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Literature summary for 5.1.3.7 extracted from

  • Shin, S.; Choi, J.; Di Luccio, E.; Lee, Y.; Lee, S.; Lee, S.; Lee, S.; Lee, D.
    The structural basis of substrate promiscuity in UDP-hexose 4-epimerase from the hyperthermophilic Eubacterium Thermotoga maritima (2015), Arch. Biochem. Biophys., 585, 39-51 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in in Escherichia coli Thermotoga maritima
gene galE, sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), recombinant expression in and functional complementation of enzyme-deficient Escherichia coli strain BW25113 DELTAgalE Thermotoga maritima

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, crystal structure of the enzyme (TMGalE) and the enzyme bound to UDP-glucose is determined at resolutions of 1.9 A and 2.0 A, respectively Thermotoga maritima
purified recombinant unbound enzyme TMGalE, and TMGalE bound to UDP-Glc, X-ray diffraction structure determination and analysis at 1.9-2.0 A resolution Thermotoga maritima

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Thermotoga maritima
1.06
-
UDP-N-acetyl-alpha-D-galactosamine pH 7.0, 80°C Thermotoga maritima
1.06
-
UDP-N-acetyl-alpha-D-glucosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima
2.4
-
UDP-N-acetyl-alpha-D-glucosamine pH 7.0, 80°C Thermotoga maritima
2.4
-
UDP-N-acetyl-alpha-D-galactosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima
5.53
-
UDP-alpha-D-galactose pH 7.0, 80°C Thermotoga maritima
12.9
-
UDP-alpha-D-glucose pH 7.0, 80°C Thermotoga maritima

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme is not specifically dependent on divalent metal ions for its catalytic activity, negligible effects by Co2+, Mn2+, Mg2+, Zn2+, Ca2+, Cu2+, Fe2+, or Ni2+ Thermotoga maritima

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
70000
-
gel filtration Thermotoga maritima
70000
-
recombinant His6-tagged enzyme, gel filtration Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-N-acetyl-alpha-D-glucosamine Thermotoga maritima
-
UDP-N-acetyl-alpha-D-galactosamine
-
r

Organism

Organism UniProt Comment Textmining
Thermotoga maritima Q9WYX9
-
-
Thermotoga maritima ATCC 43589 / MSB8 / DSM 3109 / JCM 10099 Q9WYX9
-
-
Thermotoga maritima DSM 3109 Q9WYX9
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermotoga maritima
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Thermotoga maritima

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
48.7
-
purified recombinant enzyme, pH 7.0.80°C, substrate UDP-N-acetyl-glucosamine Thermotoga maritima
93.7
-
purified recombinant enzyme, pH 7.0.80°C, substrate UDP-N-acetyl-galactosamine Thermotoga maritima

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information enzyme TMGalE also has high activity for epimerization of UDP-Gal to UDP-Glc, EC 5.1.3.2. The catalytic efficiency (kcat/Km) for UDP-Gal is approximately 1.2 times higher than that for UDP-Glc, indicating that this enzyme might have a preference for UDP-Gal over UDP-Glc. The catalytic efficiencies of TMGalE for UDP-GalNAc and UDP-GlcNAc are approximately 25fold and 10fold higher than those for UDP-Gal and UDP-Glc, respectively Thermotoga maritima ?
-
?
UDP-alpha-D-galactose
-
Thermotoga maritima UDP-alpha-D-glucose
-
r
UDP-alpha-D-glucose epimerization at 36.2% compared to the activity with UDP-alpha-D-galactose Thermotoga maritima UDP-alpha-D-galactose
-
r
UDP-N-acetyl-alpha-D-galactosamine epimerization at 48.5% compared to the activity with UDP-alpha-D-galactose Thermotoga maritima UDP-N-acetyl-alpha-D-glucosamine
-
r
UDP-N-acetyl-alpha-D-glucosamine
-
Thermotoga maritima UDP-N-acetyl-alpha-D-galactosamine
-
r
UDP-N-acetyl-alpha-D-glucosamine epimerization at 25.3% compared to the activity with UDP-alpha-D-galactose Thermotoga maritima UDP-N-acetyl-alpha-D-galactosamine
-
r

