Cloning and heterologous expression of a rape cDNA encoding UDP-glucose:sinapate glucosyltransferase.
A cDNA encoding a UDP-glucose:sinapate glucosyltransferase (SGT) that catalyzes the formation of 1-O-sinapoylglucose, was isolated from cDNA libraries constructed from immature seeds and young seedlings of rape (Brassica napus L.). The open reading frame encoded a protein of 497 amino acids with a calculated molecular mass of 55,970 Da and an isoelectric point of 6.36. The enzyme, functionally expressed in Escherichia coli, exhibited broad substrate specificity, glucosylating sinapate, cinnamate, ferulate, 4-coumarate and caffeate. Indole-3-acetate, 4-hydroxybenzoate and salicylate were not conjugated. The amino acid sequence of the SGT exhibited a distinct sequence identity to putative indole-3-acetate glucosyltransferases from Arabidopsis thaliana and a limonoid glucosyltransferase from Citrus unshiu, indicating that SGT belongs to a distinct subgroup of glucosyltransferases that catalyze the formation of 1-O-acylglucosides (beta-acetal esters).
Milkowski
C
C
Leibniz-Institut für Pflanzenbiochemie, Abteilung Sekundärstoffwechsel, Halle (Saale), Germany.
Baumert
A
A
Strack
D
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000235:genetics*; D002384:Catalysis; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D005964:Glucosyltransferases; C420119:UDP-glucose sinapate glucosyltransferase, Brassica napus