Cloned (Comment) | Organism |
---|---|
gene UGE1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the enzyme fused to thioredoxin and His6 in Escherichia coli strain BL21(DE3) | Pisum sativum |
recombinant protein is expressed in Escherichia coli | Pisum sativum |
recombinant protein is expressed in Escherichia coli with C-terminal His-tag | Arabidopsis thaliana |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Hg2+ | complete inhibition, recombinan t enzyme | Pisum sativum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis | Pisum sativum | |
0.057 | - |
UDP-D-galactose | recombinant enzyme UGE4, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-Glc dehydrogenase | Arabidopsis thaliana | |
0.068 | - |
UDP-D-galactose | recombinant enzyme UGE3, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-Glc dehydrogenase | Arabidopsis thaliana | |
0.087 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme UGE1, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-Glc dehydrogenase | Arabidopsis thaliana | |
0.095 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme UGE2, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDPGlc dehydrogenase | Arabidopsis thaliana | |
0.15 | - |
UDP-D-xylose | pH 8.6, 25°C | Pisum sativum | |
0.15 | - |
UDP-xylose | pH 8.6, 25°C | Pisum sativum | |
0.15 | - |
UDP-D-galactose | recombinant enzyme UGE5, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana | |
0.16 | - |
UDP-L-arabinose | pH 8.6, 25°C | Pisum sativum | |
0.29 | - |
UDP-galactose | pH 8.6, 25°C | Pisum sativum | |
0.29 | - |
UDP-D-galactose | 50 mM Tris-HCl buffer (pH 8.6), 0.1 mM NAD+, 1 mM UDP-sugar and enzyme | Pisum sativum | |
0.31 | - |
UDP-glucose | pH 8.6, 25°C | Pisum sativum | |
0.31 | - |
UDP-D-glucose | 50 mM Tris-HCl buffer (pH 8.6), 0.1 mM NAD+, 1 mM UDP-sugar and enzyme | Pisum sativum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Arabidopsis thaliana | 5829 | - |
cytosol | - |
Pisum sativum | 5829 | - |
cytosol | the bifunctional UDP-Glc 4-epimerase/UDP-Xyl 4-epimerase, encoded by UGE1, in the cytosol is distinct from the UDP-Xyl 4-epimerase in the Golgi apparatus | Pisum sativum | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Al3+ | enzyme activity is 28.6 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Ba2+ | enzyme activity is 54.6 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Ca2+ | enzyme activity is 47.8 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Co2+ | enzyme activity is 44.7 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Cu2+ | enzyme activity is 21.8 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Fe3+ | enzyme activity is 20.3 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Hg2+ | 1 mM Hg2+ abolishes enzyme function entirely | Pisum sativum | |
Li+ | enzyme activity is 43.7 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Mg2+ | enzyme activity is 51.5 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
Mn2+ | enzyme activity is 43.2 U/mg protein at a metal concentration of 1 mM | Pisum sativum | |
additional information | no metal ion requirement of the recombinant protein for either UDP-Glc 4-epimerase or UDP-Xyl 4-epimerase activity | Pisum sativum | |
Zn2+ | enzyme activity is 17.2 U/mg protein at a metal concentration of 1 mM | Pisum sativum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39000 | - |
native Pisum sativum enzyme determined by SDS-PAGE | Pisum sativum |
41000 | - |
recombinant enzyme after purification and thrombin digestion | Pisum sativum |
56000 | - |
recombinant enzyme after purification on a chelating column | Pisum sativum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-glucose | Pisum sativum | - |
UDP-alpha-D-galactose | - |
r | |
UDP-D-glucose | Arabidopsis thaliana | - |
UDP-D-galactose | - |
