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Literature summary extracted from

  • Offen, W.; Martinez-Fleites, C.; Yang, M.; Kiat-Lim, E.; Davis, B.G.; Tarling, C.A.; Ford, C.M.; Bowles, D.J.; Davies, G.J.
    Structure of a flavonoid glucosyltransferase reveals the basis for plant natural product modification (2006), EMBO J., 25, 1396-1405.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.91 gene VvGT1, expression of His-tagged enzyme in Escherichia coli Vitis vinifera

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.91 purified recombinant detagged enzyme bound to UDP-2-fluoroglucose and kaempferol, or with UDP and quercetin, hanging drop method, 4.8 mg/ml protein in 5 mM Tris (2-carboxyethyl) phosphine HCl, 10 mM Tris–Cl, pH 8.3, diluted 1:1 with mother liquor which contains 18–22% polyethylene glycol 10 000, 0.1M Bis-Tris propane, pH 7.0, and 0–0.5% v/v Pluronic F-68, crystals appear within 24 h, X-ray diffraction structure determination and analysis at 2.2 A resolution Vitis vinifera

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.91 D374A site-directed mutagenesis, inactive mutant Vitis vinifera
2.4.1.91 H20A site-directed mutagenesis, inactive mutant Vitis vinifera
2.4.1.91 Q375H site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Vitis vinifera
2.4.1.91 Q375N site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Vitis vinifera
2.4.1.91 Q375N/T141A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Vitis vinifera
2.4.1.91 T141A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Vitis vinifera

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.91 0.031
-
 quercetin
-
 Vitis vinifera
2.4.1.91 0.042
-
 kaempferol
-
 Vitis vinifera

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.91 UDP-alpha-D-glucose + kaempferol Vitis vinifera
-
 UDP + kaempferol 3-O-beta-D-glucoside
-
 ?
2.4.1.91 UDP-alpha-D-glucose + quercetin Vitis vinifera
-
 UDP + quercetin 3-O-beta-D-glucoside
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.91 Vitis vinifera
-
 gene VvGT1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.91 recombinant His-tagged enzyme from Escherichia coli to homogeneity, the His-tag is cleaved off Vitis vinifera

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.91 dTDP-glucose + quercetin
-
 Vitis vinifera dTDP + quercetin 3-O-beta-D-glucoside
-
 ?
2.4.1.91 dTDP-xylose + quercetin low activity Vitis vinifera dTDP + quercetin 3-O-beta-D-xylose
-
 ?
2.4.1.91 GDP-glucose + quercetin low activity Vitis vinifera GDP + quercetin 3-O-beta-D-glucoside
-
 ?
2.4.1.91 additional information the enzyme catalyzes the addition of glucose from a UDP-glucose donor to the 3-O position, GT1 can also harness a wide panel of different UDP-sugars: UDP-5SGlc, UDP-Xyl, UDP-Man, UDP-Gal and UDP-GlcNAc (Table I), as well as GDP-Glc and dTDP-Xyl, substrate specificity of wild-type and mutant enzymes, overview, substrate binding structure, overview Vitis vinifera ?
-
 ?
2.4.1.91 UDP-5-thio-glucose + quercetin low activity Vitis vinifera UDP + quercetin 3-O-5-thio-beta-D-glucoside
-
 ?
2.4.1.91 UDP-alpha-D-glucose + kaempferol
-
 Vitis vinifera UDP + kaempferol 3-O-beta-D-glucoside
-
 ?
2.4.1.91 UDP-alpha-D-glucose + quercetin
-
 Vitis vinifera UDP + quercetin 3-O-beta-D-glucoside
-
 ?
2.4.1.91 UDP-galactose + quercetin low activity Vitis vinifera UDP + quercetin 3-O-beta-D-galactoside
-
 ?
2.4.1.91 UDP-mannose + quercetin very low activity Vitis vinifera UDP + quercetin 3-O-beta-D-mannoside
-
 ?
2.4.1.91 UDP-N-acetylglucose + quercetin low activity Vitis vinifera UDP + quercetin 3-O-N-acetylglucoside
-
 ?
2.4.1.91 UDP-xylose + quercetin low activity Vitis vinifera UDP + quercetin 3-O-beta-D-xylose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.91 GT1
-
 Vitis vinifera
2.4.1.91 UDPglucose:flavonoid 3-O-glycosyltransferase
-
 Vitis vinifera

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.91 37
-
 assay at Vitis vinifera

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.91 7.8
-
 assay at Vitis vinifera