Isolation and Characterization of a UDPGlucose: Flavonol O3-Glucosyltransferase from Illuminated Red Cabbage (Brassica oleracea cv Red Danish) Seedlings
Open Access
- 1 February 1991
- journal article
- Published by Oxford University Press (OUP) in Plant Physiology
- Vol. 95 (2) , 570-576
- https://doi.org/10.1104/pp.95.2.570
Abstract
A UDPGlc:flavonol O3-glucosyltransferase (EC 2.4.1.91) that catalyzes the formation of quercetin and kaempferol O3-glucosides has been purified about 1450-fold from illuminated red cabbage (Brassica oleracea cv Red Danish) seedlings with a 3.3% yield. Purification of the enzyme was achieved by (NH4)2SO4-precipitation, gel-filtration, ion-exchange chromatography on DEAE-Bio-Gel and Q-Sepharose, chromatofocusing, and electrophoresis in nondenaturing polyacrylamide (10%) gels. The enzyme preparation had a pH optimum between 5.8 and 6.2, isoelectric point in the pH range 4.25 to 4.55, a Mr of 59,000, and it was composed of two similar subunits of Mr 29,500. The glucosyltransferase reached half substrate saturation at 180 micromolar (UDPGlc) and 7 micromolar (quercetin) concentrations. Kaempferol, which was glucosylated at a relative rate of 87%, had a lesser affinity for the enzyme (Km~12 micromolar). Flavanones, flavanols, flavones, dihydroflavonols, and anthocyanidins were not readily utilized as substrates, suggesting that the enzyme is specific for flavonol glucoside biosynthesis.Keywords
This publication has 10 references indexed in Scilit:
- Biochemical, immunological, and immunocytochemical evidence for the association of chalcone synthase with endoplasmic reticulum membranes.Proceedings of the National Academy of Sciences, 1987
- Endoplasmic Reticulum as a Site of Phenylpropanoid and Flavonoid Metabolism in HippeastrumPlant Physiology, 1984
- Partial Purification and Characterization of UDPG: t -Cinnamate Glucosyltransferase in the Root of Sweet Potato, Ipomoea batatas LamThe Journal of Biochemistry, 1984
- Purification and Properties of Flavonol-Ring B Glucosyltransferase from Chrysosplenium americanumPlant Physiology, 1983
- Isolation and partial characterization of three glucosyl transferases involved in the biosynthesis of flavonol triglucosides in Pisum sativum LArchives of Biochemistry and Biophysics, 1982
- UDP-glucose:Isoflavone 7-O-glucosyltransferase from roots of chick pea (Cicer arietinum L.)Archives of Biochemistry and Biophysics, 1981
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- Free reversibility of the UDP-glucose: Flavonol 3-O-glucosyltransferase reactionArchives of Biochemistry and Biophysics, 1975
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Untersuchungen über Isorhamnetinglykoside, II. Isolierung und Konstitutionsaufklärung neuer Glykoside von Brassica napus L. und Sinapis arvensis L.European Journal of Inorganic Chemistry, 1967