthesis of gibberellins from (À) ent-kaurenoic acid in
cultures of Gibberella fujikuroi. Phytochemistry 5, 933–947.
Graebe, J., 1987. Gibberellin biosynthesis and control. Ann. Rev. Plant
Physiol. 38, 419–465.
Rademacher, W., 1994. Gibberellin formation in microorganisms. J. Plant
Growth Regul. 15, 303–314.
Rojas, M.C., Hedden, P., Gaskin, P., Tudzynski, B., 2001. P450-1 gene of
Gibberella fujikuroi encodes a multifunctional enzyme in gibberellin
biosynthesis. Proc. Natl. Acad. Sci. USA 98, 5838–5843.
Rojas, M.C., Urrutia, O., Cruz, C., Gaskin, P., Tudzynski, B., Hedden, P.,
2004. Kaurenolides and fujenoic acids are side products of the P450-1
monooxygenase in Gibberella fujikuroi. Phytochemistry 65, 821–830.
Schenkman, J.B., Jansson, I., 2003. The many roles of cytochrome b5.
Pharmacol. Therapeut. 97, 139–152.
Guenguerich, F.P., Johnson, W.W., 1997. Kinetics of ferric cytochrome
P450 reduction by NADPH–cytochrome P450 reductase: rapid reduc-
tion in the absence of substrate and variations among cytochrome
P450 systems. Biochemistry 36, 14741–14750.
Shao, W., Im, S., Zuiderweg, E.R.P., Waskell, L., 2003. Mapping the
interface of the cytochrome b5-cytochrome c complex by nuclear
magnetic resonance. Biochemistry 42, 14774–14784.
Hedden, P., 1997. The oxidases of gibberellin biosynthesis: their function
and mechanism. Physiol. Plantarum 101, 709–719.
Sibbessen, O., Koch, B., Halkier, B.A., Moller, B.L., 1995. Cytochrome
P450TYR is a multifunctional heme-thiolate enzyme catalyzing the
conversion of L-tyrosine to p-hydroxyphenylacetaldehyde oxime in the
biosynthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor
(L.) moench. J. Biol. Chem. 270, 3506–3511.
Tudzynski, B., Ho¨lter, K., 1998. Gibberellin biosynthetic pathway in
Gibberella fujikuroi: evidence for a gene cluster. Fungal Genet. Biol. 25,
157–170.
Hedden, P., MacMillan, J., Phinney, B.O., 1974. Fungal products. Part
XII. Gibberellin A14 aldehyde, an intermediate in gibberellin biosyn-
thesis in Gibberella fujikuroi. J. Chem. Soc., Perkin Trans. I, 587–592.
Hedden, P., Phillips, A.L., Rojas, M.C., Carrera, E., Tudzynski, B., 2002.
Gibberellin biosynthesis in plants and fungi: a case of convergent
evolution? J. Plant Growth Regul. 20, 319–331.
Helliwell, C.A., Chandler, P.M., Poole, A., Dennis, E.S., Peacock, W.J.,
2001. The CYP88A cytochrome P450, ent-kaurenoic acid oxidase,
catalyzes three steps of the gibberellin biosynthesis pathway. Proc.
Natl. Acad. Sci. USA 98, 2065–2070.
Tudzynski, B., Sharon, A., 2002. Biosynthesis, biological role and
application of fungal phytohormones. In: Osiewacz, H.D. (Ed.), The
Mycota, Industrial Application, vol. 10. Springer, Berlin, Heidelberg,
NY.