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Literature summary for 5.1.3.2 extracted from

  • Kotake, T.; Takata, R.; Verma, R.; Takaba, M.; Yamaguchi, D.; Orita, T.; Kaneko, S.; Matsuoka, K.; Koyama, T.; Reiter, W.D.; Tsumuraya, Y.
    Bifunctional cytosolic UDP-glucose 4-epimerases catalyse the interconversion between UDP-D-xylose and UDP-L-arabinose in plants (2009), Biochem. J., 424, 169-177.
    View publication on PubMed
Search for more literature for 5.1.3.2

Cloned(Commentary)

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

Inhibitors Comment Organism Structure
Hg2+ complete inhibition, recombinan t enzyme Pisum sativum

KM Value [mM]

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

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

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 [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/ Products (Substrates)

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

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 (Commentary)

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

Source Tissue Comment Organism Textmining
sprout
-
 Pisum sativum
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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 [°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 [°C]

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 [1/s]

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

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 Range

pH Minimum pH Maximum Comment Organism
8.5 9 maximum activity of recombinant PsUGE1 Pisum sativum

Cofactor

Cofactor Comment Organism Structure
additional information an NAD+ binding motif is GGXGXXG, not required for activity Pisum sativum

General Information

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 [mM/s]

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