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Literature summary for 5.1.3.2 extracted from

  • Dalrymple, S.A.; Ko, J.; Sheoran, I.; Kaminskyj, S.G.; Sanders, D.A.
    Elucidation of substrate specificity in Aspergillus nidulans UDP-galactose-4-epimerase (2013), PLoS ONE, 8, e76803.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold Aspergillus nidulans

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme, microbatch method, 4°C, mixing of equal volumes of protein and precipitant solution, the latter containing 20% w/v PEG 3350, 0.1 M Bis-Tris propane pH 7.5, 0.2 M sodium fluoride, cryoprotection with 25% glycerol, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement Aspergillus nidulans

Protein Variants

Protein Variants Comment Organism
K160V site-directed mutagenesis of the key residue responsible for anchoring of the co-factor, inactive mutant Aspergillus nidulans
L320C site-directed mutagenesis of a gate-keeper residue, the L320C mutant enzyme is also active in UDPGlcpNAc/UDP-GalpNAc interconversion Aspergillus nidulans
L320Y site-directed mutagenesis of a gate-keeper residue, the L320Y mutant enzyme is not active in UDPGlcpNAc/UDP-GalpNAc interconversion Aspergillus nidulans
Y156F site-directed mutagenesis of the key residue serving as the active site base, inactive mutant Aspergillus nidulans

Inhibitors

Inhibitors Comment Organism Structure
UDP-N-acetylgalactosamine competitive, binding study Aspergillus nidulans
UDP-N-acetylglucosamine competitive, binding study Aspergillus nidulans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic analyis of wild-type and mutant enzymes, overview Aspergillus nidulans
0.06
-
UDP-alpha-D-glucose mutant L320Y, pH 8.5, 25°C Aspergillus nidulans
0.1 1 UDP-alpha-D-glucose wild-type enzyme, pH 8.5, 25°C Aspergillus nidulans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40600
-
x * 40600 Aspergillus nidulans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-alpha-D-glucose Aspergillus nidulans via 4-ketose intermediate UDP-alpha-D-galactose
-
r

Organism

Organism UniProt Comment Textmining
Aspergillus nidulans C8VAU8
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Gold by nickel affinity chromatography and ultrafiltration Aspergillus nidulans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-alpha-D-glucose via 4-ketose intermediate Aspergillus nidulans UDP-alpha-D-galactose
-
r
UDP-alpha-D-glucose via 4-ketose intermediate, UDP-glucose is bound within the active site cleft, substrate binding structure, overview Aspergillus nidulans UDP-alpha-D-galactose
-
r

Subunits

Subunits Comment Organism
? x * 40600 Aspergillus nidulans

Synonyms

Synonyms Comment Organism
GalE
-
Aspergillus nidulans
UDP-galactose-4-epimerase
-
Aspergillus nidulans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Aspergillus nidulans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5
-
UDP-alpha-D-glucose mutant L320Y, pH 8.5, 25°C Aspergillus nidulans
12.8
-
UDP-alpha-D-glucose wild-type enzyme, pH 8.5, 25°C Aspergillus nidulans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Aspergillus nidulans

Cofactor

Cofactor Comment Organism Structure
NAD+ bound within the active site cleft, binding structure, overview Aspergillus nidulans

General Information

General Information Comment Organism
additional information ligand binding structures, docking study, overview Aspergillus nidulans
physiological function the enzyme produces the precursor UDP-galactopyranose required for galactofuranose synthesis Aspergillus nidulans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
26.8
-
UDP-alpha-D-glucose mutant L320Y, pH 8.5, 25°C Aspergillus nidulans
116.4
-
UDP-alpha-D-glucose wild-type enzyme, pH 8.5, 25°C Aspergillus nidulans