r | |
UDP-D-glucose | Pisum sativum | - |
UDP-D-galactose | - |
r | |
UDP-D-xylose | Pisum sativum | - |
UDP-L-arabinose | - |
r | |
UDP-galactose | Pisum sativum | - |
UDP-glucose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | Q42605 | - |
- |
Arabidopsis thaliana | Q8LDN8 | - |
- |
Arabidopsis thaliana | Q9C7W7 | - |
- |
Arabidopsis thaliana | Q9SN58 | - |
- |
Arabidopsis thaliana | Q9T0A7 | - |
- |
Pisum sativum | B0M3E8 | - |
- |
Pisum sativum | B0M3E8 | gene UGE1 | - |
Purification (Comment) | Organism |
---|---|
purification of the native enzyme from pea sprouts by ion exchange chromatography and gel filtration, further purification on a hydroxyapatite column, purification of recombinant enzymes by ion exchange chromatography | Pisum sativum |
purification of the recombinant Arabidopsis enzyme by Ni-NTA affinity chromatography | Arabidopsis thaliana |
purification of the recombinant enzyme by Ni-NTA affinity chromatography | Arabidopsis thaliana |
recombinant fusion enzyme 2.33fold from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, native UGE1 334fold from sprouts by ammonium sulfate fractionation and a chromatographic procedure | Pisum sativum |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
sprout | - |
Pisum sativum | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.7 | - |
native protein, substrate is UDP-glucose | Pisum sativum |
1.7 | - |
purified native enzyme, substrate UDP-Glc | Pisum sativum |
4.6 | - |
purified recombinant detagged enzyme, substrate UDP-Glc | Pisum sativum |
4.6 | - |
recombinant Pisum protein, substrate is UDP-glucose | Pisum sativum |
6.67 | - |
purified native enzyme, substrate UDP-D-Xyl | Pisum sativum |
6.7 | - |
purified native enzyme, substrate UDP-L-Ara | Pisum sativum |
16.7 | - |
specific activity of recombinant enzyme in cell lysate after expression in Escherichia coli | Pisum sativum |
19.8 | - |
native protein, substrate is UDP-galactose | Pisum sativum |
19.8 | - |
purified native enzyme, substrate UDP-Gal | Pisum sativum |
21.1 | - |
purified recombinant detagged enzyme, substrate UDP-L-Ara | Pisum sativum |
38.9 | - |
purified recombinant detagged enzyme, substrate UDP-D-Xyl | Pisum sativum |
38.9 | - |
specific activity of recombinant enzyme after purification by a DEAE-sepharose FF column | Pisum sativum |
49 | - |
specific activity of recombinant enzyme after purification by a Ni-sepharose colum | Pisum sativum |
53.3 | - |
purified recombinant detagged enzyme, substrate UDP-Gal | Pisum sativum |
53.3 | - |
recombinant Pisum protein, substrate is UDP-galactose | Pisum sativum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the bifunctional UDP-Glc 4-epimerase/UDP-Xyl 4-epimerase in the cytosol is distinct from the UDP-Xyl 4-epimerase in the Golgi apparatus, the two activities of UGE1 occur at the same catalytic site. No activity with UDP-N-acetylglucosamine or UDP-glucuronic acid, UGE1 substrate specificity, overview | Pisum sativum | ? | - |
? | |
UDP-alpha-D-glucose | - |
Pisum sativum | UDP-alpha-D-galactose | - |
r | |
UDP-alpha-D-glucose | the apparent equilibrium constant for UDP-Gal formation from UDP-Glc is 0.24 | Pisum sativum | UDP-alpha-D-galactose | - |
r | |
UDP-D-glucose | - |
Arabidopsis thaliana | UDP-D-galactose | - |
r | |
UDP-D-glucose | - |
Pisum sativum | UDP-D-galactose | - |
r | |
UDP-D-xylose | - |
Pisum sativum | UDP-L-arabinose | - |
r | |
UDP-D-xylose | the apparent equilibrium constant for UDP-Ara formation from UDP-Xyl is 0.89 | Pisum sativum | UDP-L-arabinose | - |
r | |
UDP-galactose | - |
Pisum sativum | UDP-glucose | - |
r | |
UDP-galactose | the apparent equilibrium constant for UDP-Gal formation from UDP-Glc is 0.24 | Pisum sativum | UDP-glucose | - |
r | |
