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Literature summary extracted from

  • Hefner, T.; Stockigt, J.
    Probing suggested catalytic domains of glycosyltransferases by site-directed mutagenesis (2003), Eur. J. Biochem., 270, 533-538.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.218 DNA and amino acid sequence determination and analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain M15 Rauvolfia serpentina

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.218 A204V site-directed mutagenesis, mutant shows slightly reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 E368A site-directed mutagenesis, mutant shows reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 E368D site-directed mutagenesis, mutant shows reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 H360E site-directed mutagenesis, mutant shows about 4000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 H360K site-directed mutagenesis, mutant shows about 1000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 H360R site-directed mutagenesis, mutant shows about 1000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 L86I site-directed mutagenesis, mutant shows slightly reduced activity compared to the wild-type enzyme Rauvolfia serpentina
2.4.1.218 additional information generation of the typical E-E motif of the enzyme does not convert a NRD1beta enzyme into a NRD1alpha enzyme Rauvolfia serpentina

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.218 0.398
-
 Vanillin pH 7.5, 50°C, recombinant mutant E368A Rauvolfia serpentina
2.4.1.218 0.411
-
 Vanillin pH 7.5, 50°C, recombinant mutant E368D Rauvolfia serpentina
2.4.1.218 0.44
-
 Vanillin pH 7.5, 50°C, recombinant wild-type enzyme Rauvolfia serpentina
2.4.1.218 0.452
-
 Vanillin pH 7.5, 50°C, recombinant mutant L86I Rauvolfia serpentina
2.4.1.218 0.706
-
 Vanillin pH 7.5, 50°C, recombinant mutant A204V Rauvolfia serpentina

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.218 UDP-glucose + hydroquinone Rauvolfia serpentina
-
 UDP + hydroquinone-O-beta-D-glucopyranoside
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.218 Rauvolfia serpentina Q9AR73
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.218 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography Rauvolfia serpentina

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.218 UDP-glucose + hydroquinone = UDP + hydroquinone-O-beta-D-glucopyranoside reaction mechanism, the NRD domain with residue Glu368 is required for activity, His360 is crucial for activity Rauvolfia serpentina
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.4.1.218 cell suspension culture
-
 Rauvolfia serpentina
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.218 additional information
-
 substrate specificity of wild-type and E368A mutant enzymes, overview Rauvolfia serpentina
2.4.1.218 0.003
-
 purified recombinant mutant H360E, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 0.01
-
 purified recombinant mutant H360K, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 0.013
-
 purified recombinant mutant H360R, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 0.054
-
 purified recombinant mutant E368D, substrate vanillin Rauvolfia serpentina
2.4.1.218 0.066
-
 purified recombinant mutant E368A, substrate vanillin Rauvolfia serpentina
2.4.1.218 0.79
-
 purified recombinant mutant A204V, substrate vanillin Rauvolfia serpentina
2.4.1.218 1.07
-
 purified recombinant mutant E368D, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 1.22
-
 purified recombinant mutant L86I, substrate vanillin Rauvolfia serpentina
2.4.1.218 1.44
-
 purified recombinant mutant E368A, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 1.45
-
 purified recombinant wild-type enzyme, substrate vanillin Rauvolfia serpentina
2.4.1.218 8.34
-
 purified recombinant mutant A204V, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 10.82
-
 purified recombinant mutant L86I, substrate hydroquinone Rauvolfia serpentina
2.4.1.218 12.17
-
 purified recombinant wild-type enzyme, substrate hydroquinone Rauvolfia serpentina

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.218 additional information substrate specificity of wild-type and E368A mutant enzymes, overview Rauvolfia serpentina ?
-
 ?
2.4.1.218 UDP-glucose + 3-methoxyphenol 11.8% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + 3-methoxyphenyl-beta-D-glucoside
-
 ?
2.4.1.218 UDP-glucose + 4-hydroxybenzaldehyde 7.3% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + 4-beta-D-glucopyranosyloxy-benzaldehyde
-
 ?
2.4.1.218 UDP-glucose + 8-hydroxyquinoline 11.9% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + 8-hydroxyquinolin-beta-D-glucoside
-
 ?
2.4.1.218 UDP-glucose + beta-naphthol 18.4% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + 2-naphthyl-beta-D-glucopyranoside
-
 ?
2.4.1.218 UDP-glucose + eugenol 4.1% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + (4-allyl-2-methoxyphenyl)-beta-D-glucopyranoside
-
 ?
2.4.1.218 UDP-glucose + hydroquinone
-
 Rauvolfia serpentina UDP + hydroquinone-O-beta-D-glucopyranoside
-
 ?
2.4.1.218 UDP-glucose + hydroquinone highly preferred substrate Rauvolfia serpentina UDP + hydroquinone-O-beta-D-glucopyranoside
-
 ?
2.4.1.218 UDP-glucose + resorcinol 7.8% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + (3-hydroxyphenyl)-beta-D-glucopyranoside
-
 ?
2.4.1.218 UDP-glucose + scopoletin 3.5% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + scopoletin-beta-D-glucoside
-
 ?
2.4.1.218 UDP-glucose + umbelliferone 5.1% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + umbelliferyl beta-D-glucoside
-
 ?
2.4.1.218 UDP-glucose + vanillin 10.6% of the activity with hydroquinone, wild-type enzyme Rauvolfia serpentina UDP + vanillin-O-beta-D-glucopyranoside
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.218 arbutin synthase
-
 Rauvolfia serpentina
2.4.1.218 AS
-
 Rauvolfia serpentina
2.4.1.218 More the enzyme belongs to the NRD1beta family of glycosyltransferases Rauvolfia serpentina

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.218 50
-
 assay at Rauvolfia serpentina

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.218 0.08
-
 Vanillin pH 7.5, 50°C, recombinant mutant E368A Rauvolfia serpentina
2.4.1.218 0.12
-
 Vanillin pH 7.5, 50°C, recombinant mutant E368D Rauvolfia serpentina
2.4.1.218 1.3
-
 Vanillin pH 7.5, 50°C, recombinant mutant A204V Rauvolfia serpentina
2.4.1.218 1.84
-
 Vanillin pH 7.5, 50°C, recombinant mutant L86I Rauvolfia serpentina
2.4.1.218 2.59
-
 Vanillin pH 7.5, 50°C, recombinant wild-type enzyme Rauvolfia serpentina

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.218 7.5
-
 assay at Rauvolfia serpentina