Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.1.218 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)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
A204V site-directed mutagenesis, mutant shows slightly reduced activity compared to the wild-type enzyme Rauvolfia serpentina
E368A site-directed mutagenesis, mutant shows reduced activity compared to the wild-type enzyme Rauvolfia serpentina
E368D site-directed mutagenesis, mutant shows reduced activity compared to the wild-type enzyme Rauvolfia serpentina
H360E site-directed mutagenesis, mutant shows about 4000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
H360K site-directed mutagenesis, mutant shows about 1000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
H360R site-directed mutagenesis, mutant shows about 1000fold reduced activity compared to the wild-type enzyme Rauvolfia serpentina
L86I site-directed mutagenesis, mutant shows slightly reduced activity compared to the wild-type enzyme Rauvolfia serpentina
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Rauvolfia serpentina Q9AR73
-
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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

Source Tissue

Source Tissue Comment Organism Textmining
cell suspension culture
-
Rauvolfia serpentina
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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