UDP-L-arabinose | the apparent equilibrium constant for UDP-Ara formation from UDP-Xyl is 0.89 | Pisum sativum | UDP-D-xylose | - |
r |
Synonyms | Comment | Organism |
---|---|---|
AtUGE1 | - |
Arabidopsis thaliana |
AtUGE2 | - |
Arabidopsis thaliana |
AtUGE3 | - |
Arabidopsis thaliana |
AtUGE4 | - |
Arabidopsis thaliana |
AtUGE5 | - |
Arabidopsis thaliana |
More | PsUGE1 belongs to plant UGE I family | Pisum sativum |
PsUGE1 | - |
Pisum sativum |
UDP-Glc 4-epimerase | - |
Arabidopsis thaliana |
UDP-Glc 4-epimerase | - |
Pisum sativum |
UDP-glucose 4-epimerase | - |
Arabidopsis thaliana |
UDP-glucose 4-epimerase | - |
Pisum sativum |
UDP-Xyl 4-epimerase | - |
Pisum sativum |
UDP-xylose 4-epimerase | - |
Pisum sativum |
UGE1 | - |
Pisum sativum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | - |
recombinant enzyme | Pisum sativum |
15 | - |
maximum activity of recombinant PsUGE1 | Pisum sativum |
30 | - |
native enzyme | Pisum sativum |
30 | - |
maximum activity of native enzyme | Pisum sativum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
purified recombinant enzyme, complete inactivation above | Pisum sativum |
40 | - |
recombinant Pisum enzyme loses both activities completely at temperatures higher than 40°C | Pisum sativum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9 | - |
UDP-glucose | pH 8.6, 25°C | Pisum sativum | |
9 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme | Arabidopsis thaliana | |
17 | - |
UDP-D-xylose | pH 8.6, 25°C | Pisum sativum | |
17 | - |
UDP-xylose | pH 8.6, 25°C | Pisum sativum | |
20 | - |
UDP-D-galactose | recombinant enzyme UGE4 | Arabidopsis thaliana | |
23 | - |
UDP-L-arabinose | pH 8.6, 25°C | Pisum sativum | |
27 | - |
UDP-D-galactose | recombinant enzyme UGE3 | Arabidopsis thaliana | |
55 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme UGE2 | Arabidopsis thaliana | |
64 | - |
UDP-galactose | pH 8.6, 25°C | Pisum sativum | |
64 | - |
UDP-D-galactose | recombinant enzyme UGE5 | Arabidopsis thaliana |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | 9 | recombinant enzyme | Pisum sativum |
8.6 | - |
assay at | Arabidopsis thaliana |
8.6 | - |
assay at | Pisum sativum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
8.5 | 9 | maximum activity of recombinant PsUGE1 | Pisum sativum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | an NAD+ binding motif is GGXGXXG, not required for activity | Pisum sativum |
General Information | Comment | Organism |
---|---|---|
physiological function | bifunctional cytosolic UDP-glucose 4-epimerase catalyses the interconversion between UDP-D-xylose and UDP-L-arabinose in plants | Arabidopsis thaliana |
physiological function | bifunctional cytosolic UDP-glucose 4-epimerase catalyses the interconversion between UDP-D-xylose and UDP-L-arabinose in plants | Pisum sativum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
28 | - |
UDP-glucose | pH 8.6, 25°C | Pisum sativum | |
103 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme UGE1, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana | |
115 | - |
UDP-D-xylose | pH 8.6, 25°C | Pisum sativum | |
115 | - |
UDP-xylose | pH 8.6, 25°C | Pisum sativum | |
141 | - |
UDP-L-arabinose | pH 8.6, 25°C | Pisum sativum | |
220 | - |
UDP-galactose | pH 8.6, 25°C | Pisum sativum | |
350 | - |
UDP-D-galactose | recombinant enzyme UGE4, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana | |
397 | - |
UDP-D-galactose | recombinant enzyme UGE3, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana | |
435 | - |
UDP-D-galactose | recombinant enzyme UGE5, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana | |
578 | - |
UDP-D-galactose | recombinant Arabidopsis thaliana enzyme UGE2, 100 mM glycine/NaOH (pH 8.6), 0.015-1 mM UDP-Gal, 2 mM NAD+ and 40 mU of UDP-glucose dehydrogenase | Arabidopsis thaliana |