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 | 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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-glucose + hydroquinone | Rauvolfia serpentina | - |
UDP + hydroquinone-O-beta-D-glucopyranoside | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rauvolfia serpentina | Q9AR73 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography | Rauvolfia serpentina |
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 | Comment | Organism | Textmining |
---|---|---|---|
cell suspension culture | - |
Rauvolfia serpentina | - |
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 | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
assay at | Rauvolfia serpentina |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Rauvolfia serpentina |