PMID: 2505672Sep 1, 1989Paper

Phytoalexin synthesis in soybean: purification and reconstitution of cytochrome P450 3,9-dihydroxypterocarpan 6a-hydroxylase and separation from cytochrome P450 cinnamate 4-hydroxylase

Archives of Biochemistry and Biophysics
G Kochs, H Grisebach

Abstract

Elicitor-challenged soybean (Glycine max) cell cultures were used for detergent solubilization and purification of cytochrome P450 3,9-dihydroxypterocarpan 6a-hydroxylase (D6aH). D6aH was purified to electrophoretic homogeneity from such cells by a five-step procedure. It could be separated from cytochrome P450 cinnamate 4-hydroxylase on hydroxyapatite. This is the first report on separation of two cytochrome P450 enzymes from a higher plant. On sodium dodecyl sulfate polyacrylamide gels D6aH migrated with a Mr about 55,000. For reconstitution experiments soybean NADPH:cytochrome P450 (cytochrome c) reductase was purified to homogeneity. Reconstitution of D6aH in the presence of NADPH was dependent on cytochrome P450 D6aH, the reductase, and lipid. Dilauroylphosphatidylcholine gave higher D6aH activity than soybean lipids (asolectin). The reconstituted D6aH system showed a much higher temperature stability than the microsomal system.

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Citations

Nov 1, 1992·Plant Physiology·M Simontacchi, S Puntarulo
Jul 1, 1991·Plant Physiology·R P Donaldson, D G Luster
Jan 1, 1995·Drug Metabolism and Drug Interactions·D Werck-Reichhart
Jan 31, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jian-Qiang KongZhi-Biao Wang
Nov 18, 2003·The Plant Journal : for Cell and Molecular Biology·Chang-Jun LiuRichard A Dixon
Jul 1, 1991·Archives of Biochemistry and Biophysics·B GabriacF Durst
Oct 29, 1998·Biochemical and Biophysical Research Communications·T AkashiS Ayabe
Jan 22, 1998·Archives of Biochemistry and Biophysics·A RoscoT M Kutchan
Dec 1, 1994·Phytochemistry·G P BolwellA Zimmerlin
Feb 11, 1992·Trends in Biochemical Sciences·H Sandermann
May 31, 1991·Biochemical and Biophysical Research Communications·I BenvenisteF Durst
Feb 1, 2008·Journal of Natural Products·Rahul S Khupse, Paul W Erhardt

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