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  • Identification of the glucosyltransferase that mediates direct flavone C-glucosylation in Gentiana triflora.

Identification of the glucosyltransferase that mediates direct flavone C-glucosylation in Gentiana triflora.

FEBS letters (2014-12-06)
Nobuhiro Sasaki, Yuzo Nishizaki, Eri Yamada, Fumi Tatsuzawa, Takashi Nakatsuka, Hideyuki Takahashi, Masahiro Nishihara
ABSTRACT

The major flavonoids accumulated in leaves of Japanese gentian (Gentiana triflora) were determined as isoorientin (luteolin 6-C-glucoside) and isoorientin 4'-glucoside. A cDNA (GtUF6CGT1) was isolated that encoded the UDP-glucose-dependent glucosyltransferase that is involved in C-glycosylflavone biosynthesis. The recombinant GtUF6CGT1 protein could transfer a glucose group to the C6 position of a flavone skeleton through C-linkage, using UDP-glucose as the glucosyl donor. These C-glycosylflavones also accumulated in petals. A good correlation was observed between GtUF6CGT1 expression and C-glycosylflavone accumulation in leaves and petals. GtUF6CGT1 is the first reported C-glucosyltransferase that mediates direct C-glucosylation of the flavone skeleton.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
DL-Dithiothreitol solution, BioUltra, Molecular Biology, ~1 M in H2O
Sigma-Aldrich
Isoorientin, ≥98% (HPLC)
Supelco
DL-Dithiothreitol solution, 1 M in H2O
Sigma-Aldrich
Potassium phosphate tribasic, reagent grade, ≥97%
Isoorientin, primary reference standard
Sigma-Aldrich
Phosphorylase b from rabbit muscle, For use as a marker in SDS-PAGE
Sigma-Aldrich
Phosphorylase b from rabbit muscle, lyophilized powder, ≥20 units/mg protein, 2× crystallization
Sigma-Aldrich
Luteolin, ≥98% (TLC), powder
Supelco
Luteolin, analytical standard
Luteolin, primary reference standard