Subunits

Subunits Comment Organism
homodimer 2 * 35000, SDS-PAGE Thermotoga maritima
homodimer 2 * 34899, calculated from sequence Thermotoga maritima
homodimer 2 * 35000, recombinant His6-tagged enzyme, SDS-PAGE, 2 * 34899, sequence calculation Thermotoga maritima
More enzyme structure analysis, overview Thermotoga maritima

Synonyms

Synonyms Comment Organism
GalE
-
Thermotoga maritima
More cf. EC 5.1.3.2 Thermotoga maritima
TM0509
-
Thermotoga maritima
TMGalE
-
Thermotoga maritima
UDP-hexose 4-epimerase
-
Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
Thermotoga maritima

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 100 over 50% of maximal activity within this range Thermotoga maritima
30 100 30°C: 53% of the maximal activity, 100°C: 90% of maximal activity Thermotoga maritima

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
3 h, no significant decrease in activity Thermotoga maritima
70
-
purified recombinant His6-tagged enzyme, no significant decrease in enzyme activity after 3 h Thermotoga maritima
80
-
3 h, the enzyme retains more than 85% of its original activity Thermotoga maritima
80
-
purified recombinant His6-tagged enzyme, retains more than 85% of its original activity under standard assay conditions after 3 h Thermotoga maritima
90
-
40 min, almost 50% of its original activity Thermotoga maritima
90
-
purified recombinant His6-tagged enzyme, retains 50% of its original activity under standard assay conditions after 40 min Thermotoga maritima

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7757
-
UDP-alpha-D-galactose pH 7.0, 80°C Thermotoga maritima
14677
-
UDP-alpha-D-glucose pH 7.0, 80°C Thermotoga maritima
27671
-
UDP-N-acetyl-alpha-D-glucosamine pH 7.0, 80°C Thermotoga maritima
27671
-
UDP-N-acetyl-alpha-D-galactosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima
38295
-
UDP-N-acetyl-alpha-D-galactosamine pH 7.0, 80°C Thermotoga maritima
38295
-
UDP-N-acetyl-alpha-D-glucosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Thermotoga maritima

pH Range

pH Minimum pH Maximum Comment Organism
5 10 over 50% of maximal activity within this range Thermotoga maritima
5 10 pH 5.0: about 50% of maximal activity, pH 10: about 55% of maximal activity Thermotoga maritima
6 8 over 95% of maximal activity within this range Thermotoga maritima

Cofactor

Cofactor Comment Organism Structure
NAD+ enzyme-bound, exogenous NAD+ does not appear to be a strong activator of enzyme activity, there is no significant difference in enzyme activity regardless of the presence and absence of NAD+ Thermotoga maritima

pI Value

Organism Comment pI Value Maximum pI Value
Thermotoga maritima sequence calculation
-
5.72

General Information

General Information Comment Organism
metabolism the enzyme is involved in D-galactose metabolism Thermotoga maritima
additional information dinucleotide-binding pocket in the active site, and conformational changes in the active site of TMGalE, ligand binding sites of TMGalE in complex with NAD+and UDP-Glc Thermotoga maritima
physiological function UDP-galactose 4-epimerase (GalE) catalyzes the interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal), which is a pivotal step in the Leloir pathway for D-galactose metabolism Thermotoga maritima

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1134
-
UDP-alpha-D-glucose pH 7.0, 80°C Thermotoga maritima
1406
-
UDP-alpha-D-galactose pH 7.0, 80°C Thermotoga maritima
11465
-
UDP-N-acetyl-alpha-D-glucosamine pH 7.0, 80°C Thermotoga maritima
11465
-
UDP-N-acetyl-alpha-D-galactosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima
36120
-
UDP-N-acetyl-alpha-D-galactosamine pH 7.0, 80°C Thermotoga maritima
36120
-
UDP-N-acetyl-alpha-D-glucosamine recombinant enzyme, pH 7.0, 80°C Thermotoga maritima