Plant biochemistry of xenobiotics: isolation and characterization of a soybean O-glucosyltransferase of DDT metabolism.
The insecticide DDT is metabolized in soybean and wheat cell cultures to the acylglucoside of 2,2-bis-(4-chlorophenyl)-acetic acid (DDA) (M. Arjmand and H. Sandermann, 1985, Pesticide Biochem. Physiol. 23, 389-397). An enzyme catalyzing the conjugation reaction has been highly purified from the soluble enzyme fraction of cultured soybean cells. After the initial ammonium sulfate fractionation, quercetin and pentachlorophenol were preferentially glucosylated. In the course of 367-fold purification, DDA became the preferred substrate. The purified enzyme was unstable. A molecular weight of approximately 50 kDa was estimated for the native enzyme (gel permeation chromatography) as well as the denatured protein (sodium dodecyl sulfate-gel electrophoresis). The isoelectric point for the enzyme was near pH 4.9. Apparent Km values of about 170 microM were determined for UDP-glucose as well as DDA. The maximal velocity was 257 microkat/kg protein, corresponding to a conjugation capacity of 855 micrograms DDA/h/g fresh weight of cells.
GSF-Forschungszentrum für Umwelt und Gesundheit GmbH, Institut für Biochemische Pflanzenpathologie, Oberschleissheim, FRG.
A
A
Wetzel
H
H
Sandermann
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D003634:DDT / Q000378:metabolism*; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism*; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D013025:Glycine max / Q000201:enzymology*; D015262:Xenobiotics / Q000378:metabolism*
D015262:Xenobiotics; D003634:DDT; C090277:2,2-bis(4-chlorophenyl)acetate O-glucosyltransferase; D005964:Glucosyltransferases